Effects of cell-based therapy for treating tympanic membrane perforations in mice

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Abstract

Objective 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. Study Design Prospective animal study. Setting Experimental. Subjects and Methods 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. Results 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. Conclusions The topical application of bone marrow-derived MSCs enhances the healing of TMPs in this animal model and is a promising alternative to tympanoplasty.

Original languageEnglish (US)
Pages (from-to)1106-1114
Number of pages9
JournalOtolaryngology - Head and Neck Surgery (United States)
Volume154
Issue number6
DOIs
StatePublished - Jun 1 2016

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Tympanic Membrane Perforation
Cell- and Tissue-Based Therapy
Mesenchymal Stromal Cells
Bone Marrow
Polyvinyl Alcohol
Green Fluorescent Proteins
Keratin-19
Tympanoplasty
Blister
Confocal Microscopy
General Anesthesia
Needles
Swine
Animal Models
Epithelial Cells
Phosphates
Prospective Studies
Control Groups
Wounds and Injuries
Therapeutics

Keywords

  • alternative therapy
  • bone marrow-derived stem cells
  • differentiation
  • hyaluronate-based biomaterials
  • porcine-derived collagen scaffold
  • stem cells
  • tympanic membrane perforations
  • wound healing

ASJC Scopus subject areas

  • Otorhinolaryngology
  • Surgery

Cite this

@article{9e530bc3cf894dbaa2944deb80af1eb6,
title = "Effects of cell-based therapy for treating tympanic membrane perforations in mice",
abstract = "Objective 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. Study Design Prospective animal study. Setting Experimental. Subjects and Methods 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. Results 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. Conclusions The topical application of bone marrow-derived MSCs enhances the healing of TMPs in this animal model and is a promising alternative to tympanoplasty.",
keywords = "alternative therapy, bone marrow-derived stem cells, differentiation, hyaluronate-based biomaterials, porcine-derived collagen scaffold, stem cells, tympanic membrane perforations, wound healing",
author = "Stefania Goncalves and {Bas Infante}, Esperanza and Goldstein, {Bradley J} and Angeli, {Simon I}",
year = "2016",
month = "6",
day = "1",
doi = "10.1177/0194599816636845",
language = "English (US)",
volume = "154",
pages = "1106--1114",
journal = "Otolaryngology - Head and Neck Surgery (United States)",
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number = "6",

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TY - JOUR

T1 - Effects of cell-based therapy for treating tympanic membrane perforations in mice

AU - Goncalves, Stefania

AU - Bas Infante, Esperanza

AU - Goldstein, Bradley J

AU - Angeli, Simon I

PY - 2016/6/1

Y1 - 2016/6/1

N2 - Objective 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. Study Design Prospective animal study. Setting Experimental. Subjects and Methods 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. Results 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. Conclusions The topical application of bone marrow-derived MSCs enhances the healing of TMPs in this animal model and is a promising alternative to tympanoplasty.

AB - Objective 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. Study Design Prospective animal study. Setting Experimental. Subjects and Methods 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. Results 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. Conclusions The topical application of bone marrow-derived MSCs enhances the healing of TMPs in this animal model and is a promising alternative to tympanoplasty.

KW - alternative therapy

KW - bone marrow-derived stem cells

KW - differentiation

KW - hyaluronate-based biomaterials

KW - porcine-derived collagen scaffold

KW - stem cells

KW - tympanic membrane perforations

KW - wound healing

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