Comparison of Therapeutic Effects of Three Human-Derived Materials in A Mouse Corneal Alkali Burn Model

Abstract

To compare the effect of amniotic membrane (AM) suspension, umbilical cord serum (UCS), and peripheral blood serum (PBS) in a mouse alkali burn model. The right eyes of 32 BALB/c mice were injured using 0.2 N NaOH. After alkali injury, the mice were divided into 4 groups and topically administered one of the following solutions four times daily: 200 μg/mL AM suspension, 20% UCS, 20% PBS, or normal saline. The epithelial defect areas, degrees of opacity, and corneal neovascularization (NV) areas were evaluated. Histologic examination and mRNA expression levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, MMP-8, and MMP-9 were also evaluated. The epithelial defect areas were smaller in all three treatment groups compared to the control group on day 2 (all p<0.05). However, only the epithelial defect areas in the UCS group were smaller than those in the control and PBS groups on day 3 (all p<0.05). The degrees of opacity were lower in all three treatment groups than that in the control group on days 2 and 7 (all p<0.05), and they were lower in the UCS and AM suspension groups than the control group on days 3 (p<0.05). Corneal NV areas were significantly smaller in UCS and AM suspension groups (all p<0.05). The expression levels of TNF-α, IL-6, MMP-8, and MMP-9 mRNA were down-regulated in the UCS and AM suspension groups compared to the control group (all p<0.05). The infiltration of the inflammatory cells in all treatment groups were lesser in all treatment groups than in the control group, and those in the PBS group was lesser than the control group, but more than those in the UCS and AM suspension groups (all p<0.05). The UCS and AM suspension eye drops promoted corneal wound healing and reduced opacity and inflammation in a mouse corneal alkali burn model. On the other hand, PBS eye drops showed limited therapeutic effects than the UCS and AM suspension eye drops.;쥐 각막 알칼리 화상 모델에서 양막현탁액, 제대혈청, 말초혈청의 치료적 효과를 비교해 보고자 한다. 32마리의 BALB/c 쥐의 우안에 0.2 N NaOH로 알칼리 화상을 입힌 후 네 군으로 나누어 200 μg/mL 양막현탁액, 20% 제대혈청, 20% 말초혈청, 생리식염수 중 한 가지를 하루 4회씩 점안하였다. 각막상피결손 면적, 혼탁 정도, 신생혈관 면적을 분석하였다. 조직학적인 검사와 tumor necrosis factor (TNF)-α, interleukin (IL)-6, vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP)-2, MMP-8, MMP-9의 mRNA 발현 정도를 분석하였다. 각막상피결손 면적은 손상 2일 째 모든 치료군에서 대조군보다 작았다 (all p<0.05). 그러나 손상 3일 째에는 오직 제대혈청군의 각막상피결손면적이 대조군과 제대혈청군보다 작았다 (all p<0.05). 각막 혼탁의 정도는 손상 2일 째 와 7일 째 모든 치료군에서 대조군보다 적었지만, 손상 3일째는 제대혈청군과 양막현탁액군에서만 대조군보다 적었다 (all p<0.05). 각막 신생혈관의 면적은 손상 7일 째 제대혈청군과 양막현탁액군에서 대조군보다 적었다 (all p<0.05). 모든 치료군에서 TNF-α, IL-6, MMP-8, MMP-9 mRNA 발현이 대조군보다 감소되어 있었으며, 제대혈청군과 양막현탁액군의 TNF-α와 MMP-9 mRNA 발현은 말초혈청군보다 낮았다 (all p<0.05). 염증성 세포의 침윤 정도는 모든 치료군에서 대조군에 비해 현저하게 적었으며, 말초혈청군의 염증성 세포의 침윤 정도는 대조군에 비해서는 적었고 제대혈청군과 양막현탁액군에 비해서는 많았다 (all p<0.05). 제대혈청과 양막현탁액은 쥐 알칼리 화상 모델에서 각막창상치유를 증진시키고 혼탁과 염증의 정도를 현저하게 줄였다. 반면에, 말초혈청은 제대혈청과 양막현탁액에 비해 제한된 치료 효과를 보였다.