| Worldwide,the number of patients with bone damage and bone deformity caused by trauma,congenital dysplasia,occupying lesions and bone diseases is as high as tens of millions each year,and the number of patients is increasing every year.Therefore,a reasonable repair method and a large amount of bone repair scaffold material are urgently needed.At present,the research of bone repair scaffold materials is mainly focused on solving the consistency between the inductive activity of the scaffold material and the autologous bone activity,so as to quickly repair the bone defect,promote the generation of new bone,and restore the morphology and function of the bone defect.Therefore,in the process of bone defect repair,the scientific application of repair methods and composite scaffold materials is extremely important.Objective: This paper is to explore the effects of hydroxyapatite(HA)mineralization on calcined bone surface on mechanical properties,osteogenic ability of bone marrow mesenchymal stem cells(BMSCs),and osteogenesis in vivo.It will provide new ideas for the exploration of bone scaffold materials.Methods: 1.Bovine cancellous bone is subjected to high-temperature calcination treatment to obtain a calcined bone material,namely ?-tricalcium phosphate material(?-TCP).After soaking in SBF,the mineralized calcined bone material with HA on its surface was obtained.The calcined bone material,namely ?-TCP/HA material,is used to detect the pore size,porosity,main components,and mechanical properties of the composite scaffold material through energy spectroscopy(EDX)and universal tester,to obtain the basic characteristics of the composite scaffold material.Finally,data support is provided for subsequent experiments;2.BMSCs were isolated directly from rat femur using rat lymphocyte isolate solution.Cell morphology and growth trends were observed to obtain high-purity BMSCs;3.In vitro cytotoxicity of the material was detected by live-dead fluorescence and MTT box;4.In vitro adhesion experiments between materials and cells were used to detect biocompatibility;5.ALP kit and QRT-PCR method detect to early osteogenic protein ALP and m RNA expression of osteogenic gene,revealing the ability of composite scaffolds to induce osteogenesis;6.Bone graft materials were implanted into animal models of bone defects to observe the repairing ability of bone defects,content of new bone and biodegradation rate of scaffold material to investigate the effect of composite scaffolds combined with BMSCs on osteogenesis.Result: 1.The main component of calcined bone is ?-TCP.The main component of calcined bone after surface mineralization is ?-TCP/HA.Its pore size is 100~400 ?m,and the porosity is(71.52±0.18)% and(70.86.±0.22)%,there is no significant difference(P>0.05).The mechanical properties of mineralized calcined bone material are significantly higher than that of calcined bone material(P<0.05).2.High purity BMSCs were obtained.3.Calcined bone and mineralized modified calcined bone materials were biocompatible.4.BMSCs and mineralized calcined bone materials have good combination.5.Compared with calcined bone material,the expression of ALP in mineralized calcined bone material was significantly increased(P<0.01),and the expression level of osteogenic COL ? gene was significantly increased(P<0.05).There was no significant change in OPN gene m RNA,and the expression level of OCN gene m RNA was significant increased(P<0.05).6.In order to study the osteogenesis effect of the composite of mineralized calcined bone and BMSCs,the repairing ability,the content of new bone and the biodegradation rate of the scaffold were observed.Conclusion: Mineralized calcined bone scaffolds have good cell compatibility,osteoinductive activity,physical and chemical properties,which can improve the osteogenic ability of cells.In addition,the mineralized bone scaffold material with BMSCs can be implanted into the defect site,which can quickly promote formation of new bone and shorten the healing time of bone defects. |
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