In vitro comparison of human plasma-based and self-assembled tissue-engineered skin substitutes: two different manufacturing processes for the treatment of deep and difficult to heal injuries.

IF 6.3 1区 医学 Q1 DERMATOLOGY
Burns & Trauma Pub Date : 2023-10-30 eCollection Date: 2023-01-01 DOI:10.1093/burnst/tkad043
Álvaro Sierra-Sánchez, Brice Magne, Etienne Savard, Christian Martel, Karel Ferland, Martin A Barbier, Anabelle Demers, Danielle Larouche, Salvador Arias-Santiago, Lucie Germain
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Abstract

Background: The aim of this in vitro study was to compare side-by-side two models of human bilayered tissue-engineered skin substitutes (hbTESSs) designed for the treatment of severely burned patients. These are the scaffold-free self-assembled skin substitute (SASS) and the human plasma-based skin substitute (HPSS).

Methods: Fibroblasts and keratinocytes from three humans were extracted from skin biopsies (N = 3) and cells from the same donor were used to produce both hbTESS models. For SASS manufacture, keratinocytes were seeded over three self-assembled dermal sheets comprising fibroblasts and the extracellular matrix they produced (n = 12), while for HPSS production, keratinocytes were cultured over hydrogels composed of fibroblasts embedded in either plasma as unique biomaterial (Fibrin), plasma combined with hyaluronic acid (Fibrin-HA) or plasma combined with collagen (Fibrin-Col) (n/biomaterial = 9). The production time was 46-55 days for SASSs and 32-39 days for HPSSs. Substitutes were characterized by histology, mechanical testing, PrestoBlue™-assay, immunofluorescence (Ki67, Keratin (K) 10, K15, K19, Loricrin, type IV collagen) and Western blot (type I and IV collagens).

Results: The SASSs were more resistant to tensile forces (p-value < 0.01) but less elastic (p-value < 0.001) compared to HPSSs. A higher number of proliferative Ki67+ cells were found in SASSs although their metabolic activity was lower. After epidermal differentiation, no significant difference was observed in the expression of K10, K15, K19 and Loricrin. Overall, the production of type I and type IV collagens and the adhesive strength of the dermal-epidermal junction was higher in SASSs.

Conclusions: This study demonstrates, for the first time, that both hbTESS models present similar in vitro biological characteristics. However, mechanical properties differ and future in vivo experiments will aim to compare their wound healing potential.

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基于人血浆和自组装组织工程皮肤替代品的体外比较:两种不同的制造工艺,用于治疗深度和难以愈合的损伤。
背景:这项体外研究的目的是比较用于治疗严重烧伤患者的两种并排的人类双层组织工程皮肤替代品(hbTESS)模型。这些是无支架的自组装皮肤替代品(SASS)和基于人血浆的皮肤替代物(HPSS) = 3) 并且使用来自同一供体的细胞来产生两种hbTESS模型。对于SASS的制造,将角质形成细胞接种在包括成纤维细胞和它们产生的细胞外基质(n = 12) ,而对于HPSS的生产,角质形成细胞是在水凝胶上培养的,水凝胶由成纤维细胞组成,成纤维细胞包埋在血浆中作为独特的生物材料(纤维蛋白),血浆与透明质酸结合(纤维蛋白HA)或血浆与胶原结合(纤维素Col)(n/生物材料 = 9) 。SASS的生产时间为46-55天,HPSS的生产时间则为32-39天。通过组织学、机械测试、PrestoBlue对替代品进行了表征™-免疫荧光(Ki67,Keratin(K)10,K15,K19,Loricrin,IV型胶原)和Western印迹(I型和IV型胶原蛋白) p值 + 在SASS中发现了细胞,尽管它们的代谢活性较低。表皮分化后,K10、K15、K19和Loricrin的表达没有显著差异。总的来说,SASS中I型和IV型胶原的产生以及真皮-表皮连接处的粘附强度更高。结论:本研究首次证明,两种hbTESS模型都具有相似的体外生物学特性。然而,机械性能各不相同,未来的体内实验将旨在比较它们的伤口愈合潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Burns & Trauma
Burns & Trauma 医学-皮肤病学
CiteScore
8.40
自引率
9.40%
发文量
186
审稿时长
6 weeks
期刊介绍: The first open access journal in the field of burns and trauma injury in the Asia-Pacific region, Burns & Trauma publishes the latest developments in basic, clinical and translational research in the field. With a special focus on prevention, clinical treatment and basic research, the journal welcomes submissions in various aspects of biomaterials, tissue engineering, stem cells, critical care, immunobiology, skin transplantation, and the prevention and regeneration of burns and trauma injuries. With an expert Editorial Board and a team of dedicated scientific editors, the journal enjoys a large readership and is supported by Southwest Hospital, which covers authors'' article processing charges.
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