Response Of Osteoblast-like Cells Cultured On Zirconia To Transforming Growth Factor ?1

[Objective]The aim of this study was to MC3T3Elosteoblast behavior on zirconia under conditions cultured with low concentration Transforming growth factor ?1(4ng/L) And explore response of MC3T3E1 osteoblast was effected on zirconia, and low conce ntration TGF?1 on the MC3T3 E1 cell adhesion in the surface of zirconium oxide, prol iferation differentiation, mineralization osteogenesis.[Methods]Mouse preosteoblastic cell line(MC3T3E1) were cultured on sandblasted zirconia and discs will be divided into 4 groups:1) the normal control group(Cont);2) TGF-beta 1 group(TG);3) zirconium oxide group(Zr);4) TGF-beta 1 and zirconium oxidegroup(T GZ). At 24 hours after seeding MC3T3E1 incubated in media containing:1?Cell proliferation was assessed by MTT at 1?2?3?4?5?6?7 day.2?Alkaline phosphatase (ALP) assay kit to detect the ALP activity in cell at 3?7 day.3?Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) method was used to examine the expression of OPG, Runx2, BGP and BMP2. Collecting cells, extracting total mRNA, reverse transcription cDNA, using real-time PCR method to detect each group of osteoblasts osteogenesis in the expression of related genes at 3? 7day4?LSCM observe cell skeleton and microstructure after 3?5?7 hour.[Result]MTT test result that after loading the TGF ?1, cellular proliferation on the zirconia disc s was increased significantly after day 3rd (P<0.05); TGZ cell proliferation ability and TG group were significantly higher than that of Zr group and the control group, the difference is statistically significant (P<0.05)Osteocalcin, Runx-2,Bmp-2and osteoprotegerin expression, that of cells on TGZ group was much more highly increased at day 3 (P<0.05). The gene expression level of cells cultured on TG group was higher than that on group with Zr group at day 3(P <0.05). The gene expression level of cells cultured on TGZ was higher than that on group TG at day 7 (P<0.05). In BMP2, osteocalcin, Runx-2and osterixgene expression, that of cells on TGZ was much more highly increased at day 7. Differences between groups in more obvious.ALPase activity:compared with control group, TG, TGZ group and Zr can effectively enhance MC3T3 El-cells of alkaline phosphatase (ALP) activity, and enhance the trend change significantly over time, and then influence the osteoblast mineralization. Comparison shows that the ALP activity of TGZ group was obviously higher than that of TG and Zr group, the difference statistically significant (P< 0.05). LSCM observed that promote the spread of F-actin cytoskeleton. TGZ group cell morphological rules, F-actin crisscross is arranged with surrounding cells, cell morphology of ductility is more apparent.[Conclusions]1?The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were increased in vitro.2?Zirconia own good biocompatibility.3?With the stimulation of TGF ?1, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells.