Ti–xZr (x = 5, 15, 25, 35, 45% wt%) alloys with low elastic modulus and high mechanical strength were fabricated as a novel implant material. The biocompatibility of the Ti–xZr alloys was evaluated by osteoblast-like cell line (MG63) in terms of cytotoxicity, proliferation, adhesion, and osteogenic induction using CCK-8 and live/dead cell assays, electron microscopy, and real-time PCR. The Ti–xZr alloys were non-toxic and showed superior biomechanics compared to commercially pure titanium (cpTi). Ti–45Zr had the optimum strength/elastic modulus ratio and osteogenic activity, thus is a promising to used as dental implants.
Backgroundp66Shc, a Src homologue and collagen homologue (Shc) adaptor protein, mediates oxidative stress signaling. The p66Shc-null mice have increased lifespan and enhanced resistance to oxidative stress. Studies have also indicated its potential role in inner ear aging, which can lead to deafness.ObjectiveThe aim of this study was to determine the effects of p66Shc down-regulation on the marginal cells (MCs) of the inner ear stria vascularis.MethodsPrimary MCs were isolated from neonatal rats and treated with glucose oxidase to induce oxidative stress. The cells were transduced with adenovirus expressing siRNA, and the knockdown was verified by Western blotting. The reactive oxygen species (ROS) levels and apoptosis were analyzed using the DCFH-DA probe and Annexin-V/7-AAD staining respectively. The ultrastructure of the differentially-treated cells was examined by transmission electron microscopy (TEM).Results: The in vitro oxidative stress model was established successfully in rat MCs. Knockdown of p66Shc alleviated the high ROS levels and apoptosis in the glucose oxidase-treated cells. In addition, glucose oxidase significantly increased the number of peroxisomes in the MCs, which was decreased by p66Shc inhibition.ConclusionOxidative stress increases p66Shc levels in the marginal cells of the inner ear, which aggravates ROS production and cellular injury. Blocking p66Shc expression can effectively reduce oxidative stress and protect the MCs.
Tantalum (Ta) and zirconium (Zr) alloys were found to had low elastic modulus and similar biomechanical characteristics as the human bone. However, the biocompatibility and osteogenic potential of Ta-xZr alloyswith different proportions (20, 30, 40 and 50% Zr by atom) remains to be investigated. In this study, the biocompatibility of Ta-xZr alloys and commercially pure titanium (cpTi) was evaluated in vitro by cell counting kit-8 assay. The adhesion of MG63 osteoblasts to the surface of the alloys was observed by fluorescence microscopy, and their morphology was analyzed by scanning electron microscopy (SEM). The expressions of alkaline phosphatase (ALP), Ki67, osteocalcin (OC), collagen-I (Col-I) and Integrin β1 mRNA in the cultured cells were determined by RT-PCR. As a result, Ta-xZr (x = 20, 30, 40 and 50 at%) alloys were non-toxic and supported proliferation of the MG63 cells. The osteoblasts adhered to the Ta-xZr alloys, and subsequently spread and proliferated rapidly. Furthermore, the cells grown on Ta-20Zr and Ta-30Zr expressed high levels of ALP, Col I and OC, indicating that the Ta-xZr alloys can induce osteogenesis. In conclusion, Ta-xZr alloys promoted the adhesion, proliferation and osteogenic differentiation of MG63 cells. The Ta-xZr composites with a higher proportion of Ta exhibited superior osteogenic activity, and Ta-30Zr is therefore a promising alternative for Ti implants.
The aim of this study was to evaluate the biocompatibility and osteogenic potential of 50%Nb–xTi–Zr (NTZ, x = 20%, 30%, 40% by weight) alloys as compared with dental commercial pure titanium (cpTi). Cell cytotoxicity assay, fluorescence microscopy and electron microscopy were used to measure the in vitro biocompatibility of NTZ. The expression of alkaline phosphatase (ALP), integrin β1, osteocalcin (OC), Ki67 and collagen-I (Col-I) at the mRNA level was measured by real-time reverse transcription-polymerase chain reaction. Osseointegration ability was determined using x-ray evaluation and histological analysis in vivo. Compared with the MG63 cells grown on cpTi on day 3, the viability, adherence and proliferation rates of cells cultured on NTZ alloys were significantly improved (p < 0.05). Furthermore, similar expression levels of Ki67, Col-I, OC and ALP were found in the MG63 cells grown on NTZ alloys and those grown on cpTi. The Cbfα1 level was significantly higher for the 50%Nb–30%Ti–Zr (NTZ3) than for the cpTi group on day 6 (p < 0.01), indicating that NTZ alloys can induce osteogenesis. A considerable amount of new bone formation and osseointegration was observed around NTZ3 implants compared with cpTi implants in vivo. Collectively, NTZ3 showed superior biocompatibility and osteogenic activity; therefore, NTZ3 may be an excellent replacement for dental Ti implants.
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