In this article, we propose a simple scheme of using berberine (BBR) to modify porous calcium phosphate ceramics (named PCPC). These BBR molecules regulate the crystallization of hydroxyapatite nanorods on PCPC. We found that these nanorods and the adsorbed BBR changed the interface micro-environment of PCPC by SEM images. The microenvironment of PCPC surface is essential for promoting BMSCs’ proliferation and differentiation. These results demonstrated that PCPC/BBR markedly improved the bone regeneration of osteoporosis rats. Moreover, PCPC/BBR had significantly increased the expression levels of ALP, osteocalcin and bone morphogenetic protein2 and RUNX2 in BMSCs originated from osteoporosis rats.

• BMSCs
• berberine
• osteoporosis
• porous ceramics

• age-related cataract
• anterior lens capsule
• basement membrane
• human lens epithelial cell
• laminin α4
• senescence

• GPNMB
• autoimmunity
• lupus
• osteoactivin
• vasculitis

• Autophagy
• Catalase
• Differentiation
• Hydrogen peroxide
• Lipid accumulation
• Peroxisome

• Bioactive factors
• Cell membrane
• Cell-free therapy
• Inflammatory bowel diseases
• Mesenchymal stem cells

• ATP1A1
• ERK/AKT
• GSCs
• Src
• proliferation
• survival

Bufadienolides are cardiotonic steroids (CTS) identified in mammals. Besides Na⁺/K⁺-ATPase inhibition, they activate signal transduction via protein–protein interactions. Diversity of endogenous bufadienolides and mechanisms of action may indicate the presence of functional selectivity and unique cellular outcomes. We evaluated whether the bufadienolides telocinobufagin and marinobufagin induce changes in proliferation or viability of pig kidney (LLC-PK1) cells and the mechanisms involved in these changes. In some experiments, ouabain was used as a positive control. CTS exhibited an inhibitory IC₅₀ of 0.20 (telocinobufagin), 0.14 (ouabain), and 3.40 μM (marinobufagin) for pig kidney Na⁺/K⁺-ATPase activity and concentrations that barely inhibited it were tested in LLC-PK1 cells. CTS induced rapid ERK1/2 phosphorylation, but corresponding proliferative response was observed for marinobufagin and ouabain instead of telocinobufagin. Telocinobufagin increased Bax:Bcl-2 expression ratio, sub-G0 cell cycle phase and pyknotic nuclei, indicating apoptosis. Src and MEK1/2 inhibitors blunted marinobufagin but not telocinobufagin effect, which was also not mediated by p38, JNK1/2, and PI3K. However, BIO, a GSK-3β inhibitor, reduced proliferation and, as telocinobufagin, phosphorylated GSK-3β at inhibitory Ser9. Combination of both drugs resulted in synergistic antiproliferative effect. Wnt reporter activity assay showed that telocinobufagin impaired Wnt/β-catenin pathway by acting upstream to β-catenin stabilization. Our findings support that mammalian endogenous bufadienolides may exhibit functional selectivity.

• ERK1/2
• GSK-3β
• Na+/K+-ATPase
• Wnt/β-catenin pathway
• bufadienolides
• cardiotonic steroids
• functional selectivity

Ovarian clear cell carcinoma (OCCC) is an aggressive neoplasm with a high recurrence rate that frequently develops resistance to platinum-based chemotherapy. There are few prognostic biomarkers or targeted therapies exist for patients with OCCC. Here, we identified that FXYD2, the modulating subunit of Na⁺/K⁺-ATPases, was highly and specifically expressed in clinical OCCC tissues. The expression levels of FXYD2 were significantly higher in advanced-stage of OCCC and positively correlated with patients' prognoses. Silencing of FXYD2 expression in OCCC cells inhibited Na⁺/K⁺-ATPase enzyme activity and suppressed tumor growth via induction of autophagy-mediated cell death. We found that high FXYD2 expression in OCCC was transcriptionally regulated by the transcriptional factor HNF1B. Furthermore, up-regulation of FXYD2 expression significantly increased the sensitivity of OCCC cells to cardiac glycosides, the Na⁺/K⁺-ATPase inhibitors. Two cardiac glycosides, digoxin and digitoxin, had a great therapeutic efficacy in OCCC cells in vitro and in vivo. Taken together, our results demonstrate that FXYD2 is functionally upregulated in OCCC and may serve as a promising prognostic biomarker and therapeutic target of cardiac glycosides in OCCC.

• FXYD2
• Na+/K+-ATPase
• cardiac glycoside
• ovarian clear cell carcinoma

• Adipose Tissue/cytology
• B7-H1 Antigen/genetics
• B7-H1 Antigen/immunology
• B7-H1 Antigen/metabolism
• Cell-Derived Microparticles/immunology
• Cell-Derived Microparticles/ultrastructure
• Cells, Cultured
• Gene Expression/drug effects
• Immunomodulation/immunology
• Inflammation Mediators/immunology
• Inflammation Mediators/metabolism
• Interferon-gamma/pharmacology
• Lymphocyte Activation/drug effects
• Lymphocyte Activation/immunology
• Mesenchymal Stem Cells/immunology
• Mesenchymal Stem Cells/metabolism
• Microscopy, Electron, Transmission
• Monocytes/immunology
• Monocytes/metabolism
• Particle Size
• T-Lymphocyte Subsets/immunology
• T-Lymphocyte Subsets/metabolism

Bone defects caused by trauma, severe infection, tumor resection and skeletal abnormalities are common osteoporotic conditions and major challenges in orthopedic surgery, and there is still no effective solution to this problem. Consequently, new treatments are needed to develop regeneration procedures without side effects. Exosomes secreted by mesenchymal stem cells (MSCs) derived from human induced pluripotent stem cells (hiPSCs, hiPSC-MSC-Exos) incorporate the advantages of both MSCs and iPSCs with no immunogenicity. However, there are no reports on the application of hiPSC-MSC-Exos to enhance angiogenesis and osteogenesis under osteoporotic conditions. HiPSC-MSC-Exos were isolated and identified before use. The effect of hiPSC-MSC-Exos on the proliferation and osteogenic differentiation of bone marrow MSCs derived from ovariectomized (OVX) rats (rBMSCs-OVX) in vitro were investigated. In vivo, hiPSC-MSC-Exos were implanted into critical size bone defects in ovariectomized rats, and bone regeneration and angiogenesis were examined by microcomputed tomography (micro-CT), sequential fluorescent labeling analysis, microfil perfusion and histological and immunohistochemical analysis. The results in vitro showed that hiPSC-MSC-Exos enhanced cell proliferation and alkaline phosphatase (ALP) activity, and up-regulated mRNA and protein expression of osteoblast-related genes in rBMSCs-OVX. In vivo experiments revealed that hiPSC-MSC-Exos dramatically stimulated bone regeneration and angiogenesis in critical-sized calvarial defects in ovariectomized rats. The effect of hiPSC-MSC-Exos increased with increasing concentration. In this study, we showed that hiPSC-MSC-Exos effectively stimulate the proliferation and osteogenic differentiation of rBMSCs-OVX, with the effect increasing with increasing exosome concentration. Further analysis demonstrated that the application of hiPSC-MSC-Exos+β-TCP scaffolds promoted bone regeneration in critical-sized calvarial defects by enhancing angiogenesis and osteogenesis in an ovariectomized rat model.

• angiogenesis.
• hiPSC-MSC-Exos
• osteogenesis
• osteoporotic bone defects

• adult stem cell differentiation
• bone marrow stem cells
• canine
• cytotoxicity
• enterocyte like cells
• essential oils
• in vitro
• mesenchymal stem cells

• Acrolein/analogs & derivatives
• Acrolein/toxicity
• Animal Feed/toxicity
• Animals
• Bone Marrow/drug effects
• Citric Acid/toxicity
• Clove Oil/toxicity
• Cytotoxins/toxicity
• Dogs
• Enterocytes/drug effects
• Eugenol/toxicity
• Zingiber officinale/toxicity
• Guanosine Monophosphate/toxicity
• Inosine Monophosphate/toxicity
• Mesenchymal Stem Cells/drug effects
• Oils, Volatile/toxicity
• Pets
• Plant Oils/toxicity
• Plant Roots/toxicity
• Sorbose/toxicity
• Thymol/toxicity

Bone is the most common site of distant relapse in breast cancer, leading to severe complications which dramatically affect the patients’ quality of life. It is believed that the crosstalk between metastatic breast cancer cells and osteoclasts is critical for breast cancer-induced osteolysis. In this study, the effects of dihydroartemisinin (DHA) on osteoclast formation, bone resorption, osteoblast differentiation and mineralization were initially assessed in vitro, followed by further investigation in a titanium-particle-induced osteolysis model in vivo. Based on the proved inhibitory effect of DHA on osteolysis, DHA was further applied to MDA-MB-231 breast cancer-induced mouse osteolysis model, with the underlying molecular mechanisms further investigated. Here, we verified for the first time that DHA suppressed osteoclast differentiation, F-actin ring formation and bone resorption through suppressing AKT/SRC pathways, leading to the preventive effect of DHA on titanium-particle-induced osteolysis without affecting osteoblast function. More importantly, we demonstrated that DHA inhibited breast tumor-induced osteolysis through inhibiting the proliferation, migration and invasion of MDA-MB-231 cells via modulating AKT signaling pathway. In conclusion, DHA effectively inhibited osteoclastogenesis and prevented breast cancer-induced osteolysis.

Cell therapies hold great promise as the next major advance in medical treatment. To enable safe, effective ex vivo culture whilst maintaining cell phenotype, growth media constituents must be carefully controlled. We have used a chemically defined mesenchymal stromal cell culture medium to investigate the influence of different preparations of human serum albumin. We examined two aspects of cell culture, growth rate as measured by population doubling time and colony forming ability which is a representative measure of the stemness of the cell population. Albumin preparations showed comparative differences in both of these criteria. Analysis of the albumin bound fatty acids also showed differences depending on the manufacturing procedure used. We demonstrated that octanoate, an additive used to stabilize albumin during pasteurization, slows growth and lowers colony forming ability during ex vivo culture. Further to this we also found the level of Na⁺/K⁺ ATPase, a membrane bound cation pump inhibited by octanoate, is increased in cells exposed to this compound. We conclude that the inclusion of human serum albumin in ex vivo growth media requires careful consideration of not only the source of albumin, but also the associated molecular cargo, for optimal cell growth and behavior.