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Toll-like receptor activation regulates the paracrine effect of adipose-derived mesenchymal stem cells on reversing osteoarthritic phenotype of chondrocytes

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Abstract

Background

The therapeutic efficacy of intra-articular mesenchymal stem cells (MSCs) injection for patients with osteoarthritis (OA) currently exhibits inconsistency, and the underlying mechanism remains elusive. It has been postulated that the immunomodulatory properties and paracrine activity of MSCs might be influenced by the inflammatory micro-environment within osteoarthritic joints, potentially contributing to this observed inconsistency.

Methods

Adipose-derived MSCs (ADSCs) were isolated from SD rats and pre-treated with Toll-like receptor 3 (TLR3) agonist Poly I:C or Toll-like receptor 4 (TLR4) agonist LPS. The pre-treated ADSCs were then co-cultured with IL-1β-induced osteoarthritic chondrocytes using a Transwell system to analyze the paracrine effect of ADSCs on reversing the osteoarthritic phenotype of chondrocytes.

Results

RT-PCR and Western blot analysis revealed that Poly I:C and LPS pre-treatments up-regulated the expression of IL-10 and IL-6 in ADSCs, respectively. Furthermore, only Poly I:C-preconditioned ADSCs significantly promoted proliferation while inhibiting apoptosis in IL-1β-treated chondrocytes. Additionally, Poly I:C-preconditioned ADSCs downregulated MMP13 expression while upregulating aggrecan and collagen II expression levels in IL-1β-treated chondrocytes.

Conclusions

TLR3 activation polarizes ADSCs into an immunomodulatory phenotype distinct from TLR4 activation, exerting differential effects on reversing the osteoarthritic phenotype of chondrocytes; thus indicating that MSCs’ paracrine effect regulated by TLRs signaling impacts the efficacy of intra-articular MSCs injection.

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Data availability

The original contributions presented in the study are included in the article, further inquiries can be directed to the corresponding authors.

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Funding

The authors’ research work is funded by Chongqing Key Laboratory of Precision Medicine in Joint Surgery (grant: 425Z2138), Chongqing Excellent Scientist Project (grant: 425Z2W21), Chongqing Natural Science Foundation (grant: cstc2021jcyj-msxmX0135), and Chongqing Postdoctoral Research Project Special Fund (grant: 2021XM3033).

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Authors and Affiliations

  1. Center for Joint Surgery, Southwest Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038, China

    Zu Wan, Xin Wang, Zhenlan Fu, Yanming Ma, Gang Dai, Xiaoyuan Gong, Guangxing Chen & Liu Yang

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  1. Zu Wan
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  2. Xin Wang
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  3. Zhenlan Fu
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  8. Liu Yang
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Contributions

Z.W, Z.F. and Y.M. collected data, Z.W. and Z.F. wrote the main manuscript text, Z.W., G.D. and X.W. prepared all figures, and X.G., G.C and L.Y. Supervised this study. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Xiaoyuan Gong, Guangxing Chen or Liu Yang.

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The authors declare no competing interests.

Ethical statement

All animal procedures were approved by the Laboratory Animal Welfare and Ethics Committee of Army Medical University (the Third Military Medical University) (AMUWEC20223809). Our experimental design, procedures, and methods of animal study are in accordance with the "General Principles for Experimental Animal Welfare" (GB/T 42011–2022). We have strictly followed the aforementioned regulations during our experimental procedures.

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Wan, Z., Wang, X., Fu, Z. et al. Toll-like receptor activation regulates the paracrine effect of adipose-derived mesenchymal stem cells on reversing osteoarthritic phenotype of chondrocytes. Mol Biol Rep 51, 550 (2024). https://doi.org/10.1007/s11033-024-09499-1

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