Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.
Department of Biochemistry, Usmanu Danfodiyo University, Sokoto, Nigeria.
PLoS One. 2020 Sep 4;15(9):e0238449. doi: 10.1371/journal.pone.0238449. eCollection 2020.
Treatment of osteoarthritis (OA) is still a major clinical challenge due to the limited inherent healing capacity of cartilage. Recent studies utilising stem cells suggest that the therapeutic benefits of these cells are mediated through the paracrine mechanism of bioactive molecules. The present study evaluates the regenerative effect of stem cells from human exfoliated deciduous teeth (SHED) conditioned medium (CM) on OA chondrocytes. The CM was collected after the SHED were cultured in serum-free medium (SFM) for 48 or 72 h and the cells were characterised by the expression of MSC and pluripotency markers. Chondrocytes were stimulated with interleukin-1β and treated with the CM. Subsequently, the expression of aggrecan, collagen type 2 (COL 2), matrix metalloproteinase-13 (MMP-13), nuclear factor-kB (NF-kB) and the level of inflammatory and anti-inflammatory markers were evaluated. SHED expressed mesenchymal stromal cell surface proteins but were negative for haematopoietic markers. SHED also showed protein expression of NANOG, OCT4 and SOX2 with differential subcellular localisation. Treatment of OA chondrocytes with CM enhanced anti-inflammation compared to control cells treated with SFM. Furthermore, the expression of MMP-13 and NF-kB was significantly downregulated in stimulated chondrocytes incubated in CM. The study also revealed that CM increased the expression of aggrecan and COL 2 in OA chondrocytes compared to SFM control. Both CM regenerate extracellular matrix proteins and mitigate increased MMP-13 expression through inhibition of NF-kB in OA chondrocytes due to the presence of bioactive molecules. The study underscores the potential of CM for OA treatment.
由于软骨的内在修复能力有限，骨关节炎 (OA) 的治疗仍然是一个主要的临床挑战。最近利用干细胞的研究表明，这些细胞的治疗益处是通过生物活性分子的旁分泌机制介导的。本研究评估了人脱落乳牙（SHED）条件培养基（CM）对 OA 软骨细胞的再生作用。在 SHED 在无血清培养基（SFM）中培养 48 或 72 小时后收集 CM，并通过 MSC 和多能性标志物的表达对细胞进行表征。用白细胞介素-1β刺激软骨细胞，并用 CM 处理。随后，评估聚集蛋白聚糖、胶原类型 2（COL 2）、基质金属蛋白酶-13（MMP-13）、核因子-kB（NF-kB）和炎症和抗炎标志物的水平。SHED 表达间充质基质细胞表面蛋白，但对造血标志物呈阴性。SHED 还显示出 NANOG、OCT4 和 SOX2 的蛋白表达，具有不同的亚细胞定位。与用 SFM 处理的对照细胞相比，用 CM 处理 OA 软骨细胞可增强抗炎作用。此外，在 CM 孵育的刺激软骨细胞中，MMP-13 和 NF-kB 的表达明显下调。该研究还表明，与 SFM 对照相比，CM 增加了 OA 软骨细胞中聚集蛋白聚糖和 COL 2 的表达。由于存在生物活性分子，CM 通过抑制 NF-kB 来再生细胞外基质蛋白并减轻 OA 软骨细胞中 MMP-13 表达的增加。该研究强调了 CM 在 OA 治疗中的潜力。
