Chen Guangzi, Xu Tao, Gao Ran, Liu Wenbin, Li Weigang, Zeng Delu, Li Jian, Fang Xuan, Sheng Gaohong, Zhao Hongqi, Liu Chaoxu
Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, P.R. China.
Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, P.R. China.
Stem Cell Res Ther. 2025 Apr 28;16(1):212. doi: 10.1186/s13287-025-04307-4.
Periosteum plays an important role in bone defect repair due to its rich vascular network and cells. However, natural periosteum is difficult to meet clinical requirements due to its low availability. Therefore, it is necessary to develop a tissue engineering strategy of biomimetic periosteum for bone defect repair.
Poly-ε-caprolactone/chitosan/Whitlockite electrospun bionic membrane (PCL/CS/WH) was prepared using electrospinning technology, then it was conjugated with an E7 peptide as PCL/CS/WH/E7 bionic membrane. The physical properties of the membranes were evaluated by TEM, FTIR and tensile strength testing. In vitro, LIVE/DEAD staining and Cell Counting Kit-8 assay of bone marrow mesenchymal stem cells (BMSCs) and Endothelial progenitor cells (EPCs) are used to assess the biocompatibility of bionic membranes. Matrigel was applied to evaluate the ability of the different composite nanofibers samples to promote angiogenesis. Mineralized nodule and collagen formation in the BMSCs was detected by alizarin red staining and sirius red staining respectively. The expression of osteogenesis related genes and angiogenesis associated genes were detected using quantitative real-time polymerase chain reaction (qRT-PCR). In vivo, the ability of PCL/CS/WH/E7 membrane to promote bone regeneration and angiogenesis was assessed by Micro-CT and associated staining.
The addition of chitosan (CS) and E7 peptide (E7) enhanced the hydrophilicity and cytocompatibility of pure PCL membranes. Additionally, CS, E7 and Mg released from Whitlockite (WH) had a synergistic effect to promote angiogenesis and osteogenic differentiation. Three weeks after implantation, the membrane successfully bridged the bone defect and significantly promoted the formation of new bone and blood vessels.
The PCL/CS/WH/E7 membrane to achieve efficient repair of bone tissue and enriches clinical solutions for the treatment of critical bone defects.
骨膜因其丰富的血管网络和细胞，在骨缺损修复中发挥着重要作用。然而，天然骨膜由于其来源有限，难以满足临床需求。因此，有必要开发一种用于骨缺损修复的仿生骨膜组织工程策略。
采用静电纺丝技术制备聚-ε-己内酯/壳聚糖/硅钙石电纺仿生膜（PCL/CS/WH），然后将其与E7肽偶联形成PCL/CS/WH/E7仿生膜。通过透射电子显微镜（TEM）、傅里叶变换红外光谱（FTIR）和拉伸强度测试对膜的物理性能进行评估。在体外，使用活/死染色和细胞计数试剂盒-8（Cell Counting Kit-8）检测法对骨髓间充质干细胞（BMSCs）和内皮祖细胞（EPCs）进行检测，以评估仿生膜的生物相容性。应用基质胶评估不同复合纳米纤维样品促进血管生成的能力。分别通过茜素红染色和天狼星红染色检测BMSCs中矿化结节和胶原蛋白的形成。使用定量实时聚合酶链反应（qRT-PCR）检测成骨相关基因和血管生成相关基因的表达。在体内，通过微型计算机断层扫描（Micro-CT）和相关染色评估PCL/CS/WH/E7膜促进骨再生和血管生成的能力。
壳聚糖（CS）和E7肽（E7）的加入增强了纯PCL膜的亲水性和细胞相容性。此外，从硅钙石（WH）释放的CS、E7和镁具有协同作用，可促进血管生成和成骨分化。植入三周后，该膜成功桥接了骨缺损，并显著促进了新骨和血管的形成。
PCL/CS/WH/E7膜实现了骨组织的高效修复，丰富了治疗严重骨缺损的临床解决方案。
