温敏性bFGF/Dex脂质体-水凝胶复合支架的神经化骨再生评价  

Evaluation of neurogenic bone regeneration of thermosensitive bFGF/Dex liposome hydrogel composite scaffold

作　　者：楚晨 宗倩[2] 寻兴祥 赵谦 袁荣涛 许晓 CHU Chen;ZONG Qian;XUN Xing-xiang;ZHAO Qian;YUAN Rong-tao;XU Xiao(School of Stomatology,Qingdao University,Qingdao 266003;Stomatology Center,Qingdao Municipal Hospital,Qingdao 266071,Shandong Province,China)

机构地区：[1]青岛大学口腔医学院,山东青岛266003 [2]青岛市市立医院口腔医学中心,山东青岛266071

出　　处：《中国口腔颌面外科杂志》2024年第4期322-328,共7页China Journal of Oral and Maxillofacial Surgery

基　　金：山东省自然科学基金(ZR2022QC034)。

摘　　要：目的:利用脂质体同时包载具有促进神经分化作用的碱性成纤维细胞生长因子(bFGF)和成骨诱导小分子(Dex),与海藻酸钠溶液混合,构建一种具有温敏性和可注射性的bFGF/Dex脂质体-水凝胶复合支架,评价复合支架促进神经分化和新骨再生的作用。方法:通过薄膜分散法制备bFGF/Dex脂质体,与RGD多肽活化后的海藻酸钠溶液混合,通过自由基聚合反应合成温敏性bFGF/Dex脂质体-水凝胶复合支架。使用Zeta粒径仪检测脂质体的粒径、多分散度和Zeta电位,扫描电镜观察支架的微观结构,利用万能材料试验机测定支架的机械强度,通过RT-PCR和免疫荧光染色检测bFGF促进hBMSCs成神经分化相关基因和蛋白的表达。构建兔颅骨缺损模型,通过micro-CT及免疫组织化学染色,评价复合支架在体内促进神经化骨再生的作用。采用SPSS 26.0软件包对数据进行统计学分析。结果:成功构建温敏性bFGF/Dex脂质体-水凝胶复合支架,该支架在常温下(25℃)呈溶液状态且具有流动性,大于临界温度(32℃)时可迅速转变为凝胶状态。支架表面呈疏松多孔的蜂窝状,能够承担一定应力。体外成神经相关基因和蛋白表达提示,bFGF在诱导hBMSCs成神经分化中具有重要作用。动物实验结果表明,复合支架具有高效促进神经化骨再生的作用。结论:具有温敏性、可注射性和突出生物功能的智能bFGF/Dex脂质体-水凝胶复合支架,为颌骨缺损修复提供了新的材料。PURPOSE:Liposomes were used to simultaneously encapsulate basic fibroblast growth factor(b FGF) that can promote nerve differentiation and osteogenic small molecule(Dex).Then,a temperature sensitive and injectable b FGF/Dex liposome-hydrogel composite sustained-release scaffold was constructed by mixing it with sodium alginate solution.METHODS:b FGF/Dex liposomes were prepared by film dispersion method,mixed with sodium alginate solution activated by RGD polypeptide,and thermosensitive b FGF/Dex liposome hydrogel composite scaffolds were synthesized by free radical polymerization.The particle size,polydispersity,and Zeta potential of liposomes were detected by Zeta particle size analyzer.Microstructure of the scaffold was observed under scanning electron microscopy,and mechanical strength of the scaffold was measured using a universal material testing machine.Real time fluorescence quantitative PCR(RT-PCR) and immunofluorescence staining were used to detect the expression of b FGF promoting neural differentiation related genes and proteins in h BMSCs.Rabbit skull defect models were constructed and the effect of the composite scaffold on promoting neurogenic bone regeneration in vivo was evaluated through micro-CT and immunohistochemical staining.SPSS 26.0 software package was used for statistical analysis.RESULTS:Thermosensitive b FGF/Dex liposome hydrogel composite scaffold was successfully constructed.It was in a solution state and had fluidity at normal temperature(25 ℃),and quickly changed to a gel state when higher than the critical temperature(32 ℃).The surface of scaffold was loose and porous,which could bear certain stress.The expression of genes and proteins related to neurogenesis suggested that b FGF had a strong promoting effect on neurogenesis in h BMSCs.The results of animal experiments showed that the composite had a good neurogenic bone regeneration effect.CONCLUSIONS:The intelligent b FGF/Dex liposome hydrogel composite scaffold with temperature sensitivity and outstanding biological function

关 键 词：颌骨缺损 骨移植材料 神经化骨再生 温敏性复合支架 

分 类 号：R782[医药卫生—口腔医学]