Álvaro Sierra-Sánchez, Brice Magne, Etienne Savard, Christian Martel, Karel Ferland, Martin A Barbier, Anabelle Demers, Danielle Larouche, Salvador Arias-Santiago, Lucie Germain
{"title":"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。","authors":"Álvaro Sierra-Sánchez, Brice Magne, Etienne Savard, Christian Martel, Karel Ferland, Martin A Barbier, Anabelle Demers, Danielle Larouche, Salvador Arias-Santiago, Lucie Germain","doi":"10.1093/burnst/tkad043","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The aim of this <i>in vitro</i> 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).</p><p><strong>Methods: </strong>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).</p><p><strong>Results: </strong>The SASSs were more resistant to tensile forces (<i>p-</i>value < 0.01) but less elastic (<i>p-</i>value < 0.001) compared to HPSSs. A higher number of proliferative Ki67<sup>+</sup> 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.</p><p><strong>Conclusions: </strong>This study demonstrates, for the first time, that both hbTESS models present similar <i>in vitro</i> biological characteristics. However, mechanical properties differ and future <i>in vivo</i> experiments will aim to compare their wound healing potential.</p>","PeriodicalId":9553,"journal":{"name":"Burns & Trauma","volume":"11 ","pages":"tkad043"},"PeriodicalIF":6.3000,"publicationDate":"2023-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10615253/pdf/","citationCount":"0","resultStr":"{\"title\":\"<i>In vitro</i> 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.\",\"authors\":\"Álvaro Sierra-Sánchez, Brice Magne, Etienne Savard, Christian Martel, Karel Ferland, Martin A Barbier, Anabelle Demers, Danielle Larouche, Salvador Arias-Santiago, Lucie Germain\",\"doi\":\"10.1093/burnst/tkad043\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The aim of this <i>in vitro</i> 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).</p><p><strong>Methods: </strong>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).</p><p><strong>Results: </strong>The SASSs were more resistant to tensile forces (<i>p-</i>value < 0.01) but less elastic (<i>p-</i>value < 0.001) compared to HPSSs. A higher number of proliferative Ki67<sup>+</sup> 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.</p><p><strong>Conclusions: </strong>This study demonstrates, for the first time, that both hbTESS models present similar <i>in vitro</i> biological characteristics. However, mechanical properties differ and future <i>in vivo</i> experiments will aim to compare their wound healing potential.</p>\",\"PeriodicalId\":9553,\"journal\":{\"name\":\"Burns & Trauma\",\"volume\":\"11 \",\"pages\":\"tkad043\"},\"PeriodicalIF\":6.3000,\"publicationDate\":\"2023-10-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10615253/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Burns & Trauma\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1093/burnst/tkad043\",\"RegionNum\":1,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q1\",\"JCRName\":\"DERMATOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Burns & Trauma","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1093/burnst/tkad043","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/1/1 0:00:00","PubModel":"eCollection","JCR":"Q1","JCRName":"DERMATOLOGY","Score":null,"Total":0}
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.
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.
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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