负载去铁胺聚乳酸-羟基乙酸/羟基磷灰石复合支架的血管化成骨  

Desferrioxamine-loaded polylactic-co-glycolic acid/hydroxyapatite composite scaffold: vascularization and osteogenesis

作　　者：张圣敏 曹长红[1] 王宁宁 王静[1] 李章一[2] Zhang Shengmin;Cao Changhong;Wang Ningning;Wang Jing;Li Zhangyi(Department of Stomatology,Cangzhou Medical College,Cangzhou 061001,Hebei Province,China;Department of Stomatology,Tianjin Fifth Central Hospital,Tianjin 300450,China)

机构地区：[1]沧州医学高等专科学校口腔系,河北省沧州市061001 [2]天津市第五中心医院口腔科,天津市300450

出　　处：《中国组织工程研究》2022年第34期5413-5418,共6页Chinese Journal of Tissue Engineering Research

基　　金：沧州市科技计划项目(183302068),项目负责人:张圣敏;沧州医学高等专科学校博士指导科研项目(19Z013),项目负责人:张圣敏。

摘　　要：背景:3D打印的聚乳酸-羟基乙酸/羟基磷灰石支架因其个性化的外型、优异的生物相容性和骨传导性,被广泛应用于临界骨缺损修复,然而由于支架内部缺乏早期的血管供应,植入后对骨组织再生会产生不利影响。目的:验证负载去铁胺的聚乳酸-羟基乙酸/羟基磷灰石复合支架的成骨与成血管作用。方法:采用低温3D打印技术分别构建聚乳酸-羟基乙酸/羟基磷灰石复合支架(对照支架)与负载去铁胺的聚乳酸-羟基乙酸/羟基磷灰石复合支架(实验支架),表征支架的微观形貌与体外降解性能,同时检测负载去铁胺的聚乳酸-羟基乙酸/羟基磷灰石复合支架的去铁胺缓释性能。将大鼠脂肪干细胞分别接种于两组支架上,以接种培养板的细胞为对照,分别进行细胞骨架观察、细胞增殖实验、活死细胞染色、qRT-PCR检测与免疫荧光分析。结果与结论:(1)扫描电镜下可见,两种支架具有均匀分布的大孔结构,表观形貌无明显差异,支架孔径约510μm;(2)体外降解实验显示,两组支架均缓慢稳定的降解,第8周时的降解率均小于10%;两种支架周围的pH值稳定在7.1-7.4;(3)体外缓释实验显示,实验支架浸泡在PBS中前2周时去铁胺释放较快,后期释放较为平缓,在8周内释放了75%左右;(4)荧光倒置显微镜下可见,两种支架上的脂肪干细胞均呈纺锤形,铺展较大;扫描电镜下可见,两组支架上均有很多细胞伪足,细胞在材料表面有较大的铺展;(5)CCK-8实验显示,3组细胞增殖无差异(P>0.05);(6)qRT-PCR检测显示,与对照组及对照支架组比较,实验组支架组的血管内皮生长因子、骨钙素基因表达升高(P<0.01);免疫荧光分析显示,与对照组及对照支架组比较,实验支架组的血管内皮生长因子、骨钙素蛋白表达升高(P<0.01);(7)结果表明,3D打印乳酸-羟基乙酸/羟基磷灰石/去铁胺支架有望发挥良好的成骨和血管化双重作用。BACKGROUND:Three-dimensional printed polylactic-co-glycolic acid/hydroxyapatite scaffold is widely used in bone defect repair,because of personalised shapes,excellent osteoconductivity and biocompatibility.However,implanted scaffolds have an adverse effect on bone tissue regeneration due to their poor ability to induce neovascularisation.OBJECTIVE:To verify the osteogenic and angiogenic effects of the desferrioxamine-loaded polylactic-co-glycolic acid/hydroxyapatite composite scaffold.METHODS:The polylactic-co-glycolic acid/hydroxyapatite composite scaffold(control scaffold)and desferrioxamine-loaded polylactic-co-glycolic acid/hydroxyapatite composite scaffold(experimental scaffold)could be fabricated using a low-temperature three-dimensional-printing technique.The microstructure and in vitro degradation of the scaffold were characterized.At the same time,the sustained-release properties of the deferriamine from the desferrioxamine-loaded polylactic-co-glycolic acid/hydroxyapatite composite scaffold were detected.The rat derived adipose stem cells were seeded on the two groups of scaffolds respectively,and the cells inoculated on the culture plate were used as the control group.Cytoskeleton observation,cell proliferation test,staining of live and dead cells,qRT-PCR detection,and immunofluorescence analysis were performed.RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the control scaffold and experimental scaffold had uniformly distributed macroporous structure,and there was no obvious difference.The pore size of the scaffolds was about 510μm.(2)In vitro degradation experiments showed that the degradation rate of the control scaffold and experimental scaffold was less than 10%at 8 weeks,and the pH value remained relatively stable and always remained at 7.1 to 7.4.(3)In vitro sustained-release experiments showed that the deferoxamine had a burst release from the experimental scaffolds within 2 weeks and sustained a steady level,at about 75%within 8 weeks.(4)It can be seen under a fluorescent i

关 键 词：聚乳酸-羟基乙酸 羟基磷灰石 骨再生 血管化 3D打印 去铁胺 脂肪干细胞 复合支架 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]