토끼 태자에 형성시킨 구순열상의 치유과정에서 세포외기질 분포에 관한 연구 = A STUDY ON THE EXTRACELLULAR MATRIX IN THE ARTIFICIALLY CREATED CLEFT LIP WOUND HEALING OF RABBIT FET

국문 초록 (Abstract)

태아와 성체의 상처치유과정에서 다른 점으로는 태아 상처가 염증반응과 반흔조직의 생성이 없이 신속히 치유됨을 들수 있다. 태자상처의 비반흔성 치유와 세포외기질의 역할을 연관지어 해석하려는 노력이 진행중에 있으나, 각 실험 동물종간의 차이 및 세포외기질 구성요소의 다양성 등 많은 변수가 아직 미결의 과제로 남아 있다. 본 연구의 목적은 토끼 태자의 상처 치유과정에서 반흔조직의 형성과 관련이 있는 세포외 기질의 주요성분인 교원질 Ⅰ,Ⅲ,Ⅳ,Ⅴ형, fibronectin, laminin의 시기적 출현 및 분포양상을 조사하기 위한 것이다.
뉴질랜드산 흰 토끼를 임신 3기의 중반인 24일 째에 자궁절개술을 시행하고 태자에 인위적으로 구순열상을 만든 후, 봉합한 군(봉합군)과 봉합하지 않은 군(비봉합군), 정상대조군(sham operation group)으로 나누고 이들을 각각 수술 후 1,2,3,5,7일째에 희생시켜 상처치유에 대한 육안관찰 소견과 H&F염색소견 및 교원질Ⅰ,Ⅲ,Ⅳ,Ⅴ형, fibronectin, laminin의 면역조직화학적 염색 소견을 관찰하여 다음과 같은 결론을 얻었다.

1. 봉합군과 비봉합군에서 전기간동안 염증반응은 없었다.
2. 비봉합군의 재상피화가 봉합군에 비해 다소 느렸다.
3. 교원질 Ⅰ,Ⅲ,Ⅴ형은 봉합군과 비봉합군에서 2일군까지는 발견되지 않았고 3일군 이후 존재하였으나 염색정도가 인접정상조직이나 정상대조군에 비하여 증가하지 않았다.
4. 상피 기저막에서 교원질 Ⅳ형의 염색소견은 봉합군과 비봉합군에서 3일군이 다소 증가하였으나 그후 감소하여 인접정상조직이나 정상조직이나 정상대조군과 차이가 없었고, laminin의 염색소견은 봉합군과 비봉합군에서 5일군이후 발견되었고 염색정도의 증감없이 7일군까지 유지되었고 인접정상조직이나 정상대조군과 차이가 없었다.
5. 혈관내피세포 기저막에서 laminin과 교원질 Ⅳ형의 염색소견은 인접정상조직이나 정상대조군에 비해서 증가하지 않았다.
6. Fibronectin은 봉합군과 비봉합군에서 fibrin clot과 창상 기저부 및 창상연을 따라 3일군까지 염색소견이 증가하였다가 그후 감소하여 인접정상조직이나 정상대조군과 차이가 없었다.

다국어 초록 (Multilingual Abstract)

Adult wound healing is accompanied with inflammation and eventual scar formation, whereas fetal wounds heal rapidly by mesenchymal prpliferation without significant inflammatory cell participation and with minimal or no scar formation. The cellular mechanisms underlying these differing forms of wound healing are unknown but the extracellular matrix, through its effects on cell function, may play a key role. Therefore the purpose of this study is to investigate the spatial and temporal deposition of several component of extracellular matrix, which are known to be involved with scar formation, in the artificially created cleft lip wound healing of fetuses.
The author had undergone hysterotomy and created cleft lip-like defects on fetuses of New Zealand White Rabbit in mid-third trimester(24 days). Fetuses were divided into the repaired group, the unrepaired group and the sham-operated control group. At 1,2,3,5,7 days after procedure, fetuses were obtained by Caeserean section. After documenting the viability of fetuses, they were photographed to compare size and facial morphology and sectioned for histological examination by H&E stain and spatial and temporal deposition of collagen typeⅠ,Ⅲ,Ⅳ,Ⅴand fibronectin, laminin by immunohistochemical method.
The findings are summarized as follows
1. There were lack of inflammation in the repaired and the unrepaired group during experimental periods.
2. The reepithelialization of the unrepaired group was slower than that of the repaired group.
3. Collagen Ⅰ,Ⅲ,Ⅴ were found from post-op. third day. There were no difference of distribution in the control, the repaired and the unrepaired group. Collagen typesⅠ,Ⅲ,Ⅴ were present in all groups with restoration of the normal collagen pattern in the fetus. This implies that lack of scarring in fetal wounds is due to the difference of collagen organization pattern within wound and not simply lack of collagen formation.
4. Collagen Ⅳ was slightly increased at post-op. third day and decreased after post-op. fifth day. Eventually there were no difference in control, the repaired and the unrepaired group. Laminin was found at post-op. fifth day and maintained staining density until post-op. seventh day. There were no differences in the control, the repaired and the unrepaired group. According to staining of laminin and collagen typeⅣ in epithelial basement membrane, formation of epithelial basement membrane was not completed until reepithelialization was finished.
5. According to staining of laminin and collagen typeⅣ, there were no increase of neovascularity in the repaired and the unrepaired group.
6. Fibronectin was increased until post-op. third day at fibrin clot, wound base and margin and decreased after post-op. fifth day. Eventually, there were no differences in the control, the repaired and the unrepaired group. So it implies fibronectin plays a role as provisional matrix for fetal wound healing.