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목적 : To evaluate the effect of exogenous recombinant human BMP-7 (rhBMP-7) on transforming growth factor-β (TGF-β) induced epithelial mesenchymal transition (EMT) of human corneal epithelial cells in vitro and corneal fibrosis during wound healing after keratectomy in a rabbit in vivo model.
방법 : Human corneal epithelial cells (HCECs) were used for the EMT in vitro study. To evaluate the feasibility of ocular surface application of rhBMP-7, cytotoxicity and proliferation were evaluated. The effects of rhBMP-7 on TGF-β induced EMT were evaluated by fibronectin, α-smooth muscle actin (α-SMA), E-cadherin, and matrix metaloproteinase-9 (MMP-9) expressions. The effect of topically applied anti-fibrotic effect of rhBMP-7 was evaluated by immunohistochemical staing for α-SMA.
결과 : The number of metabolically active cells was significantly higher in the cells cultured with rhBMP-7 alone and rhBMP-7 with 0.1% hyaluronic acid (HA). (P < 0.05) Increased fibronectin level with TGF-β stimulation in vitro HCECs was attenuated by both rhBMP-7 (31.6%, P < 0.05) and rhBMP-7+HA (35.7%, P < 0.01) containing media. HCECs cultured in the media containing rhBMP-7 (72.2%; P < 0.01) and rhBMP-7 with HA (69.3%; P < 0.01) demonstrated statistically significant decreased α-SMA protein levels. E-cadherin expression was enhanced in the media containing rhBMP-7 (289.7%, P < 0.01) and containing rhBMP-7 with HA (290.2%, P < 0.01) MMP-9 expression was attenuated in the media containing HCECs rhBMP-7 (23.6%, P < 0.05) and rhBMP-7 with HA (27.4%, P < 0.05). Topical application of 2.5 and 25 ㎍/㎖ rhBMP-7 with 0.1 HA significantly decreased α-SMA+ cells as compared to corneas that received 0.1% HA and/or 0.25 ㎍/㎖ rhBMP-7 (P < 0.05).
결론 : Exogenous rhBMP-7 modulates wound healing and attenuates fibrosis in vivo and in vitro model.
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