Multiple umbilical cord-derived MSCs administrations attenuate rat osteoarthritis progression via preserving articular cartilage superficial layer cells and inhibiting synovitis
Multiple umbilical cord-derived MSCs administrations attenuate rat osteoarthritis progression via preserving articular cartilage superficial layer cells and inhibiting synovitis
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多次脐带间充质干细胞给药通过保留关节软骨表层细胞和抑制滑膜炎来减轻大鼠骨关节炎的进展
DOI：
10.1016/j.jot.2020.03.007
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发表时间：
2020-07-01
影响因子：
6.6
通讯作者：
Tian, Hongtao
中科院分区：
文献类型：
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作者：
Tong, Wei;Zhang, Xiaoguang;Tian, Hongtao
背景/目的：关节软骨侵蚀可能在骨关节炎（OA）的发生和发展中起重要作用。研究表明，脐带来源的间充质干细胞（UCMSCs）可以延缓软骨细胞凋亡，改善OA患者的进展，但详细的机制在很大程度上是未知的。本研究旨在研究UCMSCs对碘乙酸钠（MIA）诱导的大鼠OA模型的影响，并探讨其细胞学机制。方法：在200 g体重的雄性Sprague-Dawley大鼠左膝关节内注射0.3 mg MIA/50 μ L生理盐水，诱导大鼠膝关节OA。分别在MIA后一天给予单剂量的2.5 × 10（5）未分化的UCMSC或在第1、7和14天给予三次关节内注射2.5 × 10（5）UCMSC。MIA后4周，收获关节并进行石蜡切片。采用番红O染色、苏木精-伊红染色及MMP-13、ADAMTS-5、Col-2、CD 68、CD 4免疫组化染色观察软骨及滑膜的侵蚀情况。在体外研究中，通过transwell实验评估UCMSCs对软骨表面层细胞（SFCs）的迁移能力。实时荧光定量PCR检测MIA诱导的UCMSCs诱导的SFCs的Col-X和BCL-2基因的变化。CCK-8法检测UCMSCs条件培养液对SFCs增殖能力的影响。结果：在本研究中，我们将具有高度增殖性和非侵入性可收集性的人UCMSCs局部注射到MIA诱导的大鼠膝关节OA中。一个重要的发现是，与单次注射相比，在MIA注射后重复注射UCMSC明显改善了软骨侵蚀并降低了OA Mankin评分，与媒介物相比，这两者都减弱了OA进展。有趣的是，我们观察到关节软骨表面上的SFCs数量显著增加，这可能与UCMSCs条件培养基体外处理培养的SFCs的增殖、动员和抑制的catalytic标记物：Col-X和BCL-2基因表达有关。除了独特的SFC的变化，ADAMTS-5和MMP-13的catalysts标志物在整个软骨层软骨细胞中也显著上调。值得注意的是，MIA诱导了CD 4 + Th细胞和CD 68+巨噬细胞上的炎性细胞浸润以及滑膜增生，这通过反复注射UCMSCs而减轻。我们的研究表明，在OA进展期间，重复UCMSC给药对保护SFC功能、软骨结构和抑制滑膜炎具有关键作用，本文的翻译潜力：UCMSCs是一种相对“年轻”的干细胞，可无创性采集。在我们的研究中，我们清楚地表明，它可以有效地延缓OA进展，可能是通过保留SFCs功能和抑制滑膜炎。因此，经过进一步的临床试验，它可能是一种新的有希望的治疗OA的细胞来源。
Background/objectives: Articular cartilage erosion probably plays a substantial role in osteoarthritis (OA) initiation and development. Studies demonstrated that umbilical cord-derived mesenchymal stem cells (UCMSCs) could delay chondrocytes apoptosis and ameliorate OA progression in patients, but the detailed mechanisms are largely uncharacterised. In this study, we aimed to study the effects of UCMSCs on monosodium iodoacetate (MIA)induced rat OA model, and explore the cellular mechanism of this effect.Methods: Intra-articular injection of 0.3 mg MIA in 50 mu L saline was performed on the left knee of the 200 g weight male Sprague-Dawley rat to induce rat knee OA. A single dose of 2.5 x 10(5) undifferentiated UCMSCs one day after MIA or three-time intra-articular injection of 2.5 x 10(5) UCMSCs on Days 1, 7 and 14 were given, respectively. Four weeks after MIA, joints were harvested and processed for paraffin sections. Safranine-O staining, haematoxylin and eosin staining and immunohistochemistry of MMP-13, ADAMTS-5, Col-2, CD68 and CD4 were performed to observe cartilage erosion and synovium. For in vitro studies, migration ability of cartilage superficial layer cells (SFCs) by UCMSCs were accessed by transwell assay. Furthermore, catabolism change of MIA-induced SFCs by UCMSCs was performed by real-rime polymerase chain reaction of Col-X and BCL-2 genes. CCK-8 assay was performed to check proliferation ability of SFCs by UCMSCs-conditioned media.Result: In this study, we locally injected human UCMSCs, which is highly proliferative and noninvasively collectible, into MIA-induced rat knee OA. An important finding is on obviously ameliorated cartilage erosion and decreased OA Mankin score by repeated UCMSCs injection after MIA injection compared with single injection, both of which attenuated OA progression compared with vehicle. Interestingly, we observed significantly increased number of SFCs on the articular cartilage surface, probably related to elevated proliferation, mobilisation and inhibited catabolism marker: Col-X and BCL-2 gene expression of cultured SFCs by UCMSCs-conditioned media treatment in vitro. In addition to the change of unique SFCs, catabolism markers of ADAMTS-5 and MMP-13 were substantially upregulated in the whole cartilage layer chondrocytes as well. Strikingly, MIAinduced inflammatory cells infiltration, on both CD4+ Th cells and CD68+ macrophages, and hyperplasia of the synovium, which was alleviated by repeated UCMSCs injection.Conclusion: Our study demonstrated a critical role of repeated UCMSCs dosing on preserving SFCs function, cartilage structure and inhibiting synovitis during OA progression, and thus provided mechanistic proof of evidence for the use of UCMSCs on OA patients in the future.The translational potential of this article: UCMSCs are a relatively "young" stem cell, and noninvasively collectible. In our study, we clearly demonstrated that it could effectively delay OA progression, possibly through reserving SFCs function and inhibiting synovitis. Therefore, it could be a new promising therapeutic cell source for OA after further clinical trials.