J Pediatr Adolesc Gynecol. 2018 May 21. pii: S1083-3188(18)30216-X. doi: 10.1016/j.jpag.2018.05.003. [Epub ahead of print]

Creation of an acellular vaginal matrix for potential vaginal augmentation and cloacal repair.

Greco KV¹, Jones LG¹, Obiri-Yeboa I¹, Ansari T².

Abstract

STUDY OBJECTIVE:

our aim was to use porcine vagina to create a vaginal matrix and test its cellular biocompatibility.

DESIGN:

vagina was harvested from pigs and de-cellularised (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 (AD-MSC) were harvested from abdominal fat together with vaginal epithelial cells (VEC) and seeded onto the mucosal aspect of the vaginal scaffold. Both cells populations were seeded individually and assessed histologically at days 3 and 10. MAIN OUTCOMES/RESULTS: the combination of enzymes and detergents resulted in a totally acellular matrix with very low DNA amount (control= 97.5ng/μl ± 10.8 vs DC= 40.1 ng/μl ±0.33 p=0.02). The extra cellular matrix (ECM) showed retention of collagen fibres and elastin and a 50% retention in glycosaminoglycan content; (control= 1.18μg/mg ± 0.28 DC = 1.35μg/mg ± 0.1 p=0.03) and an intact basement membrane (positive for both laminin and collagen IV). Seeded scaffolds showed cell attachment with both AD-MSC and VEC at days 3 and 10.

CONCLUSIONS:

it is possible to generate an acellular porcine vaginal matrix capable of supporting cells to reconstruct the vagina for future pre-clinical testing, and holds promise for creating clinically relevant sized tissue for human application.