Study On Preparation Of New Bioengineered Bone Scaffolds By Partial Deproteinized Bone And Type Ⅰ Collagen

Objective:In this study,partial deproteinized bone(PDPB)prepared by cationic surfactant,silane coupling agent and soaking drying were combined with type I collagen(COL I)to prepare PDPB/COL I composite scaffold.Then,rat bone marrow mesenchymal stem cells(BMSCs)were cultured and inoculated on the composite scaffold.To explore the optimal conditions and methods for the construction of tissue engineered bone and cell complex in vitro,and to provide theoretical basis for the construction of tissue engineered bone in vivo.Methods:① PDPB was prepared according to the literature method,and 5 PDPB were randomly selected and observed by naked eye and scanning electron microscope.PDPB was randomly divided into three groups.The soaking drying method was used as the control group.The cationic surfactant and silane coupling agent were used as the experimental group to make surface modification and modification of PDPB.The universal biomechanical tester detects the three-point bending resistance,compression resistance and other mechanical indexes of the material;②Bone marrow was extracted from rats,BMSCs were isolated and purified,cultured and identified in vitro,and their biological characteristics were studied;③BMSCs were inoculated into PDPB/COL I composite scaffolds treated by the three methods for co-culture.The proliferation of BMSCs on the composite scaffold was detected by flow cytometry,and the adhesion and growth of cells on the composite scaffold were observed under scanning electron microscope.Results:①The prepared PDPB was white to the naked eye,hard and brittle,and had a large number of interconnected pores on the surface,which were honeycombed.Under scanning electron microscope,the PDPB had primitive bone trabecula and loose mesh structure,and the pore wall was smooth.The protein content of the composites prepared by the three methods increased with time,and the protein content of the composites treated with silane coupling agent was significantly higher than that of the other two groups(P<0.05).In the three-point bending and compression tests,the yield strength and maximum stress of PDPB/CoL I composites treated with silane coupling agent for 72 hours were the highest compared with the other two groups(P<0.05);②Bone marrow was extracted from rats,BMSCs were isolated and purified,cultured and identified in vitro,and their biological characteristics were studied;③BMSCs were seeded into PDPB/COL I composite scaffolds treated by the three methods for co-culture.The proliferation of BMSCs on the composite scaffolds was detected by flow cytometry,and the adhesion and growth of cells on the composite scaffolds were observed under scanning electron microscopy.Conclusions:① Compared with cationic surfactant and soaking drying,PDPB treated with silane coupling agent is more favorable to the adhesion of type I collagen,and the mechanical properties of PDPB/type I collagen composite prepared by silane coupling agent are better;②The cells isolated in this experiment were BMSCs with stable proliferation,which were suitable for bone tissue engineering seed cells;③The PDPB/I collagen composite scaffolds prepared by the three methods did not cause obvious cellular immune response,and the cellular compatibility was good,among which the composite materials treated with silane coupling agent were the most favorable for cell adhesion and proliferation.