Therapeutic Effects of Lyophilized Conditioned-Medium Derived from Corneal Mesenchymal Stromal Cells on Corneal Epithelial Wound Healing

ABSTRACT

Objectives: The conditioned-medium derived from corneal mesenchymal stromal cells (cMSCs) has been shown to have wound healing and immunomodulatory effects in corneal injury models. Here, the therapeutic effects of lyophilized cMSC conditioned-medium were compared with fresh conditioned-medium.

Methods: The epithelial wound healing effects of fresh and lyophilized cMSC conditioned-medium were compared with conditioned-medium from non-MSC cells (corneal epithelial cells) using scratch assay. To evaluate the anti-inflammatory effects of fresh and lyophilized cMSC conditioned-media, macrophages were stimulated by a Toll-Like Receptor (TLR) ligand followed by treatment with the conditioned-media and measuring the expression of inflammatory genes. In vivo wound healing effects of fresh and lyophilized cMSC conditioned-media were assessed in a murine model of cornea epithelial injury.

Results: Both fresh and lyophilized cMSCs-derived conditioned-medium induced significantly faster closure of in vitro epithelial wounds compared to conditioned-medium from non-MSC cells (P < .0001). Treating stimulated macrophages with fresh or lyophilized cMSCs-derived conditioned-media significantly decreased the expression of inflammatory genes compared to control (P < .0001). Murine corneal epithelial wounds were healed by 87.6 ± 2.7% and 86.2 ± 4.6% following treatment with fresh and lyophilized cMSC conditioned-media, respectively, while the control was healed by 64.7 ± 16.8% (P < .05).

Conclusion: Lyophilized cMSC-derived conditioned-medium is as effective as fresh conditioned-medium in promoting wound healing and modulating inflammation. The results of this study support the application of lyophilized cMSCs-derived conditioned-medium, which allows for more extended storage, as a promising non-invasive option in the treatment of corneal wounds.

KEYWORDS:

• Human cornea
• mesenchymal stromal cell
• wound healing
• conditioned media
• conditioned-medium
• lyophilization

Author Contribution

S.J., A.R.D.: conceptualization, final approval of manuscript; S.J, G.Y., L.N.K., E.C.: manuscript writing, collection and/or assembly of data, data analysis, data interpretation; K.N.A.: review and editing; S.J., G.Y., L.N.K., K.K., M.G., P.H., A.R.D.: writing—review and editing; A.R.D.: funding acquisition.

Contributor Information

Ali R. Djalilian, Email: [email protected]

Disclosure of Potential Conflicts of Interest

The authors indicated no potential conflicts of interest.

Additional information

Funding

This work was supported by R01 EY024349 (ARD), Clinical Scientist Development Program Award K12EY021475 (KK), Core Grant for Vision Research EY01792 (MIR) from NEI/NIH; Unrestricted Grant to the department and Physician-Scientist Award both from Research to Prevent Blindness; and Eversight (providing both seed funding and human corneal research tissue). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.