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목적 : Brain-derived neurotrophic factor (BDNF), a well-known neuroprotective cytokine, has been found to play an important role in neurogenesis. As one of the most innervated tissue, however, the role of BDNF in cornea epithelium remains unclear. This study was to explore the roles and mechanisms of BDNF in the activation of corneal limbal progenitor cells and wound healing of mouse corneal epithelium.
방법 : The entire corneal epithelium including limbal region in C57BL/6 mice was scraped with brush and exogenous BDNF was applied once in a day during 5 days. The defects of corneal epithelium were visualized at 24, 48, and 72 hours by instilling 0.25% fluorescein sodium. Mouse or human corneal limbal cells was harvested and cultured with or without BDNF in keratinocyte serum-free medium (KSFM) supplemented with bovine pituitary extract or human limbal specific media.
결과 : In mice subjecting to mechanical removal of corneal epithelium, the limbal progenitor cells activation and wound healing were promoted by exogenous BDNF application. In cultured mouse limbal cells, BDNF enhanced the colony-forming efficiency, stimulated the mitogenic proliferation, and upregulated the expression levels of limbal markers (ABCG2 and P63). Furthermore, in ex vivo culture of human limbal tissue, supplement of BDNF promoted colony-forming efficiency, limbal cell proliferation, and increased expression of limbal markers.
결론 : Thus, the capability of expanding corneal limbal progenitor cells and promoting corneal epithelial wound healing indicates the potential application of BDNF in ameliorating limbal stem cell deficiency.
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