title: Effects of Cell-Based Therapy for Treating Tympanic Membrane Perforations in Mice.
abstract: To investigate the effectiveness of scaffold-embedded mesenchymal stem cells (MSCs) as a topical treatment for healing tympanic membrane perforations (TMPs) in a mouse model.
  
  
  
  Prospective animal study.
  
  
  
  Experimental.
  
  
  
  In vitro: under sterile conditions, porcine-derived (Gelita-Spon [GS]), hyaluronate-derived (EpiDisc [ED]), and polyvinyl alcohol (PVA) scaffolds were cut into small pieces and cocultured with murine bone marrow-derived MSCs (BM-MSCs) expressing green fluorescent protein (GFP) for 72 hours. The cultures were either analyzed by confocal microscopy or used for subsequent in vivo experiments. In vivo: 26 mice were divided into 3 groups (ie, control [n = 9], GS [n = 8], ED [n = 9]). Under general anesthesia, TMPs of equal sizes were performed bilaterally using a sterile 27-gauge needle under a surgical microscope. The BM-MSCs embedded within GS or ED scaffolds were soaked in phosphate-buffered saline and then topically applied on right TMPs, and scaffolds alone were applied on left TMPs 6 to 8 hours after injury. Control mice did not receive treatment. On day 7, animals were euthanized and bullae were harvested for histological analysis.
  
  
  
  In vitro: BM-MSCs grew well on both GS (P = .0012) and ED (P = .0001) scaffolds compared with PVA. In vivo: 100% of untreated (control) TMPs remained open after 7 days. Animals treated with MSC-embedded ED scaffolds had a higher percentage of TMP closure (P = .016) and a thicker neotympanum (P = .0033) than control animals. The experimentally applied BM-MSCs engrafted and differentiated into epithelial cells suggested by the colocalized expression of cytokeratin-19 and GFP.
  
  
  
  The topical application of bone marrow-derived MSCs enhances the healing of TMPs in this animal model and is a promising alternative to tympanoplasty.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, University of Miami Miller School of Medicine, University of Miami Ear Institute, Miami, Florida, USA.
authors.forename: Stefania
authors.initials: S
authors.lastname: Goncalves
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, University of Miami Miller School of Medicine, University of Miami Ear Institute, Miami, Florida, USA.
authors.forename: Esperanza
authors.initials: E
authors.lastname: Bas
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, University of Miami Miller School of Medicine, University of Miami Ear Institute, Miami, Florida, USA Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, Florida, USA.
authors.forename: Bradley J
authors.initials: BJ
authors.lastname: Goldstein
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, University of Miami Miller School of Medicine, University of Miami Ear Institute, Miami, Florida, USA sangeli@med.miami.edu.
authors.forename: Simon
authors.initials: S
authors.lastname: Angeli
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D018920:Coculture Techniques; D004195:Disease Models, Animal; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D018613:Microscopy, Confocal; D011446:Prospective Studies; D054457:Tissue Scaffolds; D018058:Tympanic Membrane Perforation / Q000628:therapy*; D014945:Wound Healing
keywords: alternative therapy; bone marrow–derived stem cells; differentiation; hyaluronate-based biomaterials; porcine-derived collagen scaffold; stem cells; tympanic membrane perforations; wound healing