Use of versican variant V3 and versican antisense expression to engineer cultured human skin containing increased content of insoluble elastin.
Use of versican variant V3 and versican antisense expression to engineer cultured human skin containing increased content of insoluble elastin.
复制标题
使用多功能蛋白聚糖变体 V3 和多功能蛋白聚糖反义表达来工程化培养的人类皮肤，使其含有增加的不溶性弹性蛋白含量。
DOI：
10.1002/term.1913
复制
发表时间：
2017
影响因子：
3.3
通讯作者：
Wight,ThomasN
中科院分区：
文献类型：
--
作者：
Merrilees,MervynJ;Falk,BenA;Zuo,Ning;Dickinson,MichelleE;May,BarnabyCH;Wight,ThomasN
用于修复皮肤伤口的皮肤替代品缺乏功能性弹性纤维。我们报告说，在培养的人皮肤的真皮中的不溶性弹性蛋白的含量可以增加，通过使用两种方法，增强皮肤成纤维细胞的弹性，迫使表达的多功能蛋白聚糖变体V3，它缺乏糖胺聚糖（GAG）链，和迫使表达的多功能蛋白聚糖反义降低水平的多功能蛋白聚糖变体V1与GAG链。将用V3或抗versican转导的人真皮成纤维细胞在标准条件下培养4周以产生真皮片，在前3周通过多次接种来增强生长。将在补充培养基中培养的人角质形成细胞添加到4周真皮片中，并将皮肤层再培养一周。5周时，角质形成细胞呈多层结构并分化，在鳞状细胞上层有桥粒连接和角蛋白沉积。真皮层由胶原束细胞外基质包围的分层成纤维细胞组成，在对照培养物中，有小的分散的弹性蛋白沉积物。V3和多功能蛋白聚糖反义的强制表达减缓了生长，降低了多功能蛋白聚糖V1表达，增加了原弹性蛋白表达和/或真皮层中不溶性弹性蛋白的大聚集体的沉积，并且增加了组织硬度，如通过纳米压痕所测量的。皮肤片也在Endoform Dermal Template™上培养，Endoform Dermal Template ™是由绵羊前肠固有层制成的可生物降解伤口敷料。皮肤结构和通过V3和抗多功能蛋白聚糖的强制表达而增强的弹性蛋白沉积被保留在该支持性基底上。版权所有© 2014约翰威利父子有限公司.
Skin substitutes for repair of dermal wounds are deficient in functional elastic fibres. We report that the content of insoluble elastin in the dermis of cultured human skin can be increased though the use of two approaches that enhance elastogenesis by dermal fibroblasts, forced expression of versican variant V3, which lacks glycosaminoglycan (GAG) chains, and forced expression of versican antisense to decrease levels of versican variant V1 with GAG chains. Human dermal fibroblasts transduced with V3 or anti‐versican were cultured under standard conditions over a period of 4 weeks to produce dermal sheets, with growth enhanced though multiple seedings for the first 3 weeks. Human keratinocytes, cultured in supplemented media, were added to the 4‐week dermal sheets and the skin layer cultured for a further week. At 5 weeks, keratinocytes were multilayered and differentiated, with desmosome junctions thoughout and keratin deposits in the upper squamous layers. The dermal layer was composed of layered fibroblasts surrounded by extracellular matrix of collagen bundles and, in control cultures, small scattered elastin deposits. Forced expression of V3 and versican antisense slowed growth, decreased versican V1 expression, increased tropoelastin expression and/or the deposition of large aggregates of insoluble elastin in the dermal layer, and increased tissue stiffness, as measured by nano‐indentation. Skin sheets were also cultured on Endoform Dermal Template™, the biodegradable wound dressing made from the lamina propria of sheep foregut. Skin structure and the enhanced deposition of elastin by forced expression of V3 and anti‐versican were preserved on this supportive substrate. Copyright © 2014 John Wiley & Sons, Ltd.