Primary fabrication of a tissue-engineered mesh for pelvic floor reconstruction and research in vivo

Abstract

To explore the fabrication of a tissue-engineered mesh fabricated with autologous adipose-derived mesenchymal stem cells (ADSC) and silk fibroin scaffold for female pelvic reconstruction. Sprague-Dawley rat adipose tissue was obtained from inguinal region. Mesenchymal stem cells were isolated and proliferated in vitro. The sericin of silk was removed after knitting in to web. Then a compound scaffold freeze-dried of silk fibroin web and silk fibroin solution was prepared for ADSC seeding. after 1-week culturing, the cellular morphology and extracellular matrix secretion on scaffold were observed by scanning electronic microscopy. The tissue-engineered meshes were implanted subcutaneously into the homologous rats while silk fibroin scaffolds served as control. Then the biomechanics, collagen matrix and local tissue response to tissue-engineered meshes in vivo with that to silk fibroin scaffolds were estimated. ADSC were fibroblast-like and proliferated well in vitro. SEM showed that the scaffold had an interwoven structure with a smooth surface and the silk sponges in scaffold had a uniform and porous structure. ADSC adhered firmly to the scaffold, secreted extracellular matrix and formed cell sheets on scaffold. In vivo studies showed that the tissue matrix of tissue-engineered meshes was better organized than silk fibroin scaffolds. They also had higher failure force (3.081 ± 0.121 vs 2.167 ± 0.148 N) and Young's modulus (3.191 ± 0.146 vs 2.263 ± 0.213 MPa) and more collagen content (18.648 ± 0.867 vs 14.123 ± 0.989 µg/mg) than silk fibroin scaffolds (P < 0.05). As a suitable cell type for tissue engineering, ADSC may be successfully isolated and stably proliferated in vitro. The tissue-engineered meshes have an excellent biocompatibility and appropriate properties for pelvic floor reconstruction.