可注射GelMA水凝胶搭载CD271与PDGF修复骨缺损的实验研究

Experimental study of injectable GelMA hydrogel loaded with CD271 and PDGF in bone defect repair

目的:探讨负载CD271与血小板衍生生长因子(PDGF)的可注射甲基丙烯酸明胶(GelMA)水凝胶支架在SD大鼠颅骨缺损修复中的效果。方法:利用5%的GelMA水凝胶与PDGF共同构建得到GelMA/PDGF支架,然后将CD271抗体对支架进行修饰。实验共分为GelMA、GelMA/PDGF、GelMA/CD271、GelMA/PDGF/CD2714组。通过水凝胶支架与雄性SD大鼠骨髓间充质干细胞(BMSCs)在体外共同培养,检测细胞的生物相容性及成骨相关蛋白的表达情况;颅骨缺损模型使用的动物为质量约350 g的SD雄性大鼠,并在骨缺损处植入水凝胶支架;大鼠颅骨在术后8周接受Micro-CT检测以评估修复效果。结果:BMSCs与负载CD271与PDGF的GelMA水凝胶支架具有良好的生物相容性,可提高成骨相关蛋白(碱性磷酸酶、Ⅰ型胶原蛋白、骨钙素和骨桥蛋白)的表达(P<0.05),并可促进修复体内颅骨的缺损。结论:负载CD271与PDGF的混合水凝胶支架具有良好的生物相容性,在骨组织工程中极具应用潜力,能促进BMSCs的成骨分化。

Objective:To investigate the effect of an injectable gelatin methacrylamide(GelMA)hydrogel scaffold loaded with CD271 and PDGF in the repair of skull defects in SD rats.Methods:The GelMA/PDGF scaffold was jointly constructed by using a 5%GelMA hydrogel and PDGF,and then the scaffold was modified with the CD271 antibody.The experiments were divided into four groups:GelMA,GelMA/PDGF,GelMA/CD271,and GelMA/PDGF/CD271.The biocompatibility of the cells and the expression of osteogenic-related proteins were examined by co-culturing hydrogel scaffolds with male SD rat bone marrow mesenchymal stem cells(BMSCs)in vitro.The skull defect model was established in SD male rats weighing about 350 g and implanted with hydrogel scaffolds.Micro-CT of the rat skull was performed at 8 weeks after surgery to assess repair efficacy.Results:BMSCs and GelMA hydrogel scaffold loaded with CD271 and PDGF had good biocompatibility,could upregulate the expression of osteogenic-related proteins(alkaline phosphatase,collagenⅠ,osteocalcin,and osteopontin),and promoted the repair of cranial defects in vivo.Conclusions:Mixed hydrogel scaffold loading CD271 and PDGF has good biocompatibility and promotes the osteogenic differentiation of BMSCs,and has great application potential in bone tissue engineering.