功能化生物活性玻璃支架促眶骨再生  

Functionalized bioactive glass scaffold promotes orbital bone regeneration

作　　者：刘治蓉 任晓斌 姚清清[1,2] 吴文灿 LIU Zhirong;REN Xiaobin;YAO Qingqing;WU Wencan(School of Ophthalmology&Optometry,Biomedical Engineering,Wenzhou Medical University,Wenzhou 325035,China;Eye Hospital of Wenzhou Medical University,Wenzhou 325027,China)

机构地区：[1]温州医科大学眼视光学院(生物医学工程学院),浙江温州325035 [2]温州医科大学附属眼视光医院,浙江温州325027

出　　处：《温州医科大学学报》2024年第9期689-700,共12页Journal of Wenzhou Medical University

基　　金：浙江省自然科学基金项目(LY22H120009);国家重点研发计划项目(2021YFA1101200)。

摘　　要：目的:通过金属离子掺杂增强生物活性玻璃(BG)支架诱导干细胞成骨分化功能,并在支架表面负载缺氧诱导处理的脂肪间充质干细胞外泌体(h Exo)以增强支架的新生血管、抗炎性能,最终实现眶骨缺损修复。方法:通过碱性磷酸酶(ALP)蛋白定量和PCR分析成骨相关基因表达实验比较骨形态发生蛋白-2(BMP2)、胰岛素样生长因子-1(IGF-1)、hExo、Zn、Sr对兔脂肪间充质干细胞(rADSCs)成骨作用大小,找到最适成骨诱导剂并对其最佳作用浓度进行探索;将hExo负载于BG/Zn支架并对其新生血管、干细胞募集、抗炎功能进行体外探索。结果:本研究使用3D打印技术成功制备了Zn离子掺杂和hExo功能化的BG支架。体外实验结果发现Zn离子相对Sr离子可作为最适成骨掺杂剂,且将Zn离子掺杂到BG支架中的实验结果证明10 mmol/L掺杂量可显著促进ALP蛋白以及OPN、RUNX2、ALP、OCN和BMP2成骨相关基因表达(P<0.01);缺氧诱导的外泌体可显著促进血管生成相关因子HIF-1α、VEGF以及趋化因子SDF-1的表达(P<0.05),且hExo修饰到BG/Zn支架后,上述促血管功能保留(P<0.05)。最后分别将BG、BG/Zn、BG/Zn-hExo支架植入新西兰大白兔眶骨缺损部位进行体内实验,CT结果表明BG/Zn-hExo支架显著促进眶骨再生。结论:Zn离子与h Exo可作为一种安全有效的添加剂,使用它们对BG支架功能化后,可增强支架的成骨、成血管性能,实现骨组织工程移植材料的优化。Objective:To enhance the osteogenic differentiation function of stem cells by doping metal ions into bioactive glass(BG)scaffolds and loading hypoxia-induced adipose-derived mesenchymal stem cell-derived exosomes(hExo)on the scaffold surface to enhance the scaffold’s angiogenic and anti-inflammatory properties,ultimately achieving orbital bone defect repair.Methods:Through ALP protein quantification and PCR analysis of osteogenesis-related gene expression,we compared the osteogenic effect of BMP2,IGF-1,hExo,Zn,and Sr on rADSCs to identify the most suitable osteogenic inducer and explore its optimal concentration.With hExo loaded onto BG/Zn scaffolds,their effects on angiogenesis,stem cell recruitment,and anti-inflammatory functions were investigated in vitro.Results:This study successfully fabricated Zn-doped and hExo-functionalized BG scaffolds using 3D printing technology.In vitro experimental results showed that Zn ions,compared with Sr ions,could serve as the most suitable osteogenic dopant,and the experiment of Zn ions incorporated into the BG scaffolds,the results demonstrated that a doping concentration of 10 mmol/L significantly promoted the expression of ALP protein and osteogenic-related genes OPN,RUNX2,ALP,OCN and BMP2 as well(P<0.01).Hypoxia-induced exosomes significantly promoted the expression of angiogenic factors HIF-1α,VEGF and chemokine SDF-1(P<0.05),with the above pro-angiogenic function retained after hExo modification on the BG/Zn scaffold(P<0.05).Finally,BG,BG/Zn,and BG/Zn-hExo scaffolds were implanted into the orbital bone defect of New Zealand white rabbits for in vivo experiments,and the CT results demonstrated that the BG/Zn-hExo scaffold could significantly promote orbital bone regeneration.Conclusion:Zn ions and hExo,as safe and effective additives,can be utilized to functionalize BG scaffolds,which can enhance the scaffold’s osteogenic and angiogenic properties and optimize bone tissue engineering implant materials.

关 键 词：生物活性玻璃 成骨分化 缺氧诱导干细胞外泌体 新生血管 

分 类 号：R783.1[医药卫生—口腔医学] R77[医药卫生—临床医学] R68