可注射水凝胶包埋骨髓间充质干细胞可有效促进损伤软骨的修复

doi:10.16352/j.issn.1001-6325.2025.01.0044

基础医学与临床 ›› 2025, Vol. 45 ›› Issue (1): 44-50.doi: 10.16352/j.issn.1001-6325.2025.01.0044

可注射水凝胶包埋骨髓间充质干细胞可有效促进损伤软骨的修复

刘德国, 陈宏, 张颉鸿, 郑宇翔, 刘振刚, 陈宣辰, 侯振海^*

中国人民解放军联勤保障部队第九〇三医院骨三科, 浙江杭州 310000

收稿日期:2024-08-26 修回日期:2024-11-19 出版日期:2025-01-05 发布日期:2024-12-25
通讯作者: ^*zhenhai_hou@163.com
基金资助:
浙江省教育厅高校一般科研项目(Y202352253)

Injectable hydrogel loaded with bone marrow- mesenchymal stem cells promotes the repair of cartilage defect

LIU Deguo, CHEN Hong, ZHANG Jiehong, ZHENG Yuxiang, LIU Zhengang, CHEN Xuanchen, HOU Zhenhai^*

Department of Orthopedics, the 903rd Hospital of the Joint Logistics Support Force of the People's Liberation Army of China, Hangzhou 310000 ,China

Received:2024-08-26 Revised:2024-11-19 Online:2025-01-05 Published:2024-12-25
Contact: ^*zhenhai_hou@163.com

摘要/Abstract

摘要： 目的构建可注射水凝胶(IH)包埋骨髓间充质干细胞(BM-MSCs)体系,探究其促进关节软骨修复的作用及机制。方法使用全骨髓贴壁法培养小鼠原代BM-MSCs。采用茜素红(ARS)、油红O(ORO)及阿尔新蓝(AB)染色鉴定其分化潜能;采用流式细胞测量术鉴定其表面CD44、CD90、CD105、CD34、CD45的表达。制备可注射水凝胶包埋BM-MSC(BM-MSC-IH),采用流变学测试检测BM-MSC-IH的流变性能。构建小鼠膝关节软骨全层损伤模型,采用随机数字表法将小鼠分为盐水组、BM-MSC组和BM-MSC-IH组,每组8只。采用关节注射法给药,试验组小鼠注射BM-MSC-IH 0.2 mL,BM-MSCs组注射等量细胞悬液,盐水组注射等量0.9%氯化钠溶液。分别于注射药物6周及12周后,使用番红-固绿染色观察小鼠膝关节软骨的修复情况,使用免疫组织化学染色观察小鼠膝关节Ⅱ型胶原表达情况。结果原代培养的小鼠BM-MSCs阳性表达CD44、CD90、CD105,而CD34、CD45表达呈阴性,且具有成骨、成脂、成软骨分化潜能;构建的BM-MSCs-IH体系为类固体胶体,流变性能符合水凝胶的基本特征;BM-MSC-IH可有效促进小鼠膝关节软骨再生,并增加局部Ⅱ型胶原的表达,效果较单独使用BM-MSCs更佳。结论可注射水凝胶包埋骨髓间充质干细胞可有效促进损伤软骨的修复,该作用可能是通过促进软骨细胞分化、促进Ⅱ型胶原的表达实现的。

关键词: 水凝胶, 骨髓间充质干细胞, 软骨损伤修复, 软骨再生

Abstract: Objective To construct an injectable hydrogel (IH) entrapment system of bone marrow mesenchymal stem cells(BM-MSCs) and to explore its effect of promoting articular cartilage repair and underlying mechanism . Methods The primary BM-MSCs of mice were cultured by whole bone marrow adherent method. The differentiation potential was identified by alizarin red (ARS), oil red O (ORO) and Alcian blue (AB) staining, and the expression of CD44, CD90, CD105, CD34 and CD45 on the surface was detected by flow cytometry. BM-MSCs loaded with IH(BM-MSC-IH) was prepared, and the rheological properties of BM-MSC-IH were tested by rheological test. The model of full-thickness injury of knee cartilage in mice was established, and the mice were randomly divided into normal saline group ,BM-MSC group and BM-MSC-IH group with 8 in each. The mice were injected with BM-MSC-IH 0.2 mL, cell suspension 0.2 mL and 0.9% NaCl solidion, independently. After 6 and 12 weeks of injection, the repair of knee cartilage in mice was observed by safranin-fast green staining, and the expression of type Ⅱ collagen in knee joint was examined by immunohistochemical staining microscopy. Results The primary cultured cells showed positive expression of CD44, CD90 and CD105, but negative expression of CD34 and CD45, and had the potential of osteogenic, adipogenic and chondrogenic differentiation. The constructed BM-MSC-IH system was solid colloid, and the rheological properties were consistent with the basic characteristics of hydrogel. BM-MSC-IH effectively promoted the regeneration of mouse knee cartilage and increase the local expression of type Ⅱ collagen. Conclusions Bone marrow-mesenchymal stem cells embedded with injectable hydrogel effectively promote the repair of injured cartilage and the underlying mechanism is potentially promoting the differentiation of chondrocytes and the expression of type Ⅱ collagen. Its effect is better than application of BM-MSCs alone.

Key words: hydrogel, bone marrow-mesenchymal stem cell, cartilage injury repair, cartilage regeneration

中图分类号:

R684

刘德国, 陈宏, 张颉鸿, 郑宇翔, 刘振刚, 陈宣辰, 侯振海. 可注射水凝胶包埋骨髓间充质干细胞可有效促进损伤软骨的修复[J]. 基础医学与临床, 2025, 45(1): 44-50.

LIU Deguo, CHEN Hong, ZHANG Jiehong, ZHENG Yuxiang, LIU Zhengang, CHEN Xuanchen, HOU Zhenhai. Injectable hydrogel loaded with bone marrow- mesenchymal stem cells promotes the repair of cartilage defect[J]. Basic & Clinical Medicine, 2025, 45(1): 44-50.

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可注射水凝胶包埋骨髓间充质干细胞可有效促进损伤软骨的修复

Injectable hydrogel loaded with bone marrow- mesenchymal stem cells promotes the repair of cartilage defect

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