中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (8): 1903-1911.doi: 10.12307/2026.501

• 水凝胶材料Hydrogel materials • 上一篇    下一篇

血管内皮生长因子165基因转染骨髓间充质干细胞构建血管化两亲性肽凝胶模块

蒋星海1，宋玉林2，李德津2，邵建敏1，徐军志1，刘华凯1，吴应国1，沈岳辉1，冯思诚1   

  1. 1景德镇市第三人民医院，江西省景德镇市  333000；2南昌大学第二附属医院，江西省南昌市  330000
  • 收稿日期:2024-10-24 接受日期:2024-12-17 出版日期:2026-03-18 发布日期:2025-07-14
  • 通讯作者: 宋玉林，博士，主任医师，南昌大学第二附属医院，江西省南昌市 330000
  • 作者简介:蒋星海，男，1991年生，江西省上饶市人，汉族，2018年南昌大学毕业，硕士，主治医师，主要从事骨组织工程材料研究。
  • 基金资助:
    景德镇市科技计划项目(2022SFZC012)，项目负责人：蒋星海

Vascular endothelial growth factor 165 genes transfected into bone marrow mesenchymal stem cells to construct a vascularized amphiphilic peptide gel module

Jiang Xinghai1, Song Yulin2, Li Dejin2, Shao Jianmin1, Xu Junzhi1, Liu Huakai1, Wu Yingguo1, Shen Yuehui1, Feng Sicheng1   

  1. Jiang Xinghai, MS, Attending physician, The Third People’s Hospital of Jingdezhen, Jingdezhen 333000, Jiangxi Province, China
  • Received:2024-10-24 Accepted:2024-12-17 Online:2026-03-18 Published:2025-07-14
  • Contact: Song Yulin, MD, Chief physician, The Second Affiliated Hospital of Nanchang University, Nanchang 330000, Jiangxi Province, China
  • About author:Jiang Xinghai, MS, Attending physician, The Third People’s Hospital of Jingdezhen, Jingdezhen 333000, Jiangxi Province, China
  • Supported by:
    Jingdezhen Science and Technology Plan Project, No. 2022SFZC012 (to JXH)

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摘要：

文题释义：
树枝状两亲性肽：是近年来发展起来的一类新型高分子肽支架材料，由亲水性肽列及疏水性脂肪族长烷基链构成，亲水性的肽列可以连接活性结构，自组装成超分子纳米结构，形成扭曲的条带状和高纵横比的纳米纤维状，自组装多肽序列的活性区域是水溶性的，可暴露于水溶液中，形似伸展的树枝状。
感染复数：传统的感染复数概念起源于噬菌体感染细菌的研究，是感染时噬菌体与细菌的数量比值，也就是平均每个细菌感染噬菌体的数量。噬菌体的数量单位为pfu。一般认为感染复数是一个比值，没有单位。后来感染复数被普遍用于病毒感染细胞的研究中，含义是感染时病毒与细胞数量的比值。通过摸索合适的感染复数值使病毒转染达到满意的效率。

背景：组织工程模块为股骨头坏死的治疗提供了新的思路，将血管化支架移植于坏死区域，在生长因子及支架的作用下诱导骨髓间充质干细胞向血管内皮细胞分化，促进微血管生成改善局部血运能够促进坏死区域的修复。
目的：探讨体外构建诱导骨髓间充质干细胞向血管内皮细胞分化的树枝状两亲性肽自组装凝胶模块的可行性。
方法：分离提取4-6周龄SD大鼠骨髓间充质干细胞，采用流式细胞术进行鉴定，体外定向成脂、成骨诱导分化检测细胞多向分化潜能。采用固相法合成肽           KGGGGAAAA(K)-C16H31O，调配质量浓度为10 mg/mL多肽溶液，加入生理盐水后触发多肽自组装，形成半透明的水凝胶，透射电子显微镜下观察凝胶结构。用携带血管内皮生长因子165基因的腺病毒转染大鼠骨髓间充质干细胞，然后将10 mg/mL多肽溶液和细胞悬液混合，细胞分布于凝胶内部形成三维培养体系，设置为实验组；将转染空病毒的骨髓间充质干细胞与10 mg/mL多肽溶液混合形成三维培养体系，设置为对照组。体外培养7 d后采用免疫荧光及RT-qPCR检测血管内皮细胞分化情况。
结果与结论：①流式细胞术结果显示间质干细胞表面分子CD29、CD44表达≥95%，造血干细胞表面分子CD34、CD45表达≤2%；在体外能够向成骨、成脂诱导分化；②树枝状多肽溶液在离子触发下形成的凝胶呈透明状，透射电镜下观察呈形态均一的小尺寸纳米胶束；③免疫荧光及RT-qPCR结果显示，实验组血管内皮生长因子165表达水平高于对照组，且血管内皮特异性标记物CD31、CD34表达水平高于对照组。结果表明，血管内皮生长因子165基因转染骨髓间充质干细胞种植于多肽凝胶构建组织工程模块，能够诱导骨髓间充质干细胞向血管内皮细胞分化。
https://orcid.org/0000-0002-0469-4148(蒋星海)

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料；口腔生物材料；纳米材料；缓释材料；材料相容性；组织工程

关键词: 树枝状两亲性肽">, 骨髓间充质干细胞">, 基因转染">, 血管内皮生长因子165">, 三维培养">, 血管化">, 股骨头坏死

Abstract: BACKGROUND: The tissue engineering module provides a new idea for the treatment of femoral head necrosis. A vascularized scaffold was constructed and transplanted into the necrotic area, and the bone marrow mesenchymal stem cells were induced to differentiate into vascular endothelial cells under the action of growth factors and scaffolds, which promoted the generation of microvessels and improved local blood vascularity, thereby promoting the repair of the necrotic area. 
OBJECTIVE: To investigate the feasibility of constructing a dendritic amphiphilic peptide self-assembly gel module with the ability to induce the differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells in vitro. 
METHODS: Bone marrow mesenchymal stem cells were isolated and extracted from SD rats aged 4-6 weeks and identified by flow cytometry. In vitro directed adipogenic and osteogenic induced differentiation was used to detect the multidirectional differentiation potential of cells. Peptides            KGGGGAAAA(K)-C16H31O were synthesized by solid-phase method. The concentration was 10 mg/mL peptide solution. After the addition of normal salt solution, the peptide self-assembly was triggered, forming a translucent hydrogel. Transmission electron microscopy was utilized to observe the gel structure. In the experimental group, rat bone marrow mesenchymal stem cells were transfected with adenovirus carrying vascular endothelial growth factor 165 gene. The 10 mg/mL peptide solution was mixed with the cell suspension, and the cells were distributed inside the gel to form a three-dimensional culture system. In the control group, bone marrow mesenchymal stem cells transfected with empty virus were mixed with 10 mg/mL peptide solution to form a three-dimensional culture system. Immunofluorescence and RT-qPCR were used to detect the differentiation of vascular endothelial cells after 7 days of in vitro culture.
RESULTS AND CONCLUSION: (1) Flow cytometry results showed that the expression of CD29 and CD44 on the surface of mesenchymal stem cells was ≥95%, and the expression of CD34 and CD45 on the surface of hematopoietic stem cells was ≤2%. Bone marrow mesenchymal stem cells were successfully differentiated by osteogenic induction and adipogenic induction in vitro. (2) The dendritic peptide solution was triggered by ions to form a transparent gel, and the small-sized nanomicelles with uniform morphology were observed under electron microscopy. (3) The results of immunofluorescence and RT-qPCR showed that the expression level of vascular endothelial growth factor 165 in the experimental group was higher than that in the control group, and the expression levels of CD31 and CD34 were higher than those in the control group. These results suggest that vascular endothelial growth factor 165 gene transfected bone marrow mesenchymal stem cells implanted in a tissue engineering module constructed by peptide gel can induce the differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells. 

Key words: dendritic amphiphilic peptide">, bone marrow mesenchymal stem cell">, gene transfection">, vascular endothelial growth factor 165">, three-dimensional culture">, vascularization">, femoral head necrosis

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