Experiment Study On Novel Acellular Bone Extracellular Matrix/Chitosan Scaffold For Bone Tissue Engineering

Objective To fabricate novel acellular bone extracellular matrix and acellular bone extracellular matrix/chitosan scaffold for bone tissue engineering and evaluate the physicochemical properties and biocompatibility, and to investigate the feasibility as a natural scaffold of bone tissue engineering.Methods (1)Fresh porcine fumer were treated with Triton X-100 and sodium dodecyl sulfate(SDS). The acellular bone matrix-chitosan scaffold for bone tissue engineering was prepared by the solution-mixed and freeze-drying method. The ABECM/CS scaffold were investigated by histological staining, the biochemical composition, morphology, SEM observation, porosity measurement, and mechanical properties analysis. (2)BMSCs were isolated from rabbit bone marrow aspirated. Biocompatibility were evaluated using BMSCs by cell-scaffold cultured. MTT test was done to assess cytotoxicity of the extract fluid of the ABECM/CS scaffods. Test of hemolysis was done to assess the percentage of hemolysis of the extract fluid of the scaffods. Test of pyrogen was done to assess the change of temperature of the extract fluid of the scaffods.Results (1)The histological staining showed that there are no cellular debris in the scaffolds. Anti-collagen I and anti-BMP-2 immunohistochemistry staining were positive. SEM scanning showed that scaffold had a porous structure with pore diameter ranging 50-200μm. The porosity assay showed the average porosity were (53.50±4.23) %. The mechanical assay showed that the compress module of dried scaffold were (33.23±14.45) MPa and the compress of wetted scaffold were (2.27±1.13) MPa. (2) The cell adhesion test showed BMSCs grew well on the scaffold. Test for in vitro cytotoxicity of scaffold showed OD scores of three groups were not significant differences(P>0.05) at every time interval. Test of hemolysis showed the hemolysis percentage of the scaffold was 3.0%; the scaffold had no obvious pyrogenic characteristics.Conclusion The acellular bone matrix/chitosan scaffold reserves most of bone extracellular matrix after thoroughly decellularization and has good pore diameter and porosity. The ABECM/CS scaffods has good biocompatibility and to make it a novel cell-carrier for bone tissue engineering.