中国实用口腔科杂志

中国实用口腔科杂志

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骨髓间充质干细胞促进皮肤创伤愈合及细胞因子在愈合过程表达研究

丁    越，张    辉，孙晓菊   

  1. 辽宁省人民医院口腔科，沈阳  110016
  • 出版日期:2015-02-15 发布日期:2015-04-09
  • 基金资助:

    恶性肿瘤转化医学研究中心及协同研究网络建设（2013225021）

DING Yue，ZHANG Hui ，SUN Xiao-ju   

  • Online:2015-02-15 Published:2015-04-09

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摘要：

目的    探讨骨髓间充质干细胞（bone marrow mesenchymal stem cells， BMSCs）促进皮肤创伤愈合作用及细胞因子在愈合过程中的表达变化。方法    本研究于2013年7月至2014年10月在沈阳万类生物技术实验室完成。体外分离、培养大鼠BMSCs，镜下观察细胞生长形态，取第3代BMSCs进行实验。流式细胞仪检测细胞表面标记物鉴定BMSCs。选择健康雄性SD大鼠48只，随机分成2组。于大鼠尾静脉注射PKH-26标记的BMSCs，1周后进行皮肤创伤模型制备，实验组于大鼠背部正中线划长3 cm的纵行切口，对照组不做创伤处理。荧光染色观察不同时间点正常皮肤及创面组织BMSCs数量、分布情况，HE染色观察不同时间点正常皮肤及创面组织病理变化情况。ELISA检测大鼠尾静脉血中不同时间点基质细胞衍生因子（SDF-1）、肿瘤坏死因子（TNF-α）、白细胞介素（IL-1β）、碱性成纤维细胞生长因子（bFGF）、血小板衍生生长因子（PDGF）的含量变化。结果    镜下观察培养第三代的BMSCs细胞形态为均一梭形，贴壁生长。BMSCs移植能够加速创面愈合，ELISA结果显示，BMSCs移植后血清中SDF-1α、TNF-α、IL-1β、bFGF和PDGF的含量随着创伤愈合进程呈先上升后下降的趋势。结论    BMSCs能促进大鼠皮肤创伤愈合。

关键词: 骨髓间充质干细胞, 皮肤创伤愈合, 细胞因子

Abstract:

Objective    To explore bone marrow mesenchymal stem cells（BMSCs）in the acceleration of skin wound healing and the expression of cytokines during wound healing. Methods    BMSCs in rats were isolated and cultured in vitro. The morphological changes were analyzed by the microscopy. The third generation of BMSCs were chosen to subsequent experiments. Flow cytometry was conducted to detect cell surface markers of BMSCs. Another 48 healthy male SD rats were divided into two groups at random；BMSCs labeled with PKH-26 were injected into the rat caudal vein. Wound healing model was established after one week. On the center of rat back approximately 3 cm incisor was made in the experiment group，in control group trauma not dealed with. The number and distribution of BMSCs were observed by the fluorescent staining at different time points. The pathological changes of wound in the normal skin and trauma were observed by HE staining. The expression of SDF-1， TNF-α，IL-1β，bFGF and PDGF in the rat caudal vein were detected by ELISA at different time points. Results    The third generation of BMSCs appeared fusiform shape and adherence growth. Transplanted BMSCs obviously enhanced the healing of the wound. The after-rising downward trends of SDF-1，TNF-α，IL-1β，bFGF and PDGF in the rat caudal vein were observed after BMSCs transplantation. Conclusion    BMSCs can accelerate the wound healing.

Key words: bone marrow mesenchymal stem cells, skin wound healing, cytokines