南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (8): 1183-1190.doi: 10.12122/j.issn.1673-4254.2021.08.09

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负载成骨生长肽的静电纺丝 PLGA 支架可加速大鼠颅骨缺损再生

王 聪，桑伟林，陈燕敏，宋滇文   

  1. 南京医科大学附属上海一院临床医学院骨科，上海 201620；上海市第一人民医院教育处，上海 201620
  • 出版日期:2021-08-20 发布日期:2021-09-07

Electrospun PLGA scaffold loaded with osteogenic growth peptide accelerates cranial bone repair in rats

WANG Cong, SANG Weilin, CHEN Yanmin, SONG Dianwen   

  1. Department of Orthopedics, Shanghai General Hospital of Nanjing Medical University, Shanghai 201620, China; Department of Education, Shanghai First People's Hospital, Shanghai 201620, China
  • Online:2021-08-20 Published:2021-09-07

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摘要： 目的 从体内外水平评价一种负载了成骨生长肽（OGP）的聚乳酸-羟基乙酸共聚物（PLGA）纤维支架作为新型骨组织工程支架的可行性。方法 采用静电纺丝法制备支架，一共有4组。对照组：纯PLGA支架（不含OGP的 PLGA支架）；实验组：0.1%OGP@PLGA（电纺含有0.1%OGP的PLGA溶液制得的支架）、0.2%OGP@PLGA（电纺含有0.2%OGP的PLGA溶液制得的支架）、0.4%OGP@PLGA（电纺含有0.4%OGP的PLGA溶液制得的支架）。通过扫描电子显微镜（SEM）观察支架的微观结构，将材料浸泡在PBS中观察支架中OGP的释放规律，CCK-8和活死细胞染色实验评估支架的体外生物相容性，ALP活性检测和ARS染色评估支架上大鼠骨髓间充质干细胞（rBMSCs）的体外成骨分化水平，在雄性SD大鼠上制备直径为5 mm大小的颅骨缺损模型，将支架植入8周后利用Micro-CT检测、HE染色和Masson染色分析缺损处的骨修复情况。结果 SEM结果显示，支架具有类细胞外基质（ECM）的纤维结构，负载的OGP能持续从支架内缓释长达1月以上，将细胞与支架共培养4、7 d后，负载高浓度OGP（OGP浓度大于2%）的PLGA支架细胞增殖率显著高于纯PLGA支架（P<0.01），ALP活性检测结果显示第14天时，在负载0.4%含量OGP的PLGA支架上rBMSCs的ALP活性最高（P<0.01）。ARS染色结果显示，细胞14 后在负载0.4% OGP的PLGA支架上分泌的钙化结节最多。Micro-CT扫描结果发现，负载0.4% OGP的PLGA组材料周围较其他两组有更多的新骨生成（P<0.01）。此外组织学HE和Masson染色结果和以上结果类似。结论 负载OGP的静电纺丝PLGA支架有效模拟了体内细胞外基质，具有良好的生物相容性及促成骨分化能力，是一种具有潜在应用价值的新型骨组织工程支架。

关键词: 静电纺丝；成骨生长肽；PLGA；骨组织工程

Abstract: Objective To study the feasibility of electrospun poly(D,L-lactide-co-glycolide) (PLGA) scaffold loaded with osteogenic growth peptide (OGP) for bone tissue engineering. Methods PLGA scaffolds were prepared by electrospinning PLGA solution without OGP (control group) or with 0.1%, 0.2% and 0.4% OGP (0.1% OGP@PLGA, 0.2%OGP@PLGA, and 0.4% OGP@PLGA scaffolds, respectively). The microstructure of the scaffolds was observed by scanning electron microscopy (SEM). The scaffolds were soaked in PBS to confirm the release pattern of OGP. The biocompatibility of the scaffolds was evaluated using CCK-8 assay and live/dead staining after a 7-day coculture with rat bone marrow-derived mesenchymal stem cells (BMSCs). ALP assay and ARS staining were used to evaluate osteoinduction capacity of the scaffolds co-cultured with rat BMSCs for 14 days. In a male SD rat model of skull defect (5 mm in diameter), bone defect repair was evaluated 8 weeks after implantation of the scaffolds using Micro-CT, HE and Masson staining. Results The electrospun scaffolds had a fibrous structure similar to extracellular matrix (ECM) and were capable of sustained release of OGP for at least one month. Co-culture with 0.2% OGP@PLGA and 0.4% OGP@PLGA scaffolds, as compared with pure PLGA scaffold, significantly promoted the growth of rat BMSCs ((P<0.01). The cells co-cultured with 0.4%OGP@PLGA scaffold showed the highest ALP activity and the greatest number of calcium nodules, indicating its strong osteoinduction ability (P<0.01). Micro-CT and HE and Masson staining results showed that 0.4%OGP@PLGA scaffold had significantly better ability for promoting bone repair than the other two OGP-loaded scaffolds (P<0.01). Conclusion The electrospun PLGA scaffold loaded with OGP effectively mimics the structure of ECM and has a good biocompatibility and osteoinduction ability, suggesting its potential as a new bone tissue engineering scaffold for bone defect repair.

Key words: electrospinning; osteogenic growth polypeptide; PLGA; bone tissue engineering