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Ex-vivo Evaluation of Cellular Scaffolds Using Growth Hormone (IGF-1): A Novel Therapeutic Approaches for Volumetric Muscle loss
Mohamed Awad MD2, Anand Kumar MD1,2
1 University Hospitals, Department of Plastic Surgery, Cleveland, OH
2 Case Western Reserve University School of Medicine, Plastic Surgery Department, Cleveland, OH
University Hospitals, Department of Plastic Surgery, Cleveland, OH
2021-02-15
Presenter: Mohamed Awad
Affidavit:
Mohamed Awad
Director Name: Anand Kumar
Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Background: Muscle-derived stem cells (MDSCs) are multipotent cells with regenerative capacity. Our lab has previously determined, in mice, the ability of cell enhanced collagen scaffold to regenerate significant muscle healing in volumetric muscle loss. Our study aim is to evaluate the effect of growth hormone (IGF-1) on different cellular enhanced scaffolds.
Methods: Porcine Muscle samples were harvested. MDSCs were isolated using a modified preplate technique. Cells were transfected, expanded and same number of cells (8.65 million cells seeded on 5x5mm collagen and amniotic scaffold (with and without growth hormone),. On Day 8 assays were imaged using bioluminescence In Vivo Imaging System (IVIS). Collagen Scaffolds cross sectioned, embedded in paraformaldehyde and stained for Myosin heavy chain (Green) and DAPI (Blue) by Immunohistochemistry. Images were analyzed using ImageJ.
Results: Bioluminescence showed significant cellular population in amniotic scaffold with IGF-1 compared to collagen scaffold with IGF-1 and collagen scaffold (2245 vs 1061 vs 481 total luminescence; p = 0.05). but was not significantly different between amniotic scaffold without IGF-1 (2245 vs 1684 total luminescence; p=0.9). Collagen scaffold with IGF-1 was significant compared to collagen scaffold without IGF-1 (1061 vs 481 total luminescence p=0.04). These results were also confirmed by Immunofluorescence (amniotic scaffold with IGF-1 vs collagen scaffold with IGF-1 vs collagen scaffold), (1700 vs 900 vs 650; P=0.03)
Conclusion: Growth hormone augment cellular population in collagen and amniotic scaffold. using IGF-1 muscle-derived stem cell–enriched amniotic scaffold could have higher augmentation of muscle regeneration in a volumetric muscle loss compared to collagen scaffold.
