Plastic and Reconstructive Surgery Research, Manchester Interdisciplinary Biocentre, University of Manchester, Manchester, UK.
Exp Dermatol. 2012 May;21(5):376-81. doi: 10.1111/j.1600-0625.2012.01476.x.
Keloid disease (KD) is a common fibroproliferative disorder of unknown aetiopathogenesis, with highly unsatisfactory treatment. Therefore, it is crucial to have a robust and clinically relevant model for studying KD pathobiology as well as preclinical testing of potential KD therapeutics. However, the unique occurrence of KD in human skin and the corresponding lack of animal models pose a major challenge in KD research. Therefore, we developed a simplified assay for the serum-free, long-term organ culture of KD tissue that facilitates quantitative analyses of major KD read-out parameters. Four millimetre KD punches embedded in a collagen matrix and organ-cultured at the epidermis air-liquid interphase (ALI) in supplemented William's E medium showed optimal tissue, cell and RNA preservation for up to 6 weeks (as measured by H & E and Pyronin Y histochemistry as well as by MTT assay, lactate dehydrogenase release and quantitative Ki67/TUNEL immunohistomorphometry). The keloid phenotype persisted well during this period, as shown by collagen-I and -III synthesis (Herovici's histochemistry staining and ELISA), and analysis of the expression of significant KD markers (CD3, CD20, CD31, CD34, CD56, tryptase, Langerin, vimentin, neutrophil elastase, CTGF and Collagen). To functionally evaluate whether this assay can test the response to candidate therapeutics, dexamethasone, a glucocorticosteroid often used in KD therapy, was administered. Indeed, dexamethasone significantly reduced the keloid volume and cellularity plus induced epidermal shrinkage. Therefore, this novel assay provides a quantitative, clinically relevant model system for studying KD pathobiology and response to treatment.
瘢痕疙瘩病（KD）是一种常见的纤维组织增生性疾病，其病因和发病机制尚不清楚，治疗效果极不理想。因此，建立一个稳健且与临床相关的模型来研究 KD 的病理生物学以及对潜在 KD 治疗方法进行临床前测试至关重要。然而，KD 仅发生于人类皮肤，且相应的动物模型缺乏，这给 KD 研究带来了重大挑战。因此，我们开发了一种简化的无血清、长期器官培养 KD 组织的方法，该方法便于对主要 KD 读出参数进行定量分析。将 4 毫米的 KD 皮瓣嵌入胶原基质中，并在补充有威廉氏 E 培养基的表皮气液界面（ALI）进行器官培养，可使组织、细胞和 RNA 得到最佳保存，保存时间长达 6 周（通过 H&E 和派罗宁 Y 组织化学以及 MTT 测定、乳酸脱氢酶释放和 Ki67/TUNEL 免疫组织形态计量学定量分析来测量）。在此期间，瘢痕疙瘩表型保持良好，这表现为 I 型和 III 型胶原的合成（Herovici 组织化学染色和 ELISA），以及显著 KD 标志物（CD3、CD20、CD31、CD34、CD56、胰蛋白酶、朗格汉斯细胞、波形蛋白、中性粒细胞弹性蛋白酶、CTGF 和胶原蛋白）表达的分析。为了功能评估该方法是否可以测试候选治疗药物的反应，我们给予了地塞米松，这是一种常用于 KD 治疗的糖皮质激素。事实上，地塞米松显著减小了瘢痕疙瘩的体积和细胞密度，并诱导了表皮收缩。因此，这种新的测定方法为研究 KD 的病理生物学和对治疗的反应提供了一个定量的、与临床相关的模型系统。
