中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (33): 5274-5279.doi: 10.3969/j.issn.2095-4344.2017.33.005

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓基质干细胞与负载血管内皮生长因子/骨形态发生蛋白2聚乳酸-羟基乙酸共聚物/明胶纳米纤维支架的生物相容性

安 刚1，鲁 斌2，张文博3，蒋玉东1   

  1. 1黑龙江省医学科学院，哈尔滨医科大学附属第一医院，黑龙江省哈尔滨市 150086；2芜湖市第一人民医院骨科，安徽省芜湖市 241000；3山东航天电子技术研究所，山东省烟台市 264000
  • 修回日期:2017-07-18 出版日期:2017-11-28 发布日期:2017-12-01
  • 通讯作者: 安刚，博士，主治医师，黑龙江省医学科学院，哈尔滨医科大学附属第一医院，黑龙江省哈尔滨市 150086
  • 作者简介:安刚，男，1982年生，黑龙江省海林市人，汉族，2017年哈尔滨医科大学毕业，博士，主治医师，主要从事骨组织工程骨缺损修复研究。
  • 基金资助:

    黑龙江省科研计划项目(2015001)

Biocompatibility of bone marrow stromal stem cells with vascular endothelial growth factor/bone morphogenetic protein 2-poly(lactic-co-glycolic acid) copolymer/gelatin nanofibrous scaffold

An Gang1, Lu Bin2, Zhang Wen-bo3, Jiang Yu-dong1   

  1. 1First Affiliated Hospital of Harbin Medical University, Harbin 150086, Heilongjiang Province, China; 2Department of Orthopedics, Wuhu First People’s Hospital, Wuhu 241000, Anhui Province, China; 3Shandong Institute of Space Electronic Technology, Yantai 264000, Shandong Province, China
  • Revised:2017-07-18 Online:2017-11-28 Published:2017-12-01
  • Contact: An Gang, First Affiliated Hospital of Harbin Medical University, Harbin 150086, Heilongjiang Province, China
  • About author:An Gang, M.D., Attending physician, First Affiliated Hospital of Harbin Medical University, Harbin 150086, Heilongjiang Province, China
  • Supported by:

    the Science and Technology Research Plan of Heilongjiang Province, No. 2015001

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摘要：

文章快速阅读：

文题释义：
复合纳米纤维支架：具备孔隙大、高孔隙率、纤维直径小、比表面积大、生物相容性好等特性，并且其三维立体结构与细胞外基质的三维结构相类似，能够更好地促进种子细胞的黏附、增殖及分化，所以成为目前骨组织工程支架材料的研究热点。
负载双因子：血管内皮生长因子和骨形态发生蛋白2两种生长因子分别复合于纳米纤维支架内外层，实现两种因子的顺序释放，早期释放血管内皮生长因子可促进支架内血管生成，实现生物支架的血管化，促进营养物质、生长因子及细胞进入支架内部。顺序释放成骨生长因子骨形态发生蛋白2可促进支架内细胞的成骨分化，并且两种因子促进血管生成及成骨存在着协同作用。

 

摘要
背景：应用骨组织工程学方法进行骨修复重建是治疗骨缺损的理想方法，负载生长因子的生物支架与种子细胞的结合是关键。
目的：将大鼠骨髓基质干细胞种植于负载血管内皮生长因子(vascular endothelial growth factor，VEGF)/骨形态发生蛋白2(bone morphogenetic protein-2，BMP-2)的聚乳酸-羟基乙酸共聚物(poly(lactic-co-glycolic acid)，PLGA)/明胶纳米纤维支架，观察二者的生物相容性。
方法：实验分为5组：空白支架组、BMP-2组、VEGF组、VEGF/BMP-2组(1.2∶1)、VEGF/BMP-2组(0.8∶1)、VEGF/BMP-2组(0.4∶1)，将各组纳米纤维支架放入细胞培养孔板底部，然后将大鼠骨髓基质干细胞接种于培养孔内，扫描电镜观察细胞在各组支架上的黏附情况，CCK-8法检测细胞增殖能力，RT-PCR检测成骨分化基因RUNX-2，ALP，OCN的表达。
结果与结论：①在扫描电镜下可见细胞与支架黏附，并进入支架内部，VEGF/BMP-2(0.4︰1)组的细胞黏附生长状态最好，细胞与支架紧密结合成为支架细胞复合体；②负载生长因子的纳米纤维支架上的细胞增殖及成骨分化标记物ALP、RUNX-2和OCN表达量高于空白支架组，并且VEGF/BMP-2浓度比例为0.4︰1时成骨分化标记物表达量最高；③结果表明，负载VEGF/BMP-2的PLGA/明胶纳米纤维支架可促进大鼠骨髓基质干细胞的黏附、增殖及成骨分化，具备良好的细胞相容性，且VEGF/BMP-2浓度比例0.4∶1为最佳比例。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0001-7993-2430(安刚)

关键词: 干细胞, 骨髓干细胞, 骨髓基质干细胞, 聚乳酸-羟基乙酸共聚物, 明胶, 纳米纤维支架, 骨形态发生蛋白2, 血管内皮生长因子, 骨组织工程

Abstract:

BACKGROUND: Using bone tissue engineering methods for reconstruction of bone repair is an ideal treatment for bone defects, and it is crucial to combine biological scaffolds carrying growth factors with seed cells.
OBJECTIVE: To observe the biocompatibility of rat bone marrow stromal stem cells (BMSCs) seeded onto vascular endothelial growth factor (VEGF)/bone morphogenetic protein 2 (BMP-2)-poly(lactic-co-glycolic acid) (PLGA) copolymer/ gelatin nanofibrous scaffolds.
METHODS: Rat BMSCs were divided into five groups: blank scaffold group, BMP-2 group, VEGF group, VEGF/BMP-2 groups (1.2:1, 0.8:1, 0.4:1). In each group, the nanofibrous scaffold was placed at the bottom of the cell culture plate, and then rat BMSCs were seeded into the culture plate. Cell adhesion was observed under scanning electron microscope; cell proliferation was detected by cell counting kit-8; and ALP, RUNX-2 and OCN expression was determined by RT-PCR.
RESULTS AND CONCLUSION: Under the scanning electron microscope, the BMSCs adhered to the scaffold and further grew into the scaffold. The adherent cells grew best in the 0.4:1 group, in which the cells were tightly integrated with the scaffold to form a cell-scaffold complex. Compared with the blank scaffold group, the cell proliferation and expression of ALP, RUNX-2 and OCN were both higher in the other groups, which was highest in the 0.4:1 group. To conclude, the VEGF/BMP-2-loaded PLGA copolymer/gelatin nanofibrous scaffold can promote cell adhesion, proliferation, osteogenic differentiation of rat BMSCs because of a good cytocompatibility. Moreover, the concentration ratio of VEGF/BMP-2 is optimized at 0.4:1. 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Morphogenetic Proteins, Vascular Endothelial Growth Factors, Tissue Scaffolds, Bone Marrow, Mesenchymal Stem Cells, Tissue Engineering

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