Maxillofacial Plastic and Reconstructive Surgery

  • 제27권6호
  • /
  • Pages.510-522
  • /
  • 2005
  • /
  • 2288-8101(pISSN)
  • /
  • 2288-8586(eISSN)
대한악안면성형재건외과학회 (Korean Association of Maxillofacial Plastic and Reconstructive Surgeons)
권호 표지

인간 무세포성 진피기질 위에 배양한 가토 구강각화상피세포의 중충화와 기저막 형성에 관한 연구

FORMATION OF BASEMENT MEMBRANE AND STRATIFICATION OF RABBIT ORAL KERATINOCYTES CULTURED ON HUMAN ACELLULAR DERMAL MATRIX

  • 김용덕 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 안강민 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 염학렬 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 정헌종 (건국대학교 의과대학 소아과) ;
  • 김성민 (강릉대학교 치과대학 구강악안면외과학교실) ;
  • 장정원 (연세대학교 의과대학 약리학교실) ;
  • 성미애 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 박희정 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 황순정 (서울대학교 치과대학 구강악안면외과학교실) ;
  • 이종호 (서울대학교 치과대학 구강악안면외과학교실)
  • Kim, Yong-Deok (Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Ahn, Kang-Min (Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Yum, Hak-Yeol (Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Chung, Hun-Jong (Department of Pediatrics, Chung-Ju Medical Hospital, Kon-Kuk University) ;
  • Kim, Soung-Min (Department of Oral and Maxillofacial Surgery, College of Dentistry, Kang-Nung National University) ;
  • Jang, Jeong-Won (Department of Pharmacology, Yonsei School of Medicine) ;
  • Sung, Mi-Ae (Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Park, Hee-Jung (Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Hwang, Soon-Jung (Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University) ;
  • Lee, Jong-Ho (Department of Oral and Maxillofacial Surgery, College of Dentistry, Seoul National University)
  • 발행 : 2005.12.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

To assess the clinical applicability of bio-artificial mucosa which was made with autologous oral keratinocytes and human acellular dermal matrix, the formation of basement membrane and stratification of oral keratinocytes were evaluated. Six New Zealand white rabbits (around 2kg in weight) were anesthetized and its buccal mucosa was harvested (1.0 $\times$ 0.5cm size). Oral keratinicytes were extracted and cultured primarily with the feeder layer of pretreated NIH J2 3T3 fibroblast. These confluent cells were innoculated on the human acellular dermal matrix and cultured in multiple layer by air-rafting method. After 3, 5, 7, 10, 14 days of culture, each cultured bio-artificial mucosa was investigated the number of epthelial layer of by H&E stain and toluidine blue stain. The immuhohistochemical methods were used to evaluate the cell division capacity, the formation of basement membrane, and it's property of specific cells (PCNA, cytokeratin 14, laminin). Transmission electromicroscopy was used for the attachment between cells and matrix with the number of hemidesmosome. In result, the numbers of layer of stratified growth of oral keratinocyte cultured on the human acellular dermal matrix and the number of hemidesomal attachment between epithelial cells and human acellular dermal matrix were similar to the layers of normal oral mucosa after 10 days of culture. The cell division rate, basement membrane formation and proliferation rate increased as culture period increased. With these results, bio-artificial mucosa with autologous oral epithelial cells cultured on the acellular dermal matrix had clinically adaptable properties after 10 days' culture and this new bio-artificial mucosa model with relatively short culture time can be expected clinical applicability.

키워드

참고문헌

  1. McGregor IA, Morgan G : Axial and random pattern flaps. Br J Plast Surg 26 : 202, 1973 https://doi.org/10.1016/0007-1226(73)90003-9
  2. Shepherd NS, Maloney PL, Doku HC : Fenestrated palatal mucosal grafts for vestibulaoplasty. J Oral Surg 33 : 34, 1975
  3. Sharzer LA, Horton CE, Adamson JE et al : Intraoral reconstruction in head and neck cancer surgery. Clin Plast Surg 3 : 495, 1976
  4. Wells MD, Edwards AL, Luce EA : Intraoral reconstructive techniques. Clin Plast Surg 22 : 91, 1995
  5. Brown JS : T2 tongue : Reconstruction of surgical defect. Br J Oral Maxillofac Surg 37 : 194, 1999 https://doi.org/10.1054/bjom.1999.0033
  6. Nehrer-Tairych GV, Millesi W, Schuried O et al : A comparison of the donor-site morbidity after using the prelaminated fasciomucosal flap and the fasciocutaneous radial forearm flap for intraoral reconstruction. Br J Plast Surg 55 : 198, 2002 https://doi.org/10.1054/bjps.2002.3812
  7. Rheinwald JG, Green H : Serial cultivation of strains of human epidermal keratinocytes : the formation of keratinizing colonies from single cells. Cell 6 : 331, 1975 https://doi.org/10.1016/S0092-8674(75)80001-8
  8. Carsin P. Ainaud H. Le Bever J et al : Cultured epithelial autografts in extensive burn coverage of severely traumatized patients: a five year single-center experience with 30 patients. Burns 26 : 379, 2000 https://doi.org/10.1016/S0305-4179(99)00143-6
  9. Clugston PA, Snelling CF, Macdonald IB et al : Cultured epithelial autografts: three years of clinical experience with eighteen patients. J Burn Care Rehabil 12 : 533, 1991 https://doi.org/10.1097/00004630-199111000-00008
  10. Parenteau NL, Nolte CM, Bilbo P et al : Epidermis generated in vitro: practical considerations and applications. J Cell Biochem 45 : 245, 1991 https://doi.org/10.1002/jcb.240450304
  11. Kangesu T, Navsaria HA, Manek S et al : Kerato-dermal grafts: the importance of dermis for the in vivo growth of cultured keratinocytes. Br J Plast Surg 46 : 401, 1993 https://doi.org/10.1016/0007-1226(93)90046-E
  12. Compton CC, Hickerson W, Nadire K et al : Acceleration of skin regeneration from cultured epithelial autografts by transplantation to homograft dermis. J Burn Care Rehabil 14 : 653, 1993 https://doi.org/10.1097/00004630-199311000-00010
  13. Gallico GG, O'Connor NE : Engineering a skin replacement. Tissue Eng 1 : 231, 1995 https://doi.org/10.1089/ten.1995.1.231
  14. Bell E, Ehrlich HP, Buttle KF : Livining tussue formed in vitro and accepted as skin-equivalent tissue of full tickness. Scince 211 : 1052, 1981 https://doi.org/10.1126/science.7008197
  15. Hull BE, Sher SE, Rosen S et al : Structural integration of skin equivalents grafted to Lewis and Sprague-Dawley rat. J Invest Dermatol 81 : 429, 1983 https://doi.org/10.1111/1523-1747.ep12522599
  16. Burke JF, Yannas IV, Quinby WC et al : Successful use of a physiologically acceptable artificial skin in the treatment of extensive burn injury. Ann Surg 194 : 413, 1981 https://doi.org/10.1097/00000658-198110000-00005
  17. Langdon J, Williams PM, Navsaria H et al : Autologous keratinocyte grafting. A new technique for intra-oral reconstruction, Br Dental J 171 : 87, 1991 https://doi.org/10.1038/sj.bdj.4807616
  18. Lauer G, Schimming R : Tissue-engineered mucosa graft for reconstruction of the intraoral lining after freeing of the tongue : A clinical and immunohistologic study. J Oral Maxillofac Surg 59 : 169, 2001 https://doi.org/10.1053/joms.2001.20489
  19. Ueda M, Ebata K, Kaneda T : In vitro fabrication of bioartificial mucosa for reconstruction of oral mucosa: basic research and clinical application. Ann Plast Surg 27 : 540, 1991 https://doi.org/10.1097/00000637-199112000-00005
  20. Miki H, Ando N, Ozawa S, Sato M et al : An artificial esophagus constructed of cultured human esophageal epithelial cells, fibroblasts, polyglycolic acid mesh, and collagen. ASAIO J 45 : 502, 1999 https://doi.org/10.1097/00002480-199909000-00025
  21. Nakamura T, Endo K, Cooper LJ et al : The successful culture and autologous transplantation of rabbit oral mucosal epithelial cells on amniotic membrane, Invest Ophthalmol Vis Sci 44 : 106, 2003 https://doi.org/10.1167/iovs.02-0195
  22. Park HJ : Development of bioartificial oral buccal mucosa model using human amnoitic membrane, collagen sponge and cultured oral mucosa epithelial cells. Dongguk University (thesis: master's degree) 2003
  23. Cuono DB, Langdon R, Birchall N et al : Composite autologous- allogeneic skin replacement : development and clinical application. Plast Reconst Surg 80 : 626, 1987 https://doi.org/10.1097/00006534-198710000-00029
  24. Izumi K, Feinberg SE : Skin and oral mucosal substitutes; in MJ, Kellelr JC (eds): Oral and Maxillofacial Clinics of North America, Philadelphia, Saunders 4 : 61, 2002
  25. Wainwright DJ : Use of an acellular allograft dermal matrix (AlloDerm) in the management of full-thicknes burns. Bum 21 : 243, 1995
  26. Rennekampff HO, Hansbrough JF, Woods V Jr et al : Integrin and matrix molecule expression in cultured skin replacements. J Burn Care Rehabil 17 : 213, 1996 https://doi.org/10.1097/00004630-199605000-00006
  27. Livesey SA, Herndon DN, Hollyoak MA, et al : Transplanted acellular allograft dermal matrix. Transplantation 60 : 1, 1995 https://doi.org/10.1097/00007890-199507150-00001
  28. Medalie DA, Tompkins RG, Morgan JR : Evaluation of acellular human dermis as a dermal analog in a composite skin graft. ASAIO J 42 : 455, 1996 https://doi.org/10.1097/00002480-199609000-00030
  29. Tark KC, Chung S, Shin KS et al : Skin flap preprbrication using acellular dermal matrix and cultured keratinocytes in a porcine model. Ann Plast Surg 44 : 392, 2000 https://doi.org/10.1097/00000637-200044040-00007
  30. Wilke B, Bandemir B : Methodologic aspects in cultivating human keratinocytes. Dermatol Monatsschr 175 : 635, 1989
  31. Gallico GG 3rd, O'Connor NE, Compton CC et al : Permanent coverage of large burn wounds with autologous cultured human epithelium. N Engl J Med 311 : 448, 1984 https://doi.org/10.1056/NEJM198408163110706
  32. Ponchio L, Duma L. Oliviero B et al : Mitomycin C as an alternative to irradiation to inhibit the feeder layer growth in long-term culture assays. Cytotherapy 2 : 281, 2000 https://doi.org/10.1080/146532400539215
  33. Johnson MC, Meyer AA, deSerres S et al : Persistence of fetal bovine serum proteins in human keratinocytes. J Burn Care Rehabil 11 : 504, 1990
  34. Shin YM, Chung HJ, Ahn KM et al : Farbrication of myomucosal flap using cultured oral epithelium in rabbit model. J Korean Maxillofac Plast Reconstr Surg 27 : 259, 2005
  35. Williamson JS, Snelling CF, Clugston P et al : Cultured epithelial autograft: five years of clinical experience with twenty-eight patients. J Trauma 39 : 309, 1995 https://doi.org/10.1097/00005373-199508000-00020
  36. Myers S, Navsaria H, Sanders R et al : Transplantation of keratinocytes in the treatment of wounds. Am J Surg 170 : 75, 1995 https://doi.org/10.1016/S0002-9610(99)80258-X
  37. Srivastava A, DeSagun EZ, Jennings LJ et al : Use of porcine acellular dermal matrix as a dermal substitute in rats. Ann Surg 233 : 400, 2001 https://doi.org/10.1097/00000658-200103000-00015
  38. Izumi K, Takacs G, TErashi H et al : Ex vivo development of a composite human oral mucosal equivalent. J Oral Maxilofac Surg 57 : 571, 1999 https://doi.org/10.1016/S0278-2391(99)90077-0
  39. Lee JJ, Kuo MY, Cheng SJ, Chiang CP et al : Higher expressions of p53 and proliferating cell nuclear antigen (PCNA) in atrophic oral lichen planus and patients with areca quid chewing, Oral Surg Oral Med Oral Pathol Oral Radiol Endod 99 : 471, 2005 https://doi.org/10.1016/j.tripleo.2004.10.018
  40. Izumi K, Terashi H, Marcelo CL et al : Development and characterization of a tissue-engineered human oral mucosa equivalent produced in a serum-free culture system. J Dent Res 79 : 798, 2000 https://doi.org/10.1177/00220345000790030301
  41. Timpl R, Brown JC : the laminins. Matrix Biol 14 : 275, 1994 https://doi.org/10.1016/0945-053X(94)90192-9
  42. Schultze-Mosgau S, Lee BK, Ries J et al : In vitro cultured autologous pre-confluent oral keratinocytes for experimental prefabrication of oral mucosa. Int J Oral Maxillofac Surg 33 : 476, 2004 https://doi.org/10.1016/j.ijom.2003.12.005
  43. Banks SS, Harris CC : Aberrant expression of keratin proteins and cross-linked envelipes in human esophageal carcinomas. Cancer Res 44 : 1153, 1984
  44. Lauer G, Wiedmann-Al-Ahmad M, Otten JE et al : Immunohisto-chemical study during healing of free palatal mucosa grafts on plastic embedded samples. J Oral Pathol Med 30 : 104, 2001 https://doi.org/10.1034/j.1600-0714.2001.300207.x
  45. Reis-Filho JS, Simpson PT, Martins A et al : Distribution of p63, cytokeratin 5/6 and cytokeratin 14 in 51 normal and 400 neoplastic human tissue samples using TARP-4 multi-tumor tissue microarray. Virchows Arch 443 : 122, 2003 https://doi.org/10.1007/s00428-003-0859-2
  46. Izumi K, Song J, Feinberg SE : Development of a tissueengineered human oral mucosa: from the bench to the bed side. Cells Tissues Organs 176 : 134, 2004 https://doi.org/10.1159/000075034