摘要
目的通过聚L-丙交酯-己内酯电纺纤维(PLCL)负载二甲基草酰甘氨酸(DMOG),研究其在低氧和常氧成骨诱导过程中对大鼠骨髓间充质干细胞(BMSC)的血管化和体外成骨分化的作用。方法本研究经静电纺丝技术制备PLCL电纺纤维(P)和负载DMOG的PLCL电纺纤维(PD),通过扫描电镜观察电纺纤维形貌;通过细胞骨架染色观察BMSC在不同电纺纤维上的黏附和生长状态;通过碱性磷酸酶和茜素红染色检测在不同电纺纤维上的BMSC经常氧和低氧成骨诱导7 d后的碱性磷酸酶表达和14 d时的钙沉积情况;通过RT-PCR检测在不同电纺纤维上的BMSC经常氧和低氧成骨诱导7 d和14 d时的成骨相关基因(ALP、Runx2、Coll和OCN)和促血管化相关基因(VEGF)的表达情况。结果扫描电镜结果表明,P和PD具有纤维状形态并呈现为多孔结构。细胞实验表明,BMSC可在P和PD表面黏附生长,且低氧条件下在PD上表现出更好的形态。与常氧条件相比,在低氧条件下,P和PD在7 d时的碱性磷酸酶表达减少。但低氧条件成骨诱导14 d时PD仍能促进钙的沉积。在常氧条件下,电纺纤维P可上调ALP、Runx2、Coll、OCN和VEGF的表达,但低氧条件下其对上述基因的上调作用不明显。而电纺纤维PD在常氧和低氧条件下均可促进ALP、Runx2、Coll、OCN和VEGF的表达。结论本研究制备的负载DMOG的PLCL电纺纤维在低氧条件下具有良好的体外促血管化和促成骨分化性能,预期可作为一种较好的成骨修复材料。
Objective To load dimethyloxallyl glycine(DMOG)in electrospun poly(L-lactide-co-caprolactone)(PLCL)fibers and explore its effects on the vascularization and osteogenic differentiation of rat bonemarrowmesenchymal stem cells(BMSCs)during hypoxia and normoxic osteogenic induction cultures.Methods PLCL electrospun fiber(P)and PLCL electrospun fiber loaded with DMOG(PD)were prepared by electrospinning.The morphology of electrospun fiber was observed by scanning electron microscope.The cell adhesion and growth of BMSCs on different electrospun fibers were observed by cytoskeleton staining.The osteogenic activity of BMSCs on different electrospun fibers was detected by alkaline phosphatase staining at day 7 and alizarin red staining at day 14 during hypoxia and normoxic osteogenic induction culture.The expression of osteogenic related genes(ALP,Runx2,Col1 and OCN)and vascularization related genes(VEGF)were detected by RT-PCR at 7 and 14 days after hypoxia and normoxic osteogenic induction culture.Results The scanning electron microscope images show that P and PD exhibited fibrous morphology and porous structure.Cell experiments showed that BMSCs could adhere to the surface of P and PD,and show better morphology on PD under hypoxia condition.Compared with normoxic osteogenic induction culture,the expression of alkaline phosphatase of P and PD decreases at 7 days under hypoxic osteogenic induction condition.However,PD can still promote calcium deposition under hypoxic osteogenic induction culture at 14 days.Under normoxic osteogenic induction culture,P electrospun fibers could upregulate the expression of ALP,Runx2,Col1,OCN and VEGF,but the upregulation of these genes is not obvious under hypoxic condition.However,the expression of ALP,Runx2,Col1,OCN and VEGF in PD group is enhanced in both normoxia and hypoxia conditions.Conclusion The PLCL electrospun fiber loaded with DMOG prepared in this study could promote vascularization and bone differentiation in vitro under hypoxic conditions,which is expected to be a good cand
作者
李家颖
杨隆君
花艳丽
陈起新
孟庆琛
李斌
韩凤选
Li Jiaying;Yang Longjun;Hua Yanli;Chen Qixin;Meng Qingchen;Li Bin;Han Fengxuan(Orthopaedic Institute of Soochow University,Suzhou Jiangsu,215006;The First Affiliated Hospital of Soochow University,Suzhou Jiangsu,215006,China)
出处
《生物骨科材料与临床研究》
CAS
2021年第3期13-17,23,共6页
Orthopaedic Biomechanics Materials and Clinical Study
基金
国家杰出青年科学基金项目(81925027)
国家自然科学基金(31872748)。
