Skin Engineering Laboratory, Adult Burns Centre, Royal Adelaide Hospital, Adelaide, South Australia, Australia.
Adult Burns Centre, Royal Adelaide Hospital, Adelaide, South Australia, Australia.
Wound Repair Regen. 2023 Jan;31(1):87-98. doi: 10.1111/wrr.13059. Epub 2022 Dec 12.
This study compared three acellular scaffolds as templates for the fabrication of skin substitutes. A collagen-glycosaminoglycan (C-GAG), a biodegradable polyurethane foam (PUR) and a hybrid combination (PUR/C-GAG) were investigated. Scaffolds were prepared for cell inoculation. Fibroblasts and keratinocytes were serially inoculated onto the scaffolds and co-cultured for 14 days before transplantation. Three pigs each received four full-thickness 8 cm × 8 cm surgical wounds, into which a biodegradable temporising matrix (BTM) was implanted. Surface seals were removed after integration (28 days), and three laboratory-generated skin analogues and a control split-thickness skin graft (STSG) were applied for 16 weeks. Punch biopsies confirmed engraftment and re-epithelialisation. Biophysical wound parameters were also measured and analysed. All wounds showed greater than 80% epithelialisation by day 14 post-transplantation. The control STSG displayed 44% contraction over the 16 weeks, and the test scaffolds, C-GAG 64%, Hybrid 66.7% and PUR 67.8%. Immunohistochemistry confirmed positive epidermal keratins and basement membrane components (Integrin alpha-6, collagens IV and VII). Collagen deposition and fibre organisation indicated the degree of fibrosis and scar produced for each graft. All scaffold substitutes re-epithelialised by 4 weeks. The percentage of original wound area for the Hybrid and PUR was significantly different than the STSG and C-GAG, indicating the importance of scaffold retainment within the first 3 months post-transplant. The PUR/C-GAG scaffolds reduced the polymer pore size, assisting cell retention and reducing the contraction of in vitro collagen. Further investigation is required to ensure reproducibility and scale-up feasibility.
本研究比较了三种去细胞支架作为制造皮肤替代物的模板。研究了胶原蛋白-糖胺聚糖（C-GAG）、可生物降解的聚氨酯泡沫（PUR）和混合组合（PUR/C-GAG）。为细胞接种制备了支架。将成纤维细胞和角质形成细胞连续接种到支架上，并在移植前共培养 14 天。每只猪接受四个 8cm×8cm 的全层手术伤口，将可生物降解的临时基质（BTM）植入其中。整合后（28 天）去除表面密封物，并应用三个实验室生成的皮肤类似物和一个对照的全厚皮片移植物（STSG）16 周。通过打孔活检证实了植入和再上皮化。还测量和分析了生物物理伤口参数。所有伤口在移植后 14 天内的上皮化率均大于 80%。对照 STSG 在 16 周内收缩了 44%，而测试支架的 C-GAG 收缩了 64%，混合支架收缩了 66.7%，PUR 支架收缩了 67.8%。免疫组织化学证实了表皮角蛋白和基底膜成分（整合素 alpha-6、IV 型和 VII 型胶原蛋白）呈阳性。胶原蛋白沉积和纤维组织表明了每个移植物产生的纤维化和疤痕的程度。所有支架替代品在 4 周内重新上皮化。Hybrid 和 PUR 的原始伤口面积百分比与 STSG 和 C-GAG 有显著差异，表明在移植后前 3 个月内支架保留的重要性。PUR/C-GAG 支架减小了聚合物孔的大小，有助于细胞保留并减少体外胶原蛋白的收缩。需要进一步研究以确保可重复性和放大可行性。
