Abstract

Knee osteoarthritis (KOA) is a degenerative joint illness which leads to knee pain and functional limitation. In this study, we combined microfracture surgery with kartogenin (KGN), a small bioactive molecule used to promote the differentiation of mesenchymal stem cells (MSCs), and explored its impact on cartilage repair and possible latent mechanisms of action. The research offers a brand-new idea for the clinical cure of KOA. The microfracture technique in combination with KNG treatment was performed on a rabbit model of KOA. Animal behaviour was evaluated after the intra-articular injection of miR-708-5p and Special AT-rich sequence binding protein 2 (SATB2) lentiviruses. Later, the expression of the tumour necrosis factor α (TNF-α) and interleukin- 1 (IL-1), the pathology of synovial tissue and cartilage tissue, and the positive cartilage type II collagen, MMP-1, MMP-3 and TIMP-1 were detected. Finally, a luciferase assay was conducted to verify the interaction of miR-708-5p and SATB2. Our results showed that miR-708-5p was elevated in the rabbit KOA model; however, the expression of SATB2 was reduced. Meanwhile, the microfracture technology combined with MSCs inducer KGN drove cartilage repair and regeneration in rabbit KOA by repressing the miR-708-5p expression. We also found that miR-708-5p directly targeted the SATB2 mRNA to regulate its expression. Furthermore, our data urged that elevating miR-708-5p or restraining SATB2 may reverse the therapeutic effect of the microfracture technique combined with MSCs inducer on rabbit KOA. Microfracture technique combined with MSCs inducer represses miR-708-5p to target SATB2 to drive cartilage repair and regeneration in rabbit KOA. This indicates that the microfracture technique combined with MSCs inducers is supposed to be an effective latent method for osteoarthritis cure.

摘要

膝骨关节炎（KOA）是一种退行性关节疾病，会导致膝关节疼痛和功能限制。在这项研究中，我们将微骨折手术与卡托细胞因子（KGN）（一种用于促进间充质干细胞（MSC）分化的生物活性小分子）相结合，并探讨了其对软骨修复的影响和可能的潜在作用机制。该研究为KOA的临床治疗提供了全新的思路。微骨折技术结合 KNG 治疗在 KOA 兔模型上进行。关节内注射 miR-708-5p 和富含特殊 AT 序列结合蛋白 2 (SATB2) 慢病毒后评估动物行为。随后观察肿瘤坏死因子α（TNF-α）和白细胞介素-1（IL-1）的表达情况，滑膜组织和软骨组织的病理变化，以及软骨II型胶原蛋白、MMP-1、MMP-3和MMP-1、MMP-3的阳性结果。检测到TIMP-1。最后，进行荧光素酶测定以验证 miR-708-5p 和 SATB2 的相互作用。我们的结果表明，miR-708-5p 在兔 KOA 模型中升高；然而，SATB2的表达却降低了。同时，微骨折技术结合间充质干细胞诱导剂KGN，通过抑制miR-708-5p表达，驱动兔KOA软骨修复和再生。我们还发现miR-708-5p直接靶向SATB2 mRNA来调节其表达。此外，我们的数据表明，升高miR-708-5p或抑制SATB2可能会逆转微骨折技术联合MSCs诱导剂对兔KOA的治疗效果。微骨折技术联合 MSCs 诱导剂抑制 miR-708-5p 靶向 SATB2 驱动兔 KOA 软骨修复和再生。这表明微骨折技术联合MSCs诱导剂有望成为治疗骨关节炎的有效潜在方法。