This review focuses on dental pulp stem cells (DPSCs) which are mesenchymal stem cells (MSCs) and originating from the neural crest. These cells possess a high capacity for self-renewal and multilineage differentiation. Because of these traits, they represent promising sources for tissue engineering, regenerative medicine, and clinical applications. The objective of this study was to assess the extrinsic and intrinsic factors influencing DPSC characteristics and their potential in tissue engineering. This review discusses the external and internal factors affecting DPSC properties, including proliferation, migration, differentiation, and gene expression post extraction. Additionally, it explores the impact of the microenvironment-its composition and physical properties-and genetic and epigenetic regulation on DPSC behavior. Variations in the microenvironment and genetic regulation play pivotal roles in modulating DPSC functions, including their proliferation and differentiation potential. Intrinsic and extrinsic factors are key barriers to realizing the full therapeutic potential of DPSCs. A deeper understanding of the extrinsic and intrinsic factors affecting DPSC behavior is critical for optimizing their use in regenerative medicine, particularly for dental and craniofacial applications. Although DPSCs hold significant promise, challenges remain, and this review provides insights into the current limitations and future directions for DPSC-based therapies. Researchers and clinicians are offered a comprehensive resource for advancing the field.

This in vitro study assessed how shade changes induced by endodontic medicaments affect the transmission of single and multiples wavelengths of infrared light through enamel and dentin.

Eighteen extracted single-rooted permanent teeth were prepared, removing all extrinsic staining, and cementum. Tooth slices were treated for 4 weeks with UltraCal XS, Ledermix, or were untreated controls. Light transmission through enamel-dentin and dentin regions was assessed using diode lasers (660, 808, 904 nm) and a multi-wavelength light-emitting diode (LED) light source (700-1100 nm). Absorption studies were performed to evaluate light absorption by components of medicaments.

Samples treated with Ledermix showed the greatest shade change, with a corresponding reduction in visible and near infrared light transmission (P < .001) in dentin, whereas UltraCal XS had a milder effect (P < .01). Across different substrates, the greatest light transmission was seen for the multi-wavelength LED light source, followed by 904 nm, 808 nm, and finally 660 nm. Spectrophotometric studies revealed light absorption by turbid and saturated calcium hydroxide solutions.

This study shows that medicaments may influence transmission of visible red and near infrared light. Photobiomodulation protocols used in regenerative endodontics should take this effect into account, by incorporating longer near infrared wavelengths (up to 1100 nm) and using multi-wavelength light sources to account for this absorption.

Photobiomodulation (PBM) therapy is a continuously growing approach to stimulating healing and reducing inflammation and pain. However, its effects in the fields of regenerative medicine and tissue engineering are still under investigation. Studying PBM effects on the regenerative capacity of zebrafish can allow the application of novel clinical approaches where the impact of PBM will be cross-linked with the stem-cell therapeutic approaches. This study was done to establish an in-vivo experimental setup for studying the effects of laser and ultraviolet therapy on zebrafish caudal-fin regeneration and vascularization. Thirty zebrafish were randomly and equally allocated into three groups. The caudal-fins of all zebrafish were amputated under anaesthesia. In the first control group, the caudal-fin was only monitored until fully regenerated. In the second group, the amputated-fin was irradiated with ultraviolet. Finally, in the third group, the amputated-fin was irradiated with laser. Caudal-fin regeneration and vascularization were assessed at days 0, 3, 6, 9, 12, and 15 in all fish. In terms of regeneration, the results indicated that it is possible to discriminate the regenerative effect of laser with the experimental setup as laser therapy showed a statistically significant difference when compared to control-group. It was also found that regenerative stimulation of the group that received ultraviolet therapy showed significant difference when compared to the control group. In terms of vascularization, there was a statistically significant difference in all groups of the study, which may suggest that laser as well as ultraviolet have limited effects in terms of improving vascularization. This study presented a novel, simple and inexpensive method for the assessment of PBM effects on zebrafish. Laser and ultraviolet therapy appeared to act as regenerative stimulators for caudal-fin regeneration of zebrafish. However, laser therapy results were, to some extent, better than ultraviolet therapy. This novel in-vivo design of the experiment led to more rapid and reproducible results than in-vitro experiments.

Meniscus-derived stem cells (MeSCs), a unique type of MSC, have outstanding advantages in meniscal cytotherapy and tissue engineering, but the effects and molecular mechanisms of PBM on MeSCs are still unclear. We used 660-nm LED light with different energy densities to irradiate six human MeSC samples and tested their proliferation rate via cell counting, chondrogenic differentiation capacity via the DMMB assay, mitochondrial activity via the MTT assay, and gene expression via qPCR. The proliferation ability, chondrogenic capacity and mitochondrial activity of the 18 J/cm2 group were greater than those of the 4 J/cm2 and control groups. The mRNA expression levels of Akt, PI3K, TGF-β3, Ki67 and Notch-1 in the 18 J/cm2 group were greater than those in the other groups in most samples. After chondrogenic induction, the expression of Col2A1, Sox9 and Aggrecan in the 18 J/cm2 group was significantly greater than that in the 4 J/cm2 and control groups in most of the samples. The variation in the MTT values and Src, PI3K, Akt, mTOR and GSK3β levels decreased with time. The results showed that 660-nm LED red light promoted proliferation and chondrogenic differentiation and affected the gene expression of MeSCs, and the effects on gene expression and mitochondrial activity decreased with time.

Wounds represent a growing global issue demanding increased attention. To expedite wound healing, technologies are under development, and light emitting diode (LED) devices of varying wavelengths are being explored for their stimulating influence on the healing process. This article presents a systematic literature review aiming to compile, organize, and analyze the impacts of LED devices on wound healing. This review is registered on the PROSPERO platform [CRD42023403870]. Two blinded authors conducted searches in the Pubmed, Web of Science, Scopus, Embase, and ScienceDirect databases. In vitro and in vivo experimental studies assessing LED utilization in the wound healing process were included. The search yielded 1010 studies, of which 27 were included in the review. It was identified that LED stimulates different healing pathways, promoting enhanced cell proliferation and migration, angiogenesis stimulation, increased collagen deposition, and modulation of the inflammatory response. Thus, it can be concluded that the LED stimulates cellular and molecular processes contingent on the utilized parameters. The effects depend on the standards used. Cell migration and proliferation were better influenced by green and red LED. The extracellular matrix components and angiogenesis were regulated by all wavelengths and the modulation of inflammation was mediated by green, red, and infrared LEDs.

Photobiomodulation therapy (PBMT) is the application of a low-level laser device to generate physiological changes and provide therapeutic effects. Till now, the effects of PBMT on the neural differentiation of mesenchymal stem cells have been rarely reported. Herein, the potential effect and mechanism of PBMT on the neural differentiation of dental pulp stem cells (DPSCs) were preliminarily investigated in our research. The optimal dose of 3.75 J/cm2 was first screened for use in the following neural-inducing studies. Then, DPSCs were cultured in neural induction medium and treated with laser irradiation for 7 days. From the results of morphology and immunofluorescence, we found that irradiation promoted the formation of neural stem cell-like spheroids derived from DPSCs and enhanced potential neural differentiation. Furthermore, neural differentiation gene expressions of Nestin, microtubule-associated protein-2, and neural cell adhesion molecule were increased after PBMT irradiation. The protein expressions of class III β-tubulin and neurogenic differentiation factor 1 were also improved. Meanwhile, the involvement of extracellular signal-regulated kinase (ERK1/2) was investigated by western blot. Our study showed that the neural differentiation of DPSCs was promoted by PBMT, and the underlying mechanism in this process was associated with activating the ERK1/2 signaling pathway.

Background: This investigation set out to compare the impacts of low-level diode laser (LLDL) and red light-emitting diode (LED) on the survival of human dental pulp stem cells (hDPSCs) and osteogenic/odontogenic differentiation. Methods and materials: In this ex vivo experimental study, the experimental groups underwent the irradiation of LLDL (4 J/cm2 energy density) and red LED in the osteogenic medium. Survival of hDPSCs was assessed after 24 and 48 h (n = 9) using the methyl thiazolyl tetrazolium (MTT) assay. The assessment of osteogenic/odontogenic differentiation was conducted using alizarin red staining (ARS; three repetitions). The investigation of osteogenic and odontogenic gene expression was performed at two time points, specifically 24 and 48 h (n = 12). This analysis was performed utilizing real-time reverse-transcription polymerase chain reaction (RT-PCR). The groups were compared at each time point using SPSS version 24. To analyze the data, the Mann-Whitney U test, analysis of variance, Tukey's test, and t-test were utilized. Results: The MTT assay showed that LLDL significantly decreased the survival of hDPSCs after 48 h, compared with other groups (p < 0.05). The qualitative results of ARS revealed that LLDL and red LED increased the osteogenic differentiation of hDPSCs. LLDL and red LED both upregulated the expression of osteogenic/odontogenic genes, including bone sialoprotein (BSP), alkaline phosphatase (ALP), dentin matrix protein 1 (DMP1), and dentin sialophosphoprotein (DSPP), in hDPSCs. The LLDL group exhibited a higher level of gene upregulation (p < 0.0001). Conclusions: The cell survival of hDPSCs was reduced, despite an increase in osteogenic/odontogenic activity. Clinical relevance: Introduction of noninvasive methods in regenerative endodontic treatments.

The aim of this study was to determine the effect of low-level laser therapy (LLLT) on cell proliferation, mitochondrial membrane potential changes (∆Ψm), reactive oxygen species (ROS), and osteoblast differentiation of human dental pulp stem cells (hDPSCs). These cells were irradiated with 660- and 940-nm lasers for 5 s, 50 s, and 180 s. Cell proliferation was assessed using the resazurin assay, cell differentiation by RUNX2 and BMP2 expression, and the presence of calcification nodules using alizarin-red S staining. ROS was determined by the dichlorofluorescein-diacetate technique and changes in ∆Ψm by the tetramethylrhodamine-ester assay. Data were analyzed by a Student's t-test and Mann-Whitney U test. The 940-nm wavelength for 5 and 50 s increased proliferation at 4 days postirradiation. After 8 days, a significant decrease in proliferation was observed in all groups. Calcification nodules were evident in all groups, with a greater staining intensity in cells treated with a 940-nm laser for 50 s, an effect that correlated with increased RUNX2 and BMP2 expression. ROS production and Δψm increased independently of irradiation time. In conclusion, photobiomodulation (PBM) with LLLT induced morphological changes and reduced cell proliferation rate, which was associated with osteoblastic differentiation and increased ROS and Δψm, independent of wavelength and time.

Dental regeneration benefits from improving the features of dental derived stem cells. Gallium-aluminum-arsenide laser had a significant role in modification of cell behavior in different cell lines and culture conditions. Hence, exploring its mechanism and effect on dental derived stem cells would benefit prospective regenerative dental therapies.

To assess the impact of photo biomodulation by Low-Level-Laser on isolated Dental Pulp derived Stem Cells and Periodontal Ligament derived Stem Cells regarding their proliferation and osteogenic differentiation.

Isolated DPSCs and PDLSCs from impacted third molars were subjected to Gallium-aluminum-arsenide laser for 12 sec and 3.6 J/cm2. The proliferative capacity was evaluated via 3-(4,5-dimethylthiazol-2-yl),2,5-diphenyltetrazolium bromide (MTT) Assay and Trypan blue stain. Cell osteogenic differentiation potentials were assessed by alkaline phosphatase assay and alizarin red stain, polymerase chain reaction was performed to quantify Nuclear factor Kappa gene expression.

DPSCs subjected to laser bio-stimulation showed the best results regarding cell viability (MTT) and osteogenic differentiation (ALP assay), and calcium deposition at 3 intervals (3, 7, 14 days), meanwhile, PDLSCs subjected to laser bio-stimulation showed better result than control but less than DPSCs. While NF-KB gene expression was proven to be approximately comparable for both groups. Generally, the Photo-bio modulated groups showed better results than their control groups.

Low-level laser bio-stimulation (LLL) therapy improves DPSC and PDLSC osteogenic differentiation and proliferation via the activation of the NF-KB pathway. Also, the DPSCs outperformed PDLSCs in terms of performance.

These results can be beneficial information and a reference database for more research in tissue engineering, dental therapy, and regeneration.

Photobiomodulation therapy involves exposing tissues to light sources, including light-emitting diodes or low-level lasers, which results in cellular function modulation. The molecular mechanism of this treatment is revealed, demonstrating that depending on the light settings utilized, it has the potential to elicit both stimulatory and inhibitory reactions.

The current systematic review aimed to evaluate the impact of photobiomodulation therapy on dental stem cells and provide an evidence-based conclusion in this regard.

This systematic review was performed and reported based on the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) revised guidelines. PICO(S) components were employed to define the inclusion criteria. Web of Science, Scopus, Medline as well as grey literature, and google scholar were searched up to September 2021 to retrieve relevant papers.

Photobiomodulation therapy showed promising effects on the proliferation, viability, and differentiation of dental stem cells. This finding was based on reviewing related articles with a low risk of bias.

Despite the positive benefits of photobiomodulation therapy on dental stem cells, the current data do not provide a definitive conclusion on the best physical parameters for enhancing cell viability, proliferation, and differentiation.

The application of blue light (400-480 nm) in photobiotherapy remains controversial. This systematic review aimed to collect and analyze the biological effects of blue light-emitting diode (LED) on mesenchymal stem cells (MSCs). Inclusion and exclusion criteria were formulated, and relevant English articles from January 1982 to September 2022 were searched in PubMed, Scopus, and Web of Science. Nine articles with a medium (n = 4) to low (n = 5) risk of bias were included. Most of the MSCs reported were derived from human tissue; only one article used MSCs derived from mouse. The wavelength of the LED used was in the 400-480 nm range, and the irradiation modes were continuous (n = 8) and pulse waves (n = 1). A chiral polarizer was used in one such study in which the irradiance was 14 mW/cm2 and the irradiation time was 24 h. The energy densities used in other studies were between 0.378 and 72 J/cm2, and the irradiation times were between 10 and 3600 s. Blue LED light can inhibit proliferation and promote differentiation of MSCs in an appropriate energy density range, which may be related to the activation of transient receptor potential vanilloid 1 (TRPV1). Additionally, polarized light may reduce the toxic effects of blue light on MSCs. However, the heterogeneity of the design schemes and LED parameters, as well as the small number of studies, limited the conclusiveness of the review. Therefore, further studies are needed to determine the optimal irradiation strategy for promoting MSC function.

Reconstruction of lost tooth structures and the periodontium with the help of tissue engineering has found a special place in dentistry in recent years with reports of great therapeutic success. Stem cells from the periodontal ligament have the potential for high differentiation into the bone and periodontal ligament cells and are therefore a suit candidate for regenerative therapies of the periodontium and other tissues. In this regard, the use of photobiomodulation on these cells by light irradiation can be effective in increasing the efficiency of these regenerative methods. The effect of red and near-infrared lasers was investigated in pulsed and continuous modes on the cell viability, ROS production and the cell cycle of Periodontal Ligament Stem cells (PDLSCs) using MTT assay and flowcytometry techniques. The result shows that both red and near-infra-red (NIR) irradiations at 3 J/cm2 maintain cell viability. ROS generation assay indicated that in PDL stem cells irradiated with NIR laser (940 nm), ROS production was greater than in the red (660 nm) irradiated groups. Cell cycle analysis revealed that NIR irradiation can enhance the proportion of S-phase cells and declinedecline the proportion of G1-phase cells compared to the red laser irradiation groups. Moreover, this enhancement was greater in the pulsed group compared to the continuous mode group. Overall, the current study results showed that photobiomodulation can support the cell viability of PDLSCs and could affect the ROS production and cell cycle. This effect was more with 940 nm (NIR) irradiation pulsed mode compared to 660 nm (red).Communicated by Ramaswamy H. Sarma.

This experimental study aimed to assess the effect of irradiation of red light-emitting diode (LED) and Diode low-level laser (LLL) on osteogenic/odontogenic differentiation of stem cells from the apical papilla (SCAPs).

SCAPs were isolated from the human tooth root. The experimental groups were subjected to 4 J/cm² diode low level laser and red LED irradiation in osteogenic medium. The control group did not receive any irradiation. Cell viability/proliferation of SCAPs was assessed by the methyl thiazolyl tetrazolium (MTT) assay on days 1 and 2 (n = 9). Osteogenic differentiation was evaluated by alizarin red staining (ARS) (n = 3), and expression of osteogenic genes by real-time polymerase chain reaction (RT-PCR) (n = 12) on days 1 and 2. SPSS version 18 was used for data evaluation. The Kruskal-Wallis and Mann-Whitney tests were used to compare the groups at each time point.

The MTT assay showed no significant difference in cell viability/proliferation of SCAPs in the low level laser, red LED, and control groups at 24 or 48 h (P < 0.001). The ARS assessment showed that low level laser and red LED irradiation enhanced osteogenic differentiation of SCAPs. low level laser and red LED irradiation both induced over-expression of osteogenic/dentinogenic genes including alkaline phosphatase (ALP), dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP-1), and bone sialoprotein (BSP) in SCAPs. Up-regulation of genes was significantly greater in low level laser irradiation group than red LED group (P < 0.001).

Diode low level laser irradiation with 4 J/cm² energy density and red LED irradiation enhanced osteogenic differentiation of SCAPs without adversely affecting cell viability.

Human dental pulp stem cells (hDPSCs) have attracted tremendous attention in tissue regeneration engineering due to their excellent multidirectional differentiation potential. Photobiomodulation (PBM) using low-level light-emitting diodes (LEDs) or lasers has been proved to promote the osteogenesis of mesenchymal stem cells. However, the effect of LEDs on osteogenic differentiation of hDPSCs has little published data. In this work, the effect of blue LEDs with different energy densities of 2, 4, 6, 8, 10 J/cm² on osteogenic differentiation of hDPSCs was examined by using in vitro ALP staining, ALP activity, mineralization, and real-time PCR. The results showed that compared with the control group, osteogenic differentiation was significantly enhanced in blue LEDs treated groups. As the energy density increased, the level of osteogenesis initially increased and then decreased reaching the highest level at 6 J/cm². Transient receptor potential vanilloid 1 (TRPV1), a Ca²⁺ ion channel, was believed to be a potential player in osteogenesis by photobiomodulation. By immunofluorescence assay, calcium influx assay, PCR, and ALP staining, it was shown that blue LEDs irradiation can increase the activity of TRPV1 and intracellular calcium levels similarly to the agonist of TRPV1 capsaicin. Additionally, pretreatment with capsazepine, a selective TRPV1 inhibitor, was able to abrogate the osteogenic effect of blue LEDs. In conclusion, these findings proposed that blue LEDs can promote the osteogenesis of hDPSCs within the appropriate range (4-8 J/cm²) during culture of osteogenic medium, and TRPV1/Ca²⁺ may be an essential signaling pathway involved in blue LEDs-induced osteogenesis, providing new insights for the use of hDPSCs in tissue regeneration engineering.

Objective: In recent years, fractionated irradiation protocols, rather than a simple plan of exposure, have been proposed as a more effective method in the field of tissue regeneration. Thus, this study aimed at a comparative analysis of single versus double irradiation of an 808-nm diode laser, in terms of dental pulp stem cells' (DPSCs) viability and proliferation in vitro. Methods: Subcultured DPSCs were either irradiated, or not (control group), with energy densities of 3, 7, and 12 J·cm^-2 in a single- or double-session manner (24 h apart). On 0, 12, 24, 48, and 72 h postirradiation, cell viability and proliferation were evaluated through Trypan Blue and alamarBlue assays, respectively. Results: During the first 48 h postirradiation, the highest rates of DPSC proliferation were assigned to double irradiation at 3 or single exposure to 7 J⋅cm^-2, with no cytotoxic effects on cell viability. Inversely, single irradiation at 12, or a double session of exposure to 7 or 12 J⋅cm^-2, led to a significant descent in the rates of proliferation and cell viability. Conclusions: Within the limitations of this study, evidence suggests a positive impact on the biological responses of DPSCs following double session of exposure to lower energy densities as well as a single irradiation at a higher energy dosage.

The development of protocols for laser-assisted therapy demands strict compliance with comprehensive operating parametry. The purpose of this investigation was to examine the accuracy of correlation between laser control panel and fibre emission power values in a selection of diode dental lasers. Through retrospective analysis using successive systematic review and meta-analysis, it is clear that there is inconsistency in the details, and possible inaccuracies in laser power applied and associated computed data. Through a multi-centre investigation, 38 semi-conductor ("diode") dental laser units were chosen, with emission wavelengths ranging from 445 to 1064 nm. Each unit had been recently serviced according to manufacturer's recommendations, and delivery fibre assembly checked for patency and correct alignment with the parent laser unit. Subject to the output capacity of each laser, four average power values were chosen using the laser control panel-100 mW, 500 mW, 1.0 W, and 2.0 W. Using a calibrated power meter, the post-fibre emission power value was measured, and a percentage power loss calculated. For each emission, a series of six measurements were made and analysed to investigate sources of power losses along the delivery fibre, and to evaluate the precision of power loss determinations. Statistical analysis of a dataset comprising % deviations from power setting levels was performed using a factorial ANOVA model, and this demonstrated very highly significant differences between devices tested and emission power levels applied (p < 10-142 and < 10-52 respectively). The devices × emission power interaction effect was also markedly significant (p < 10-66), and this confirmed that differences observed in these deviations for each prior power setting parameter were dependent on the device employed for delivery. Power losses were found to be negatively related to power settings applied. Significant differences have emerged to recommend the need to standardize a minimum set of parameters that should form the basis of comparative research into laser-tissue interactions, both in vitro and in vivo.

This study aims to evaluate the impact of red LED irradiation on the viability, proliferation, colonogenic potential, markers expression along with osteogenic and chondrogenic differentiation of dental pulp stem cells. DPSCs were isolated from sound human permanent teeth using enzymatic digestion method and seeded with regular culture media. Cells at P4 were irradiated using red LED Light (627 nm, 2 J/cm²) and examined for growth kinetics, and multilineage differentiation using the appropriate differentiation media. The irradiated groups showed an increase in cellular growth rates, cell viability, clonogenic potential, and decrease in population doubling time compared to the control group. Cells of the irradiated groups showed enhanced differentiation towards osteogenic and chondrogenic lineages as revealed by histochemical staining using alizarin red and alcian blue stains. Photobiomodulation is an emerging promising element of tissue engineering triad besides stem cells, scaffolds, and growth factors.

The purpose of this study is to analyze the influence of InGaAlP diode laser (660 nm) with or without an odontogenic medium (OM) in the functional activity of OD-21 cells. Undifferentiated OD-21 pulp cells were cultivated with or without OM and divided into four groups (n = 5): nonirradiated control (C -), nonirradiated + OM (C +), irradiated (L -), and irradiated + OM (L +). Laser application was performed in two sessions of a 24-h interval with an irradiance of 11.3 mW/cm2, energy density of 1 J/cm², and total cumulative energy/well of 4.6 J. Cell proliferation, VEGF-164 expression, mineralization, and expression of Alp, Runx2, and Dmp1 genes, as well as immunolocalization of RUNX2 and MEPE proteins, were evaluated. Data were analyzed by statistical tests (α = 0.05). All studied groups showed a similar increase in cell proliferation with or without OM. After 7 and 10 days, a significatively higher concentration of VEGF-164 in L - group when compared to C - group was observed. A significant increase in mineralized nodules in the L + was noted when compared to C + in the same conditions. Photobiomodulation upregulated significantly Runx2 and Dmp1 expression after 10 days in L - and after 7 days in L + , with downregulation of Dmp1 after 10 days in L + group. Immunolocalization of RUNX2 and MEPE was expressive after 7 days of culture in the cytoplasm adjacent to the nucleus with a decrease after 10 days, regardless of the presence of OM. Photobiomodulation enhances metabolism associated with angiogenesis, gene expression, and mineralization regardless of the odontogenic medium in OD-21 cells.

Systematic reviews (SRs) and meta-analyses (MAs) are commonly conducted to evaluate and summarize medical literature. This is especially useful in assessing in vitro studies for consistency. Our study aims to systematically review all available quality assessment (QA) tools employed on in vitro SRs/MAs.

A search on four databases, including PubMed, Scopus, Virtual Health Library and Web of Science, was conducted from 2006 to 2020. The available SRs/MAs of in vitro studies were evaluated. DARE tool was applied to assess the risk of bias of included articles. Our protocol was developed and uploaded to ResearchGate in June 2016.

Our findings reported an increasing trend in publication of in vitro SRs/MAs from 2007 to 2020. Among the 244 included SRs/MAs, 126 articles (51.6%) had conducted the QA procedure. Overall, 51 QA tools were identified; 26 of them (51%) were developed by the authors specifically, whereas 25 (49%) were pre-constructed tools. SRs/MAs in dentistry frequently had their own QA tool developed by the authors, while SRs/MAs in other topics applied various QA tools. Many pre-structured tools in these in vitro SRs/MAs were modified from QA tools of in vivo or clinical trials, therefore, they had various criteria.

Many different QA tools currently exist in the literature; however, none cover all critical aspects of in vitro SRs/MAs. There is a need for a comprehensive guideline to ensure the quality of SR/MA due to their precise nature.

Blood extracts containing platelet products are gaining popularity in promoting healing and pulp regeneration. This study was designed to evaluate the effect of platelet-rich plasma (PRP) and gallium-aluminum-arsenide (GaAlAs) laser on proliferation and differentiation of human dental pulp stem cells (hDPSCs). In this ex vivo study, hDPSCs isolated from impacted mandibular third molars were cultured in Dulbecco's Modified Eagle's medium )DMEM(with 10% fetal bovine serum (FBS). After reaching the desired confluence, the cells were distributed into 4 groups, namely, control, PRP, laser, and PRP+laser for MTT assay and alkaline phosphatase (ALP) test. In the PRP and PRP+laser groups, 10% PRP was added to each well on the plate. In the laser and PRP+laser groups, as for the proliferation test, laser irradiation was carried out for 45 s, while 135 s was designated for ALP test. After 1, 3, and 5 days, cell proliferation and ALP activity were assessed using MTT and ALP colorimetric assay, respectively. Two-way ANOVA was utilized to analyze data. In PRP and PRP+laser groups, cell proliferation and viability increased until day 3 but began to decline afterwards until the 5th day. In the laser group, the increase in proliferation and viability was observed till day 5 which was less than the control group. Laser and control groups exhibited significantly higher cell viability and proliferation than both PRP and PRP+laser groups. ALP activity was more pronounced in PRP+laser, PRP, and laser in descending order; however, all were less than that of the control group. Only in the control group did the ALP activity augment during the 5-day period. Laser irradiation could induce pulp cell proliferation and demonstrated a better performance than PRP in this regard.