Abstract
Purpose :
Lumican is a keratan sulfate proteoglycan core protein that is involved in the regulation of collagen fiber alignment, epithelial healing, and local inflammation in the cornea. The presenter reported that Lumican gene deletion (KO) mice have delayed wound healing of whole corneal incisions (ARVO2019). To elucidate the role of Lumican in the wound healing process of corneal parenchymal incisions and its mechanism, we examined growth factor expression using ocular fibroblasts from Lumican knockout mice.
Methods :
Ocular fibroblasts were cultured from the eyeballs of 1-day-old wild-type (WT) and KO mice. The obtained ocular fibroblasts were adjusted to 3 × 106 cells/ml and mixed with Collagen-Based Cell Contraction Assay Kit, and the progress with and without TGFβ1 addition was observed every 12 hours for a total of 96 hours, and area fraction (%) = [(initial surface area of gel The degree of gel shrinkage was analyzed over time by calculating [-Final surface area of gel/Initial surface area of gel] × 100. Ocular fibroblasts were collected at a certain time after addition of TGFβ1 (1 ng/ml), and the expression of αSMA, collagen I, fibronectin, Smad3, and p-smad3 in WT and KO cells were compared by immunohistological analysis.
Results :
After 24 hours from the addition of TGF-β1, its contraction was significantly inhibited in the gel mixed with KO cells. Immunohistological examination showed staining differences between WT and KO cells in αSMA and collagen1. Fibronectin showed no difference in staining between WT and KO cells
Conclusions :
Lumican is involved in the expression of αSMA and type 1 collagen in mesenchymal cells, which may be a possible mechanism for the delayed corneal parenchymal wound healing in KO mice.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.