Effect Of Calcium Silicate Incorporation On Collagen-chitosan Scaffold

Objective:Different percentage of calcium silicate scaffolds were prepared to explore the effects of physicochemical properties and biological properties of collagen-chitosan scaffolds while incorporated with different percentage of calcium silicate,and to find the best percentage of calcium silicate scaffolds to provide a more suitable choice for bone tissue engineering.Methods:1.The scaffolds with 0%,1%,2%,5%and 8%(w/v)calcium silicate were prepared by freeze-drying method.The 0%(w/v)calcium silicate group was the control group,and the 1%,2%,5%,and 8%(w/v)calcium silicate group was the experimental group.2.The physicochemical properties of the scaffolds were tested by scanning electron microscopy and universal testing machine.3.Electron-Coupled Plasma Atomic Emission Spectrometry(ICP-AES)was used to measure the concentration of Ca,P and Si ions in each group of scaffold extracts,and compare the changes of ion concentrations.4.The in vitro mineralization ability of different mass fraction calcium silicate-collagen-chitosan scaffolds was investigated by in vitro simulated mineralization experiments and scanning electron microscopy and energy dispersive spectroscopy.5.Mouse osteoblasts(MC-3T3)were cultured in vitro,cell viability was detected by CCK-8 kit,and ALP(Alkaline Phosphatase)expression was detected by alkaline phosphatase kit.To explore the change of the biological properties of calcium silicate-collagen-chitosan scaffolds with different percentag.Results:1.Calcium silicate-collagen-chitosan scaffolds with different percentage of 0%,1%,2%,5%and 8%(w/v)were successfully prepared.Scanning electron microscopy showed no significant difference in the appearance of the scaffolds.And the scaffolds have uniform pore and the suitable size.2.Compared with the control group(0%w/v calcium silicate),as the calcium silicate content increased,the mechanical properties of the scaffold increased,and the compressive elastic modulus of the scaffold increased gradually(P<0.05),and the porosity decreased(P<0.05).3.The results of ICP-AES showed that there was no significant change in the concentration of P ions in the extracts of the scaffolds.The concentration of Ca and Si ions increased with the increase of calcium silicate content,and when the content of calcium silicate reached 2%,the ion concentration does not change obviously.4.The results of CCK-8 experiment in mouse osteoblasts showed that the composite scaffolds incorporating calcium silicate could promote the proliferation of mouse osteoblasts,and the cell viability was higher when the calcium silicate content was 2%.Alkaline phosphatase experiments showed that the composite scaffold with calcium silicate could promote the expression of ALP in mouse osteoblasts(P<0.05).5.The composite scaffolds incorporating calcium silicate have varying degrees of hydroxyapatite crystal deposition,while the control group(without calcium silicate)has almost no hydroxyapatite crystal deposition.Conclusion:Appropriate amount of calcium silicate can enhance the mechanical properties of the collagen-chitosan scaffold,promote the growth of osteoblasts,and facilitate osteogenesis and in vitro mineralization.The inclusion of proper amount of calcium silicate in the scaffold can increase the concentration of Ca and Si ions in the microenvironment.We speculate that this is conducive to the growth of blood vessels,thereby increasing the survival rate of artificial implantation,and is expected to become a scaffold material for bone tissue engineering.