Hautier A, Sabatier F, Stellmann P, Andrac L, Nouaille De Gorce Y, Dignat-George F, Magalon G
Service de Chirurgie Plastique, Centre des Grands Brûlés, Hôpital de La Conception, 147 boulevard Baille, Marseille, France.
Cell Tissue Bank. 2008 Mar;9(1):19-29. doi: 10.1007/s10561-007-9042-3. Epub 2007 Apr 26.
Human skin allografts are used in the treatment of severe burns and their preservation is therefore critical for optimal clinical benefit. Current preservation methods, such as 4 degrees C storage or cryopreservation, cannot prevent the decrease of tissue viability. The aim of this study was to assess viability and function of skin allografts in a new skin organ culture model, allowing conservation parameters as close as possible to physiological conditions: 32 degrees C, air-liquid interface and physiological skin tension.
Twelve skin samples, harvested from 6 living surgical donors, were conserved 35 days in two conditions: conservation at 4 degrees C and organ culture. Viability and function of skin samples were investigated at Day 0, 7, 14, 21, 28 and 35 using cell culture methods (trypan blue exclusion, Colony Forming Efficiency and Growth Rate), histopathological and histoenzymological studies (Ki67 immunostaining).
In the two conditions, fibroblast and keratinocyte viability was progressively affected by storage, with a significant decrease observed after 35 days. No statistical difference could be observed between the two conditions. The two methods were also comparable regarding alterations of fibroblast and keratinocyte culture parameters, which were respectively significantly reduced at Day 7 and 21, compared to fresh skin. By contrast, histopathological and histoenzymological studies revealed a better preservation of skin architecture and proliferative potential at 4 degrees C, as compared to organ culture.
These results indicate that skin organ culture does not provide significant advantages for skin allograft preservation. However, its potential use as an experimental model to study skin physiology and wound healing should be further evaluated.
人同种异体皮肤移植用于严重烧伤的治疗，因此其保存对于获得最佳临床疗效至关重要。目前的保存方法，如4℃储存或冷冻保存，无法防止组织活力下降。本研究的目的是在一种新的皮肤器官培养模型中评估同种异体皮肤的活力和功能，该模型能使保存参数尽可能接近生理条件：32℃、气液界面和生理皮肤张力。
从6名活体手术供体获取12个皮肤样本，在两种条件下保存35天：4℃保存和器官培养。在第0、7、14、21、28和35天，使用细胞培养方法（台盼蓝排斥试验、集落形成效率和生长速率）、组织病理学和组织酶学研究（Ki67免疫染色）来研究皮肤样本的活力和功能。
在两种条件下，成纤维细胞和角质形成细胞的活力均逐渐受到储存的影响，35天后观察到显著下降。两种条件之间未观察到统计学差异。在成纤维细胞和角质形成细胞培养参数的改变方面，两种方法也具有可比性，与新鲜皮肤相比，这些参数在第7天和第21天分别显著降低。相比之下，组织病理学和组织酶学研究显示，与器官培养相比，4℃时皮肤结构和增殖潜能的保存更好。
这些结果表明，皮肤器官培养在同种异体皮肤移植保存方面没有显著优势。然而，其作为研究皮肤生理学和伤口愈合的实验模型的潜在用途应进一步评估。
