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Photobiomodulation at 660 nm promotes collagen synthesis via downregulation of HIF-1α expression without photodamage in human scleral fibroblasts in vitro in a hypoxic environment

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Abstract

Purpose

The increasing prevalence of myopia is a global public health issue. Because of the complexity of myopia pathogenesis, current control methods for myopia have great limitations. The aim of this study was to explore the effect of photobiomodulation (PBM) on human sclera fibroblasts (HSFs) under hypoxia, in the hope of providing new ideas for myopia prevention and control.

Methods

Hypoxic cell model was established at 0, 6, 12, and 24 h time points to simulate myopia microenvironment and explore the optimal time point. Control, hypoxia, hypoxia plus light, and normal plus light cell models were set up for the experiments, and cells were incubated for 24 or 48 h after PBM (660 nm, 5 J/cm2), followed by evaluation of hypoxia-inducible factor 1α (HIF-1α) and collagen I a1 (COL1A1) proteins using Western blotting and immunofluorescence, and photo damage was detected by CCK-8, scratch test, and flow cytometry assays. We also used transfection technology to further elucidate the regulatory mechanism.

Results

The change of target proteins is most obvious when hypoxia lasts for 24 h (p < 0.01). PBM at 660 nm increased extracellular collagen content (p < 0.001) and downregulated expression of HIF-1α (p < 0.05). This treatment did not affect the migration and proliferation of cells (p > 0.05), and effectively inhibited apoptosis under hypoxia (p < 0.0001). After overexpression of HIF-1α, the effect of PBM was attenuated (p > 0.05).

Conclusions

Photobiomodulation at 660 nm promotes collagen synthesis via downregulation of HIF-1α expression without photodamage.

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Data availability

The datasets supporting the conclusions of this article will be available from the corresponding author on reasonable request.

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Authors and Affiliations

  1. Department of Ophthalmology, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, 200092, China

    Pengbo Zhang & Huang Zhu

  2. Department of Ophthalmology, Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan, China

    Xibo Zhang

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  1. Pengbo Zhang
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Contributions

Pengbo Zhang conceived, designed, and performed the experiments. Xibo Zhang and Huang Zhu designed and modified this article.

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Correspondence to Xibo Zhang or Huang Zhu.

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All named authors meet the International Committee of Medical Journal Editors (ICMJE) criteria for authorship for this article, take responsibility for the integrity of the work as a whole, and have given their approval for this version to be published.

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The study was performed in accordance with the Helsinki Declaration of 1964, and its later amendments. Informed consent to include potentially identifiable data was acquired from all the participants.

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Zhang, P., Zhang, X. & Zhu, H. Photobiomodulation at 660 nm promotes collagen synthesis via downregulation of HIF-1α expression without photodamage in human scleral fibroblasts in vitro in a hypoxic environment. Graefes Arch Clin Exp Ophthalmol 261, 2535–2545 (2023). https://doi.org/10.1007/s00417-023-06066-5

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