Rebuild, restore, reinnervate: do human tissue engineered dermo-epidermal skin analogs attract host nerve fibers for innervation?
Rebuild, restore, reinnervate: do human tissue engineered dermo-epidermal skin analogs attract host nerve fibers for innervation?
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DOI：
10.1007/s00383-012-3208-1
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发表时间：
2013-01-01
影响因子：
1.8
通讯作者：
Meuli, Martin
中科院分区：
文献类型：
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作者：
Biedermann, Thomas;Boettcher-Haberzeth, Sophie;Meuli, Martin
组织工程皮肤替代品是覆盖大面积皮肤缺损的有前途的工具，但人们对移植物的神经重新支配知之甚少。在这项实验研究中，我们在大鼠模型中分析了宿主周围神经纤维向人体组织工程真皮-表皮皮肤替代品的向内生长。使用表皮室中不同的细胞类型，我们想要评估表皮细胞类型对替代物神经支配的影响。我们从人体皮肤活检中分离出角质形成细胞、黑素细胞、成纤维细胞和小汗腺细胞。扩增后，将表皮细胞按如下方式接种在含有人真皮成纤维细胞的 I 型胶原水凝胶上：(1) 仅角质形成细胞，(2) 角质形成细胞与黑素细胞，(3) 汗腺细胞。这些替代物被移植到免疫缺陷大鼠背部的全层皮肤伤口中，并在 3 周和 8 周后进行分析。使用 PGP9.5、NF-200 和 NF-145 等标记物检查组织学切片，了解有髓鞘和无髓鞘神经纤维向内生长。3 周后，所有三种表皮细胞变体的皮肤替代品均未显示神经元从宿主向移植物内生长。 8 周后，我们可以检测到所有三种皮肤替代品的神经支配。然而，神经纤维仅限于真皮室，我们在表皮中找不到任何无髓神经纤维。此外，用于生成表皮的不同细胞类型产生的构建体之间没有明显差异。我们的人体组织工程真皮-表皮皮肤替代品早在移植后 8 周就证明了真皮区室的宿主衍生神经支配。因此，我们的替代品显然有能力吸引邻近宿主组织的神经纤维，这些神经纤维也长成移植物，从而有可能恢复皮肤敏感性。
Tissue engineered skin substitutes are a promising tool to cover large skin defects, but little is known about reinnervation of transplants. In this experimental study, we analyzed the ingrowth of host peripheral nerve fibers into human tissue engineered dermo-epidermal skin substitutes in a rat model. Using varying cell types in the epidermal compartment, we wanted to assess the influence of epidermal cell types on reinnervation of the substitute.We isolated keratinocytes, melanocytes, fibroblasts, and eccrine sweat gland cells from human skin biopsies. After expansion, epidermal cells were seeded on human dermal fibroblast-containing collagen type I hydrogels as follows: (1) keratinocytes only, (2) keratinocytes with melanocytes, (3) sweat gland cells. These substitutes were transplanted into full-thickness skin wounds on the back of immuno-incompetent rats and were analyzed after 3 and 8 weeks. Histological sections were examined with regard to myelinated and unmyelinated nerve fiber ingrowth using markers such as PGP9.5, NF-200, and NF-145.After 3 weeks, the skin substitutes of all three epidermal cell variants showed no neuronal ingrowth from the host into the transplant. After 8 weeks, we could detect an innervation of all three types of skin substitutes. However, the nerve fibers were restricted to the dermal compartment and we could not find any unmyelinated fibers in the epidermis. Furthermore, there was no distinct difference between the constructs resulting from the different cell types used to generate an epidermis.Our human tissue engineered dermo-epidermal skin substitutes demonstrate a host-derived innervation of the dermal compartment as early as 8 weeks after transplantation. Thus, our substitutes apparently have the capacity to attract nerve fibers from adjacent host tissues, which also grow into grafts and thereby potentially restore skin sensitivity.