Vienna Burn Centre, Division of Plastic and Reconstructive Surgery, Department of Surgery, Medical University Vienna, Vienna, Austria.
Burns. 2011 Jun;37(4):626-30. doi: 10.1016/j.burns.2010.07.016. Epub 2010 Sep 23.
After deep excision of burn eschar down to the muscle fascia patients have a non-reversible loss of the skin and underlying subcutaneous tissue. These patients would benefit from the development of a sufficient epidermal, dermal, and hypodermal tissue-engineered replacement provided by new technologies of tissue engineering. The aim of the present study was to determine whether keratinocytes and preadipocytes grow simultaneously on a bovine-derived collagen-elastin matrix under in vitro conditions in order to obtain a multi-layer skin substitute.
Human keratinocytes as well as human preadipocytes were seeded onto a collagen-elastin matrix (Matriderm®). Human preadipocytes were isolated from human subcutaneous adipose tissue and seeded onto the scaffold directly after isolation. Keratinocytes were isolated from fresh human split-thickness skin harvests and seeded onto the surface of the scaffold after 4 days of proliferation. Twenty one days after seeding all scaffolds were histologically evaluated, using hematoxylin eosin, immunohistochemical staining with collagen IV as well as immunofluorescence labeling with anti-Ki67 antibody and DAPI (4',6-diamidino-2-phenylindole).
Simultaneous growth of keratinocytes and preadipocytes could be observed on the collagen-elastin matrix. Keratinocytes adhered well to the surface of the matrix and formed a confluent epidermis-like layer. Preadipocytes adhered well and also penetrated into the deeper layers of the matrix.
In this study, a collagen-elastin matrix served as a suitable scaffold for simultaneous culturing of preadipocytes and keratinocytes. Preadipocytes showed good penetration into deeper layers of the scaffold, whereas keratinocytes attached only to the uppermost surface of the matrix. This approach towards a multi-layered skin substitute might be a useful asset for future reconstructive surgery.
深度切除烧伤焦痂直至肌肉筋膜后，患者会不可逆转地失去皮肤和皮下组织。这些患者将受益于新技术的组织工程开发出足够的表皮、真皮和皮下组织工程替代物。本研究的目的是确定在体外条件下，角质形成细胞和前脂肪细胞是否可以同时在牛源胶原-弹性蛋白基质上生长，以获得多层皮肤替代物。
将人角质形成细胞和人前脂肪细胞接种到胶原-弹性蛋白基质（Matriderm®）上。人前脂肪细胞从人皮下脂肪组织中分离出来，并在分离后直接接种到支架上。角质形成细胞从新鲜的人全厚皮采集物中分离出来，并在增殖 4 天后接种到支架表面。接种后 21 天，所有支架均通过苏木精-伊红染色、胶原 IV 免疫组织化学染色以及抗 Ki67 抗体和 DAPI（4'，6-二脒基-2-苯基吲哚）免疫荧光标记进行组织学评估。
可以观察到角质形成细胞和前脂肪细胞在胶原-弹性蛋白基质上同时生长。角质形成细胞很好地附着在基质表面，并形成了连续的表皮样层。前脂肪细胞很好地附着并也渗透到基质的深层。
在这项研究中，胶原-弹性蛋白基质作为同时培养前脂肪细胞和角质形成细胞的合适支架。前脂肪细胞很好地渗透到支架的深层，而角质形成细胞仅附着在基质的最上层。这种多层皮肤替代物的方法可能对未来的重建手术有帮助。
