CAJ | 학술논문

目的探讨β1转化生长因子(transforming growth factor β1,TGF-β1)诱导人表皮干细胞(human epidermal stem cells,hESCs)分化与瘢痕形成之间存在的关联.方法将包皮环切术后的包皮用中性蛋白水解酶消化,采用改良的Ⅳ型胶原选择黏附法分离、培养,传代至第3代时经过hESCs表面标志物β1整合素和CK19检测,证实为hESCs后接种于24孔板,随机分为3组:3 d组、7 d组、空白对照组,前两组分别加入梯度浓度的TGF-β1(0.1、5.0、10.0 ng/ml)诱导,采用HE常规染色及Masson胶原特殊染色、免疫组织化学染色等手段检测量波形蛋白表达和羟脯氨酸试剂盒法测量各组上清液中胶原含量.结果 hESCs在TGF-β1诱导下,形态由圆形转变为梭形类成纤维细胞样,Masson胶原染色阳性,释放到细胞上清液中的胶原浓度,均显著高于对照组,抗-波形蛋白染色阳性率各组都至少在(95.00±1.20)%以上,大于对照组的(5.70±0.20)%(P<0.05).结论结果表明,hESCs与病理性瘢痕发生关系极其密切,在TGF-β1体外诱导下向成纤维细胞分化,分泌胶原,提示在病理性瘢痕的发生中,hESCs可能是活化的成纤维细胞的另一个来源,hESCs可能参与瘢痕增生发生过程.
목적 탐토β1전화생장인자(transforming growth factor β1,TGF-β1)유도인표피간세포(human epidermal stem cells,hESCs)분화여반흔형성지간존재적관련.방법 장포피배절술후적포피용중성단백수해매소화,채용개량적Ⅳ형효원선택점부법분리、배양,전대지제3대시경과hESCs표면표지물β1정합소화CK19검측,증실위hESCs후접충우24공판,수궤분위3조:3 d조、7 d조、공백대조조,전량조분별가입제도농도적TGF-β1(0.1、5.0、10.0 ng/ml)유도,채용HE상규염색급Masson효원특수염색、면역조직화학염색등수단검측량파형단백표체화간포안산시제합법측량각조상청액중효원함량.결과 hESCs재TGF-β1유도하,형태유원형전변위사형류성섬유세포양,Masson효원염색양성,석방도세포상청액중적효원농도,균현저고우대조조,항-파형단백염색양성솔각조도지소재(95.00±1.20)%이상,대우대조조적(5.70±0.20)%(P<0.05).결론 결과표명,hESCs여병이성반흔발생관계겁기밀절,재TGF-β1체외유도하향성섬유세포분화,분비효원,제시재병이성반흔적발생중,hESCs가능시활화적성섬유세포적령일개래원,hESCs가능삼여반흔증생발생과정.
Objective To investigate the correlation between human epidermal stem cell (hESCs) and hypertrophic scar or keloid. Methods Improved collagen Ⅳ-coated adhesion methods was used to isolate and culture the epidermal stem cells after neutral protease selectively digested the dermo-epidermal junctions. After the cells were cultured and expanded in vitro, and passage 3 hESCs were induced by different concentrations of TGF-β1 (0.1, 5.0, and 10.0 ng/ml). Morphological fea-tures and identification of these cells were meseasured by HE, Masson, immunohistochemical staining on the days 3 and 7, respectively. Results After induced by TGF-β1 for 3 and 7 days, the morpholo-gy of the epidermal stem cell (hESCs) was changed into fusiform shape, similar to fibroblasts. 70 % ofthe cell which was induced by TGF-β1 were blue stained in the cytoplasm by Masson stain, which is the distinctive method for collagen, suggesting collagen appeared or increased in the cells. The collagen concentrations in supernatants of hESCs were 0.4150±0.0014, 0.3380±0. 0020, and 0.3870±0.0020, much higher than that in control group (0.0780±0.0025) and normal skin fibro-blast group (0.15004±0.0051) (P<0.05). Immunohistochemical staining revealed that positive rates of these cells for anti-vimentin staining were more than (95.00±1.20)% in experiments and (5.70±0.20)% in control group. Conclusion The differentiantion of hESCs induced by TGF-β1 into fibro-blasts indicates that hESCs may play a role in the pathogenesis of hypetrophic scar and keloid.