局部皮质切开术对骨髓抽吸浓缩物中间充质干细胞和生长因子的影响

doi:10.19439/j.sjos.2022.06.005

摘要/Abstract

摘要： 目的: 探讨局部皮质切开术对骨髓抽吸浓缩物（bone marrow aspirate concentrate, BMAC）中所含的间充质干细胞（mesenchymal stem cells, MSC）和促再生生长因子的影响。方法: 纳入5头4~5个月家猪（Sus Scrofa）,不分性别。每头猪在随机选择的胫骨上进行皮质切开术,形成2个1 cm长的伤口（皮质切开侧）,而另一根胫骨则保持完整（对照侧）。术后第14天,在两侧胫骨手术区域附近相同部位抽吸出骨髓,处理成为BMAC样品,从中分离出MSC和血浆,评估BMAC样品中所含MSC的数量、增殖和成骨分化特性以及再生生长因子的含量。采用SPSS 25.0 软件包对数据进行统计学分析。结果: 皮质切开术、骨髓抽吸术以及之后的伤口愈合均无异常。成纤维细胞集落生成单位测定（colony-forming fibroblast unit assay, CFU-F）和流式细胞术评估发现,MSC数量在皮质切开侧相较于对照侧显著增加（P<0.05）。尽管实时反转录聚合酶链技术（RT-PCR）结果显示,只有骨钙蛋白（osteocalcin, OCN）的mRNA表达量显著增加（P<0.05）,但皮质切开侧的MSCs增殖速度显著增加（P<0.05）,显示出更强的成骨分化潜能。皮质切开侧 BMAC中TGF-β、BMP2和PDGF等生长因子的浓度高于对照侧,但无显著差异。结论: 局部皮质切开术可以增强BMAC中所含MSC的数量以及增殖和成骨分化特性。

关键词: 损伤, 骨髓浓缩提取物, 间充质干细胞, 生长因子, 组织工程

Abstract: PURPOSE: This study investigated the impact of local corticotomies on mesenchymal stem cells (MSCs) and pro-regenerative growth factors contained in bone marrow aspirate concentrate(BMAC). METHODS: Five domestic pigs (Sus Scrofa), 4-5 months of age, either sex, were included. Each pig underwent operations for creation of two 1cm-long corticotomies at one randomly chosen tibia (as experimental), with the other tibia left intact (as control). At postoperative day-14, bone marrow was aspirated from both tibiae and processed into BMAC samples, from which MSCs and plasmas were separated. MSCs quantity, proliferative and osteogenic differentiation properties, as well as several regenerative growth factors contained in BMAC samples, were assessed and compared between the two sides. Statistical analysis was performed with SPSS 25.0 software package. RESULTS: Creation of corticotomy, bone marrow aspiration and corticotomy healing all occurred uneventfully. The quantity of MSCs assessed by colony-forming fibroblast unit assay and flow cytometry was significantly higher on the corticotomy side (P<0.05). MSCs from the corticotomy side also proliferated significantly faster (P<0.05) and showed a tendency of stronger osteogenic differentiation potential although only mRNA expression of osteocalcin reached statistical significance(P<0.05). The concentration of TGF-β, BMP2 and PDGF contained in BMAC tended to be higher on corticotomy side than the control side but the difference did not reach statistical significance. CONCLUSIONS: Local corticotomies are able to boost the quantity and proliferative/osteogenic differentiation property of MSC contained in BMAC.

Key words: Injury, Bone marrow aspirate concentrate, Mesenchymal stem cell, Growth factors, Tissue engineering

中图分类号:

R780.2

石磊, 汪俊. 局部皮质切开术对骨髓抽吸浓缩物中间充质干细胞和生长因子的影响[J]. 上海口腔医学, 2022, 31(6): 588-596.

SHI Lei, WANG Jun. Effect of local corticotomies on mesenchymal stem cells and growth factors of bone marrow aspirate concentrate[J]. Shanghai Journal of Stomatology, 2022, 31(6): 588-596.

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