Optimizing Decellularization Protocols for the Production of Porcine Acellular Muscle Matrix Scaffolds
College
College of Arts and Sciences
Department
Biology
Faculty Mentor
Dr. Matthew Stern
Abstract
Skeletal muscle tissue is one of the most common sites of traumatic injury in the human body. A variety of biomaterials that facilitate muscle regeneration are in development; however, few are able to provide the structural and biochemical cues present in the tissue’s native scaffolding, its extracellular matrix. We hypothesized that the process of decellularization, which removes the cellular content of a tissue or organ while leaving the extracellular matrix intact, could be used to produce biomaterial scaffolds of a clinically relevant size from porcine skeletal muscle tissue. To test this hypothesis, we systematically evaluated the effectiveness of ten decellularization protocols, each of which used a different combination and/or order of decellularization agents. Qualitative histological examination, scanning electron microscopy, and quantification of DNA content of the different forms of the material produced revealed a spectrum of effectiveness among the methods tested. Each protocol yielded a different combination of a) removal of cellular content and b) retention of extracellular matrix content and architecture. At least two protocols appear to produce scaffolds that are completely decellularized while retaining extracellular matrix elements and architecture. Future work will seek to quantify histological differences among and mechanical properties of the different forms of the material. Those forms exhibiting sufficient decellularization and retention of extracellular matrix will be termed Porcine Acellular Muscle Matrix (PAMM) and will be used in subsequent studies testing their ability to support the growth and differentiation of different populations of myogenic cells.
Previously Presented/Performed?
National Conference on Undergraduate Research (NCUR), Asheville, North Carolina, April 2016
South Carolina Academy of Science Annual Meeting, Winthrop University, April 2016
Grant Support?
Supported by grants from the National Institutes of Health IDeA Networks for Biomedical Research Excellence (NIH INBRE) and the Winthrop University Research Council
Start Date
22-4-2016 3:00 PM
End Date
22-4-2016 3:15 PM
Optimizing Decellularization Protocols for the Production of Porcine Acellular Muscle Matrix Scaffolds
West Center,Room 219
Skeletal muscle tissue is one of the most common sites of traumatic injury in the human body. A variety of biomaterials that facilitate muscle regeneration are in development; however, few are able to provide the structural and biochemical cues present in the tissue’s native scaffolding, its extracellular matrix. We hypothesized that the process of decellularization, which removes the cellular content of a tissue or organ while leaving the extracellular matrix intact, could be used to produce biomaterial scaffolds of a clinically relevant size from porcine skeletal muscle tissue. To test this hypothesis, we systematically evaluated the effectiveness of ten decellularization protocols, each of which used a different combination and/or order of decellularization agents. Qualitative histological examination, scanning electron microscopy, and quantification of DNA content of the different forms of the material produced revealed a spectrum of effectiveness among the methods tested. Each protocol yielded a different combination of a) removal of cellular content and b) retention of extracellular matrix content and architecture. At least two protocols appear to produce scaffolds that are completely decellularized while retaining extracellular matrix elements and architecture. Future work will seek to quantify histological differences among and mechanical properties of the different forms of the material. Those forms exhibiting sufficient decellularization and retention of extracellular matrix will be termed Porcine Acellular Muscle Matrix (PAMM) and will be used in subsequent studies testing their ability to support the growth and differentiation of different populations of myogenic cells.
