백서에서 자가 구강점막세포와 혈소판 농축 혈장의 이식에 의한 점막 근 피판의 조직공학적 제작 = FABRICATION OF TISSUE ENGINEERED MYO-MUCOSAL FLAP BY GRAFTING THE COMPLEX OF AUTOLOGOUS ORAL KERATINOCYTES AND PLATELET RICH PLASMA (PRP) IN A RAT MODEL

다국어 초록 (Multilingual Abstract)

Backgrounds: To overcome limited amount of autogenous mucosa for the reconstruction of various mucosal defect including oral mucosal defect, tissue engineered mucosa has been recently introduced. However, introduced conventional technique of tissue engineered mucosa still have serious pitfalls such as long fabrication time, fragility of the reconstructed mucosa, and complexity of the technique.
Aim of the study: To examine whether the complex of preconfluent autologous keratinocytes and autologous PRP (Platelet rich plasma) can reconstruct oral mucosa on the muscular flap with easier and faster way compared to conventional mucosal tissue engineering technique.
Materials and methods: One day before the operation, oral mucosa (3mm in diameter) were taken and treated for extraction of oral keratinocytes according to the routine manner. The day of operation, oral keratinocytes were prepared in the laboratory and then moved to the operating theater.
Autologous PRP was also prepared and then mixed with oral keratinocytes just before grafting on the prepared muscular flap. After keratinocyte-PRP complex was seated, then a sterilized rubber sheet was placed on the graft and the elevated skin flap was replaced and sutured. Biopsies were proceeded at 3,5,7,14 and 21 days. Tissue samples were evaluated clinically, histologically, and immunohistochemically.
Results: All of the oral keratinocyte-PRP complexes were successfully grafted on the recipient sites (100%). On 3 days after the operation, 1-2 continuous epithelial layer and many inflammatory cells were observed. On 5 days after the operation, increase of layers of keratinocyte was observed with less inflammatory response. Thickness of the layers was gradually increased from 7 to 21 days after the operation. Cytokeratin confirms epithelium
in every specimen.
Conclusions: Preconfluent graft of autogenous oral keratinocytes mixed with autogenous PRP have successfully reconstructed myo-mucosal flap.
This technique could be a useful alternative for oral mucosal reconstruction in the near future.

참고문헌 (Reference)

1 "an autologous alternative to fibrin glue with applications in oral and maxillofacial surgery" 55 : 1294-1299, 1997

2 "Transplantation of cultured mucosal epithelium: an experimental study" 24 : 352-359, 1996

3 "The use of a new bilayer artificial dermis for vestibular extension" 36 : 457-459, 1998

4 "The effect of detachment of cultured epithelial sheets on cellular ultrastructure and organisation" 4 : 43-51, 1995

5 "The alpha 6 beta 4 integrin is a receptor for both laminin and kalinin. Exp Cell Res 1994" 360-367,

6 "Take percentage and conditions of cultured epithelium were improved when basement membrane of the graft was maintained and anchoring mesh was added" 107 : 105-115, 2001

7 "Signaling vascular morphogenesis and maintenance" 277 : 48-50, 1997

8 "Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells" 331-343,

9 "Reconstruction of human skin from glycerol-preserved allodermis and cultured keratinocyte sheets. Burns 1994" 201-, 19-22.

10 "Preparation of growth factor enriched autologous platelet gel" April26,2001

11 "Pre-confluent keratinocyte grafting: the future for cultured skin replacements" 24 : 591-593, 1998

12 "Involvement of platelets in stimulating osteogenic activity" 13 : 655-, 1995

13 "Integrin alpha v beta 3 antagonists promote tumor regression by inducing apoptosis of angiogenetic blood vessels. Cell 1994" 1157-1164,

14 "Integrin alpha 6/beta 4 complex is located in hemidesmosomes suggesting a major role in epidermal cell-basement membrane adhesion. J Cell Biol 1991" 907-917,

15 "Insulinlike growth factor binding protein-3 is present in the granules of platelets. Endocrinology 1993" 996-,

16 "InVitro cultured autologous pre-confluent oral keratinocytes for experimental prefabrication of oral mucosa" 33 : 476-, 2001

17 "Free transplanted, microvascular reanastomosed forearm flap for reconstruction of the mouth cavity and oropharynx" 616-623, 1996

18 "Epidermal regulation of dermal fibroblast activity" 102 : 135-139, 1998

19 "Early embryonic development" 31 : 34-45, 1996

20 "Cultured keratinocyte sheets enhance spontaneous re-epithelialization in a dermal explant model of partialthickness wound healing. J Invest Dermatol 1990" 341-346,

21 "Cultured human keratinocytes on type I collagen membranes to reconstitute the epidermis" 6 : 53-67, 2000

22 "Clinical results of cultured epithelial cell grafting in the oral and maxillofacial region" 25 : 4-8, 1997

23 "Clinical application of free microvascular flaps in reconstructive surgery of the oral cavity. Acta Otorrinolaringol. Esp 1994" Garcia-Tapia R 457-460,

24 "An air-liquid interface promotes the differentiation of gastric surface mucous cells (GSM06) in culture" 271 : 741-746, 2000

25 "A newly developed collagen/silicone bilayer membrane as a mucosal substitute: a preliminary report" 35 : 85-91, 1997