title: Induction of endometrial mesenchymal stem cells into tissue-forming cells suitable for fascial repair.
abstract: Pelvic organ prolapse is a major hidden burden affecting almost one in four women. It is treated by reconstructive surgery, often augmented with synthetic mesh. To overcome the growing concerns of using current synthetic meshes coupled with the high risk of reoperation, a tissue engineering strategy has been developed, adopting a novel source of mesenchymal stem cells. These cells are derived from the highly regenerative endometrial lining of the uterus (eMSCs) and will be delivered in vivo using a new gelatin-coated polyamide scaffold. In this study, gelatin properties were optimized by altering the gelatin concentration and extent of crosslinking to produce the desired gelation and degradation rate in culture. Following cell seeding of uncoated polyamide (PA) and gelatin-coated meshes (PA+G), the growth rate of eMSCs on the PA+G scaffolds was more than that on the PA alone, without compromising cell shape. eMSCs cultured on the PA+G scaffold retained their phenotype, as demonstrated by W5C5/SUSD2 (eMSC-specific marker) immunocytochemistry. Additionally, eMSCs were induced to differentiate into smooth muscle cells (SMC), as shown by immunofluorescence for smooth muscle protein 22 and smooth muscle myosin heavy chain. eMSCs also differentiated into fibroblast-like cells when treated with connective tissue growth factor with enhanced detection of Tenascin-C and collagen type I as well as new tissue formation, as seen by Masson's trichrome. In summary, it was demonstrated that the PA+G scaffold is an appropriate platform for eMSC delivery, proliferation and differentiation into SMC and fibroblasts, with good biocompatibility and the capacity to regenerate neo-tissue.
authors.affiliation: CSIRO Manufacturing Flagship, Bayview Avenue, Clayton, Victoria 3169, Australia.
authors.forename: Kai
authors.initials: K
authors.lastname: Su
authors.affiliation: CSIRO Manufacturing Flagship, Bayview Avenue, Clayton, Victoria 3169, Australia.
authors.forename: Sharon L
authors.initials: SL
authors.lastname: Edwards
authors.affiliation: MIMR-PHI Institute, 27-31 Wright Street, Clayton, Victoria 3168, Australia.
authors.forename: Ker S
authors.initials: KS
authors.lastname: Tan
authors.affiliation: CSIRO Manufacturing Flagship, Bayview Avenue, Clayton, Victoria 3169, Australia.
authors.forename: Jacinta F
authors.initials: JF
authors.lastname: White
authors.affiliation: CSIRO Manufacturing Flagship, Bayview Avenue, Clayton, Victoria 3169, Australia.
authors.forename: Shital
authors.initials: S
authors.lastname: Kandel
authors.affiliation: CSIRO Manufacturing Flagship, Bayview Avenue, Clayton, Victoria 3169, Australia.
authors.forename: John A M
authors.initials: JAM
authors.lastname: Ramshaw
authors.affiliation: MIMR-PHI Institute, 27-31 Wright Street, Clayton, Victoria 3168, Australia.
authors.forename: Caroline E
authors.initials: CE
authors.lastname: Gargett
authors.affiliation: CSIRO Manufacturing Flagship, Bayview Avenue, Clayton, Victoria 3169, Australia. Electronic address: jerome.werkmeister@csiro.au.
authors.forename: Jerome A
authors.initials: JA
authors.lastname: Werkmeister
mesh_terms: D061249:Batch Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D004717:Endometrium / Q000166:cytology* / Q000502:physiology; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005205:Fascia / Q000166:cytology*; D000071938:Fasciotomy; D005260:Female; D005780:Gelatin / Q000737:chemistry*; D006801:Humans; D008422:Materials Testing; D045164:Mesenchymal Stem Cell Transplantation / Q000295:instrumentation*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D009757:Nylons / Q000737:chemistry*; D017695:Soft Tissue Injuries; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods; D054457:Tissue Scaffolds
chemical_list: D009757:Nylons; D005780:Gelatin
keywords: Endometrial mesenchymal stem cells; Gelatin; Pelvic organ prolapse; Polyamide; Tissue engineering