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CD44 mediates hyaluronan to promote the differentiation of human amniotic mesenchymal stem cells into chondrocytes

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Abstract

Objectives

CD44 is the major receptor for hyaluronan (HA), but its effect on HA-induced differentiation of human amnion mesenchymal stem cells into chondrocytes is unclear. This study aimed to investigate the effects and mechanisms of CD44 in HA-induced chondrogenesis.

Methods

Immunocytochemistry and toluidine blue staining were used to assess the secretion of type II collagen and aggrecan, respectively. qRT-PCR and western blotting were performed to evaluate the expression of key genes and proteins.

Results

The expression of aggrecan and type II collagen was downregulated after using the anti-CD44 antibody (A3D8). The transcriptional levels of chondrocytes‑associated genes SRY‑box transcription factor 9, aggrecan, and collagen type II alpha 1 chain were also decreased. Thus, CD44 may mediate HA-induced differentiation of hAMSCs into chondrocytes. Further investigation indicated that expression of phosphorylated (p)‑Erk1/2 and p‑Smad2 decreased following CD44 inhibition. The changes in the expression of p-Erk1/2 and p-Smad2 were consistent after using the ERK1/2 inhibitor (U0126) and agonist (EGF), respectively. After administering the p-Smad2 inhibitor, the expression levels of p-ERK1/2 and p-Smad2 appeared downregulated. The results showed crosstalk between Erk1/2 and Smad2. Moreover, inhibition of p-Erk1/2 and p-Smad2 significantly reduced the accumulation of aggrecan and type II collagen.

Conclusion

These data indicate that CD44 mediates HA-induced differentiation of hAMSCs into chondrocytes by regulating Erk1/2 and Smad2 signaling.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors are grateful for the financial support from the National Natural Science Foundation of China, PR China (grant number: 31960191), Guizhou High-Level Innovative Talent Support Program, PR China (grant number: QKHPT-RC-GCC[2022]001-1), the Science and Technology Foundation of Guizhou, PR China (grant number: QKHJC-ZK[2022]-YB-666), and the Science and Technology Foundation of Zunyi Science and Technology Bureau, PR China (grant numbers: ZSKH-HZ-2020-212, ZSKH-HZ-2021-09, ZSKH-HZ-2021-87).

Funding

Science and Technology Foundation of Zunyi Science and Technology Bureau, Grant/Award Number: ZSKH-HZ-2020-212, ZSKH-HZ-2021-09, and ZSKH-HZ-2021-87; Department of Science and Technology of Guizhou Province, Grant/Award Number: QKHJC-ZK[2022]-YB-666; Guizhou High-Level Innovative Talent Support Program, Grant/Award Number: QKHPT-RC-GCC[2022]001-1; National Natural Science Foundation of China, Grant/Award Number: 31960191.

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Authors and Affiliations

  1. Institute of Medicinal Biotechnology, Affiliated Hospital of Zunyi Medical University, 149 Dalian Road, Huichuan District, Zunyi, 563003, China

    Yan Xu, Ai-Tong Wang & Jian-Hui Xiao

  2. Department of Orthopaedics, Affiliated Hospital of Zunyi Medical University, Zunyi, China

    Yan Xu & Jian-Hui Xiao

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  1. Yan Xu
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Contributions

All authors contributed to the study conception and design. J-HX designed the experiment, interpreted the data and modified the manuscript. Material preparation, data collection and analysis were performed by YX and A-TW. The first draft of the manuscript was written by YX and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Jian-Hui Xiao.

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Competing interests

The authors have no relevant financial or non-financial interests to disclose.

Ethical approval

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of Affiliated Hospital of Zunyi Medical University (approval no. (2014) 2-085).

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Informed consent was obtained from all individual participants included in the study.

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Xu, Y., Wang, AT. & Xiao, JH. CD44 mediates hyaluronan to promote the differentiation of human amniotic mesenchymal stem cells into chondrocytes. Biotechnol Lett 45, 411–422 (2023). https://doi.org/10.1007/s10529-022-03322-2

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  • DOI: https://doi.org/10.1007/s10529-022-03322-2

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