High effect of culture media derived from adipose tissue-derived stem cells on corneal epithelial wound healing

Abstract

Purpose : This study aimed to assess the efficacy of culture media derived from adipose tissue-derived stem cells (CM-ADSCs) for corneal epithelial wound healing compared to platelet-rich plasma (PRP).

Methods : The concentrations of epidermal growth factor (EGF), transforming growth factor (TGF) β-1, and fibronectin of PRP and CM-ADSCs were measured by enzyme-linked immunosorbent assay (ELISA). The efficacies of CM-ADSCs, PRP, and phosphate-buffered saline (PBS) were compared using an in vitro human corneal epithelial wound healing assay. The in vitro human corneal epithelial wounds were 70-μm wide for each well (96-well plate) containing cultured human corneal epithelial cells (HCECs). Following wounding, culture media were replaced with PBS, PRP, and CM-ADSCs, and diluted in corneal epithelial cell basal medium. Wound images were taken every 3h, and the area of the wound was calculated using the ImageJ software.

Results : EGF levels were significantly higher in CM-ADSCs compared to PRP (4253.2 ± 73.7 pg/mL vs. 74.4 ± 2.6 pg/mL, N = 3, P < 0.001). TGF-β1 and fibronectin levels were significantly higher in PRP compared to CM-ADSCs (TGF-β1; 67.8 ± 1.3 ng/mL vs. 3.8 ± 0.1 ng/mL, fibronectin: 285.0 ± 5.4 μg/mL vs. 15.3 ± 0.6 μg/mL, N = 3, P < 0.001, respectively). In the in vitro human corneal epithelial wound healing assay, the CM-ADSCs group significantly promoted wound healing compared to the PBS group and the PRP group (N = 6, P < 0.001, P = 0.020, respectively). In addition, the PRP group significantly promoted wound healing compared to the PBS group (N = 6, P < 0.001). The mean wound closure time was significantly shortened in the CM-ADSCs group (21.0 ± 3.3 hours) compared to the PBS group (55.0 ± 3.1 hours, P < 0.001) and PRP group (26.5 ± 3.5 hours, P <0.001). Wound closure time was also significantly reduced in the PRP group compared to the PBS group (P = 0.003).

Conclusions : CM-ADSCs promoted human corneal epithelial wound healing compared to PRP, suggesting that CM-ADSCs may meet an unmet medical need in the treatment of refractory corneal epithelial disorders.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.