Karin Vincente Greco1, Lauren Grace Jones1, Irene Obiri-Yeboa1, Tahera Ansari2. 1. Northwick Park Institute for Medical Research, Tissue Engineering & Regenerative Medicine, Harrow, United Kingdom. 2. Northwick Park Institute for Medical Research, Tissue Engineering & Regenerative Medicine, Harrow, United Kingdom. Electronic address: t.i.ansari@imperial.ac.uk.
Abstract
STUDY OBJECTIVE: Our aim was to use porcine vagina to create a vaginal matrix and test its cellular biocompatibility. DESIGN, SETTING, AND PARTICIPANTS: Vagina was harvested from pigs and decellularized (DC) using a combination of detergents (Triton X-100 and sodium deoxycholate) and enzymes (DNAse/RNAse). INTERVENTIONS: The presence of cellular material, collagen structural integrity, and basement membrane proteins were assessed histologically. To address cytocompatibility, porcine adipose-derived mesenchymal stem cells were harvested from abdominal fat together with vaginal epithelial cells and seeded onto the mucosal aspect of the vaginal scaffold. Both cell populations were seeded individually and assessed histologically at days 3 and 10. MAIN OUTCOME MEASURES AND RESULTS: The combination of enzymes and detergents resulted in a totally acellular matrix with very low DNA amount (control = 97.5 ng/μL ± 10.8 vs DC = 40.1 ng/μL ± 0.33; P = .02). The extracellular matrix showed retention of collagen fibers and elastin and a 50% retention in glycosaminoglycan content (control = 1.18 μg/mg ± 0.28; DC = 1.35 μg/mg ± 0.1; P = .03) and an intact basement membrane (positive for laminin and collagen IV). Seeded scaffolds showed cell attachment with adipose-derived mesenchymal stem cells and vaginal epithelial cells at days 3 and 10. CONCLUSION: It is possible to generate an acellular porcine vaginal matrix capable of supporting cells to reconstruct the vagina for future preclinical testing, and holds promise for creating clinically relevant-sized tissue for human application.
STUDY OBJECTIVE: Our aim was to use porcine vagina to create a vaginal matrix and test its cellular biocompatibility. DESIGN, SETTING, AND PARTICIPANTS: Vagina was harvested from pigs and decellularized (DC) using a combination of detergents (Triton X-100 and sodium deoxycholate) and enzymes (DNAse/RNAse). INTERVENTIONS: The presence of cellular material, collagen structural integrity, and basement membrane proteins were assessed histologically. To address cytocompatibility, porcine adipose-derived mesenchymal stem cells were harvested from abdominal fat together with vaginal epithelial cells and seeded onto the mucosal aspect of the vaginal scaffold. Both cell populations were seeded individually and assessed histologically at days 3 and 10. MAIN OUTCOME MEASURES AND RESULTS: The combination of enzymes and detergents resulted in a totally acellular matrix with very low DNA amount (control = 97.5 ng/μL ± 10.8 vs DC = 40.1 ng/μL ± 0.33; P = .02). The extracellular matrix showed retention of collagen fibers and elastin and a 50% retention in glycosaminoglycan content (control = 1.18 μg/mg ± 0.28; DC = 1.35 μg/mg ± 0.1; P = .03) and an intact basement membrane (positive for laminin and collagen IV). Seeded scaffolds showed cell attachment with adipose-derived mesenchymal stem cells and vaginal epithelial cells at days 3 and 10. CONCLUSION: It is possible to generate an acellular porcine vaginal matrix capable of supporting cells to reconstruct the vagina for future preclinical testing, and holds promise for creating clinically relevant-sized tissue for human application.