title: Construction and characterization of human oral mucosa equivalent using hyper-dry amniotic membrane as a matrix.
abstract: BACKGROUND
  Human amniotic membrane(HAM) as a graft material has been used in various fields. Hyper-dry amniotic membrane (HD-AM) is a novel dried amniotic membrane that is easy to handle and can be preserved at room temperature without time limitation. The purpose of this study was to investigate the useful properties of HD-AM in reconstruction of the oral mucosa.
  
  
  METHODS
  Human oral keratinocytes were isolated and seeded on HD-AM in serum-free culture system. Oral mucosa equivalent (OME) was developed and transplanted onto full-thickness wound on athymic mice. The wound healing was analyzed and the OME both before and after transplantation was analyzed with hematoxylin-eosin staining and immunohistochemical staining for Cytokines 10 (CK10), Cytokines 16 (CK16), and Ivolucrin (IVL).
  
  
  RESULTS
  Oral keratinocytes spread and proliferated well on HD-AM. Two weeks after air-lifting, OME had formed with good differentiation and morphology. We confirmed immunohistochemically that the expression of CK10 was positive in all suprabasal layers, as was CK16 in the upper layers, while IVL was present in all cell layers. Three weeks after transplantation to athymic mice, the newly generated tissue had survived well with the smallest contraction. The epithelial cells of newly generated tissue expressed CK10 throughout in all suprabasal layers, IVL was mainly in the granular layer, and CK16 positive cells were observed in all spinous layer and granular layer but were not expressed in the mouse skin, all of which were similar to native gingival mucosa.
  
  
  CONCLUSIONS
  The OME with HD-AM as a matrix revealed a good morphology and stable wound healing. This study demonstrates that HD-AM is a useful and feasible biomaterial for oral mucosa reconstruction.
authors: Department of Dentistry and Oral Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumato 390-8621, Japan.
authors: Fangfang
authors: F
authors: Qi
authors: Department of Regenerative Medicine, University of Toyama Graduate School of Medicine and Pharmaceutical Sciences, 2630 Sugitani, Toyama 930-0194, Japan.
authors: Toshiko
authors: T
authors: Yoshida
authors: Department of Dentistry and Oral Surgery, Ina Central Hospital, 1313-1 Koshiroukubo, Ina, Nagano 396-8555, Japan.
authors: Takeshi
authors: T
authors: Koike
authors: Department of Dentistry and Oral Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumato 390-8621, Japan.
authors: Hitoshi
authors: H
authors: Aizawa
authors: Department of Dentistry and Oral Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumato 390-8621, Japan.
authors: Tetsu
authors: T
authors: Shimane
authors: Department of Dentistry and Oral Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumato 390-8621, Japan.
authors: Yinghui
authors: Y
authors: Li
authors: Department of Dentistry and Oral Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumato 390-8621, Japan.
authors: Shinichi
authors: S
authors: Yamada
authors: Department of Regenerative Medicine, University of Toyama Graduate School of Medicine and Pharmaceutical Sciences, 2630 Sugitani, Toyama 930-0194, Japan.
authors: Motonori
authors: M
authors: Okabe
authors: Department of Regenerative Medicine, University of Toyama Graduate School of Medicine and Pharmaceutical Sciences, 2630 Sugitani, Toyama 930-0194, Japan.
authors: Toshio
authors: T
authors: Nikaido
authors: Department of Dentistry and Oral Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumato 390-8621, Japan. Electronic address: hkurita@shinshu-u.ac.jp.
authors: Hiroshi
authors: H
authors: Kurita
mesh_terms: D000650:Amnion / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology; D005881:Gingiva / Q000166:cytology; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009061:Mouth Mucosa / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D012867:Skin / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*; D016378:Tissue Transplantation / Q000379:methods; D014183:Transplantation, Heterologous
chemical_list: D001672:Biocompatible Materials; D016207:Cytokines
keywords: Hyper-dry amniotic membrane; Matrix; Oral keratinocytes; Oral mucosal equivalent; Reconstruction