Merrilees Mervyn, Buunk Neil, Zuo Ning, Larsen Nigel, Karimi Samaneh, Tucker Nick
Department of Anatomy and Medical Imaging, University of Auckland, Auckland 1023, New Zealand.
Electrospinz Limited, 44 Lee Street, Blenheim 7201, New Zealand.
Bioengineering (Basel). 2021 Jan 7;8(1):7. doi: 10.3390/bioengineering8010007.
This paper describes a novel method for the rapid construction of skin, using multiple layers of aligned electrospun fibers as starting scaffolds. Scaffolds were spun from biodegradable L-lactide/glycolide (molar ratio 10:90) with predominantly parallel arrays of fibers attached peripherally to thin 304 stainless steel layer frames. Each layer frame was held between two thicker support frames. Human skin cells were seeded onto multiple (three-nine) scaffolds. Dermal fibroblasts were seeded on both sides of each scaffold except for one on which keratinocytes were seeded on one side only. Following 48 h of culture, the scaffolds and layer frames were unmounted from their support frames, stacked, with keratinocytes uppermost, and securely held in place by upper and lower support frames to instantly form a multilayered "dermis" and a nascent epidermis. The stack was cultured for a further 5 days during which time the cells proliferated and then adhered to form, in association with the spun fibers, a mechanically coherent tissue. Fibroblasts preferentially elongated in the dominant fiber direction and a two-dimensional weave of alternating fiber and cell alignments could be constructed by selected placement of the layer frames during stacking. Histology of the 7-day tissue stacks showed the organized layers of fibroblasts and keratinocytes immuno-positive for keratin. Electron microscopy showed attachment of fibroblasts to the lactide/glycolide fibers and small-diameter collagen fibers in the extracellular space. This novel approach could be used to engineer a range of tissues for grafting where rapid construction of tissues with aligned or woven layers would be beneficial.
本文描述了一种快速构建皮肤的新方法，该方法使用多层排列的电纺纤维作为起始支架。支架由可生物降解的L-丙交酯/乙交酯（摩尔比10:90）纺制而成，其纤维主要呈平行排列，周边附着在薄的304不锈钢层框架上。每个层框架夹在两个较厚的支撑框架之间。将人类皮肤细胞接种到多个（三到九个）支架上。除了一个支架仅在一侧接种角质形成细胞外，在每个支架的两侧接种真皮成纤维细胞。培养48小时后，将支架和层框架从支撑框架上取下，堆叠起来，角质形成细胞在最上层，并用上下支撑框架牢固固定，以立即形成多层“真皮”和新生表皮。将堆叠物再培养5天，在此期间细胞增殖，然后与纺制纤维结合粘附形成机械连贯的组织。成纤维细胞优先沿主要纤维方向伸长，通过在堆叠过程中选择层框架的放置方式，可以构建纤维和细胞交替排列的二维编织结构。7天组织堆叠物的组织学检查显示有成纤维细胞和角质形成细胞的有组织层，对角蛋白免疫呈阳性。电子显微镜显示成纤维细胞附着在丙交酯/乙交酯纤维上，细胞外空间有小直径胶原纤维。这种新方法可用于设计一系列用于移植的组织，在这些组织中，快速构建具有排列或编织层的组织将是有益的。
