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Nanoparticle hydrogel system delivery of miR-494-3p to improve tendon healing by targeting CXXC4

Gene Therapy (2025)Cite this article

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Abstract

Due to the poor healing capacity of tendons, the healing process is slow, with a risk of re-rupture post-injury. In this study, we found that miR-494-3p was one of the miRNAs with significant expression differences after tendon injury by sequencing in the rat Achilles tendon injury model. Therefore, we hypothesized that regulating miR-494-3p expression in tendons could improve tendon healing. Considering the long healing process of the tendons and the short half-life of miRNA, we hope to achieve the best efficacy by delivering miR-494-3p using a sustained-release nanoparticle hydrogel system. In the results, with an increase in miR-494-3p, the tendon biomechanics were significantly improved after 2-week repair, and the content of collagen I (Col I) also increased. Through bioinformatics prediction, double luciferase, and immunohistochemistry experiments, we confirmed that miR-494-3p targeting CXXC finger protein 4 (CXXC4) promoted tendon healing. In conclusion, the miR-494-3p/nanoparticles hydrogel delivery system can protect and sustainedly transfer miR-494-3p into tenocytes, block the translation of CXXC4, increase the expression of Col I, and ultimately improve tendon healing.

A nanoparticle hydrogel delivery system of miRNA was constructed and applied to injured tendons. Finally, we confirmed that the miR-494-3p/nanoparticles hydrogel delivery system can protect and sustainedly transfer miR-494-3p into tenocytes, block the translation of CXXC4, increase the expression of Col I, and ultimately improve tendon healing.

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Fig. 1: miRNA sequencing analysis of the gene expression profile of tendon tissue.
Fig. 2: Characteristics of the nanoparticle hydrogel system and biomechanical experiment.
Fig. 3: Therapeutic effect of hydrogel-containing miR-494-3p/nanoparticles in vivo.
Fig. 4: The effect of miR-494-3p/nanoparticles on rat Achilles tenocytes.
Fig. 5: mRNA sequencing analysis of the gene expression profile of tendon tissue.
Fig. 6: Possible target genes of miR-494-3p.
Fig. 7: CXXC4 was regulated by miR-494-3p and the effect of CXXC4 on Col I expression.

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Data availability

The RNA sequencing data generated in this study have been deposited in the NCBI Sequence Read Archive (SRA) (https://www.ncbi.nlm.nih.gov/sra). Additional raw data are available from the corresponding author upon reasonable request.

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Funding

This study was supported by the National Natural Science Foundation of China (Nos 82102186) and the Jiangsu Provincial Medical Key Discipline (XKTJ-XK202003). This study was also supported by the Jiangsu Provincial Medical Innovation Center of Neuromicroscopy and Minimally Invasive Translational Medicine (CXZX202212) and CNNC Youth Talent Program.

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Author notes

  1. These authors contributed equally: Gu Heng Wang, Lei Wang, Lei Sheng.

Authors and Affiliations

  1. Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, PR China

    Gu Heng Wang, Lei Sheng, Hua Jian Shan & Xiao Zhong Zhou

  2. Department of Hand Surgery, Affiliated Hospital of Nantong University, Nantong, PR China

    Gu Heng Wang, Wei Gang Zhu, Ai Dong Deng & Jun Tan

  3. Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, PR China

    Lei Wang

  4. Department of Medical Informatics, School of Medicine, Nantong University, Nantong, PR China

    Ya Lan Chen

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  1. Gu Heng Wang
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Contributions

G.H.W. performed most of the experiments, wrote and revised the manuscript. L.W. partly designed and performed the experiments. L.S. partly designed the experiments and revised the manuscript. H.J.S. partly performed the experiments and revised the manuscript. W.G.Z. and Y.L.C. partly performed the experiments and data analysis. A.D.D. and J.T. partly designed and supervised the experiments. X.Z.Z. provided the initial idea of this study, designed the experiments, and revised the manuscript. G.H.W., L.W. and L.S. contributed equally to this work.

Corresponding authors

Correspondence to Ai Dong Deng, Jun Tan or Xiao Zhong Zhou.

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The authors declare no competing interests.

Ethical approval

All animal experiments involving Sprague Dawley rats were conducted in accordance with the relevant guidelines and regulations, and were approved by the Experimental Animal Care and Use Committee of Nantong University (Approval number: S20200314-016). All rats were provided by the Laboratory Animal Center of Nantong University.

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Wang, G.H., Wang, L., Sheng, L. et al. Nanoparticle hydrogel system delivery of miR-494-3p to improve tendon healing by targeting CXXC4. Gene Ther (2025). https://doi.org/10.1038/s41434-025-00543-8

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