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      Exploiting decellularized cochleae as scaffolds for inner ear tissue engineering

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          Abstract

          Background

          Use of decellularized tissues has become popular in tissue engineering applications as the natural extracellular matrix can provide necessary physical cues that help induce the restoration and development of functional tissues. In relation to cochlear tissue engineering, the question of whether decellularized cochlear tissue can act as a scaffold and support the incorporation of exogenous cells has not been addressed. Investigators have explored the composition of the cochlear extracellular matrix and developed multiple strategies for decellularizing a variety of different tissues; however, no one has investigated whether decellularized cochlear tissue can support implantation of exogenous cells.

          Methods

          As a proof-of-concept study, human Wharton’s jelly cells were perfused into decellularized cochleae isolated from C57BL/6 mice to determine if human Wharton’s jelly cells could implant into decellularized cochlear tissue. Decellularization was verified through scanning electron microscopy. Cocheae were stained with DAPI and immunostained with Myosin VIIa to identify cells. Perfused cochleae were imaged using confocal microscopy.

          Results

          Features of the organ of Corti were clearly identified in the native cochleae when imaged with scanning electron microscopy and confocal microscopy. Acellular structures were identified in decellularized cochleae; however, no cellular structures or lipid membranes were present within the decellularized cochleae when imaged via scanning electron microscopy. Confocal microscopy revealed positive identification and adherence of cells in decellularized cochleae after perfusion with human Wharton’s jelly cells. Some cells positively expressed Myosin VIIa after perfusion.

          Conclusions

          Human Wharton’s jelly cells are capable of successfully implanting into decellularized cochlear extracellular matrix. The identification of Myosin VIIa expression in human Wharton’s jelly cells after implantation into the decellularized cochlear extracellular matrix suggest that components of the cochlear extracellular matrix may be involved in differentiation.

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          Most cited references32

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          Prestin is the motor protein of cochlear outer hair cells.

          The outer and inner hair cells of the mammalian cochlea perform different functions. In response to changes in membrane potential, the cylindrical outer hair cell rapidly alters its length and stiffness. These mechanical changes, driven by putative molecular motors, are assumed to produce amplification of vibrations in the cochlea that are transduced by inner hair cells. Here we have identified an abundant complementary DNA from a gene, designated Prestin, which is specifically expressed in outer hair cells. Regions of the encoded protein show moderate sequence similarity to pendrin and related sulphate/anion transport proteins. Voltage-induced shape changes can be elicited in cultured human kidney cells that express prestin. The mechanical response of outer hair cells to voltage change is accompanied by a 'gating current', which is manifested as nonlinear capacitance. We also demonstrate this nonlinear capacitance in transfected kidney cells. We conclude that prestin is the motor protein of the cochlear outer hair cell.
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            Auditory hair cell replacement and hearing improvement by Atoh1 gene therapy in deaf mammals.

            In the mammalian auditory system, sensory cell loss resulting from aging, ototoxic drugs, infections, overstimulation and other causes is irreversible and leads to permanent sensorineural hearing loss. To restore hearing, it is necessary to generate new functional hair cells. One potential way to regenerate hair cells is to induce a phenotypic transdifferentiation of nonsensory cells that remain in the deaf cochlea. Here we report that Atoh1, a gene also known as Math1 encoding a basic helix-loop-helix transcription factor and key regulator of hair cell development, induces regeneration of hair cells and substantially improves hearing thresholds in the mature deaf inner ear after delivery to nonsensory cells through adenovectors. This is the first demonstration of cellular and functional repair in the organ of Corti of a mature deaf mammal. The data suggest a new therapeutic approach based on expressing crucial developmental genes for cellular and functional restoration in the damaged auditory epithelium and other sensory systems.
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              Atoh1 directs the formation of sensory mosaics and induces cell proliferation in the postnatal mammalian cochlea in vivo.

              Hearing impairment due to the loss of sensory hair cells is permanent in humans. Considerable interest targets the hair cell differentiation factor Atoh1 as a potential tool with which to promote hair cell regeneration. We generated a novel mouse model to direct the expression of Atoh1 in a spatially and temporally specific manner in the postnatal mammalian cochlea to determine the competency of various types of cochlear epithelial cells for hair cell differentiation. Atoh1 can generate cells in young animals with morphological, molecular, and physiological properties reminiscent of hair cells. This competency is cell type specific and progressively restricted with age. Significantly, Atoh1 induces ectopic sensory patches through Notch signaling to form a cellular mosaic similar to the endogenous sensory epithelia and expansion of the sensory mosaic through the conversion of supporting cells and nonautonomous supporting cell production. Furthermore, Atoh1 also activates proliferation within the normally postmitotic cochlear epithelium. These results provide insight into the potential and limitations of Atoh1-mediated hair cell regeneration.
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                Author and article information

                Contributors
                amellott@kumc.edu
                hsjk1@ku.edu
                jbrantley@kumc.edu
                detamore@ou.edu
                (913) 588-1227 , hstaecker@kumc.edu
                Journal
                Stem Cell Res Ther
                Stem Cell Res Ther
                Stem Cell Research & Therapy
                BioMed Central (London )
                1757-6512
                28 February 2017
                28 February 2017
                2017
                : 8
                : 41
                Affiliations
                [1 ]ISNI 0000 0001 2177 6375, GRID grid.412016.0, Department of Plastic Surgery, , University of Kansas Medical Center, ; Kansas City, KS 66160 USA
                [2 ]ISNI 0000 0001 2106 0692, GRID grid.266515.3, Microscopy and Analytical Imaging Laboratory, , University of Kansas, ; Lawrence, KS 66045 USA
                [3 ]ISNI 0000 0001 2177 6375, GRID grid.412016.0, Department of Otolaryngology, Head and Neck Surgery, , University of Kansas Medical Center, ; 3901 Rainbow Blvd, MS 3010, Kansas City, KS 66160 USA
                [4 ]ISNI 0000 0004 0447 0018, GRID grid.266900.b, Stephenson School of Biomedical Engineering, , University of Oklahoma, ; Norman, OK 73019 USA
                Article
                505
                10.1186/s13287-017-0505-6
                5330011
                28241887
                91f46953-288a-4d02-b3af-aed8a3b3fe8e
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 27 October 2016
                : 18 January 2017
                : 10 February 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100000002, National Institutes of Health;
                Award ID: AR056347
                Award Recipient :
                Funded by: State of Kansas
                Categories
                Research
                Custom metadata
                © The Author(s) 2017

                Molecular medicine
                tissue engineering,decellularization,stem cells,gene delivery,hair cells,cochlea
                Molecular medicine
                tissue engineering, decellularization, stem cells, gene delivery, hair cells, cochlea

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