Washington University, St. Louis, Missouri.
Washington University and Shriners Hospitals for Children-St. Louis, St. Louis, Missouri.
Arthritis Rheumatol. 2021 Dec;73(12):2249-2260. doi: 10.1002/art.41794. Epub 2021 Oct 28.
Recent evidence delineates an emerging role of periostin in osteoarthritis (OA), since its expression after knee injury is detrimental to the articular cartilage. We undertook this study to examine whether intraarticular (IA) knockdown of periostin would ameliorate posttraumatic OA in a murine model.
Posttraumatic OA was induced in 10-week-old male C57BL/6J mice (n = 24) by destabilization of the medial meniscus (DMM), and mice were analyzed 8 weeks after surgery. Periostin expression was inhibited by small interfering RNA (siRNA) delivered IA using a novel peptide-nucleotide polyplex. Following histologic assessment of the mouse knee cartilage, the extent of cartilage degeneration was determined using Osteoarthritis Research Society International (OARSI) cartilage damage score, and severity of synovitis was also assessed. Bone changes were measured using micro-computed tomography. The effect and mechanism of periostin silencing were investigated in human chondrocytes that had been stimulated with interleukin-1β (IL-1β) with or without the IκB kinase 2 inhibitor SC-514.
Periostin expression in mice with posttraumatic OA was significantly abolished using IA delivery of a peptide-siRNA nanoplatform. OARSI cartilage damage scores were significantly lower in mice receiving periostin siRNA (mean ± SEM 10.94 ± 0.66) compared to untreated mice (22.38 ± 1.30) and mice treated with scrambled siRNA (22.69 ± 0.87) (each P = 0.002). No differences in the severity of synovitis were observed. Subchondral bone sclerosis, bone volume/total volume, volumetric bone mineral density, and heterotopic ossification were significantly lower in mice that had received periostin siRNA treatment. Immunostaining of cartilage revealed that periostin knockdown reduced the intensity of DMM-induced matrix metalloproteinase 13 (MMP-13) expression and also diminished the phosphorylation of p65 and immunoreactivity of the aggrecan neoepitope DIPEN. Periostin knockdown also suppressed IL-1β-induced MMP-13 and ADAMTS-4 expression in chondrocytes. Mechanistically, periostin-induced MMP-13 expression was abrogated by SC-514, demonstrating a link between periostin and NF-κB.
IA delivery of the periostin-siRNA nanocomplex represents a promising clinical approach to mitigate the severity of joint degeneration in OA. Our findings may thus provide an unequivocal scientific rationale for longitudinal studies of this approach. Utilizing a cartilage-specific gene-knockout strategy will further illuminate the functional role of periostin in OA.
最近的证据表明，外骨蛋白在骨关节炎（OA）中发挥着新的作用，因为其在膝关节损伤后的表达对关节软骨有害。我们进行这项研究是为了检验关节内（IA）外骨蛋白敲低是否会改善骨关节炎模型中的创伤后 OA。
通过内侧半月板不稳定（DMM）在 10 周龄雄性 C57BL/6J 小鼠（n = 24）中诱导创伤后 OA，并在手术后 8 周进行分析。使用新型肽-核苷酸聚集体通过 IA 传递小干扰 RNA（siRNA）抑制外骨蛋白表达。在对小鼠膝关节软骨进行组织学评估后，使用骨关节炎研究协会国际（OARSI）软骨损伤评分确定软骨退变的程度，并评估滑膜炎的严重程度。使用微计算机断层扫描测量骨变化。研究了在白细胞介素 1β（IL-1β）刺激下具有或不具有 IκB 激酶 2 抑制剂 SC-514 的人软骨细胞中外骨蛋白沉默的作用和机制。
使用 IA 递送肽-siRNA 纳米平台可显著消除创伤后 OA 小鼠中外骨蛋白的表达。接受外骨蛋白 siRNA 治疗的小鼠的 OARSI 软骨损伤评分明显低于未治疗的小鼠（10.94 ± 0.66）和接受 scrambled siRNA 治疗的小鼠（22.38 ± 1.30）（均 P = 0.002）。滑膜炎的严重程度没有差异。接受外骨蛋白 siRNA 治疗的小鼠的软骨下骨硬化、骨体积/总体积、体积骨密度和异位骨化明显降低。软骨免疫染色显示，外骨蛋白敲低降低了 DMM 诱导的基质金属蛋白酶 13（MMP-13）表达的强度，并且还降低了 p65 的磷酸化和聚集蛋白 neoepitope DIPEN 的免疫反应性。外骨蛋白敲低还抑制了软骨细胞中 IL-1β诱导的 MMP-13 和 ADAMTS-4 表达。机制上，SC-514 阻断了外骨蛋白诱导的 MMP-13 表达，表明外骨蛋白和 NF-κB 之间存在联系。
IA 递送外骨蛋白-siRNA 纳米复合物代表了一种有前途的临床方法，可以减轻 OA 中关节退化的严重程度。我们的发现可能为该方法的纵向研究提供明确的科学依据。利用软骨特异性基因敲除策略将进一步阐明外骨蛋白在 OA 中的功能作用。
