Intra-articular injection of placental mesenchymal stromal cells ameliorates pain and cartilage anabolism/catabolism in knee osteoarthritis

Mengqiang Fan; Jingwen Zhang; Li Zhou; Zuxiang Chen; Ronghua Bao; Longpo Zheng; Peijian Tong; Yuhai Ma; Letian Shan

doi:10.3389/fphar.2022.983850

Intra-articular injection of placental mesenchymal stromal cells ameliorates pain and cartilage anabolism/catabolism in knee osteoarthritis

Front Pharmacol. 2022 Nov 29:13:983850. doi: 10.3389/fphar.2022.983850. eCollection 2022.

Authors

Mengqiang Fan¹, Jingwen Zhang², Li Zhou¹, Zuxiang Chen¹, Ronghua Bao³, Longpo Zheng⁴, Peijian Tong¹, Yuhai Ma⁵, Letian Shan^{1

2}

Affiliations

¹ The First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, China.
² Cell Resource Bank and Integrated Cell Preparation Center of Xiaoshan District, Hangzhou Regional Cell Preparation Center (Shangyu Biotechnology Co Ltd), Hangzhou, China.
³ Fuyang Orthopaedics and Traumatology Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, China.
⁴ Department of Orthopedics, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China.
⁵ The Department of Orthopedics, Hangzhou Hospital of Zhejiang Provincial Armed Police Corps, Hangzhou, China.

Abstract

Background: Knee Osteoarthritis (kOA), the most common joint degenerative disorder, lacks effective therapeutics. Placenta-derived mesenchymal stromal cells (PMSCs) are effective in tissue repairing and generation, which have potential in treating kOA. This study aimed to determine the anti-kOA efficacy of PMSCs and to explore its action mode. Methods: Flow cytometry and three-line differentiation were performed for identification of PMSCs. In vivo, a rat kOA model established by anterior cruciate ligament transection (ACLT) surgery was used to evaluate the efficacy of PMSCs. Histopathological HE and SO staining with Osteoarthritis Research Society International scoring were conducted, and cartilage expressions of MMP13 and Col2 were measured by immunohistochemistry. Pain behavior parameters by mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL), were measured. In vitro, wound healing and cell immunofluorescence assays were conducted to detect the proliferation and migration ability of chondrocytes treated with PMSCs conditioned medium (PMSCs-CM). Quantitative real-time PCR (qRT-PCR) and Western blot (WB) assays were applied to explore the molecular action of PMSCs on chondrocytes. Results: The results of flow cytometry indicated that the surface markers of PMSCs (CD73 > 95%, CD90 > 95%, and CD34 < 2%) were consistent with the typical mesenchymal stromal cells. The in vivo data showed that PMSCs significantly reversed the kOA progression by protection of cartilage, regulation of anabolic (Col2) and catabolic (MMP13) expressions, and relief of pain symptoms. The in vitro data showed that PMSCs promoted chondrocyte proliferation and migration and significantly restored the IL-1β-induced abnormal gene expressions of Col2, Mmp13, Adamts4, Adamts5 and Sox9 and also restored the abnormal protein expressions of Col2, Mmp13 and Sox9 of chondrocytes. The molecular actions of PMSCs on chondrocytes in nested co-culture way or in conditioned medium way were similar, confirming a paracrine-based mode of action. Conclusion: This study demonstrated PMSCs' anti-kOA efficacy and its paracrine-based action mode, providing novel knowledge of PMSCs and suggesting it as a promising cell therapy for treatment of kOA.

Keywords: conditioned medium; osteoarthritis; paracrine; placenta-derived mesenchymal stromal cells; rat.