To Evaluate The Cytocompatibility And Osteogenic Ability Of β-tricalcium Phosphate-hydroxyapatite-graphene Oxide Composites

Animal bone waste is increasing worldwide due to the increasing consumption of different meats and meat products.However,a large number of animal bones are used to process low-end products or are directly abandoned every year,which makes this resource not effectively used and causes a large amount of waste.With the rapid development of animal husbandry in recent years,it is considered to be a sustainable strategy to use waste animal bone as xenogeneic bone for bone repair materials.Xenogeneic and allogeneic bone repair materials are mainly used to remove their immunogenicity by calcination,while retaining the good osteogenic induction of bone.Under different calcination conditions,the main components are slightly different,mainly divided into high purity hydroxyapatite and β-tricalcium phosphate.Objective: To evaluate the cytocompatibility of β-tricalcium phosphate(β-TCP),hydroxyapatite(HA)and graphene oxide(GO)composite on MC3T3-E1,a mouse embryonic osteoblast precursor cell line,and its effect on osteogenic differentiation in vitro.To provide a new perspective for the exploration of xenogeneic bone repair materials.Methods: 1.Fresh bovine bovine trabecular bone was calcinated at different temperatures in a maifu furnace to obtain β-TCP material.Then β-TCP material was immersed in HA-GO composite material solution and modified by HA and GO to obtainβ-TCP material,HA-GO composite material and β-TCP/HA/GO composite material,respectively.2.The main components of the composite materials were detected by Fourier transform infrared spectroscopy(FTIR),scanning electron microscopy(SEM),transmission electron microscopy(TEM),X-ray energy spectrometer(EDX),etc.,and the basic characteristics of the composite materials were obtained to provide data support for subsequent experiments.3.MC3T3-E1 cells were cultured in vitro,and MTT kit was used to verify the cytotoxicity of the materials in vitro.The biocompatibility of the composites was tested by phalloidin staining and calculating the adhesion rate of MC3T3-E1 cells on the materials.4.The differentiation of MC3T3-E1 into osteoblasts was detected by alizarin red staining,alkaline phosphatase(ALP)kit,RT-QPCR,and Western Blot to reveal the ability of the composite to induce osteogenic differentiation.Results: Fresh bovine cancellous bone was calcinated at different temperatures in a mufter to obtain β-TCP material.After HA and GO modification,β-TCP/HA/GO composite material was finally obtained.The elastic modulus of β-TCP /HA/GO was significantly different from that of β-TCP(0.01<P<0.05).In vitro experiments on MC3T3-E1 cells showed that the materials were non-toxic and had good biocompatibility.Compared with β-TCP,the ALP activity of MC3T3-E1 cells in β-TCP/HA/GO and HA-GO was significantly increased(P < 0.01).The expression of osteogenesis-related genes in β-TCP/HA/GO was significantly higher than that in HAGO and β-TCP(P < 0.01).Conclusion: β-TCP/HA/GO is a composite material with good mechanical properties,cell compatibility and osteogenic induction.This study provides a basis for whether the β-TCP/HA/GO composite material can be used as a bone repair material,and provides a new idea for the utilization of waste animal bone.