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Cellular Dynamics in Early Healing of Mouse Tympanic Membranes

Chari, Divya A.; Frumm, Stacey M.; Akil, Omar; Tward, Aaron D.

doi: 10.1097/MAO.0000000000002060

Aim: To better elucidate the cellular dynamics by which perforations in the tympanic membrane (TM) are healed.

Background: Under normal conditions, epidermal cells are born and then migrate radially outward from the malleus in the TM. It is unknown what the relative contribution of newly proliferated cells from different lineages is in the healing of TM perforations.

Methods: Thirty-six female mice were used in this study. Ethynyl deoxyuridine, a thymidine analogue that labels newly proliferated cells, was injected intraperitoneally into each mouse and then subsequently supplied in the drinking water. Acute perforations were performed on the right TM and the left TM served as the control and remained intact. The animals were sacrificed at six time points between 2 hours and 6 days. We stained for proliferative, epithelial, mesenchymal markers, and ethynyl deoxyuridine and analyzed the distribution of cells.

Results: In control TMs, newly proliferated cells were detected around the malleus handle and then migrated radially outward. Perforated TMs had a significantly higher number of newly proliferated cells throughout the tympanic membrane with a marked proliferative response of epithelial, mesenchymal, and mucosal cells in the region of the malleus and perforation. The majority of cells in the healed perforation were newly proliferated. In the anterior TM opposite the perforation, an increased turnover of keratinocytes was noted, but not mesenchymal cells.

Conclusions: Perforation of the TM alters the cellular dynamics throughout the entire TM, rather than simply adjacent to the perforation. This argues for long distance signaling occurring in the perforated TM.

Department of Otolaryngology—Head and Neck Surgery, University of California, San Francisco, San Francisco, California

Address correspondence and reprint requests to Aaron D. Tward, M.D., Ph.D., Department of Otolaryngology, University of California, San Francisco, 2233 Post Street, 3rd floor, San Francisco, CA 94115; E-mail:

D.A.C. and S.M.F. contributed equally to this work.

This work was supported by a grant from Hearing Research Inc. to A.D.T. and a fellowship from the American Otological Society to S.M.F.

The authors disclose no conflicts of interest.

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