Tissue structure and mechanical properties associated with various skin decellularization methods

Dominguez, Dominic Daniel (2021) Tissue structure and mechanical properties associated with various skin decellularization methods. Masters thesis, Northern Arizona University.

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Abstract

Burn wounds contribute to an increase in morbidity around the world and have a financial burden on the healthcare system. Third-degree burns lead to destruction of the epidermis, dermis, and hypodermis, limiting wound healing and increasing the risk of environmental or nosocomial infections. To treat these burns, acellular skin grafts are used to cover wounds and assist with wound healing. One common type of graft is an allogeneic skin graft, recovered from cadaveric tissue. However, cadaveric tissue containing cells with intact major histocompatibility (MHC) antigens has the potential for host immune rejection. While allogeneic skin grafts can be decellularized to mitigate this, decellularization has also been shown to disrupt the architecture of the tissue. Establishing a method that retains the skin biochemical, structural, and mechanical properties will be a beneficial treatment for wound healing. Three chemical-based decellularization reagents were compared to understand the impacts each method had on the skin’s properties. The success of cellular removal, architecture of the extracellular matrix, and mechanical properties were evaluated and compared to native tissue. These data demonstrated that the SDS-treatment method resulted in the most successful removal of cellular material on skin samples but negatively altered the structure and mechanics of the tissue. Tonicity treated skin appeared to fully remove cellular material as determined visually through hematoxylin and eosin staining, but the use of PicoGreen indicated that DNA was still present in the skin samples. Triton X-100-treatment did not remove the cellular or nuclear material on skin samples and caused damage to the skin’s structure that was visible through scanning electron microscopy. Evaluation of the tissue’s mechanical properties demonstrated that SDS-treatment negatively impacted the tissue’s strength and elasticity, whereas Tonicity-treatment was comparable to the native tissue. This study determined the effects each chemical reagent had on the skin’s properties and provides an initial assessment for the future possibility of in vitro recellularization.

Item Type:	Thesis (Masters)
Publisher’s Statement:	© Copyright is held by the author. Digital access to this material is made possible by the Cline Library, Northern Arizona University. Further transmission, reproduction or presentation of protected items is prohibited except with permission of the author.
Keywords:	Burns; Decellularization; Skin Grafts; Tonicity
Subjects:	R Medicine > RD Surgery
NAU Depositing Author Academic Status:	Student
Department/Unit:	Graduate College > Theses and Dissertations College of the Environment, Forestry, and Natural Sciences > Biological Sciences
Date Deposited:	07 Feb 2022 17:14
Last Modified:	07 Feb 2022 17:14
URI:	https://openknowledge.nau.edu/id/eprint/5670

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