Abstract
There is a clinical need for skin substitutes to replace full-thickness skin loss. Our group has developed a bilayered skin substitute produced from the patient's own fibroblasts and keratinocytes referred to as Self-Assembled Skin Substitute (SASS). After cell isolation and expansion, the current time required to produce SASS is 45 days. We aimed to optimize the manufacturing process to standardize the production of SASS and to reduce production time. The new approach consisted in seeding keratinocytes on a fibroblast-derived tissue sheet before its detachment from the culture plate. Four days following keratinocyte seeding, the resulting tissue was stacked on two fibroblast-derived tissue sheets and cultured at the air–liquid interface for 10 days. The resulting total production time was 31 days. An alternative method adapted to more contractile fibroblasts was also developed. It consisted in adding a peripheral frame before seeding fibroblasts in the culture plate. SASSs produced by both new methods shared similar histology, contractile behavior in vitro and in vivo evolution after grafting onto mice when compared with SASSs produced by the 45-day standard method. In conclusion, the new approach for the production of high-quality human skin substitutes should allow an earlier autologous grafting for the treatment of severely burned patients.
Highlights
The standard surgical treatment for the permanent closure of large full-thickness skin wounds that can occur following acute trauma or surgical intervention consists in replacing tissue loss with skin autografts harvested from an uninjured donor site on the patient
The tissue engineering method previously described to produce Self-Assembled Skin Substitute (SASS)-26 was compared with two new methods (SASS-3, SASS-4) allowing for a shorter production time
The addition of the donor-matched keratinocytes onto the unanchored fibroblast-derived sheet (SASS-3) induced contraction resulting in unusable shrunk tissue
Summary
The standard surgical treatment for the permanent closure of large full-thickness skin wounds that can occur following acute trauma or surgical intervention consists in replacing tissue loss with skin autografts harvested from an uninjured donor site on the patient. Progress in tissue engineering has led to the development of technologies, allowing the production of skin substitutes. These substitutes can be either epidermal or dermal substitutes or bilayered skin substitutes. Some models of bilayered skin substitutes produced in the laboratory have been reported to allow permanent coverage of full-thickness wound, to reduce the need for harvesting autografts, and are indicated as an adjunct treatment for massive burns.[1,2,3]
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