Orthopedics and Traumatology Service, School of Medicine and University Hospital 'Dr. José Eleuterio González', Universidad Autonoma de Nuevo León, Monterrey, Nuevo León 64460, México.
Department of Biochemistry and Molecular Medicine, School of Medicine, Universidad Autonoma de Nuevo León, Monterrey, Nuevo León 64460, México.
Mol Med Rep. 2020 May;21(5):2243-2250. doi: 10.3892/mmr.2020.11004. Epub 2020 Feb 26.
Previous clinical studies have reported the clinical effectiveness of non‑animal stabilized hyaluronic acid (NASHA) and adipose‑derived mesenchymal stromal/stem cells (MSC) in the treatment of knee osteoarthritis (OA). Unlike MSC secreted mediators, in vitro anti‑inflammatory effects of NASHA have not been evaluated. We aimed to evaluate and compare the anti‑inflammatory effect of NASHA and MSC conditioned medium (stem cell‑conditioned medium; SC‑CM), in an explant‑based coculture model of OA. Cartilage and synovial membrane from seven patients undergoing total knee arthroplasty were used to create a coculture system. Recombinant IL‑1β was added to the cocultures to induce inflammation. Four experimental groups were generated: i) Basal; ii) IL‑1β; iii) NASHA (NASHA + IL‑1β); and iv) SC‑CM (SC‑CM + IL‑1β). Glycosaminoglycans (GAG) released in the culture medium and of nitric oxide (NO) production were quantified. Gene expression in cartilage and synovium of IL‑1β, matrix metallopeptidase 13 (MMP13), ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) and tissue inhibitor of metalloproteinases 1 (TIMP1) was measured by reverse transcription‑quantitative PCR. Media GAG concentration was decreased in cocultures with NASHA and SC‑CM (48 h, P<0.05; 72 h, P<0.01) compared with IL‑1β. Production of NO was significantly lower only in SC‑CM after 72 h (P<0.01). In cartilage, SC‑CM inhibited the expression of IL‑1β, MMP13 and ADAMTS5, while NASHA had this effect only in MMP13 and ADAMTS5. In synovium, SC‑CM decreased the expression level of MMP13 and ADAMTS5, while NASHA only decreased ADAMTS5 expression. Both NASHA and SC‑CM increased TIMP1 expression in cartilage and synovium. Treatments with NASHA and SC‑CM were shown to be a therapeutic option that may help counteract the catabolism produced by the inflammatory state in knee OA. The anti‑inflammatory mediators produced by MSC promote a lower expression of inflammatory targets in our study model.
先前的临床研究已经报道了非动物稳定透明质酸（NASHA）和脂肪来源间充质基质/干细胞（MSC）在治疗膝骨关节炎（OA）方面的临床疗效。与 MSC 分泌的介质不同，NASHA 的体外抗炎作用尚未得到评估。我们旨在评估和比较 NASHA 和 MSC 条件培养基（干细胞条件培养基；SC-CM）在 OA 体外共培养模型中的抗炎作用。从 7 名接受全膝关节置换术的患者中取出软骨和滑膜膜，以建立共培养系统。向共培养物中添加重组白细胞介素-1β（IL-1β）以诱导炎症。共生成了 4 个实验组：i）基础组；ii）IL-1β 组；iii）NASHA（NASHA+IL-1β）组；iv）SC-CM（SC-CM+IL-1β）组。定量分析培养物中释放的糖胺聚糖（GAG）和一氧化氮（NO）的产生。通过逆转录定量 PCR 测量软骨和滑膜中 IL-1β、基质金属蛋白酶 13（MMP13）、含血栓素样金属蛋白酶 1 基序的 ADAM 金属肽酶 5（ADAMTS5）和金属蛋白酶组织抑制剂 1（TIMP1）的基因表达。与 IL-1β 相比，NASHA 和 SC-CM（48 小时，P<0.05；72 小时，P<0.01）共培养物中的介质 GAG 浓度降低。仅在 SC-CM 后 72 小时（P<0.01）NO 的产生显著降低。在软骨中，SC-CM 抑制了 IL-1β、MMP13 和 ADAMTS5 的表达，而 NASHA 仅对 MMP13 和 ADAMTS5 有这种作用。在滑膜中，SC-CM 降低了 MMP13 和 ADAMTS5 的表达水平，而 NASHA 仅降低了 ADAMTS5 的表达。NASHA 和 SC-CM 均增加了软骨和滑膜中 TIMP1 的表达。NASHA 和 SC-CM 的治疗被证明是一种治疗选择，可能有助于对抗膝骨关节炎炎症状态引起的分解代谢。在我们的研究模型中，MSC 产生的抗炎介质促进了炎症靶点表达的降低。
