MC3T3-E1 Alkaline Phosphatase Activity Research Articles

Preparation of Absorbable Suture Biomaterial Formed by Glycolide/Lactide and Its Application in the Congenital Thumb Deformity Surgery

Conducting polymers (CPs) can regulate cell behavior and promote the regeneration of damaged tissues, which are widely applied in tissue engineering. In this study, poly(3-hexylthiophene) (PHTP) was first obtained by the catalyst + transfer poly-condensation method, and then, it was mixed with poly(lactide-co-glycolide) (PLGA) to obtain a composite material with superconductivity (PHTP/PLGA). Besides, ferric chloride (FeCl3) was selected as the dopant, and the doping time was changed, so as to obtain 5 PHTP/PLGA materials with different conductivity (10-7 S/cm, 10-6 S/cm, 10-5 S/cm, 10-4 S/cm, and 10-3 S/cm). While PHTP/PLGA was physically characterized, the cell proliferation ability and blood compatibility of this series of materials were further analyzed, and the materials were made into medical sutures for the treatment of congenital thumb deformities. In the experiment, the surface morphology, hydrophilicity, and hydrophobicity of materials with different conductivity would not affect the difference in cell behavior. Using mouse embryonic osteoblasts (MC3T3-E1) as the research objects, increasing PHTP/PLGA conductivity could promote MC3T3-E1 alkaline phosphatase activity cell mineralization, and collagen type I expression. PHTP/PLGA series materials had a low hemolysis rate and were not easily affected by platelet adhesion and activation, meaning that they would not cause coagulation. The material was applied in the direct suture method of hammer deformity, and the results showed that the patient’s deformed thumb had a knot reaction and local skin edge necrosis. After dressing change, it gradually improved. Moreover, the excellent and good rate of deformed thumb based on this material was higher than that of wire extraction method and Kirschner wire fixation method (P < 0.05).

Catalpol Promotes the Proliferation and Differentiation of Osteoblasts Induced by High Glucose by Inhibiting KDM7A.

IntroductionThe protective effect of catalpol on diabetic osteoporosis (DOP) and its mechanism remain unclear. This study aimed to explore whether catalpol enhanced the proliferation and differentiation of MC3T3 cells induced by high glucose by inhibiting the expression of KDM7A.MethodsMC3T3 cells were induced by high glucose (HG) and treated with different concentrations of catalpol. The proliferation and mineralization abilities of MC3T3-E1 cells were determined by CCK-8 assay and Alizarin Red Staining, respectively. The expression of differentiation-related osteogenic proteins, KDM7A and related proteins of Wnt/β-catenin signaling pathway was analyzed by Western blot analysis. The alkaline phosphatase (ALP) activity was detected by ALP assay kits.ResultsMC3T3-E1 cells induced by high glucose showed decreased proliferation and mineralization abilities and decreased ALP activity, which were all reversed by the treatment of catalpol. High glucose induction inhibited the expression of KDM7A, Total-β-catenin, Nuclear-β-catenin and p-GSK3β, which was reversed by the treatment of catalpol. And KDM7A interference up-regulated the expression of Total-β-catenin, Nuclear-β-catenin and p-GSK3β, which was down-regulated by KDM7A overexpression. Furthermore, the proliferation and mineralization abilities and ALP activity were improved when treated with KDM7A interference and decreased when treated with KDM7A overexpression. However, SKL2001 could improve the proliferation and mineralization abilities and ALP activity of MC3T3-E1 cells.DiscussionCatalpol promotes the proliferation and differentiation of osteoblasts induced by high glucose by regulating the Wnt/β-catenin signaling pathway through KDM7A.

Cytocompatible 3D chitosan/hydroxyapatite composites endowed with antibacterial properties: toward a self-sterilized bone tissue engineering scaffold

Bone tissue scaffolds based on bioactive polymer–hydroxyapatite composites have caused infections that seriously limit their extended application. In this study, we proposed a practical ion substitution method to synthesize in situ silver phosphate on the surface of a two-level, three-dimensional chitosan/nano-hydroxyapatite scaffold. A release test of silver ions in a phosphate buffered saline (PBS) solution was performed to demonstrate that silver ions were released continuously from the silver phosphate during the initial 6 days of the study. The antibacterial property and cytocompatibility of the scaffolds treated with different concentrations of silver nitrate solution were assessed by in vitro assays with Escherichia coli and MC3T3-E1, respectively. The ability of the silver-containing scaffolds to induce bacteriostasis was confirmed by the inhibition zone (15mm) and high bactericidal rate (>99%). Cell proliferation, morphology and the alkaline phosphatase activity of MC3T3-E1 cultured on the scaffold with low silver phosphate contents were comparable with those cultured on control samples.

Effect of safflower seeds supplementation on stimulation of the proliferation, differentiation and mineralization of osteoblastic MC3T3-E1 cells

Anti-bone resorption properties of the Korean herbal formulation, Gami-Honghwain (HJ), which comprises Carthamus tinctorius L. seed and hominis placenta, were investigated. We demonstrate that the production of PGE2 is inhibited by 20–100 μg/ml HJ in nontransformed osteoblastic cells (MC3T3-E1 cells), indicating that HJ inhibits PGE2 production. The effect of HJ on the proliferation and osteoblastic differentiation in MC3T3-E1 was also studied. HJ dose-dependently increased DNA synthesis (significant at 20–100 μg/ml), and increased alkaline phosphatase (ALP) and prolyl hydroxylase activities of MC3T3-E1 cells (20–100 μg/ml), while anti-estrogen tamoxifen eliminated the stimulation of proliferation and ALP activity of MC3T3-E1 which was induced by HJ. These results indicate that HJ directly stimulates cell proliferation and differentiation of osteoblasts. Also, when we assessed the effects of HJ on osteoblastic differentiation in MC3T3-E1, HJ enhanced ALP activity and mineralization in a dose- and time-dependent fashion. This stimulatory effect of the HJ was observed at relatively low doses (significant at 20–100 μg/ml and maximal at 100 μg/ml). Northern blot analysis showed that the HJ (60 μg/ml) increased in bone morphogenetic protein-2 as well as ALP mRNA concentrations in MC3T3-E1 cells. HJ (100 μg/ml) slightly increased in type I collagen mRNA abundance throughout the culture period, whereas it markedly inhibited the gene expression of collagenase-1 between days 15 and 20 of culture. These results indicate that HJ has anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

Stimulative effects of Ulmus davidiana Planch (Ulmaceae) on osteoblastic MC3T3-E1 cells

Ulmus davidiana Planch (Ulmaceae) has long been known to have anti-inflammatory and protective effects on damaged tissue, inflammation and bone among other functions. To treat rheumatoid arthritis (RA), a herbal medicine, Ulmus davidiana Planch (Ulmaceae) extract (UD) is being used in traditional oriental medicine. The effect of UD on the proliferation and osteoblastic differentiation in non-transformed osteoblastic cells (MC3T3-E1) was studied. UD dose-dependently increased DNA synthesis (significant at 5–20 μg/ml). UD increased alkaline phosphatase (ALP) activity and prolyl hydroxylase activity of MC3T3-E1 cells (5–20 μg/ml). Antiestrogen tamoxifen eliminated the stimulation of proliferation and ALP activity of MC3T3-E1, which was induced by UD. UD at concentrations ranged from 30 to 100 μg/ml inhibited prostaglandin E2 production in MC3T3-E1. These results indicate that UD directly stimulates cell proliferation and differentiation of osteoblasts. These results also suggest and UD is effective for bone anti-resorptive action in bone cells.

Stimulative effects of Drynariae Rhizoma extracts on the proliferation and differentiation of osteoblastic MC3T3-E1 Cells

Pharmacological factors are needed to prevent bone loss that occurs with increasing age. The chemical compounds that act on bone metabolism in herbal medicines, however, are poorly understood. Effects of traditional Korean medicine, Drynariae Rhizoma [ Drynaria fortunei (kunze) J. Sm] extract (DR), on the osteoblastic proliferation and differentiation were investigated. The effect of DR, a natural phyto herb, on the proliferation and osteoblastic differentiation in non-transformed osteoblastic cells (MC3T3-E1) was studied. DR dose-dependently increased DNA synthesis (significant at 50–150 μg/ml). DR increased alkaline phosphatase (ALP) activity and prolyl hydroxylase activity of MC3T3-E1 cells (50–150 μg/ml). Antiestrogen tamoxifen eleminated the stimulation of proliferation and ALP activity of MC3T3-E1, which were induced by DR. DR at concentrations ranged from 30–100 μg/ml inhibited prostaglandin E2 production in MC3T3-E1. These results indicate that DR directly stimulates cell proliferation and differentiation of osteoblasts. These results also suggest and DR is effective for bone anti-resorptive action in bone cells.