中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (31): 4940-4945.doi: 10.12307/2022.776

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

水凝胶搭载骨髓间充质干细胞治疗大鼠子宫内膜损伤

吕  妍1，关永格2，宋  阳1，李  玥1   

  1. 1广州中医药大学，广东省广州市   510405；2广州中医药大学第三附属医院，广东省广州市   510360
  • 收稿日期:2021-11-17 接受日期:2022-01-06 出版日期:2022-11-08 发布日期:2022-04-24
  • 通讯作者: 关永格，博士，副主任医师，广州中医药大学第三附属医院，广东省广州市 510360
  • 作者简介:吕妍，女，1996年生，广东省深圳市人，汉族，广州中医药大学在读硕士，主要从事中医药治疗女性生殖障碍及妇科疾病相关研究。
  • 基金资助:
    广东省自然科学基金(2021A1515011435)，项目负责人：宋阳；广东省自然科学基金(2020A1515010587)，项目负责人：关永格；国家自然科学基金(81873331)，项目参与人：关永格

Hydrogel combined with bone marrow mesenchymal stem cells in the treatment of damaged endometrium in rats

Lyu Yan1, Guan Yongge2, Song Yang1, Li Yue1   

  1. 1Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 2Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510360, Guangdong Province, China
  • Received:2021-11-17 Accepted:2022-01-06 Online:2022-11-08 Published:2022-04-24
  • Contact: Guan Yongge, MD, Associate chief physician, Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510360, Guangdong Province, China
  • About author:Lyu Yan, Master candidate, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Supported by:
    Natural Science Foundation of Guangdong Province, No. 2021A1515011435 (to SY); Natural Science Foundation of Guangdong Province, No. 2020A1515010587 (to GYG); the National Natural Science Foundation of China, No. 81873331 (to GYG)

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摘要：

文题释义：
泊洛沙姆407：无臭、无味，白色，相对分子质量为9 840-14 600，在水、体积分数95%乙醇、异丙醇中易溶，是具有受热反向凝胶特性的温敏型凝胶，已广泛作为药物载体和组织工程中的细胞支架应用于疾病治疗。
CM-Dil：是一种荧光染料，常用于细胞移植时的示踪，具有亲脂性特点，可标记细胞膜和细胞浆，对活细胞无毒性，其使用方便，染色速度快，标记细胞后进行消化、冻存或处理样本时固定、包埋等操作不会影响其标记效果。

背景：水凝胶作为一类组织工程的支架材料，为移植细胞在体内增殖分化提供了良好的物理支持。
目的：探讨水凝胶搭载骨髓间充质干细胞对大鼠受损子宫内膜的修复效果。
方法：将分离培养的SD大鼠骨髓间充质干细胞进行CM-Dil标记，然后制备成泊洛沙姆407-骨髓间充质干细胞复合物。取30只8周龄SD雌性大鼠随机分为5组：假手术组、模型组、水凝胶组、干细胞组、复合物组，除假手术组外，其余4组建立子宫内膜机械损伤模型，然后每侧宫角分别注射0.2 mL PBS、泊洛沙姆407水凝胶、骨髓间充质干细胞悬液或泊洛沙姆407水凝胶-骨髓间充质干细胞悬液。观察7 d后取出子宫组织，进行苏木精-伊红染色、免疫荧光染色和酶联免疫吸附检测。
结果与结论：①与假手术组相比，模型组、水凝胶组、干细胞组与复合物组内膜厚度变薄(P < 0.05)，其中水凝胶组、干细胞组和复合物组内膜厚度较模型组均有明显改善(P < 0.05)，复合物组改善最明显，但水凝胶组与干细胞组间差异无显著性意义(P > 0.05)；②与干细胞组相比，复合物组CM-Dil标记的骨髓间充质干细胞迁移至子宫内膜数量更多(P < 0.01)，并且两组细胞均具有向子宫内膜间质细胞分化潜能；③与模型组相比，水凝胶组、干细胞组和复合物组子宫内膜组织中白细胞介素1β、白细胞介素6和和肿瘤坏死因子α水平明显下降(P < 0.01)，其中，干细胞组和复合物组白细胞介素1β、白细胞介素6和肿瘤坏死因子α水平显著低于水凝胶组(P < 0.05)，干细胞组白细胞介素6和肿瘤坏死因子α水平低于复合物组(P < 0.01)，但两组白细胞介素1β水平无显著差异(P > 0.05)；④结果表明，泊洛沙姆407水凝胶搭载骨髓间充质干细胞治疗子宫内膜损伤具有可行性，为修复受损子宫内膜提供新思路。

https://orcid.org/0000-0003-3671-266X (吕妍)

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

关键词: 水凝胶, 骨髓间充质干细胞, 组织修复, 子宫内膜损伤, 组织工程

Abstract: BACKGROUND: As a scaffold material for tissue engineering, hydrogels provide a good physical support for the proliferation and differentiation of transplanted cells in vivo. 
OBJECTIVE: To investigate the effect of hydrogel combined with bone marrow mesenchymal stem cells on endometrial reproduction in rats. 
METHODS: Bone marrow mesenchymal stem cells of Sprague-Dawley rats were isolated, cultured, and labeled with CM-Dil and were prepared for Pluronic F-127-bone marrow mesenchymal stem cells complex. A total of 30 Sprague-Dawley rats aged 8 weeks old were randomly divided into sham operation group, model group, hydrogel group, stem cell group, and complex group. Except for the sham operation group, endometrial mechanical injury models were established in the other four groups and each uterine horn was injected with 0.2 mL of PBS, Pluronic F-127 hydrogel, bone marrow mesenchymal stem cell suspension, and Pluronic F-127 hydrogel-bone marrow mesenchymal stem cell suspension, respectively. After 7 days of observation, the uterus tissues were removed for hematoxylin-eosin staining, immunfluorescence staining, and enzyme-linked immunosorbent assay. 
RESULTS AND CONCLUSION: (1) Compared with the sham operation group, the endometrial thickness of the rats was thinner in the model group, hydrogel group, stem cell group, and complex group (P < 0.05). Compared with the model group, the endometrial thickness of hydrogel, stem cell, and complex groups was significantly improved (P < 0.05). The complex group had the most obvious effect, while there was no significant difference between hydrogel group and stem cell group (P > 0.05). (2) Compared with stem cell group, the number of stem cells labeled with CM-Dil migrated to the endometrium was more in the complex group (P < 0.01) and both groups had the potential to differentiate into endometrial stromal cells. (3) Compared with the model group, the levels of interleukin-1β, interleukin-6, and tumor necrosis factor α were significantly decreased in the endometrium of the hydrogel, stem cell, and complex groups (P < 0.01). The levels of interleukin-1β, interleukin-6, and tumor necrosis factor α were significantly lower in the stem cell and complex groups than those in the hydrogel group (P < 0.05). The levels of interleukin-6 and tumor necrosis factor α were lower in the stem cell group than those in the complex group (P < 0.01); however, there were no significant differences in interleukin-1β levels between the two groups (P > 0.05). (4) The results suggest that Pluronic F-127 combined with bone marrow mesenchymal stem cells is feasible for the treatment of endometrial injury and provides new ideas for endometrial repair. 

Key words: hydrogel, bone marrow mesenchymal stem cell, tissue repair, endometrial injury, tissue engineering

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