Co-culture of fibroblast-like synoviocytes with umbilical cord-mesenchymal stem cells inhibits expression of pro-inflammatory proteins, induces apoptosis and promotes chondrogenesis

Abstract

The present study aimed to investigate the effect of co‑culture of fibroblast‑like synoviocytes (FLS) with human umbilical cord‑mesenchymal stem cells (UC‑MSCs) on rheumatoid arthritis (RA) and to understand the mechanisms that mediate the induced changes. FLS and UC‑MSCs were isolated and cultured individually, FLS were then cultured with or without UC‑MSCs. The phenotype of UC‑MSCs was analyzed prior to co‑culture. The UC‑MSCs were successfully isolated and expanded, and exhibited a fibroblast‑like morphology. Enzyme‑linked immunosorbent assay (ELISA) and reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) were performed to determine the expression levels of interleukin (IL)‑1β, IL‑6, and chemokine (C‑C motif) ligand (CCL)‑2. The cell apoptosis rate was determined by flow cytometry. Furthermore, the RNAs of aggrecan and collagen type II were isolated and assessed in a chondrogenesis assay following co‑culture for 7, 14, 21 and 28 days. Protein expression levels of apoptosis‑related proteins, including B‑cell lymphoma (Bcl‑2), Bcl‑2‑associated X protein, p53 and phospho (p)‑AKT, and growth differentiation factor‑5 were analyzed by western blotting. ELISA and qRT‑PCR demonstrated that compared with FLS cultured alone, co‑culture with UC‑MSCs significantly downregulates the expression levels of IL‑1β, IL‑6 and CCL‑2. Additionally, the percentage of apoptotic cells was significantly increased in the co‑cultured cells (P<0.05), and the relative RNAs levels of aggrecan and collagen type II were increased compared with FLS alone. Furthermore, the expression levels of Bcl‑2 (P<0.05) and p‑AKT (P<0.05) were significantly decreased, whereas, p53 (P=0.001), Bax (P<0.01) and GDF‑5 (P<0.01) were increased by co‑culture of FLS with UC‑MSCs compared with FLS alone. In conclusion, co‑culture of FLS with UC‑MSCs may be important and clinically useful for the treatment of RA by inhibiting the expression of pro‑inflammatory mediators, inducing apoptosis and promoting chondrogenesis.

Intravenous Infusion of Umbilical Cord Blood-Derived Mesenchymal Stem Cells in Rheumatoid Arthritis: A Phase Ia Clinical Trial

“…They also suppress T cell activations and induced the generation of Tregs . A coculture of FLS with UCB‐MSCs inhibited the expression of pro‐inflammatory cytokines and chemokines IL‐1β, IL‐6, and the chemokine (C‐C motif) ligand‐2; it further induced FLS apoptosis and promoted chondrogenesis . hUCB‐MSCs downregulate the secretion of TNF‐α from activated macrophages and also accelerated anti‐inflammatory M2 polarizations in a paracrine manner .…”

Optimization of Human Mesenchymal Stem Cells for Rheumatoid Arthritis: Implications for Improved Therapeutic Outcomes

“…hMSCs have strong anti‐inflammatory and immunomodulatory properties; the extent to which hMSCs sense and respond to immune derangement and restore immune homeostasis is a matter of controversy ( 19 ). Studies show that treatment with hMSCs reduces the inflammatory response of peripheral blood mononuclear cells from patients with RA ex vivo ( 20 ). The ultimate goal of the studies described is to validate a potency assay for cellular products (hMSCs delivered by local injection or systemic infusion, or hMSC CM or exosomes delivered similarly) to be used therapeutically in RA and related autoimmune disorders.…”

Umbilical Cord Mesenchymal Stromal Cells for Cartilage Regeneration Applications

“…UC-MSCs have been demonstrated to secrete BMP2 in vitro and to induce the increase of endogenous BMP4, 5, and 7 levels in vivo [ 58 – 60 ]. In addition, UC-MSCs induce overexpression of GDF5/BMP14/CDMP1, promoting chondrogenic differentiation in cocultures with fibroblast-like synoviocytes, thus, suggesting their potential in cartilage repair [ 61 ]. Moreover, UC-MSCs respond to BMP6 via decapentaplegic homolog (SMAD) signaling (SMAD 1/4/5, BMPR1A, and BMPR2 receptors) enhancing osteogenic differentiation [ 62 ].…”

“…Interestingly, UC-MSCs displayed superior anti-inflammatory, immunomodulatory, and trophic effects compared to adult MSCs including articular cartilage (AC), Hoffa's fat pad (HFP), synovial membrane (SM), and maintain their immunomodulatory and anti-inflammatory properties after differentiation [ 50 , 84 , 95 ]. Moreover, coculture experiments of UC-MSCs and articular cartilage cells (ACs), fibroblast-like synoviocytes (FLSs), and nucleus pulposus cells (NPCs) showed their suitability for the treatment of arthritis, synovitis, and IVD degeneration [ 61 , 96 – 98 ]. Finally, there are several cartilage tissue engineering studies that demonstrated the osteochondral differentiation capacity of UC-MSCs in different scaffold constituted by acellular cartilage extracellular matrix (ACECM), alginate enriched in hyaluronic acid (Alg/HA), polycaprolactone/collagen (PCL/Coll), polyglycolic acid (PGA), poly L-lactide/D-lactide/glycolide (PLGA), poly-L-lactic acid (PLLA), polyvinyl alcohol-polycaprolactone (PVA-PCL), and silk fibroin/hyaluronic acid (SF/HA).…”