title: Ocular surface reconstruction with a tissue-engineered nasal mucosal epithelial cell sheet for the treatment of severe ocular surface diseases.
abstract: Severe ocular surface diseases (OSDs) with severe dry eye can be devastating and are currently some of the most challenging eye disorders to treat. To investigate the feasibility of using an autologous tissue-engineered cultivated nasal mucosal epithelial cell sheet (CNMES) for ocular surface reconstruction, we developed a novel technique for the culture of nasal mucosal epithelial cells expanded ex vivo from biopsy-derived human nasal mucosal tissues. After the protocol, the CNMESs had 4-5 layers of stratified, well-differentiated cells, and we successfully generated cultured epithelial sheets, including numerous goblet cells. Immunohistochemistry confirmed the presence of keratins 3, 4, and 13; mucins 1, 16, and 5AC; cell junction and basement membrane assembly proteins; and stem/progenitor cell marker p75 in the CNMESs. We then transplanted the CNMESs onto the ocular surfaces of rabbits and confirmed the survival of this tissue, including the goblet cells, up to 2 weeks. The present report describes an attempt to overcome the problems of treating severe OSDs with the most severe dry eye by treating them using tissue-engineered CNMESs to supply functional goblet cells and to stabilize and reconstruct the ocular surface. The present study is a first step toward assessing the use of tissue-engineered goblet-cell transplantation of nonocular surface origin for ocular surface reconstruction.
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Kobayashi
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom tnakamur@koto.kpu-m.ac.jp.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Nakamura
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Makoto
authors.initials: M
authors.lastname: Yasuda
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Yuiko
authors.initials: Y
authors.lastname: Hata
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Shoki
authors.initials: S
authors.lastname: Okura
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Miyu
authors.initials: M
authors.lastname: Iwamoto
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Maho
authors.initials: M
authors.lastname: Nagata
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Fullwood
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Noriko
authors.initials: N
authors.lastname: Koizumi
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Hisa
authors.affiliation: Department of Biomedical Engineering, Faculty of Life and Medical Sciences, and Research Center for Inflammation and Regenerative Medicine, Doshisha University, Kyoto, Japan; Departments of Ophthalmology and Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan; Biomedical and Life Sciences, School of Health and Medicine, Lancaster University, Lancaster, United Kingdom.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kinoshita
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D019573:Epithelium, Corneal / Q000601:surgery*; D020397:Goblet Cells / Q000166:cytology* / Q000637:transplantation*; D006801:Humans; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D009297:Nasal Mucosa / Q000166:cytology*; D011817:Rabbits; D023822:Tissue Engineering / Q000379:methods*
keywords: Dry eye; Goblet cell; Mucin; Nasal mucosa; Ocular surface