关节腔内注射脂肪干细胞对颞下颌关节骨关节炎模型兔关节软骨破坏的修复作用及其机制

何静,孙高,李男男,帕丽孜·阿不力克木,吴国民

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吉林大学学报(医学版) ›› 2024, Vol. 50 ›› Issue (1) : 71-78. DOI: 10.13481/j.1671-587X.20240109
基础研究

关节腔内注射脂肪干细胞对颞下颌关节骨关节炎模型兔关节软骨破坏的修复作用及其机制

  • 何静1,2,孙高1,李男男1,帕丽孜·阿不力克木1,吴国民1()
作者信息 +

Repairment effect of intra-articular adipose stem cell injection on articular cartilage destruction in rabbit model with temporomandibular joint osteoarthritis and its mechanism

  • Jing HE1,2,Gao SUN1,Nannan LI1,Palizi ABULIKEMU1,Guomin WU1()
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History +

摘要

目的 探讨关节腔内注射脂肪干细胞(ADSCs)对颞下颌关节骨关节炎(TMJOA)模型兔关节软骨破坏的修复作用,并阐明其可能的作用机制。 方法 27只兔随机分为对照组、模型组和ADSCs组。提取兔ADSCs进行培养,采用碘乙酸钠(MIA)注射法制备兔TMJOA模型。ADSCs组TMJOA模型兔颞下颌关节腔内连续2次注射1.0×106 mL-1ADSCs,对照组和模型组兔颞下颌关节腔内注射等量生理盐水,8周后取各组兔颞下颌关节行显微CT(Micro-CT)扫描,分析各组兔髁突组织的骨体积分数(BV/TV)、骨表面积/骨体积比(BS/BV)、骨小梁厚度(Tb.Th)、骨小梁间距(Tb.Sp)和骨小梁数(Tb.N),HE染色观察各组兔髁突组织病理形态表现,免疫组织化学染色法检测各组兔髁突组织中SRY相关高迁移率族盒基因9(SOX9)、基质金属蛋白酶13(MMP-13)和血管内皮生长因子(VEGF)的定位情况和蛋白表达水平,Western blotting法检测各组兔髁突组织中SOX9、MMP-13和VEGF蛋白表达水平。 结果 Micro-CT扫描,与对照组比较,模型组兔髁突组织的BV/TV、Tb.Th 和Tb.N明显降低(P<0.05),BS/BV和Tb.Sp明显升高(P<0.05);与模型组比较,ADSCs组兔髁突组织的BV/TV、Tb.Th 和Tb.N明显升高(P<0.05),BS/BV和Tb.Sp明显降低(P<0.05)。HE染色,对照组兔髁突软骨表面光滑,层次清晰,结构完整;与对照组比较,模型组兔髁突表面不规则,肥大层增厚,出现细胞缺失区和细胞簇状区;与模型组比较,ADSCs组兔髁突组织病理损伤明显减轻。免疫组织化学染色,与对照组和ADSCs组比较,模型组兔髁突组织中棕褐色颗粒增多,主要集中在肥大层,特别是骨软骨结合部位,髁突组织中SOX9、MMP-13和VEGF蛋白表达水平明显升高(P<0.05);与模型组比较,ADSCs组兔髁突组织肥大层和骨软骨结合部位棕褐色颗粒明显减少,髁突组织中SOX9、MMP-13和VEGF蛋白表达水平明显降低(P<0.05)。Western blotting法,与对照组比较,模型组兔髁突组织中SOX9、MMP-13和VEGF蛋白表达水平明显升高(P<0.05);与模型组比较,ADSCs组兔髁突组织中SOX9、MMP-13和VEGF蛋白表达水平明显降低(P<0.05)。 结论 关节腔内注射ADSCs可有效修复TMJOA的软骨破坏,减轻软骨损伤,减缓骨关节炎进展。

Abstract

Objective To discuss the repairment effect of intra-articular injection of adipose derived stem cells (ADSCs) on articular cartilage destruction in the temporomandibular joint osteoarthritis (TMJOA) model rabbits, and to clarify the possible mechanism. Methods Twenty-seven rabbits were randomly divided into control group, model group, and ADSCs group. The ADSCs of the rabbits were extracted and cultured.The rabbit TMJOA model was prepared by monosodium-iodoacetate (MIA) injection technique.The temporomandibular joint cavity of the TMJOA model rabbits in ADSCs group was given two continuous intra-articular injections of 1.0×106 mL-1 ADSCs, while the rabbits in control and model group were given sequivalent volume of saline into the temporomandibular joint cavity. After 8 weeks, Micro-CT scan was performed on the temporomandibular joints of the rabbits in various groups; the bone volume fraction (BV/TV), bone surface area/bone volume (BS/BV), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), and trabecular number(Tb.N) of condyles tissue of the rabbits in various groups were analyzed; HE staining was used to observe the pathomorphology of condyles tissue of the rabbits in various groups; immunohistochemistry was used to detect the localization and expression levels of SRY-related high mobility group box gene 9(SOX9), matrix metalloproteinase-13 (MMP-13), and vascular endothelial growth factor (VEGF) proteins in condyles tissue of the rabbits in various groups;Western blotting method was used to detect the expression levels of SOX9, MMP-13, and VEGF proteins in condyles tissue of the rabbits in various groups. Results The micro-CT scan results showed that compared with control group, the BV/TV, Tb.Th, and Tb.N of condyles tissue of the rabbits in model group were significantly decreased (P<0.05), while the BS/BV and Tb.Sp were significantly increased (P<0.05); compared with model group, the BV/TV, Tb.Th, and Tb.N in condyles tissue of the rabbits in ADSCs group were significantly increased (P<0.05), and the BS/BV and Tb.Sp were significantly decreased (P<0.05). The HE staining results showed that the condylar cartilage surface of the rabbits in control group was smooth with clear layers and intact structure; compared with control group, the surface of condyles tissue of the rabbits in model group was irregular with thickened hypertrophic layer and areas of cell depletion and clustering; compared with model group, the pathological damage of condyles tissue of the rabbits in ADSCs group was significantly decreased.The immunohistochemical staining results showed that compared with control group and ADSCs group, the number of brown granule in condyles tissue of the rabbits in model group was increased, mainly concentrated in the hypertrophic layer,especially in the bone cartilage junction site and the expression levels of SOX9, MMP-13, and VEGF proteins in condyles tissue of the rabbits in model group were significantly increased (P<0.05); compared with model group, the number of brown granule in condyles tissue of the rabbits in ADSCs group was significantly decreased,and the expression levels of SOX9, MMP-13, and VEGF proteins were significantly decreased(P<0.05). The Western blotting results showed that compared with control group, the expression levels of SOX9, MMP-13, and VEGF proteins in condyles tissue of the rabbits in model group were significantly increased (P<0.05); compared with model group, the expression levels of SOX9, MMP-13, and VEGF proteins in condyles tissue of the rabbits in ADSCs group were significantly decreased (P<0.05). Conclusion Intra-articular injection of ADSCs can effectively repair the cartilage destruction in TMJOA, alleviate the cartilage injury, and mitigate the progression of osteoarthritis.

关键词

颞下颌关节 / 骨关节炎 / 脂肪干细胞 / 软骨损伤 / 软骨修复

Key words

Temporomandibular joint / Osteoarthritis / Adipose stem cell / Cartilage destruction / Cartilage repairment

中图分类号

R782.6

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何静,孙高,李男男,帕丽孜·阿不力克木,吴国民. 关节腔内注射脂肪干细胞对颞下颌关节骨关节炎模型兔关节软骨破坏的修复作用及其机制. 吉林大学学报(医学版). 2024, 50(1): 71-78 https://doi.org/10.13481/j.1671-587X.20240109
Jing HE,Gao SUN,Nannan LI,Palizi ABULIKEMU,Guomin WU. Repairment effect of intra-articular adipose stem cell injection on articular cartilage destruction in rabbit model with temporomandibular joint osteoarthritis and its mechanism[J]. Journal of Jilin University(Medicine Edition). 2024, 50(1): 71-78 https://doi.org/10.13481/j.1671-587X.20240109

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吉林省财政厅科技项目(jcsz2020304-20)

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