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Liu J, Kong Q, Ma J, Feng P, Zhang B. Comparison of clinical efficacy between Percutaneous Endoscopic Large channels nerve decompression through Translaminar approach and Percutaneous Endoscopy Conventional channels nerve decompression through Transforaminal approach for the treatment of degenerative L4/5 spinal stenosis: a retrospective study. BMC Musculoskelet Disord 2025; 26:493. [PMID: 40389933 PMCID: PMC12087222 DOI: 10.1186/s12891-025-08623-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/01/2024] [Accepted: 04/04/2025] [Indexed: 05/21/2025] Open
Abstract
OBJECYTIVE Percutaneous endoscopic surgery via the interlaminar approach and transforaminal approach are commonly used for the treatment of degenerative lumbar spinal stenosis, and in order to compare the clinical efficacy of Percutaneous Endoscopic Large channel Translaminar approach (PEL-TL) and Percutaneous Endoscopy Conventional channels Transforaminal approach (PEC-TF) in the treatment of degenerative L4/5 spinal stenosis. METHOD A retrospective analysis was conducted on 124 patients who underwent percutaneous endoscopic single segment unilateral decompression surgery for degenerative L4/5 spinal stenosis in our hospital from January 2020 to January 2023. They were divided into PEL-TL group and PEC-TF group according to different surgical methods. Recording general information of two groups of patients, including age, gender, course of disease, and length of hospital stay. Recording the surgical time, C-arm fluoroscopy frequency, incidence and type of complications for two groups of patients. CT was used to measure the Lateral Recess Angle (LRA), and MRI was used to measure the Dural Sac Cross sectional Area (DSCA) to evaluate the degree of lateral recess stenosis and compare the neurological decompression between the two groups. Using the White Panjabi scoring system (WP) to evaluate local stability before and 3 months after surgery. Recording the Visual Analog Scale (VAS) and Oswestry Disability Index (ODI) for preoperative and postoperative hip and lower limb pain in two groups of patients. Evaluateing the efficacy using the modified Macnab criteria one year after surgery. RESULTS There was no statistically significant difference in general information between the two groups of patients (P > 0.05). The surgery time in the PEL-TL group was shorter than that in the PEC-TF group (P < 0.05). The number of C-arm fluoroscopy in the PEL-TL group was significantly lower than that in the PEC-TF group (P < 0.05). There was no statistically significant difference in the incidence of complications between the two groups of patients (11.1% in the PEL-TL group and 14.3% in the PEC-TF group) (P > 0.05). The postoperative recurrence rate of PEL-TL is lower than that of PEC-TF (P < 0.05). All enrolled patients were followed up regularly for 1 year. There was no significant difference in preoperative LRA and DSCA between the two groups of patients (P > 0.05). After 1 year of surgery, LRA and DSCA in both groups were significantly larger than before (P < 0.05). There was no statistically significant difference in postoperative DSCA between the two groups, but LRA in the PEL-TL group was more significantly larger than that in the PEC-TF group (P < 0.05). There was no statistically significant difference in preoperative and postoperative WP between the two groups of patients, and there was no significant difference in WP in two groups. The ODI scores and the VAS scores of buttock and lower limb pain at each follow-up time point after surgery in both groups of patients showed significant improvement compared to before surgery. There was no statistically significant difference in functional scores between the two groups at each follow-up time point (p > 0.05). One year after surgery, the efficacy was evaluated using the modified Macnab criteria. Among them, in the PEL-TL group, 36 cases were excellent and 14 cases were good, with an excellent and good rate of 92.6%. In the PEC-TF group, 48 cases were excellent and 16 cases were good, with an excellent and good rate of 91.4%. There was no statistically significant difference between the two groups (p > 0.05). CONCLUSION Both surgical methods can achieve satisfactory clinical efficacy in treating degenerative lumbar 4/5 spinal stenosis. PEL-TL has fewer C-arm fluoroscopy times, wider decompression range, shorter surgical time, and lower recurrence rate during surgery, while PEC-TF can be routinely performed under local anesthesia to reduce anesthesia risk.
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Affiliation(s)
- Junlin Liu
- Chengdu Office Hospital of the People's Government of the Xizang Autonomous Region, Chengdu, China
| | - Qingquan Kong
- Chengdu Office Hospital of the People's Government of the Xizang Autonomous Region, Chengdu, China.
- Spinal Surgery Department, West China Hospital of Sichuan University, Chengdu, China.
| | - Junsong Ma
- Chengdu Office Hospital of the People's Government of the Xizang Autonomous Region, Chengdu, China
| | - Pin Feng
- Chengdu Office Hospital of the People's Government of the Xizang Autonomous Region, Chengdu, China
| | - Bin Zhang
- Chengdu Office Hospital of the People's Government of the Xizang Autonomous Region, Chengdu, China
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Sun C, Janjic Rankovic M, Folwaczny M, Stocker T, Otto S, Wichelhaus A, Baumert U. Effect of Different Parameters of In Vitro Static Tensile Strain on Human Periodontal Ligament Cells Simulating the Tension Side of Orthodontic Tooth Movement. Int J Mol Sci 2022; 23:ijms23031525. [PMID: 35163446 PMCID: PMC8835937 DOI: 10.3390/ijms23031525] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/16/2021] [Revised: 01/21/2022] [Accepted: 01/26/2022] [Indexed: 02/01/2023] Open
Abstract
This study aimed to investigate the effects of different magnitudes and durations of static tensile strain on human periodontal ligament cells (hPDLCs), focusing on osteogenesis, mechanosensing and inflammation. Static tensile strain magnitudes of 0%, 3%, 6%, 10%, 15% and 20% were applied to hPDLCs for 1, 2 and 3 days. Cell viability was confirmed via live/dead cell staining. Reference genes were tested by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and assessed. The expressions of TNFRSF11B, ALPL, RUNX2, BGLAP, SP7, FOS, IL6, PTGS2, TNF, IL1B, IL8, IL10 and PGE2 were analyzed by RT-qPCR and/or enzyme-linked immunosorbent assay (ELISA). ALPL and RUNX2 both peaked after 1 day, reaching their maximum at 3%, whereas BGLAP peaked after 3 days with its maximum at 10%. SP7 peaked after 1 day at 6%, 10% and 15%. FOS peaked after 3 days with its maximum at 3%, 6% and 15%. The expressions of IL6 and PTGS2 both peaked after 1 day, with their minimum at 10%. PGE2 peaked after 1 day (maximum at 20%). The ELISA of IL6 peaked after 3 days, with the minimum at 10%. In summary, the lower magnitudes promoted osteogenesis and caused less inflammation, while the higher magnitudes inhibited osteogenesis and enhanced inflammation. Among all magnitudes, 10% generally caused a lower level of inflammation with a higher level of osteogenesis.
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Affiliation(s)
- Changyun Sun
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, 80336 Munich, Germany; (C.S.); (M.J.R.); (T.S.); (A.W.)
| | - Mila Janjic Rankovic
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, 80336 Munich, Germany; (C.S.); (M.J.R.); (T.S.); (A.W.)
| | - Matthias Folwaczny
- Department of Conservative Dentistry and Periodontology, University Hospital, LMU Munich, 80336 Munich, Germany;
| | - Thomas Stocker
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, 80336 Munich, Germany; (C.S.); (M.J.R.); (T.S.); (A.W.)
| | - Sven Otto
- Department of Oral and Maxillofacial Surgery and Facial Plastic Surgery, University Hospital, LMU Munich, 80336 Munich, Germany;
| | - Andrea Wichelhaus
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, 80336 Munich, Germany; (C.S.); (M.J.R.); (T.S.); (A.W.)
| | - Uwe Baumert
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, 80336 Munich, Germany; (C.S.); (M.J.R.); (T.S.); (A.W.)
- Correspondence:
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The effect of piezocision vs no piezocision on maxillary extraction space closure: A split-mouth, randomized controlled clinical trial. Am J Orthod Dentofacial Orthop 2021; 161:7-19.e2. [PMID: 34654603 DOI: 10.1016/j.ajodo.2021.06.015] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/01/2020] [Revised: 06/01/2021] [Accepted: 06/01/2021] [Indexed: 01/17/2023]
Abstract
INTRODUCTION The aim of this 2-arm-parallel, split-mouth trial was to investigate the effects of piezocision compared with no piezocision on maxillary canine distalization and to evaluate patient perceptions on the surgical procedure. METHODS Twenty-two participants requiring extractions of maxillary first premolars were recruited from the Department of Orthodontics (Sydney Dental Hospital) waiting list. After leveling and alignment, a minimum of 3 mm space was required for canine retraction. Piezocision cuts distal to the canines were 4 mm long and 3 mm deep into the buccal cortical plate. The canine retraction was initiated on both sides immediately after surgery, with coil springs delivering 150 g of force per side. Random assignment of piezocision or control intervention on the patient's right side was performed (www.randomisation.com) for the random number generation, and allocation concealment was accomplished with opaque, sealed envelopes. Patients were assessed every 6 weeks for coil activation and alginate impressions over 18 weeks. The primary outcome was the amount of tooth movement in mm. Secondary outcomes were canine rotation, anchorage loss measured on scanned dental models, and patient pain levels and perception on piezocision using visual analog scale questionnaires. Blinding was feasible for the dental model measurements. RESULTS Twenty patients completed the trial. The treatment × time interaction showed no statistically or clinically significant differences in maxillary extraction space closure (b = -0.02; 95% confidence interval [CI], -0.29 to 0.25; P = 0.89) canine rotation (b = -1.45; 95% CI, -4 to 1.09; P = 0.26) and anchorage loss (b = -0.02; 95% CI, -0.38 to 0.34; P = 0.92). All patients except for one had minimal pain associated with the piezocision surgery but found the procedure tolerable and would recommend it. No harm occurred during the trial. CONCLUSIONS Piezocision-assisted maxillary canine distalization was similar to distalization with conventional orthodontics with patients tolerating the procedure.
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Sun C, Janjic Rankovic M, Folwaczny M, Otto S, Wichelhaus A, Baumert U. Effect of Tension on Human Periodontal Ligament Cells: Systematic Review and Network Analysis. Front Bioeng Biotechnol 2021; 9:695053. [PMID: 34513810 PMCID: PMC8429507 DOI: 10.3389/fbioe.2021.695053] [Citation(s) in RCA: 20] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/21/2021] [Accepted: 08/10/2021] [Indexed: 01/09/2023] Open
Abstract
Orthodontic tooth movement is based on the remodeling of tooth-surrounding tissues in response to mechanical stimuli. During this process, human periodontal ligament cells (hPDLCs) play a central role in mechanosensing and mechanotransduction. Various in vitro models have been introduced to investigate the effect of tension on hPDLCs. They provide a valuable body of knowledge on how tension influences relevant genes, proteins, and metabolites. However, no systematic review summarizing these findings has been conducted so far. Aim of this systematic review was to identify all related in vitro studies reporting tension application on hPDLCs and summarize their findings regarding force parameters, including magnitude, frequency and duration. Expression data of genes, proteins, and metabolites was extracted and summarized. Studies' risk of bias was assessed using tailored risk of bias tools. Signaling pathways were identified by protein-protein interaction (PPI) networks using STRING and GeneAnalytics. According to our results, Flexcell Strain Unit® and other silicone-plate or elastic membrane-based apparatuses were mainly adopted. Frequencies of 0.1 and 0.5 Hz were predominantly applied for dynamic equibiaxial and uniaxial tension, respectively. Magnitudes of 10 and 12% were mostly employed for dynamic tension and 2.5% for static tension. The 10 most commonly investigated genes, proteins and metabolites identified, were mainly involved in osteogenesis, osteoclastogenesis or inflammation. Gene-set enrichment analysis and PPI networks gave deeper insight into the involved signaling pathways. This review represents a brief summary of the massive body of knowledge in this field, and will also provide suggestions for future researches on this topic.
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Affiliation(s)
- Changyun Sun
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, Munich, Germany
| | - Mila Janjic Rankovic
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, Munich, Germany
| | - Matthias Folwaczny
- Department of Conservative Dentistry and Periodontology, University Hospital, LMU Munich, Munich, Germany
| | - Sven Otto
- Department of Oral and Maxillofacial Plastic Surgery, Martin-Luther-University Halle-Wittenberg, Halle (Saale), Germany
| | - Andrea Wichelhaus
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, Munich, Germany
| | - Uwe Baumert
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, Munich, Germany
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Xie Y, Tang Q, Yu S, Zheng W, Chen G, Huang X, Chen L. Orthodontic Force-Induced BMAL1 in PDLCs Is a Vital Osteoclastic Activator. J Dent Res 2021; 101:177-186. [PMID: 34157911 DOI: 10.1177/00220345211019949] [Citation(s) in RCA: 9] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022] Open
Abstract
Orthodontic tooth movement (OTM) depends on periodontal ligament cells (PDLCs) sensing biomechanical stimuli and subsequently releasing signals to initiate alveolar bone remodeling. However, the mechanisms by which PDLCs sense biomechanical stimuli and affect osteoclastic activities are still unclear. This study demonstrates that the core circadian protein aryl hydrocarbon receptor nuclear translocator-like protein 1 (BMAL1) in PDLCs is highly involved in sensing and delivering biomechanical signals. Orthodontic force upregulates BMAL1 expression in periodontal tissues and cultured PDLCs in manners dependent on ERK (extracellular signal-regulated kinase) and AP1 (activator protein 1). Increased BMAL1 expression can enhance secretion of CCL2 (C-C motif chemokine 2) and RANKL (receptor activator of nuclear factor-κB ligand) in PDLCs, which subsequently promotes the recruitment of monocytes that differentiate into osteoclasts. The mechanistic delineation clarifies that AP1 induced by orthodontic force can directly interact with the BMAL1 promoter and activate gene transcription in PDLCs. Localized administration of the ERK phosphorylation inhibitor U0126 or the BMAL1 inhibitor GSK4112 suppressed ERK/AP1/BMAL1 signaling. These treatments dramatically reduced osteoclastic activity in the compression side of a rat orthodontic model, and the OTM rate was almost nonexistent. In summary, our results suggest that force-induced expression of BMAL1 in PDLCs is closely involved in controlling osteoclastic activities during OTM and plays a vital role in alveolar bone remodeling. It could be a useful therapeutic target for accelerating the OTM rate and controlling pathologic bone-remodeling activities.
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Affiliation(s)
- Y Xie
- Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,School of Stomatology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration, Wuhan, China
| | - Q Tang
- Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,School of Stomatology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration, Wuhan, China
| | - S Yu
- Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,School of Stomatology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration, Wuhan, China
| | - W Zheng
- Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,School of Stomatology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration, Wuhan, China
| | - G Chen
- Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,School of Stomatology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration, Wuhan, China
| | - X Huang
- Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,School of Stomatology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration, Wuhan, China
| | - L Chen
- Department of Stomatology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,School of Stomatology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.,Hubei Province Key Laboratory of Oral and Maxillofacial Development and Regeneration, Wuhan, China
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6
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Janjic Rankovic M, Docheva D, Wichelhaus A, Baumert U. Effect of static compressive force on in vitro cultured PDL fibroblasts: monitoring of viability and gene expression over 6 days. Clin Oral Investig 2019; 24:2497-2511. [PMID: 31728735 DOI: 10.1007/s00784-019-03113-6] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/09/2019] [Accepted: 10/07/2019] [Indexed: 12/14/2022]
Abstract
OBJECTIVES The aim was to investigate the impact of static compressive force (CF) application on human PDL-derived fibroblasts (HPDF) in vitro for up to 6 days on the expression of specific genes and to monitor cell growth and cell viability. MATERIALS AND METHODS CF of 2 g/cm2 was applied on HPDFs for 1-6 days. On each day, gene expression (cFOS, HB-GAM, COX2, IL6, TNFα, RUNX2, and P2RX2) and secretion (TNFα, PGE2) were determined by RT-qPCR and ELISA, respectively. Cell growth and cell viability were monitored daily. RESULTS In comparison with controls, significant upregulation of cFOS in compressed HPDFs was observed. HB-GAM showed no changes in expression, except on day 5 (P < 0.001). IL6 expression was significantly upregulated from day 2-5, reaching the maximum on day 3 (P < 0.001). TNFα expression was upregulated on all but day 2. COX2 showed upregulation, reaching the plateau from day 3 (P < 0.001) until day 4 (P < 0.001), and returning to the initial state till day 6. P2RX7 was downregulated on days 2 and 4 to 6 (P < 0.001). RUNX2 was downregulated on days 2 and 5 (both P < 0.001). Cells in both groups were proliferating, and no negative effect on cell viability was observed. CONCLUSION Results suggest high molecular activity up to 6 days, therefore introducing further need for in vitro studies with a longer duration that would explain other genes and metabolites involved in orthodontic tooth movement (OTM). CLINICAL RELEVANCE Extension of an established in vitro force application system for prolonged force application (6 days) simulating the initial phase of OTM.
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Affiliation(s)
- Mila Janjic Rankovic
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, Goethestrasse 70, 80336, Munich, Germany
| | - Denitsa Docheva
- Experimental Trauma Surgery, Department of Trauma Surgery, University Regensburg Medical Centre, Regensburg, Germany
| | - Andrea Wichelhaus
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, Goethestrasse 70, 80336, Munich, Germany
| | - Uwe Baumert
- Department of Orthodontics and Dentofacial Orthopedics, University Hospital, LMU Munich, Goethestrasse 70, 80336, Munich, Germany.
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Papadopoulou A, Todaro A, Eliades T, Kletsas D. Effect of hyperglycaemic conditions on the response of human periodontal ligament fibroblasts to mechanical stretching. Eur J Orthod 2019; 41:583-590. [DOI: 10.1093/ejo/cjz051] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
Abstract
Summary
Objectives
The aim of the present study was to investigate the impact of high glucose concentration on the response of human periodontal ligament fibroblasts (PDLFs) to cyclic tensile strain.
Materials and Methods
Human PDLFs were incubated under normal or high glucose conditions, and then were subjected to cyclic tensile stretching (8 per cent extension, 1 Hz). Gene expression was determined by quantitative real-time polymerase chain reaction. Intracellular reactive oxygen species (ROS) were determined by the 2’,7’-dichlorofluorescein-diacetate assay, activation of mitogen-activated protein kinase (MAPK) was monitored by western analysis and osteoblastic differentiation was estimated with Alizarin Red-S staining.
Results
Cyclic tensile stretching of PDLF leads to an immediate activation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), as well as to the increased expression of the transcription factor c-fos, known to regulate many osteogenesis-related genes. At later time points, the alkaline phosphatase and osteopontin genes were also upregulated. Hyperglycaemic conditions inhibited these effects. High glucose conditions were unable to increase ROS levels, but they increased the medium’s osmolality. Finally, increase of osmolality mimics the inhibitory effect of hyperglycaemia on MAPK activation, c-fos and osteoblast-specific gene markers’ upregulation, as well as osteogenic differentiation capacity.
Conclusion
Our findings indicate that under high glucose conditions, human PDLFs fail to adequately respond to mechanical deformation, while their strain-elicited osteoblast differentiation ability is deteriorated. The aforementioned effects are most probably mediated by the increased osmolality under hyperglycaemic conditions.
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Affiliation(s)
- Adamantia Papadopoulou
- Laboratory of Cell Proliferation & Ageing, Institute of Biosciences & Applications, National Centre for Scientific Research ‘Demokritos’, Athens, Greece
| | - Alexia Todaro
- Clinic of Orthodontics and Paediatric Dentistry, University of Zurich, Zurich, Switzerland
| | - Theodore Eliades
- Clinic of Orthodontics and Paediatric Dentistry, University of Zurich, Zurich, Switzerland
| | - Dimitris Kletsas
- Laboratory of Cell Proliferation & Ageing, Institute of Biosciences & Applications, National Centre for Scientific Research ‘Demokritos’, Athens, Greece
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Leewananthawet A, Arakawa S, Okano T, Daitoku Kinoshita R, Ashida H, Izumi Y, Suzuki T. Ozone ultrafine bubble water induces the cellular signaling involved in oxidative stress responses in human periodontal ligament fibroblasts. SCIENCE AND TECHNOLOGY OF ADVANCED MATERIALS 2019; 20:589-598. [PMID: 31258824 PMCID: PMC6586087 DOI: 10.1080/14686996.2019.1614980] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 02/19/2019] [Revised: 05/01/2019] [Accepted: 05/01/2019] [Indexed: 06/09/2023]
Abstract
Periodontitis is a chronic inflammatory disease caused by oral microorganisms in the subgingival biofilm. Stable aqueous ozone ultrafine bubble water (OUFBW) has recently begun to be used as an antiseptic in the treatment of periodontitis. The effectiveness of OUFBW is thought to depend on the bactericidal actions of dissolved ozone exerted via its oxidizing effect. On the other hand, the effects of ozone on the periodontal tissues are largely unknown. In this paper we examined the cellular responses after OUFBW treatment. Human primary periodontal ligament fibroblasts (hPDLFs) or Ca9-22 human gingival epithelial cells were treated with OUFBW or UV-inactivated OUFBW. The production of reactive oxygen species (ROS), the activation of mitogen-activated protein kinase (MAPK) and the nuclear factor-kappa B (NF-κB) activation were analyzed. The transcript profiles of hPDLFs after OUFBW treatment were also analyzed by RNA sequencing (RNA-seq). Our results showed that OUFBW induces oxidative stress by generating ROS, which, in turn, activated the MAPK pathway. OUFBW triggered activation of c-Fos, a major component of the transcription factor activator protein 1 (AP-1), and also nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2), which possessed a high sensitivity to oxidative stress. The results of RNA-seq analysis revealed that the numerous genes involved in oxidative stress responses or MAPK signaling pathway were up-regulated after OUFBW treatment. Investigation of the signaling pathways activated by OUFBW highlights another aspect of the biological roles of OUFBW, in addition to its bactericidal activity, in the treatment of periodontitis.
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Affiliation(s)
- Anongwee Leewananthawet
- Department of Bacterial Pathogenesis, Infection and Host Response, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
- Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
| | - Shinichi Arakawa
- Department of Lifetime Oral Health Care Science, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
| | - Tokuju Okano
- Department of Bacterial Pathogenesis, Infection and Host Response, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
| | - Ryo Daitoku Kinoshita
- Department of Bacterial Pathogenesis, Infection and Host Response, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
| | - Hiroshi Ashida
- Department of Bacterial Pathogenesis, Infection and Host Response, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
| | - Yuichi Izumi
- Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
- Oral Care and Perio Center, Southern TOHOKU Research Institute for Neuroscience, Fukushima, Japan
| | - Toshihiko Suzuki
- Department of Bacterial Pathogenesis, Infection and Host Response, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
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Chukkapalli SS, Lele TP. Periodontal cell mechanotransduction. Open Biol 2019; 8:rsob.180053. [PMID: 30209038 PMCID: PMC6170509 DOI: 10.1098/rsob.180053] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2018] [Accepted: 08/16/2018] [Indexed: 01/09/2023] Open
Abstract
The periodontium is a structurally and functionally complex tissue that facilitates the anchorage of teeth in jaws. The periodontium consists of various cell types including stem cells, fibroblasts and epithelial cells. Cells of the periodontium are constantly exposed to mechanical stresses generated by biological processes such as the chewing motions of teeth, by flows generated by tongue motions and by forces generated by implants. Mechanical stresses modulate the function of cells in the periodontium, and may play a significant role in the development of periodontal disease. Here, we review the literature on the effect of mechanical forces on periodontal cells in health and disease with an emphasis on molecular and cellular mechanisms.
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Affiliation(s)
- Sasanka S Chukkapalli
- Department of Oral Biology, University of Florida, College of Dentistry, Gainesville, FL 32610, USA.,Center for Molecular Microbiology, University of Florida, College of Dentistry, Gainesville, FL 32610, USA
| | - Tanmay P Lele
- Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA
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Papadopoulou A, Iliadi A, Eliades T, Kletsas D. Early responses of human periodontal ligament fibroblasts to cyclic and static mechanical stretching. Eur J Orthod 2018; 39:258-263. [PMID: 27932408 DOI: 10.1093/ejo/cjw075] [Citation(s) in RCA: 15] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/20/2023]
Abstract
Objective To compare the mechanotransduction caused by cyclic and static mechanical strains in human periodontal ligament fibroblasts (hPDLFs) cultured under identical conditions. Materials and methods hPDLFs, originating from the same donors, were exposed either to cyclic or to static tensile strain using specially designed devices and under identical culture conditions. Activation of all members of mitogen-activated protein kinases (MAPKs) was monitored by western immunoblot analysis. Expression levels of immediate/early genes c-fos and c-jun were assessed with quantitative real-time polymerase chain reaction. Results Time course experiments revealed that both types of stresses activate the three members of MAPK, that is ERK, p38, and JNK, with cyclic stress exhibiting a slightly more extended activation. Further downstream, both stresses upregulate the immediate/early genes c-fos and c-jun, encoding components of the activator protein-1 (AP-1), a key transcription factor in osteoblastic differentiation; again cyclic strain provokes a more intense upregulation. Six hours after the application of both strains, MAPK activation and gene expression return to basal levels. Finally, cells exposed to cyclic stress for longer periods are distributed approximately perpendicular to the axis of the applied strain, whereas cells exposed to static loading remain in a random orientation in culture. Conclusion The findings of the present study indicate similar, although not identical, immediate/early responses of hPDLs to cyclic and static stretching, with cyclic strain provoking a more intense adaptive response of these cells to mechanical deformation.
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Affiliation(s)
- Adamantia Papadopoulou
- Laboratory of Cell Proliferation and Ageing, Institute of Biosciences and Applications, National Centre for Scientific Research 'Demokritos', Athens
| | - Anna Iliadi
- Department of Orthodontics, School of Dentistry, University of Athens, Greece
| | - Theodore Eliades
- Clinic of Orthodontics and Paediatric Dentistry, University of Zurich, Switzerland
| | - Dimitris Kletsas
- Laboratory of Cell Proliferation and Ageing, Institute of Biosciences and Applications, National Centre for Scientific Research 'Demokritos', Athens
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Yamamoto T, Ugawa Y, Kawamura M, Yamashiro K, Kochi S, Ideguchi H, Takashiba S. Modulation of microenvironment for controlling the fate of periodontal ligament cells: the role of Rho/ROCK signaling and cytoskeletal dynamics. J Cell Commun Signal 2018; 12:369-378. [PMID: 29086204 PMCID: PMC5842188 DOI: 10.1007/s12079-017-0425-3] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/14/2017] [Accepted: 10/17/2017] [Indexed: 12/20/2022] Open
Abstract
Cells behave in a variety of ways when they perceive changes in their microenvironment; the behavior of cells is guided by their coordinated interactions with growth factors, niche cells, and extracellular matrix (ECM). Modulation of the microenvironment affects the cell morphology and multiple gene expressions. Rho/Rho-associated coiled-coil-containing protein kinase (ROCK) signaling is one of the key regulators of cytoskeletal dynamics and actively and/or passively determines the cell fate, such as proliferation, migration, differentiation, and apoptosis, by reciprocal communication with the microenvironment. During periodontal wound healing, it is important to recruit the residential stem cells into the defect site for regeneration and homeostasis of the periodontal tissue. Periodontal ligament (PDL) cells contain a heterogeneous fibroblast population, including mesenchymal stem cells, and contribute to the reconstruction of tooth-supporting tissues. Therefore, bio-regeneration of PDL cells has been the ultimate goal of periodontal therapy for decades. Recent stem cell researches have shed light on intrinsic ECM properties, providing paradigm shifts in cell fate determination. This review focuses on the role of ROCK activity and the effects of Y-27632, a specific inhibitor of ROCK, in the modulation of ECM-microenvironment. Further, it presents the current understanding of how Rho/ROCK signaling affects the fate determination of stem cells, especially PDL cells. In addition, we have also discussed in detail the underlying mechanisms behind the reciprocal response to the microenvironment.
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Affiliation(s)
- Tadashi Yamamoto
- Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan
| | - Yuki Ugawa
- Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan
| | - Mari Kawamura
- Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan
| | - Keisuke Yamashiro
- Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan
| | - Shinsuke Kochi
- Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan
| | - Hidetaka Ideguchi
- Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan
| | - Shogo Takashiba
- Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan.
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12
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Karamesinis K, Basdra EK. The biological basis of treating jaw discrepancies: An interplay of mechanical forces and skeletal configuration. Biochim Biophys Acta Mol Basis Dis 2018; 1864:1675-1683. [PMID: 29454076 DOI: 10.1016/j.bbadis.2018.02.007] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/18/2017] [Revised: 02/06/2018] [Accepted: 02/12/2018] [Indexed: 10/18/2022]
Abstract
Jaw discrepancies and malrelations affect a large proportion of the general population and their treatment is of utmost significance for individuals' health and quality of life. The aim of their therapy is the modification of aberrant jaw development mainly by targeting the growth potential of the mandibular condyle through its cartilage, and the architectural shape of alveolar bone through a suture type of structure, the periodontal ligament. This targeted treatment is achieved via external mechanical force application by using a wide variety of intraoral and extraoral appliances. Condylar cartilage and sutures exhibit a remarkable plasticity due to the mechano-responsiveness of the chondrocytes and the multipotent mesenchymal cells of the sutures. The tissues respond biologically and adapt to mechanical force application by a variety of signaling pathways and a final interplay between the proliferative activity and the differentiation status of the cells involved. These targeted therapeutic functional alterations within temporo-mandibular joint ultimately result in the enhancement or restriction of mandibular growth, while within the periodontal ligament lead to bone remodeling and change of its architectural structure. Depending on the form of the malrelation presented, the above treatment approaches, in conjunction or separately, lead to the total correction of jaw discrepancies and the achievement of facial harmony and function. Overall, the treatment of craniofacial and jaw anomalies can be seen as an interplay of mechanical forces and adaptations occurring within temporo-mandibular joint and alveolar bone. The aim of the present review is to present up-to-date knowledge on the mechano-biology behind jaw growth modification and alveolar bone remodeling. Furthermore, future molecular targeted therapeutic strategies are discussed aiming at the improvement of mechanically-driven chondrogenesis and osteogenesis.
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Affiliation(s)
- Konstantinos Karamesinis
- Department of Biological Chemistry, Cellular and Molecular Biomechanics Unit, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece
| | - Efthimia K Basdra
- Department of Biological Chemistry, Cellular and Molecular Biomechanics Unit, Medical School, National and Kapodistrian University of Athens, 11527 Athens, Greece.
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13
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Chiquet M, Katsaros C, Kletsas D. Multiple functions of gingival and mucoperiosteal fibroblasts in oral wound healing and repair. Periodontol 2000 2017; 68:21-40. [PMID: 25867977 DOI: 10.1111/prd.12076] [Citation(s) in RCA: 42] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 05/01/2014] [Indexed: 12/22/2022]
Abstract
Fibroblasts are cells of mesenchymal origin. They are responsible for the production of most extracellular matrix in connective tissues and are essential for wound healing and repair. In recent years, it has become clear that fibroblasts from different tissues have various distinct traits. Moreover, wounds in the oral cavity heal under very special environmental conditions compared with skin wounds. Here, we reviewed the current literature on the various interconnected functions of gingival and mucoperiosteal fibroblasts during the repair of oral wounds. The MEDLINE database was searched with the following terms: (gingival OR mucoperiosteal) AND fibroblast AND (wound healing OR repair). The data gathered were used to compare oral fibroblasts with fibroblasts from other tissues in terms of their regulation and function during wound healing. Specifically, we sought answers to the following questions: (i) what is the role of oral fibroblasts in the inflammatory response in acute wounds; (ii) how do growth factors control the function of oral fibroblasts during wound healing; (iii) how do oral fibroblasts produce, remodel and interact with extracellular matrix in healing wounds; (iv) how do oral fibroblasts respond to mechanical stress; and (v) how does aging affect the fetal-like responses and functions of oral fibroblasts? The current state of research indicates that oral fibroblasts possess unique characteristics and tightly controlled specific functions in wound healing and repair. This information is essential for developing new strategies to control the intraoral wound-healing processes of the individual patient.
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14
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Kawarizadeh A, Bourauel C, Götz W, Jäger A. Early Responses of Periodontal Ligament Cells to Mechanical Stimulus in vivo. J Dent Res 2016; 84:902-6. [PMID: 16183788 DOI: 10.1177/154405910508401006] [Citation(s) in RCA: 64] [Impact Index Per Article: 7.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022] Open
Abstract
Previous studies have indicated that human periodontal ligament cells undergo osteoblastic differentiation via the ERK pathway under mechanical stress in vitro. This study aimed to verify this principle in vivo. The right upper first molars of 25 anesthetized rats were loaded with constant forces of 0.1 N for up to 8 hrs. The untreated contralateral side served as a control. Paraffin-embedded sections were analyzed by immunohistochemistry for proliferating cell nuclear antigen (PCNA), runt-related transcription factor 2 (Runx2/Cbfa1), and phosphorylated extracellular signal-regulated kinases 1/2 (pERK1/2). In selected areas under tension, the proportions of Runx2-positive and pERK1/2-positive cells increased within 8 hrs of loading, whereas these proportions in selected areas under pressure were significantly lower than those in control teeth. Moreover, there were no significant changes in the number of PCNA-positive cells. Thus, mechanical stimulus up-regulates Runx2, and this regulation may be achieved via the ERK pathway.
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Affiliation(s)
- A Kawarizadeh
- Department of Orthodontics, Dental Clinic, University of Bonn, Welschnonnenstr. 17, D-53111 Bonn, Germany
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15
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Decoding the intervertebral disc: Unravelling the complexities of cell phenotypes and pathways associated with degeneration and mechanotransduction. Semin Cell Dev Biol 2016; 62:94-103. [PMID: 27208724 DOI: 10.1016/j.semcdb.2016.05.008] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/12/2016] [Accepted: 05/17/2016] [Indexed: 12/20/2022]
Abstract
Back pain is the most common cause of pain and disability worldwide. While its etiology remains unknown, it is typically associated with intervertebral disc (IVD) degeneration. Despite the prevalence of back pain, relatively little is known about the specific cellular pathways and mechanisms that contribute to the development, function and degeneration of the IVD. Consequently, current treatments for back pain are largely limited to symptomatic interventions. However, major progress is being made in multiple research directions to unravel the biology and pathology of the IVD, raising hope that effective disease-modifying interventions will soon be developed. In this review, we will discuss our current knowledge and gaps in knowledge on the developmental origin of the IVD, the phenotype of the distinct cell types found within the IVD tissues, molecular targets in IVD degeneration identified using bioinformatics strategies, and mechanotransduction pathways that influence IVD cell fate and function.
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Cyclic tensile stress of human annulus fibrosus cells induces MAPK activation: involvement in proinflammatory gene expression. Osteoarthritis Cartilage 2016; 24:679-87. [PMID: 26687822 DOI: 10.1016/j.joca.2015.11.022] [Citation(s) in RCA: 22] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/07/2015] [Revised: 11/05/2015] [Accepted: 11/24/2015] [Indexed: 02/02/2023]
Abstract
OBJECTIVE To study the role of mitogen-activated protein kinases (MAPKs) in human annulus fibrosus (AF) cells subjected to cyclic tensile stress (CTS). DESIGN An in vitro system for CTS studies was established using AF cultures on fibronectin-coated silicone dishes. MAPK phosphorylation was studied by western analysis, while gene expression was followed by qRT-PCR. DNA synthesis was assessed by both tritiated thymidine incorporation and flow cytometry, and collagen synthesis using tritiated proline incorporation and the protease-free collagenase method. RESULTS All three MAPKs studied, i.e., ERK, SAPK/JNK, and p38 were found to be phosphorylated immediately after CTS application within physiological range. A second wave of phosphorylation appeared at later time points. MAPK activation was elevated at higher CTS magnitudes, but independent of the frequency. CTS did not stimulate DNA synthesis neither extracellular matrix turnover, but it stimulated the proinflammatory genes, COX-2, IL-6, and IL-8. This stimulation was more intense at the highest magnitude (8%) tested and at the median frequency (1 Hz) and time interval (12 h). Blocking of ERK, SAPK/JNK, and p38 MAPK inhibited the CTS-induced stimulation of COX-2 and IL-8, while IL-6 expression was mediated only by SAPK/JNK and p38 MAPK. CONCLUSIONS We have described for the first time the activation of MAPKs in human AF cells in response to CTS and showed that it drives an inflammatory reaction. These observations shed light on the mechanisms of intervertebral disc (IVD) cell responses to mechanical stress, contributing to the understanding of disc pathophysiology and possibly to the design of novel therapeutic interventions.
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Yang L, Yang Y, Wang S, Li Y, Zhao Z. In vitro mechanical loading models for periodontal ligament cells: From two-dimensional to three-dimensional models. Arch Oral Biol 2015; 60:416-24. [DOI: 10.1016/j.archoralbio.2014.11.012] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/10/2014] [Revised: 11/19/2014] [Accepted: 11/20/2014] [Indexed: 02/08/2023]
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Sen S, Diercke K, Zingler S, Lux CJ, Erber R. Compression induces Ephrin-A2 in PDL fibroblasts via c-fos. J Dent Res 2015; 94:464-72. [PMID: 25604255 DOI: 10.1177/0022034514567197] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023] Open
Abstract
Ephrin-A2-EphA2 and ephrin-B2-EphB4 interactions have been implicated in the regulation of bone remodeling. We previously demonstrated a potential role for members of the Eph-ephrin family of receptor tyrosine kinases for bone remodeling during orthodontic tooth movement: compression-dependent upregulation of ephrin-A2 in fibroblasts of the periodontal ligament (PDL) attenuated osteogenesis in osteoblasts of the alveolar bone. However, factors affecting the regulation of ephrin-A2 expression upon the application of compressive forces remained unclear. Here, we report a mechano-dependent pathway of ephrin-A2 induction in PDL fibroblasts (PDLFs) involving extracellular signal-regulated kinases (ERK) 1/2 and c-fos. PDLF subjected to compressive forces (30.3 g/cm(2)) upregulated c-fos and ephrin-A2 mRNA and protein expression and displayed increased ERK1/2 phosphorylation. Inhibition of the MAP kinase kinase (MEK)/ERK1/2 pathway using the specific MEK inhibitor U0126 significantly reduced ephrin-A2 messenger RNA upregulation upon compression. Silencing of c-fos using a small interfering RNA approach led to a significant inhibition of ephrin-A2 induction upon the application of compressive forces. Interestingly, ephrin-A2 stimulation of PDLF induced c-fos expression and led also to the induction of ephrin-A2 expression. Using a reporter gene construct in murine 3T3 cells, we found that ephrin-A2 was able to stimulate serum response element (SRE)-dependent luciferase activity. As the regulation of c-fos is SRE dependent, ephrin-A2 might induce c-fos via SRE activation. Taken together, we provide evidence for an ERK1/2- and c-fos-dependent regulation of ephrin-A2 in compressed PDLF and suggest a novel pathway for ephrin-A2 induction emanating from ephrin-A2 itself. We showed previously that ephrin-A2 at compression sites might contribute to tooth movement by inhibiting osteogenic differentiation. The regulatory pathway of ephrin-A2 induction during tooth movement identified in this study might be accessible for pharmacological interventions.
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Affiliation(s)
- S Sen
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Heidelberg, Germany
| | - K Diercke
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Heidelberg, Germany
| | - S Zingler
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Heidelberg, Germany
| | - C J Lux
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Heidelberg, Germany
| | - R Erber
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Heidelberg, Germany
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Konstantonis D, Papadopoulou A, Makou M, Eliades T, Basdra E, Kletsas D. The role of cellular senescence on the cyclic stretching-mediated activation of MAPK and ALP expression and activity in human periodontal ligament fibroblasts. Exp Gerontol 2014; 57:175-80. [DOI: 10.1016/j.exger.2014.05.010] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/18/2014] [Revised: 05/07/2014] [Accepted: 05/09/2014] [Indexed: 11/26/2022]
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Spyropoulou A, Basdra EK. Mechanotransduction in bone: Intervening in health and disease. World J Exp Med 2013; 3:74-86. [DOI: 10.5493/wjem.v3.i4.74] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/28/2013] [Revised: 09/06/2013] [Accepted: 11/03/2013] [Indexed: 02/06/2023] Open
Abstract
Mechanotransduction has been proven to be one of the most significant variables in bone remodeling and its alterations have been shown to result in a variety of bone diseases. Osteoporosis, Paget’s disease, orthopedic disorders, osteopetrosis as well as hyperparathyroidism and hyperthyroidism all comprise conditions which have been linked with deregulated bone remodeling. Although the significance of mechanotransduction for bone health and disease is unquestionable, the mechanisms behind this important process have not been fully understood. This review will discuss the molecules that have been found to be implicated in mechanotransduction, as well as the mechanisms underlying bone health and disease, emphasizing on what is already known as well as new molecules potentially taking part in conveying mechanical signals from the cell surface towards the nucleus under physiological or pathologic conditions. It will also focus on the model systems currently used in mechanotransduction studies, like osteoblast-like cells as well as three-dimensional constructs and their applications among others. It will also examine the role of mechanostimulatory techniques in preventing and treating bone degenerative diseases and consider their applications in osteoporosis, craniofacial development, skeletal deregulations, fracture treatment, neurologic injuries following stroke or spinal cord injury, dentistry, hearing problems and bone implant integration in the near future.
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Senescent human periodontal ligament fibroblasts after replicative exhaustion or ionizing radiation have a decreased capacity towards osteoblastic differentiation. Biogerontology 2013; 14:741-51. [PMID: 23934584 DOI: 10.1007/s10522-013-9449-0] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2013] [Accepted: 08/02/2013] [Indexed: 01/14/2023]
Abstract
Loss of teeth increases with age or after genotoxic treatments, like head and neck radiotherapy, due to periodontium breakdown. Periodontal ligament fibroblasts represent the main cell type in this tissue and are crucial for the maintenance of homeodynamics and for its regeneration. Here, we have studied the characteristics of human periodontal ligament fibroblasts (hPDLF) that became senescent after replicative exhaustion or after exposure to ionizing radiation, as well as their ability for osteoblastic differentiation. We found that senescent hPDLF express classical markers of senescence, as well as a catabolic phenotype, as shown by the decrease in collagen type I and the increase of MMP-2 expression. In addition, we observed a considerably decreased expression of the major transcription factor for osteoblastic differentiation, i.e. Runx2, a down-regulation which was found to be p53-dependent. In accordance to the above, senescent cells have a significantly decreased alkaline phosphatase gene expression and activity, as well as a reduced ability for osteoblastic differentiation, as found by Alizarin Red staining. Interestingly, cells from both type of senescence express similar characteristics, implying analogous functions in vivo. In conclusion, senescent hPDLF express a catabolic phenotype and express a significantly decreased ability towards an osteoblastic differentiation, thus probably affecting tissue development and integrity.
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Kook SH, Jeon YM, Park SS, Lee JC. Periodontal fibroblasts modulate proliferation and osteogenic differentiation of embryonic stem cells through production of fibroblast growth factors. J Periodontol 2013; 85:645-54. [PMID: 23805819 DOI: 10.1902/jop.2013.130252] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2023]
Abstract
BACKGROUND Periodontal ligament fibroblasts (PLFs) maintain homeostasis of periodontal ligaments by producing paracrine factors that affect various functions of stem-like cells. It is hypothesized that PLFs induce proliferation and differentiation of stem cells more effectively than gingival fibroblasts (GFs) and skin fibroblasts (SFs). METHODS PLFs and GFs were isolated from extracted teeth and cultured in the presence and absence of osteogenesis-inducing factors. Mouse embryonic stem (mES) cells and SFs were purchased commercially. mES cells were incubated with culture supernatants of these fibroblasts or cocultured directly with the cells. Proliferation and mineralization in mES cells were determined at various times of incubation. Immunostaining and polymerase chain reaction were performed. The activity of mitogen-activated protein kinase and alkaline phosphatase (ALP) was also measured. RESULTS In cocultures, PLFs stimulated proliferation of mES cells more effectively than GFs or SFs. Similarly, the addition of culture supernatant of PLFs induced the most prominent proliferation of mES cells, and this was significantly inhibited by treatment with antibody against fibroblast growth factor (FGF)4 or the c-Jun N-terminal kinase inhibitor SP600125 (anthra[1,9-cd]pyrazol-6(2H)-one). Supplementation with culture supernatant from the fibroblasts induced osteogenic differentiation of mES cells in the order PLFs > GFs > SFs. These activities of PLFs were related to their potential to produce osteogenic markers, such as ALP and runt-related transcription factor-2 (Runx2), and to secrete FGF7. Pretreatment of mES cells with the extracellular signal-regulated kinase inhibitor PD98059 [2-(2-amino-3-methyoxyphenyl)-4H-1-benzopyran-4-one] or SP600125 clearly attenuated mineralization induced by culture supernatant of PLF with attendant decreases in mRNA levels of Runx2, bone sialoprotein, osteocalcin, and osteopontin. CONCLUSION PLFs regulate the proliferation and osteogenic differentiation of mES cells more strongly than GFs and SFs via the secretion of FGF through a mechanism that involves mitogen-activated protein kinase-mediated signaling.
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Affiliation(s)
- Sung-Ho Kook
- Institute of Oral Biosciences and School of Dentistry, Chonbuk National University, Jeonju, South Korea
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Neidlinger-Wilke C, Galbusera F, Pratsinis H, Mavrogonatou E, Mietsch A, Kletsas D, Wilke HJ. Mechanical loading of the intervertebral disc: from the macroscopic to the cellular level. EUROPEAN SPINE JOURNAL : OFFICIAL PUBLICATION OF THE EUROPEAN SPINE SOCIETY, THE EUROPEAN SPINAL DEFORMITY SOCIETY, AND THE EUROPEAN SECTION OF THE CERVICAL SPINE RESEARCH SOCIETY 2013; 23 Suppl 3:S333-43. [DOI: 10.1007/s00586-013-2855-9] [Citation(s) in RCA: 97] [Impact Index Per Article: 8.1] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/20/2012] [Revised: 05/10/2013] [Accepted: 06/03/2013] [Indexed: 12/24/2022]
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Li R, Wei M, Shao J. Effects of verapamil on the immediate-early gene expression of bone marrow mesenchymal stem cells stimulated by mechanical strain in vitro. Med Sci Monit Basic Res 2013; 19:68-75. [PMID: 23435320 PMCID: PMC3638684 DOI: 10.12659/msmbr.883790] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022] Open
Abstract
Background To study the effects of verapamil on the immediate-early genes (IEGs) expression of bone marrow mesenchymal stem cells (MSCs) stimulated by cyclic mechanical strain, in order to deduce the role of calcium ion channel in the cell signaling responses of MSCs to mechanical strain. Material/Methods MSCs were isolated and cultured, and the passage of 3–6 MSCs were stimulated by mechanical strain and pretreated with or without verapamil. After that, flow cytometry was used to measure the fluorescence intensity of intracellular Ca2+ immediately. The expression of early-response genes/proteins (c-fos, c-jun and c-myc) were examined by RT-PCR, immunohistochemistry and Western blot. Results Intracellular Ca2+ concentration of MSCs significantly changed when stimulated by cyclic strain, and the expression of c-fos, c-jun and c-myc remarkably increased in both mRNA and protein levels, while verapamil pre-treatment partially inhibited these effects (P<0.01). Conclusions The changes of the intracellular calcium concentration of MSCs induced by mechanical strain, dependent on the regulation of calcium channel activation, might play a role in the early response of MSCs to cyclic strain.
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Affiliation(s)
- Runguang Li
- Department of Orthopaedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou, PR China
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25
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Li Y, Li M, Tan L, Huang S, Zhao L, Tang T, Liu J, Zhao Z. Analysis of time-course gene expression profiles of a periodontal ligament tissue model under compression. Arch Oral Biol 2012; 58:511-22. [PMID: 23116693 DOI: 10.1016/j.archoralbio.2012.10.006] [Citation(s) in RCA: 37] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/05/2012] [Revised: 09/29/2012] [Accepted: 10/07/2012] [Indexed: 12/29/2022]
Abstract
OBJECTIVE We recently reported establishment of a periodontal ligament (PDL) tissue model, which may mimic the biological behaviour of human PDL under static compression in orthodontic tooth movement (OTM). In the present study, we aimed at investigating the time-course gene expression profiles of the PDL tissue model under compression. DESIGN The PDL tissue model was established through 3-D-culturing human PDL cells (PDLCs) in a thin sheet of porous poly lactic-co-glycolic acid (PLGA) scaffolds, which was subjected to 25g/cm(2) static compression for 6, 24 and 72h respectively. After that, its gene expression profiles were investigated using microarray assay, followed by signalling pathway and gene ontology (GO) analysis. Real-time RT-PCR verification was done for 15 identified genes of interest. The cell proliferation alteration was detected through EdU labelling. RESULTS (1) Among the genes identified as differentially expressed, there were numerous osteoclastogenesis inducers (including CCL20, COX-1, COX-2, RANKL, PTHrP, IL-11, IL-8, etc.), osteoclastogenesis inhibitors (including IL-1Ra, NOG, OPG, etc.), and other potential bone remodelling regulators (including STC1, CYR61, FOS, etc.). (2) According to analysis of the microarray data, the most significant pathways included Cytokine-cytokine receptor interaction (containing CCL20, RANKL, IL-11, IL-8, etc.), MAPK (containing FGF7, FOS, MAP3K8, JUN, etc.) and Cell cycle (containing CDK1, CCNA2, etc.); the most significant GOs included Cell-cell signalling (containing CCL20, STC1, FGF7, PTHrP, IL-11, IL-8, etc.), Extracellular space (containing CCL20, IL-1Ra, NOG, PTHrP, IL-11, IL-8, etc.) and Microtubule-based movement (containing KIF11, KIF23, etc.). (3) After prolonged compression, cell proliferation was significantly inhibited. CONCLUSION The present findings have expanded our understandings to the roles that PDL plays under static compression in OTM.
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Affiliation(s)
- Yu Li
- Department of Orthodontics, State Key Laboratory of Oral Diseases, West China School and Hospital of Stomatology, Sichuan University, PR China.
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Kook SH, Jang YS, Lee JC. Involvement of JNK-AP-1 and ERK-NF-κB signaling in tension-stimulated expression of Type I collagen and MMP-1 in human periodontal ligament fibroblasts. J Appl Physiol (1985) 2011; 111:1575-83. [DOI: 10.1152/japplphysiol.00348.2011] [Citation(s) in RCA: 59] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022] Open
Abstract
Type I collagen (COL I) and matrix metalloproteinase-1 (MMP-1) are the predominant matrix proteins in the extracellular matrix of the human periodontal ligament (PDL). The expression of these proteins in PDL fibroblasts (PLF) is sensitive to physiological and mechanical stress and is critical for PDL remodeling accompanied by alveolar bone remodeling. This study examined how dose tensile force regulates the expression of COL I and MMP-1 and explored the possible roles of mitogen-activated protein kinases (MAPKs) and transcription factors, such as activator protein-1 (AP-1) and nuclear factor-κB (NF-κB). Tensile force stimulated the mRNA expression of COL I and MMP-1 in the cells and also activated MAPKs including extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p38 MAPK. A pharmacological inhibitor of ERK or JNK prevented the expression of matrix genes and the nuclear translocation of c-Jun proteins in the force-applied PLF. The knockdown of c-Jun by transfecting the cells with its antisense oligonucleotides reduced the force-induced increase in matrix gene expression. In particular, the ERK inhibitor but not JNK or p38 MAPK inhibitor attenuated the force-mediated stimulation of NF-κB-DNA binding and MMP-1 expression. Overall, these results highlight the mechanotransduction pathways involved in matrix gene expression in PLF, where the tension-stimulated expression of COL I and MMP-1 is controlled by the ERK/JNK-AP-1 and ERK-NF-κB signaling pathways.
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Affiliation(s)
- Sung-Ho Kook
- Division of Hematology and Oncology, Department of Medicine, University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania
| | - Yong-Suk Jang
- Research Center of Bioactive Materials, Chonbuk National University
| | - Jeong-Chae Lee
- Research Center of Bioactive Materials, Chonbuk National University
- Cluster for Craniofacial Development and Regeneration Research, Institute of Oral Biosciences and School of Dentistry (BK21 Program), Chonbuk National University, Jeonju, South Korea
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Diercke K, Sen S, Kohl A, Lux C, Erber R. Compression-dependent Up-regulation of Ephrin-A2 in PDL Fibroblasts Attenuates Osteogenesis. J Dent Res 2011; 90:1108-15. [DOI: 10.1177/0022034511413926] [Citation(s) in RCA: 30] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/12/2023] Open
Abstract
Members of the ephrin/Eph family have recently been shown to be involved in the regulation of bone homeostasis in a murine model. The activation of the EphB4 receptor on osteoblasts by its ligand ephrin-B2 led to stimulation of osteoblastogenesis and therefore to bone formation. The activation of ephrin-A2-EphA2 signaling on osteoblasts inhibited the activation of osteoblast-specific gene expression, leading to bone resorption. Fibroblasts within the periodontal ligament periodontal ligament may be one of the first responders to orthodontic forces. Periodontal ligament fibroblasts (PDLF) are mechanoresponsive. Members of the ephrin/Eph family might link mechanical forces received by PDLF with the regulation of osteoblastogenesis on osteoblasts of the alveolar bone. To study whether ephrin-A2 is modulated upon compression, we subjected human primary PDLF to static compressive forces (30.3 g/cm2). Static compressive forces significantly induced the expression of ephrin-A2, while the expression of ephrin-B2 was significantly down-regulated. Moreover, osteoblasts of the alveolar bone stimulated with ephrin-A2 in vitro significantly suppressed their osteoblastogenic gene expression (RUNX2, ALPL) and decreased signs of osteoblastic differentiation, as demonstrated by a significantly reduced ALP activity. Together, these findings establish a role for this ligand/receptor system linking mechanical forces with the regulation of osteogenesis during orthodontic tooth movement.
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Affiliation(s)
- K. Diercke
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany
| | - S. Sen
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany
| | - A. Kohl
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany
| | - C.J. Lux
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany
| | - R. Erber
- Department of Orthodontics and Dentofacial Orthopaedics, Dental School, University of Heidelberg, Im Neuenheimer Feld 400, 69120 Heidelberg, Germany
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Asparuhova MB, Ferralli J, Chiquet M, Chiquet-Ehrismann R. The transcriptional regulator megakaryoblastic leukemia-1 mediates serum response factor-independent activation of tenascin-C transcription by mechanical stress. FASEB J 2011; 25:3477-88. [PMID: 21705668 DOI: 10.1096/fj.11-187310] [Citation(s) in RCA: 49] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/14/2023]
Abstract
The extracellular matrix protein tenascin-C (TNC) is up-regulated in processes influenced by mechanical stress, such as inflammation, tissue remodeling, wound healing, and tumorigenesis. Cyclic strain-induced TNC expression depends on RhoA-actin signaling, the pathway that regulates transcriptional activity of serum response factor (SRF) by its coactivator megakaryoblastic leukemia-1 (MKL1). Therefore, we tested whether MKL1 controls TNC transcription. We demonstrate that overexpression of MKL1 strongly induces TNC expression in mouse NIH3T3 fibroblasts and normal HC11 and transformed 4T1 mammary epithelial cells. Part of the induction was dependant on SRF and a newly identified atypical CArG box in the TNC promoter. Another part was independent of SRF but required the SAP domain of MKL1. An MKL1 mutant incapable of binding to SRF still strongly induced TNC, while induction of the SRF target c-fos was abolished. Cyclic strain failed to induce TNC in MKL1-deficient but not in SRF-deficient fibroblasts, and strain-induced TNC expression strongly depended on the SAP domain of MKL1. Promoter-reporter and chromatin immunoprecipitation experiments unraveled a SAP-dependent, SRF-independent interaction of MKL1 with the proximal promoter region of TNC, attributing for the first time a functional role to the SAP domain of MKL1 in regulating gene expression.
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Affiliation(s)
- Maria B Asparuhova
- Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation, Maulbeerstrasse 66, 4058 Basel, Switzerland
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Expression of matrix metalloproteinase-1 (MMP-1) in Wistar rat's intervertebral disc after experimentally induced scoliotic deformity. SCOLIOSIS 2011; 6:9. [PMID: 21554726 PMCID: PMC3117814 DOI: 10.1186/1748-7161-6-9] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 01/02/2011] [Accepted: 05/09/2011] [Indexed: 11/10/2022]
Abstract
Introduction A scoliotic deformity on intervertebral discs may accelerate degeneration at a molecular level with the production of metalloproteinases (MMPs). In the present experimental study we evaluated the presence of MMP-1 immunohistochemically after application of asymmetric forces in a rat intervertebral disc and the impact of the degree of the deformity on MMP-1 expression. Material-Method Thirty female Wistar rats (aged 2 months old, weighted 200 ± 10 grams) were used. All animals were age, weight and height matched. A mini Ilizarov external fixator was applied at the base of a rat tail under anaesthesia in order to create a scoliotic deformity of the intervertebral disc between the 9th and 10th vertebrae. Rats were divided into three groups according to the degree of the deformity. In group I, the deformity was 10°, in group II 30° and in group III 50°. The rats were killed 35 days after surgery. The discs were removed along with the neighbouring vertebral bodies, prepared histologically and stained immunohistochemically. Immunopositivity of disc's cells for MMP-1 was determined using a semi-quantitative scored system. Results MMP-1 immunopositivity was detected in disc cells of annulus fibrosus of all intervertebral disc specimens examined. The percentage of MMP-1 positive disc cells in annulus fibrosus in group I, II and III were 20%, 43% and 75%, respectively. MMP-1 positivity was significantly correlated with the degree of the deformity (p < 0,001). An increase of chondrocyte-like disc cells was observed in the outer annulus fibrosus and at the margin of the intervertebral disc adjacent to the vertebral end plates. The difference in the proportion of MMP-1 positive disc cells between the convex and the concave side was statistically not significant in group I (p = 0,6), in group II this difference was statistically significant (p < 0,01). In group III the concave side showed a remarkable reduction in the number of disc's cells and a severe degeneration of matrix microstructure. Conclusion The present study showed that an experimentally induced scoliotic deformity on a rat tail intervertebral disc results in over-expression of MMP-1, which is dependent on the degree of the deformity and follows a dissimilar distribution between the convex and the concave side.
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Wright V, Attia E, Bohnert K, Brown H, Bhargava M, Hannafin JA. Activation of MKK3/6, SAPK, and ATF-2/c-jun in ACL fibroblasts grown in 3 dimension collagen gels in response to application of cyclic strain. J Orthop Res 2011; 29:397-402. [PMID: 20886655 DOI: 10.1002/jor.21244] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/23/2010] [Accepted: 07/13/2010] [Indexed: 02/04/2023]
Abstract
Signal transduction pathways involved in response to cyclic tensile strain and strain deprivation in anterior cruciate ligament (ACL) fibroblasts grown in 3D collagen gels were investigated. Application of cyclic tensile strain resulted in significant activation (phosphorylation) of MKK3/6, SAPK and their downstream target transcription factors, ATF-2 and c-jun, while strain deprivation resulted in a decrease in these kinases and transcription factors. These data suggest that ACL fibroblasts cultured in 3D collagen gels respond to the mechanical environment and provide a useful system for determination of the molecular mechanisms involved in the regulation of proliferation and matrix turnover by mechanical load.
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Affiliation(s)
- Vonda Wright
- Tissue Engineering, Regeneration, and Repair Program, Research Division, Hospital for Special Surgery, 535, E 70th Street, New York, New York 10021, USA
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Early proliferation alteration and differential gene expression in human periodontal ligament cells subjected to cyclic tensile stress. Arch Oral Biol 2011; 56:177-86. [DOI: 10.1016/j.archoralbio.2010.09.009] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/27/2010] [Revised: 08/24/2010] [Accepted: 09/10/2010] [Indexed: 12/31/2022]
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Abstract
The advent of high-throughput measurements of gene expression and bioinformatics analysis methods offers new ways to study gene expression patterns. The primary goal of this study was to determine the time sequence for gene expression in a bone subjected to mechanical loading during key periods of the bone-formation process, including expression of matrix-related genes, the appearance of active osteoblasts, and bone desensitization. A standard model for bone loading was employed in which the right forelimb was loaded axially for 3 minutes per day, whereas the left forearm served as a nonloaded contralateral control. We evaluated loading-induced gene expression over a time course of 4 hours to 32 days after the first loading session. Six distinct time-dependent patterns of gene expression were identified over the time course and were categorized into three primary clusters: genes upregulated early in the time course, genes upregulated during matrix formation, and genes downregulated during matrix formation. Genes then were grouped based on function and/or signaling pathways. Many gene groups known to be important in loading-induced bone formation were identified within the clusters, including AP-1-related genes in the early-response cluster, matrix-related genes in the upregulated gene clusters, and Wnt/β-catenin signaling pathway inhibitors in the downregulated gene clusters. Several novel gene groups were identified as well, including chemokine-related genes, which were upregulated early but downregulated later in the time course; solute carrier genes, which were both upregulated and downregulated; and muscle-related genes, which were primarily downregulated.
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Affiliation(s)
- Sara M Mantila Roosa
- Department of Biomedical Engineering, Purdue University, West Lafayette, IN, USA.
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Letsiou E, Kitsiouli E, Nakos G, Lekka ME. Mild stretch activates cPLA2 in alveolar type II epithelial cells independently through the MEK/ERK and PI3K pathways. Biochim Biophys Acta Mol Cell Biol Lipids 2010; 1811:370-6. [PMID: 21185392 DOI: 10.1016/j.bbalip.2010.12.007] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/06/2010] [Revised: 12/15/2010] [Accepted: 12/16/2010] [Indexed: 10/18/2022]
Abstract
Alveolar epithelial type II cells (AT II) in which lung surfactant synthesis and secretion take place, are subjected to low magnitude stretch during normal breathing. The aim of the study was to explore the effect of mild stretch on phospholipase A(2) (PLA(2)) activation, an enzyme known to be involved in surfactant secretion. In A549 cells (a model of AT II cells), we showed, using a fluorometric assay, that stretch triggers an increase of total PLA(2) activity. Western blot experiments revealed that the cytosolic isoform cPLA(2) is rapidly phosphorylated under stretch, in addition to a modest increase in cPLA(2) mRNA levels. Treatment of A549 cells with selective inhibitors of the MEK/ERK pathway significantly attenuated the stretch-induced cPLA(2) phosphorylation. A strong interaction of cPLA(2) and pERK enzymes was demonstrated by immunoprecipitation. We also found that inhibition of PI3K pathway attenuated cPLA(2) activation after stretch, without affecting pERK levels. Our results suggest that low magnitude stretch can induce cPLA(2) phosphorylation through the MEK/ERK and PI3K-Akt pathways, independently.
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Affiliation(s)
- Eleftheria Letsiou
- Biochemistry Laboratory, Chemistry Department, University of Ioannina, 45110, Ioannina, Greece
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34
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Li Y, Tang L, Duan Y, Ding Y. Upregulation of MMP-13 and TIMP-1 expression in response to mechanical strain in MC3T3-E1 osteoblastic cells. BMC Res Notes 2010; 3:309. [PMID: 21080973 PMCID: PMC2997095 DOI: 10.1186/1756-0500-3-309] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/08/2010] [Accepted: 11/17/2010] [Indexed: 11/10/2022] Open
Abstract
BACKGROUND Mechanical strain plays a significant role in the regulation of bone matrix turnover, which is mediated in part by matrix metalloproteinase (MMP)-13 and tissue inhibitors of matrix metalloproteinase (TIMP)-1. However, little is known about the correlation between mechanical strain and osteoblastic cell activities, including extracellular matrix (ECM) metabolism. Herein, we determined the effect of different magnitudes of cyclic tensile strain (0%, 6%, 12%, and 18%) on MMP-13 and TIMP-1 mRNA and protein expression in MC3T3-E1 osteoblasts. Furthermore, we employed specific inhibitors to examine the role of distinct signal transduction pathways known to mediate cellular responses to mechanical strain. RESULTS We identified a magnitude-dependent increase in MMP-13 and TIMP-1 mRNA and protein levels in response to mechanical strains corresponding to 6%, 12%, and 18% elongation. The strain-induced increases in MMP-13 and TIMP-1 mRNA expression were inhibited by PD098059 and cycloheximide, respectively. CONCLUSIONS Our results suggest a mechanism for the regulation of bone matrix metabolism mediated by the differential expression of MMP-13 and TIMP-1 in response to increasing magnitudes of mechanical strain.
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Affiliation(s)
- Yongming Li
- Department of Orthodontics, College of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi Province 710032, PR China.
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Breshears LA, Cook JL, Stoker AM, Fox DB, Luther JK. The Effect of Uniaxial Cyclic Tensile Load on Gene Expression in Canine Cranial Cruciate Ligamentocytes. Vet Surg 2010; 39:433-43. [DOI: 10.1111/j.1532-950x.2010.00679.x] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022]
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Activation of RhoA and FAK induces ERK-mediated osteopontin expression in mechanical force-subjected periodontal ligament fibroblasts. Mol Cell Biochem 2009; 335:263-72. [PMID: 19798549 DOI: 10.1007/s11010-009-0276-1] [Citation(s) in RCA: 44] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/04/2009] [Accepted: 09/16/2009] [Indexed: 12/28/2022]
Abstract
The precise mechanism by which Rho kinase translates the mechanical signals into OPN up-regulation in force-exposed fibroblasts has not been elucidated. Human periodontal ligament fibroblasts (hPLFs) were exposed to mechanical force by centrifuging the culture plates at a magnitude of 50 g/cm(2) for 60 min. At various times of the force application, they were processed for analyzing cell viability, trypan blue exclusion, and OPN expression at protein and RNA levels. Cellular mechanism(s) of the force-induced OPN up-regulation was also examined using various kinase inhibitors or antisense oligonucleotides specific to mechanosensitive factors. Centrifugal force up-regulated OPN expression and induced a rapid and transient increase in the phosphorylation of focal adhesion kinase (FAK), extracellular signal-regulated kinase (ERK), and Elk1. Pharmacological blockade of RhoA/Rho-associated coiled coil-containing kinase (ROCK) signaling markedly reduced force-induced FAK and ERK1/2 phosphorylation. Transfecting hPLFs with FAK antisense oligonucleotide diminished ERK1/2 activation and force-induced OPN expression. Further, ERK inhibitor inhibited significantly OPN expression, Elk1 phosphorylation, and activator protein-1 (AP-1)-DNA binding activation, but not FAK phosphorylation, in the force-applied cells. These results demonstrate that FAK signaling plays critical roles in force-induced OPN expression in hPLFs through interaction with Rho/ROCK as upstream effectors and ERK-Elk1/ERK-c-Fos as downstream effectors.
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Papachristou DJ, Papachroni KK, Basdra EK, Papavassiliou AG. Signaling networks and transcription factors regulating mechanotransduction in bone. Bioessays 2009; 31:794-804. [PMID: 19444851 DOI: 10.1002/bies.200800223] [Citation(s) in RCA: 69] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/08/2023]
Abstract
Mechanical stimulation has a critical role in the development and maintenance of the skeleton. This function requires the perception of extracellular stimuli as well as their conversion into intracellular biochemical responses. This process is called mechanotransduction and is mediated by a plethora of molecular events that regulate bone metabolism. Indeed, mechanoreceptors, such as integrins, G protein-coupled receptors, receptor protein tyrosine kinases, and stretch-activated Ca(2+) channels, together with their downstream effectors coordinate the transmission of load-induced signals to the nucleus and the expression of bone-related genes. During the past decade, scientists have gained increasing insight into the molecular networks implicated in bone mechanotransduction. In the present paper, we consider the major signaling cascades and transcription factors that control bone and cartilage mechanobiology and discuss the influence of the mechanical microenvironment on the determination of skeletal morphology.
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Papachristou DJ, Papachroni KK, Papavassiliou GA, Pirttiniemi P, Gorgoulis VG, Piperi C, Basdra EK. Functional alterations in mechanical loading of condylar cartilage induces changes in the bony subcondylar region. Arch Oral Biol 2009; 54:1035-45. [PMID: 19775676 DOI: 10.1016/j.archoralbio.2009.08.010] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2008] [Revised: 08/19/2009] [Accepted: 08/26/2009] [Indexed: 12/28/2022]
Abstract
Bone remodeling is orchestrated by cells of the osteoblast lineage and involves an intricate network of cell-cell and cell-matrix interactions. This dynamic process engages systemic hormones, locally produced cytokines and growth factors, as well as the mechanical environment of the cells. In growing subjects, the mandibular condyle consists of both articular and growth components and the presence of progenitor cells is verified by their anabolic responses to growth hormones. The pathways of chondrocyte and osteoblast differentiation during endochondral bone formation are interconnected and controlled by key transcription factors. The present study was undertaken to explore the possibility and the extent by which the mechano-transduction events in chondrocytes are 'sensed' in the subchondral bony area under altered functional loading. To this end, the involvement of the JNK/ERK-AP-1/Runx2 signaling axe was investigated by immunohistochemistry in temporomandibular joints of young rats subjected to different functional mastication loads. Our results showed that mechanical load triggers differentiation phenomena through the induction of master tissue regulators, namely the expression and/or activation of the JNK-c-Jun signaling pathway components and c-Fos in subchondral osteoblasts, as well as the activation of ERK/MAPK and the cellular expression of the transcription factor Runx2 in subchondral osteoblasts.
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Kook SH, Hwang JM, Park JS, Kim EM, Heo JS, Jeon YM, Lee JC. Mechanical force induces type I collagen expression in human periodontal ligament fibroblasts through activation of ERK/JNK and AP-1. J Cell Biochem 2009; 106:1060-7. [PMID: 19206162 DOI: 10.1002/jcb.22085] [Citation(s) in RCA: 66] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/07/2022]
Abstract
Type I collagen (COL I) is the predominant collagen in the extracellular matrix of periodontal ligament (PDL), and its expression in PDL fibroblasts (PLF) is sensitive to mechanical force. However, the mechanism by which PLF induces COL I to respond to mechanical force is unclear. This study examined the nature of human PLF in mediating COL I expression in response to centrifugal force. Signal transduction pathways in the early stages of mechanotransduction involved in the force-driven regulation of COL I expression were also investigated. Centrifugal force up-regulated COL I without cytotoxicity and activated extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 kinase. ERK and JNK inhibitor blocked the expression of COL I but p38 kinase inhibitor had no effect. Centrifugal force activated activator protein-1 (AP-1) through dimerization between c-Fos and c-Jun transcription factors. ERK and JNK inhibitors also inhibited AP-1-DNA binding, c-Fos nuclear translocation, and c-Jun phosphorylation that were increased in the force-exposed PLF. Further, transfecting the cells with c-Jun antisense oligonucleotides almost completely abolished the force-induced increase of c-Jun phosphorylation and COL I induction. Our findings suggest that mechanical signals are transmitted into the nucleus by ERK/JNK signaling pathways and then stimulate COL I expression through AP-1 activation in force-exposed human PLF.
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Affiliation(s)
- Sung-Ho Kook
- Division of Biological Sciences, Chonbuk National University, Jeonju 561-756, South Korea
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40
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Rangaswami H, Marathe N, Zhuang S, Chen Y, Yeh JC, Frangos JA, Boss GR, Pilz RB. Type II cGMP-dependent protein kinase mediates osteoblast mechanotransduction. J Biol Chem 2009; 284:14796-808. [PMID: 19282289 PMCID: PMC2685661 DOI: 10.1074/jbc.m806486200] [Citation(s) in RCA: 72] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/21/2008] [Revised: 03/02/2009] [Indexed: 01/03/2023] Open
Abstract
Continuous bone remodeling in response to mechanical loading is critical for skeletal integrity, and interstitial fluid flow is an important stimulus for osteoblast/osteocyte growth and differentiation. However, the biochemical signals mediating osteoblast anabolic responses to mechanical stimulation are incompletely understood. In primary human osteoblasts and murine MC3T3-E1 cells, we found that fluid shear stress induced rapid expression of c-fos, fra-1, fra-2, and fosB/DeltafosB mRNAs; these genes encode transcriptional regulators that maintain skeletal integrity. Fluid shear stress increased osteoblast nitric oxide (NO) synthesis, leading to activation of cGMP-dependent protein kinase (PKG). Pharmacological inhibition of the NO/cGMP/PKG signaling pathway blocked shear-induced expression of all four fos family genes. Induction of these genes required signaling through MEK/Erk, and Erk activation was NO/cGMP/PKG-dependent. Treating cells with a membrane-permeable cGMP analog partly mimicked the effects of fluid shear stress on Erk activity and fos family gene expression. In cells transfected with small interfering RNAs (siRNA) specific for membrane-bound PKG II, shear- and cGMP-induced Erk activation and fos family gene expression was nearly abolished and could be restored by transducing cells with a virus encoding an siRNA-resistant form of PKG II; in contrast, siRNA-mediated repression of the more abundant cytosolic PKG I isoform was without effect. Thus, we report a novel function for PKG II in osteoblast mechanotransduction, and we propose a model whereby NO/cGMP/PKG II-mediated Erk activation and induction of c-fos, fra-1, fra-2, and fosB/DeltafosB play a key role in the osteoblast anabolic response to mechanical stimulation.
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Affiliation(s)
- Hema Rangaswami
- Department of Medicine and Cancer Center, University of California, San Diego, La Jolla, California 92093, USA
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Tan J, Kuang W, Jin Z, Jin F, Xu L, Yu Q, Kong L, Zeng G, Yuan X, Duan Y. Inhibition of NFkappaB by activated c-Jun NH2 terminal kinase 1 acts as a switch for C2C12 cell death under excessive stretch. Apoptosis 2009; 14:764-70. [DOI: 10.1007/s10495-009-0345-7] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
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Kook SH, Son YO, Hwang JM, Kim EM, Lee CB, Jeon YM, Kim JG, Lee JC. Mechanical force inhibits osteoclastogenic potential of human periodontal ligament fibroblasts through OPG production and ERK-mediated signaling. J Cell Biochem 2009; 106:1010-9. [DOI: 10.1002/jcb.22086] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
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Hou CH, Hou SM, Tang CH. Ultrasound increased BMP-2 expression via PI3K, Akt, c-Fos/c-Jun, and AP-1 pathways in cultured osteoblasts. J Cell Biochem 2009; 106:7-15. [PMID: 19009553 DOI: 10.1002/jcb.21934] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
It has been shown that ultrasound (US) stimulation accelerates fracture healing in the animal models and in clinical studies. Bone morphogenetic protein (BMP) is a crucial mediator in bone formation during fracture healing. Here we found that US stimulation increased BMP-2 expression but not other BMPs. US induced BMP-2 transcription is mediated by AP-1 element but not estrogen receptor response element and GC-rich Sp1 response element. Pretreatment of osteoblasts with phosphatidylinositol 3-kinase (PI3K) inhibitor (Ly294002) and Akt inhibitor inhibited the potentiating action of US; these results were further substantiated by transfecting with the dominant negative mutants of p85 and Akt. US stimulation increased the phosphorylation of p85 subunit of PI3K and serine 473 of Akt. Transfection of osteoblasts with c-Fos and c-Jun antisense oligonucleotide also reduced US-increased BMP-2 expression. US-increased the binding of c-Fos and c-Jun to the AP-1 element on the BMP-2 promoter and the enhancement of AP-1 luciferase activity was inhibited by Ly294002 and Akt inhibitor. Our results suggest that US increased BMP-2 expression in osteoblasts via the PI3K, Akt, c-Fos/c-Jun, and AP-1 signaling pathway.
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Affiliation(s)
- Chun-Han Hou
- Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan
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Naito K, Matsuzaka K, Ishigami K, Inoue T. Mechanical force promotes proliferation and early differentiation of bone marrow derived osteoblast-like cells in vitro. ACTA ACUST UNITED AC 2009. [DOI: 10.3353/omp.13.143] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
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Ogata T. Egr-1 mRNA induction by medium flow involves mRNA stabilization and is enhanced by the p38 inhibitor SB203580 in osteoblast-like cells. Acta Physiol (Oxf) 2008; 194:177-88. [PMID: 18485123 DOI: 10.1111/j.1748-1716.2008.01873.x] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/13/2023]
Abstract
AIM Mechanical stimuli are important for maintaining organ structure and tissue function. To elucidate signalling pathways activated by mechanical stimuli, the contribution of mRNA stabilization to induction of egr-1 mRNA by medium flow was examined and the mechanisms responsible for stabilization were analysed. An early-response gene that encodes a transcription factor, egr-1, activates transcription of several genes in response to mechanical stimuli, and was therefore selected to resolve how early-induced signals are integrated and connected to subsequent response. METHODS Mouse osteoblast-like MC3T3E1 cells were stably transfected with the chloramphenicol acetyltransferase (CAT) gene linked to the egr-1 promoter, and inductions of endogenous egr-1 and transfected CAT mRNA following medium flow were compared using real-time reverse transcriptase PCR. The mechanism of induction was examined using a transcription inhibitor and mitogen-activated protein (MAP) kinase inhibitors. Activation of MAP kinases by medium flow was investigated using western blotting. RESULTS Induction of egr-1 mRNA by medium flow was twofold higher than CAT mRNA induction. Induction of egr-1 mRNA was also observed in cells pre-treated with transcription inhibitor. The p38 inhibitor SB203580 enhanced induction of egr-1 mRNA by medium flow. Extracellular signal regulated kinase (ERK), p38 and c-Jun N-terminal kinase (JNK) were activated by medium flow. CONCLUSION A considerable part of egr-1 mRNA induction by medium flow may be due to mRNA stabilization. The p38 inhibitor SB203580 enhances induction.
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Affiliation(s)
- T Ogata
- Division of Advanced Molecular Medicine, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
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Zhao Y, Wang C, Li S, Song H, Wei F, Pan K, Zhu K, Yang P, Tu Q, Chen J. Expression of Osterix in mechanical stress-induced osteogenic differentiation of periodontal ligament cells in vitro. Eur J Oral Sci 2008; 116:199-206. [PMID: 18471237 DOI: 10.1111/j.1600-0722.2008.00533.x] [Citation(s) in RCA: 42] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2023]
Abstract
Osterix (Osx) is an osteoblast-specific transcription factor required for the differentiation of pre-osteoblasts into functional osteoblasts. This study sought to examine the changes of Osx expression in periodontal ligament cells (PDLC) subjected to mechanical force, and to investigate whether Osx is involved in the mechanical stress-induced differentiation of PDLC. Human PDLC were exposed to centrifugal force for 1-12 h. Real-time polymerase chain reaction (PCR), western blot, and immunofluorescence assays were used to examine the mRNA and protein expression of Osx and its subcellular localization. Furthermore, PDLC were transfected with the expression vector pcDNA3.1 flag-Osx and subjected to mechanical force for 6 h. The changes in alkaline phosphatase (ALP) activity and in the expression of core-binding factor alpha1 (Cbfa1), ALP, osteopontin, bone sialoprotein, osteocalcin, and collagen I were measured. After the application of mechanical force, Osx was upregulated in a time-dependent manner at both mRNA and protein levels, and Osx protein was translocated from the cytosol into the cell nuclei. Overexpression of Osx did not affect the expression of Cbfa1, but it significantly enhanced the ALP activity and the mRNA expression of all the aforementioned osteogenic marker genes, all of which increased further under mechanical stress. These results suggest that Osx might play an important role in the mechanical stress-induced osteogenic differentiation of PDLC and therefore be involved in alveolar bone remodeling during orthodontic therapy.
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Affiliation(s)
- Yanhong Zhao
- School of Stomatology, Shandong University, Jinan, Shandong, China
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Cyclic strain induces FosB and initiates osteogenic differentiation of mesenchymal cells. ACTA ACUST UNITED AC 2008. [DOI: 10.1016/j.etp.2007.11.013] [Citation(s) in RCA: 62] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/19/2022]
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Chiu YC, Huang TH, Fu WM, Yang RS, Tang CH. Ultrasound stimulates MMP-13 expression through p38 and JNK pathway in osteoblasts. J Cell Physiol 2008; 215:356-65. [PMID: 17941091 DOI: 10.1002/jcp.21322] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
It has been shown that ultrasound (US) stimulation accelerates fracture healing, bone maturation, and remodeling in the animal models and in clinical studies. One of the major factor involves in remodeling process is matrix metalloproteinases (MMPs) such as MMP-13 that has been shown to degrade the native interstitial collagens in several tissues. Here we found that US stimulation increased the secretion of MMP-13 in cultured rat osteoblasts, as shown by zymographic analysis. US stimulation also increased the mRNA level of MMP-13, c-Fos, and c-Jun. Cycloheximide (an inhibitor of protein translocation) and actinomycin D (an inhibitor of gene transcription) did not inhibit the MMP-13, c-Fos, and c-Jun mRNA expression, suggesting that such expression does not require de novo protein synthesis and not change their stabilities. p38 inhibitor, SB203580 or JNK inhibitor, SP600125 but not ERK inhibitor, PD98059 attenuated the US-induced MMP-13, c-Fos, and c-Jun expression; these results were further substantiated by transfecting with the dominant negative mutants of p38 or JNK. The binding of c-Fos and c-Jun to the AP-1 element on the MMP-13 promoter and the enhancement of AP-1 luciferase activity was enhanced by US stimulation. Taken together, our results provide evidence that US stimulation increases MMP-13 expression through p38 and JNK signaling pathway to regulate bone remodeling.
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Affiliation(s)
- Yung-Cheng Chiu
- Department of Orthopaedics, Taichung Veterans General Hospital, Taichung, Taiwan
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Maier S, Lutz R, Gelman L, Sarasa-Renedo A, Schenk S, Grashoff C, Chiquet M. Tenascin-C induction by cyclic strain requires integrin-linked kinase. BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH 2008; 1783:1150-62. [PMID: 18269918 DOI: 10.1016/j.bbamcr.2008.01.013] [Citation(s) in RCA: 43] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/18/2007] [Revised: 01/11/2008] [Accepted: 01/14/2008] [Indexed: 12/15/2022]
Abstract
Induction of tenascin-C mRNA by cyclic strain in fibroblasts depends on RhoA and Rho dependent kinase (ROCK). Here we show that integrin-linked kinase (ILK) is required upstream of this pathway. In ILK-deficient fibroblasts, RhoA was not activated and tenascin-C mRNA remained low after cyclic strain; tenascin-C expression was unaffected by ROCK inhibition. In ILK wild-type but not ILK-/- fibroblasts, cyclic strain-induced reorganization of actin stress fibers and focal adhesions, as well as nuclear translocation of MAL, a transcriptional co-activator that links actin assembly to gene expression. These findings support a role for RhoA in ILK-mediated mechanotransduction. Rescue of ILK -/- fibroblasts by expression of wild-type ILK restored these responses to cyclic strain. Mechanosensation is not entirely abolished in ILK -/- fibroblasts, since cyclic strain activated Erk-1/2 and PKB/Akt, and induced c-fos mRNA in these cells. Conversely, lysophosphatidic acid stimulated RhoA and induced both c-fos and tenascin-C mRNA in ILK -/- cells. Thus, the signaling pathways controlling tenascin-C expression are functional in the absence of ILK, but are not triggered by cyclic strain. Our results indicate that ILK is selectively required for the induction of specific genes by mechanical stimulation via RhoA-mediated pathways.
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Affiliation(s)
- Silke Maier
- Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation, CH-4058 Basel, Switzerland
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Wei F, Wang C, Zhou G, Liu D, Zhang X, Zhao Y, Zhang Y, Yang Q. The effect of centrifugal force on the mRNA and protein levels of ATF4 in cultured human periodontal ligament fibroblasts. Arch Oral Biol 2008; 53:35-43. [PMID: 17826733 DOI: 10.1016/j.archoralbio.2007.07.008] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/12/2006] [Revised: 06/11/2007] [Accepted: 07/27/2007] [Indexed: 11/30/2022]
Abstract
OBJECTIVE The aim of this study was to examine the changes of ATF4 expression in cultured human periodontal ligament fibroblasts (hPDLF) after mechanical stimuli, and to investigate whether ATF4 is essential for the mechanical stress-induced hPDLF differentiation. METHODS Reverse transcriptase polymerase chain reaction (RT-PCR) and western blotting were used to examine mRNA and protein levels of ATF4 expression in hPDLFs after application of centrifugal force. pMyc-ATF4 transfected cells were subjected to centrifugal force for 30min, and the changes of alkaline phosphatase (ALP) activity and osteocalcin (OCN), osteopontin (OPN), collagen I (COLI), bone sialoprotein (BSP) genes were measured to assess the differentiation of hPDLFs. RESULTS The mRNA and protein levels of ATF4 increased shortly and then decreased rapidly towards its pre-pressure levels. Overexpression of pMyc-ATF4 exhibited a greater increase in ALP activity and all four osteogenic genes compared to the untransfected cells in response to the centrifugal force. CONCLUSION Our results indicate that ATF4 is essential in response of hPDLFs to mechanical stress, resulting in increased differentiation of hPDLFs to osteoblast-like cells.
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Affiliation(s)
- Fulan Wei
- Department of Orthodontics, Shandong University, Shandong Province, Jinan 250012, People's Republic of China
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