Effects of calcium sulfate on proliferation of osteoblast-like MG-63 cells and osteoprotegerin/receptor activator of NF-κB ligand/receptor activator of NF-κB system / 中华创伤骨科杂志

Objective:To investigate the effects of calcium sulfate on the proliferation of osteoblast-like MG-63 cells and the osteoprotegerin/receptor activator of NF-κB ligand/receptor activator of NF-κB (OPG/RANKL/RANK) system.Methods:The extract of calcium sulfate was prepared. The osteoblast-like MG-63 cells were cultured for 24 hours in the medium containing calcium sulfate (the calcium sulfate group) and in the normal medium without calcium sulfate (the blank group), respectively. The growth of osteoblast-like MG-63 cells was observed and their proliferation detected by CCK-8. The mRNA and protein expression levels of OPG/RANKL were detected.Results:The growth of osteoblast-like MG-63 cells was fine in both groups. The CCK-8 test showed that the absorbance value at 24 h was 0.997±0.008 for the calcium sulfate group, significantly higher than that for the blank group (0.640±0.003) ( P<0.001). Respectively, the mRNA expression levels of OPG were 2.834±0.176 and 1.005±0.102 and the mRNA expression levels of RANKL 0.355±0.035 and 1.002±0.068 for the calcium sulfate group and the blank control group, showing statistically significant differences ( P<0.001). The results of Western blot showed that compared with the blank control group, the protein expression of OPG in osteoblast-like MG-63 cells was promoted but the protein expression of RANKL inhibited in the calcium sulfate group. Conclusion:Calcium sulfate may have a positive effect on bone formation, because it can promote the proliferation and activity of osteoblast-like MG-63 cells and regulate the OPG/RANKL/RANK system.

Role of TRAF6 in inflammatory responses of human osteoblast-like cells with Enterococcusfaecalis / 口腔疾病防治

Objective @#Explore the role and status of tumor necrosis factor receptor-associated factor 6 (TRAF6) in the inflammatory response of human osteoblast-like cells MG63 which was triggered by Enterococcus faecalis (E. faecalis) and its lipoteichoic acid (LTA)@*Methods@# SiRNA technology was applied to silence the TRAF6 gene of MG63 cells, Using E.faecelis and its LTA to stimulate the silence MG63 cells with different hours. After that, using real-time PCR technology to detect toll-like receptor 2 (TLR2) and TRAF6 gene expression and using ELISA assay to detect proinflammatory cytokines interleukin-1β and TNF-alpha expression levels.@*Results@#When MG63 cells was infected by E. faecalis, its LTA, TLR2 and TRAF6 gene level has increased to varying degrees (P< 0.05); interleukin-1β and TNF-alpha expression was significantly higher (P< 0.05). When TRAF6 gene of MG63 cells was silenced by siRNA, pro-inflammatory cytokines interleukin-1β, interleukin-6, interleukin -8 and TNF-alpha expression decreased significantly (P< 0.05).@*Conclusion@#E. faecalis and its toxic components is identified by MG63 cells mainly through TLR2 receptors. The major virulence factor in periapical infections caused by E. faecalis is LTA.

Screening on Active Sites and the Best Compatibility of NewYouguiyin Prescription / 世界科学技术-中医药现代化

This study was aimed to clarify the main active site and the best compatibility of the newYouguiyin prescription. The rat osteoblast-like cells was used as model cells. The investigation was made on the impact of different extraction parts of each herb and their compatibility on osteoblast proliferation. The results showed that the compatibility of butyl alcohol part fromDipsacus,Epimedium andCnidium had the most significant proliferation of osteoblast activity. It was concluded that the screening of active sites and the best compatibility of newYouguiyin prescription had laid the foundation for the development of subsequent innovation for traditional Chinese medicine (TCM).

Gene expression of MC3T3-E1 osteoblastic cells on titanium and zirconia surface

PURPOSE: This study was performed to define attachment and growth behavior of osteoblast-like cells and evaluate the gene expression on zirconia compared to titanium. MATERIALS AND METHODS: MC3T3-E1 cells were cultured on (1) titanium and (2) zirconia discs. The tetrazolium-based colorimetric assay (MTT test) was used for examining the attachment of cells. Cellular morphology was examined by scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cell differentiation rate. Mann-Whitney test was used to assess the significance level of the differences between the experimental groups. cDNA microarray was used for comparing the 20215 gene expressions on titanium and zirconia. RESULTS: From the MTT assay, there was no significant difference between titanium and zirconia (P>.05). From the SEM image, after 4 hours of culture, cells on both discs were triangular or elongated in shape with formation of filopodia. After 24 hours of culture, cells on both discs were more flattened and well spread compared to 4 hours of culture. From the ALP activity assay, the optical density of E1 cells on titanium was slightly higher than that of E1 cells on zirconia but there was no significant difference (P>.05). Most of the genes related to cell adhesion showed similar expression level between titanium and zirconia. CONCLUSION: Zirconia showed comparable biological responses of osteoblast-like cells to titanium for a short time during cell culture period. Most of the genes related to cell adhesion and signal showed similar expression level between titanium and zirconia.

The effect of several root-end filling materials on MG63 osteoblast-like cells / 대한치과보존학회지

The purpose of this study was to compare mineral trioxide aggregate (MTA; Dentsply, Tulsa Dental, Tulsa, OK, USA), which is widely used as root-end filling material, with DiaRoot BioAggregate (DB; Innovative BioCaramix Inc, Vancouver, BC, Canada), newly developed product, by using MG63 osteoblast-like cells. MTA, DB, and Intermediate Restorative Material (IRM; Dentsply Caulk, Milford, DE, USA) were used for root-end filling material while tissue culture plastic was used for control group. Each material was mixed and, the mixtures were left to set for 24 hours. MG63 cells were seeded to each group and then they were cultured for attachment for 4 hours. Following the attachment of cells to the root-end filling material, early cellular response was observed. After another 12 hours'culture, the level of attachment between cells and material was observed and in order to identify the effect of each material to bone formation, transforming growth factor beta1 (TGFbeta1) and osteocalin (OC) were estimated by using enzyme-linked immunosorbent assay (ELISA), and the amount of alkaline phosphatase (ALP) was also measured. The data were analyzed using one-way ANOVA. As a result, only at OC and the number of cells which were attached to materials, there was no statistical difference between MTA and DB. At other items, there was statistically significant difference in all groups. Although DB has not shown exactly the same cellular response like that of MTA, the number of attached cells shows that biocompatibility of the material and OC indicates bone formation rate. Therefore, if DB is used for root end filling material, it is expected to lead to similar results to MTA.

Effect of low level laser irradiation with different doses on proliferation and differentiation of human osteoblast-like cells in vitro / 吉林大学学报(医学版)

0.05). The laser group with the dose of 3 J?cm-2 showed more increase in cell survival at 24 and 48 h after being seeded as compared with control group(P

INFLUENCE OF DIFFERENT TYPES AND CONCENTRATIONS ON THE SURVIVAL OF RAPIDLY FROZEN OSTEOBLAST-LIKE CELLS: A PRELIMINARY STUDY

Effect of titanium surface roughness on cell adhesion of human osteoblast-like cells (MG63) / 대한치과보철학회지

STATEMENT OF PROBLEM: The effects of surface roughness have not or insufficiently been analyzed on earlier events such as cell adhesion though cell behavior most germane to implant performance is cell adhesion. PURPOSE: The purpose of this study was to evaluate cell adhesion of osteoblast-like cells (MG63) onto three types of titanium disks with varying roughness using the Elisa assay. MATERIALS AND METHODS: Representative disks from each group (SLA, HA, machined) were subjected to surface analysis and surface roughness was measured by the optical interferometer (Accura 2000, Intekplus Co., Seoul, Korea). Following this, MG63 cells were cultured on the titanium disks and released. Cell adhesion measurements using the Elisa assay were performed specifically at three points: after 24, 48, and 72 hours of culture. RESULTS: Among the 3 types of surface analyzed, the SLA surface was the roughest with a Ra value of 1.114 micrometer followed by HA coated surface and machined surface, consecutively. The optical density values for the SLA surface group was significantly higher than that of the machined and HA coated suface groups following 24 and 48 hours of culture. The cell culture on HA coated surface showed significantly higher values compared to the machined surface following 24, 48 and 72 hours of culture. CONCLUSION. The results suggest that surface treatment of titanium surfaces enhanced cell adhesion of human osteoblast-like cells (MG63).

Effect of estrogen receptor ? knocked down by RNA interference on osteoprotegerin expression in estrogeninduced human osteoblast-like cells / 中华内分泌代谢杂志

After estrogen receptor?(ER?)in MC,63 cell was knocked down by RNA interference, expression of osteoprotegerin(OPG)induced by 17?-estradiol was assayed by RT-PCR.17?-estradiol with various concentration obviously upregulated the expression of OPG mRNA of MG63 cell with the maxima]effect at the concentration 10~(-7)mol/L,which was not inhibited by suramin(G protein inhibitor).The present result suggested that ER?was not involved in regulation of OPG mRNA expression in MC,63 cell by 17?-estradiol.

Effects of Interleukin-1 on Cell Proliferation in UMR-106-01 Osteoblast-like cells / 대한정형외과학회잡지

Interleukin 1(IL-1), a 17.5 KD glycoprotein, is known to be associated with local bone resorption. In the present study, we examined the effects of IL-1, compared with insulin and parathyroid hormone (PTH), on DNA, protein and collagen synthesis in UMR-106-01 rat osteoblastic osteosarcoma cells. When 200 units/mL IL-1 was administered to UMR-106-01 cells, [3H]-thymidine uptake increased to 119% of the untreated control. But when 10 nM insulin was added to the cells, [3H]- thymidine uptake increased to 130% and when 1 nM PTH was added, the uptake decreased to 89% of the control. On the other hand, protein and collagen synthesis, measured by [3H]-leucine and [3H]-proline incorporation respectively, were not affected by IL-1 administration compared to the other hormones. These results indicate that IL-1 effects osteoblast-like cells, stimulating DNA synthesis via a different mechanism to the well-known cell growth factor, insulin.

Upregulatory effect of 17?-estradiol on estrogen receptor?expression in MG63 cells and human osteoblast-like cells / 中华内分泌代谢杂志

The effect of 17?-estradiol ( 17?-E2 ) on expression of estrogen receptor ?( ER?) was observed in cultured MC63 cells and human osteoblast-like (HOB) cells. The results show that the expression of ER* is in a dose-dependent manner with 17p-E2 (0-1?10-6 mol/L) in MC63 cells and has an optimal 17?-E2 concentration (1?10-8 mol/L) in HOB cells resulting in maximum expression of ERp protein.