口腔医学研究

口腔医学研究 ›› 2022, Vol. 38 ›› Issue (4): 372-378.DOI: 10.13701/j.cnki.kqyxyj.2022.04.016

• 口腔组织工程学研究 • 上一篇    下一篇

人骨髓间充质干细胞/细胞外基质复合体用于骨缺损再生修复的研究

张舟1,2, 梁想1,2, 崔浩1,2, 朱羿霏1,2, 屠军波1,2, 那思家1,2*   

  1. 1.西安交通大学口腔医院 陕西省颅颌面精准医学研究重点实验室 陕西 西安 710004;
    2.西安交通大学口腔医院口腔颌面外科 陕西 西安 710004
  • 收稿日期:2021-10-26 出版日期:2022-04-28 发布日期:2022-04-22
  • 通讯作者: *那思家,E-mail: sijiana@xjtu.edu.cn
  • 作者简介:张舟(1995~ ),女,四川广元人,硕士在读,医师,研究方向:牙周骨缺损的再生治疗。
  • 基金资助:
    国家自然科学基金地区项目(编号:81960194)陕西省自然科学基金青年项目(编号:2020JQ563)西安交通大学基本科研业务费专项科研项目(编号:xzy012020048)

Human Bone Marrow Mesenchymal Stem Cells / Extracellular Matrix Complex for Bone Defect Regeneration and Repair

ZHANG Zhou1,2, LIANG Xiang1,2, CUI Hao1,2, ZHU Yifei1,2, TU Junbo1,2, NA Sijia1,2*   

  1. 1. Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi'an Jiaotong University, Xi'an 710004, China;
    2. Department of Oral and Maxillofacial Surgery, College of Stomatology, Xi'an Jiaotong University, Xi'an 710004, China
  • Received:2021-10-26 Online:2022-04-28 Published:2022-04-22

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摘要： 目的: 评估人骨髓间充质干细胞/细胞外基质复合体(C-BMSC)的骨缺损再生修复能力。方法: 体外构建C-BMSC,通过大体观察、HE染色、扫描电镜进行组织形态学检测;TUNEL染色法检测细胞凋亡水平;免疫荧光染色和PCR法检测成骨相关因子的表达水平;大鼠颅骨缺损模型评估C-BMSC体内骨缺损再生修复能力。结果: 组织形态学显示5 d组C-BMSC的细胞外基质(ECM)最丰富,ECM将细胞完全包裹,ECM纤维粗壮,小分子物质附着;TUNEL染色结果提示3 d与5 d组细胞凋亡数量接近,而7 d时细胞凋亡显著增加(P<0.001);PCR结果表明C-BMSC对ALP、COL-1、Runx2和OCN表达均高于细胞膜片(P<0.05);免疫荧光结果显示C-BMSC中ECM对ALP、COL-1、BSP表达均呈阳性;大鼠颅骨移植实验结果表明C-BMSC组骨体积与软组织体积比显著高于空白组。结论: 本研究证明C-BMSC具有良好的成骨分化能力,这为骨缺损再生修复提供了一种新的策略。

关键词: 骨髓间充质干细胞, 干细胞/细胞外基质复合体, 细胞膜片, 骨缺损再生修复

Abstract: Objective: To evaluate the bone defect regeneration and repair ability of human bone marrow mesenchymal stem cells/extracellular matrix complex (C-BMSC). Methods: C-BMSC was constructed in vitro and examined by gross observation, HE staining, and scanning electron microscope (SEM). TUNEL staining was used to detect the level of apoptosis. The expression level of osteogenic related factors was detected by immunofluorescence staining and PCR. The rat skull defect model was used to evaluate the bone defect regeneration and repair ability of C-BMSC in vivo. Results: Histomorphology examination showed that the extracellular matrix (ECM) of C-BMSC was the most abundant in the 5-day group. The ECM completely wrapped the cells in C-BMSC. The ECM fibers were strong and small molecular substances were attached. TUNEL staining result showed that the number of apoptosis in the 3-day and 5-day groups was close, but it was increased significantly for the cell apoptosis level at 7-day (P<0.001). PCR results showed that the expressions of ALP, COL-1, Runx2, and OCN in C-BMSC was higher than those in BMSC-CS (P<0.05). Immunofluorescence results showed that ECM in C-BMSC was positive for ALP, COL-1, and BSP. C-BMSC also showed remarkable ability of bone tissue regeneration and repair in vivo (P<0.05). Conclusion: This study proves that C-BMSC has outstanding osteogenic differentiation ability, which provides a new strategy for bone defect regeneration and repair.

Key words: BMSCs, bone marrow mesenchymal stem cells/extracellular matrix complex, cell sheet, bone defect regeneration and repair