目的 (1)探究锂基生物玻璃水凝胶，赋予介孔生物玻璃材料促成骨、促血管生成和抑制脂肪生成等多重生物医学功能。(2)利用3D打印技术制作导航模板，精确定位股骨头坏死病灶区域和清除病灶，将锂基生物玻璃水凝胶精确注入，观察其修复股骨头坏死的作用。方法 利用明胶和甲基丙烯酸酐制备了甲基丙烯酰化明胶(gelatin methacryloyl，GelMA)及锂基生物玻璃水凝胶(GelMA and Li-mesoporous bioglass，GM/M-Li)。扫描电子显微镜(scanning electron microscope，SEM)、透射电子显微镜(transmission electron microscope，TEM)对材料进行表征分析;同时，通过细胞活/死染色、细胞增殖实验(cell counting kit-8，CCK-8)及细胞形态观察评估材料的生物相容性;另外，使用免疫荧光染色、碱性磷酸酯酶染色、茜素红染色、成血管实验、油红O染色评估材料的成骨分化能力、成血管能力和抑制脂肪生成的能力。体内成功建立了兔股骨头坏死模型，利用3D导航模板联合髓芯减压术将坏死骨去除后填充GM/M-Li水凝胶。通过苏木素伊红(hematoxylin eosin，HE)染色、油红O染色、免疫组化染色验证其成骨、成血管和抑制脂肪生成能力。结果 SEM结果显示GM/M-Li水凝胶材料为三维多孔的结构;TEM结果表明，M-Li材料为纳米级别;离子释放实验证明了 GM/M-Li水凝胶中的锂元素，同时具有缓释作用;细胞活/死染色、CCK-8证明了 GM/M-Li水凝胶材料具有良好的生物相容性;体外成骨、成血管、抑制脂肪生成实验证实，材料中锂离子的释放有利于骨髓间充质干细胞的成骨分化、人脐静脉血管内皮细胞的小管形成，抑制了脂肪前体细胞的脂肪形成。体内HE染色和免疫荧光验证了 GM/M-Li水凝胶有效促进了骨组织、血管再生;通过油红O染色验证了 GM/M-Li水凝胶的抑制脂肪能力。结论 利用3D打印技术制作导航模板，将GM/M-Li精确注入股骨头坏死病灶区域和清除病灶。因GM/M-Li水凝胶具有良好的生物相容性，其可通过调节骨髓间充质干细胞成骨方向分化，促进股骨头坏死中骨缺损的重建，同时促进了血管的生成，减少脂肪细胞的浸润。
Study on the Effect of Injection of Lithium Bioglass Hydrogel with Navigation Template to Repair Femoral Head Necrosis
Objective(1)To explore lithium-based bioglass hydrogels and engender mesoporous bioglass biomaterials with multiple biomedical functions such as bone promotion,angiogenesis promotion and lipogenesis inhibition.(2)3D printing technology was used to make navigation templates,accurately locate and remove the lesions of osteonecrosis of the femoral head(ONFH),and lithium-based biogalglass hydrogel was accurately injected to observe its effect on repairing ONFH.Methods GelMA and lithium bioglass hydrogel(GM/M-Li)were prepared by using Gelatin(Gelatin)and methacrylic anhydride(MA).The materials were characterized by scanning electron microscope(SEM)and transmission electron microscope(TEM).At the same time,the biocompatibility of the material was evaluated by cell alive/dead staining,cell counting kit-8(CCK-8)and cell morphology observation.In addition,immunofluorescence(IF)staining,alkaline phosphatase(ALP)staining,alizarin red staining(ARS),angiogenesis assay,and oil red O staining were used to evaluate the osteogenic differentiation ability,angiogenesis ability,and lipogenesis inhibition ability of the materials.A rabbit femoral head necrosis model was successfully established in vivo.The necrotic bone was removed by 3D navigation template combined with core decompression and then filled with GM/M-Li hydrogel.Hematoxylin eosin(HE)staining,oil red O staining and immunohistochemical staining were used to verify the osteogenesis,angiogenesis and lipogenesis inhibition.Results The results of SEM showed that the GM/M-Li hydrogel has a three-dimensional porous structure.The results of TEM showed that M-Li material was in nanometer scale.The ion release experiments show that the lithium element in GM/M-Li hydrogel has slow release effect.It was proved that GM/M-Li hydrogel had good biocompatibility by cell alive/dead staining and CCK-8.In vitro experiments on osteogenesis,angiogenesis and inhibition of adipogenesis confirmed that the release of lithium ions in the material was conducive to the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCS)and the tubule formation of human umbilical vein vascular endothelial cells(HUVECs),and inhibited the adipogenesis of adipogenic precursor cells(3T3-L1).In vivo HE staining and IF proved that GM/M-Li hydrogel effectively promoted bone tissue and blood vessel regeneration.The ability of GM/M-Li hydrogel to inhibit fat was verified by oil red O staining.Conclusion GM/M-Li can be accurately injected into the necrotic area of the femoral head and cleared the lesion with 3D printed navigation template.GM/M-Li hydrogel has good biocompatibility and can promote the reconstruction of bone defect in osteonecrosis of the femoral head by regulating BMSCS differentiation in osteogenic direction,promote the formation of blood vessels and reduce the infiltration of fat cells.
navigation templateosteonecrosis of the femoral headhydrogelbioglassosteogenesis
NETL
NSTL
万方数据
维普
