DOI QR코드

DOI QR Code

각막 내피세포 성장 거동에 대한 락타이드 글리콜라이드 공중합체 필름과 세포외 기질의 효과

Effect of Extracellular Matrix on the Growth Behavior of Corneal Endothelial Cells to Poly(lactic-co-glycolic acid) Film

  • 김은영 (전북대학교 BIN 융합공학과, 고분자나노공학과, 고분자BIN융합연구소) ;
  • 김혜민 (전북대학교 BIN 융합공학과, 고분자나노공학과, 고분자BIN융합연구소) ;
  • 송정은 (전북대학교 BIN 융합공학과, 고분자나노공학과, 고분자BIN융합연구소) ;
  • 이현수 (가톨릭대학교 의과대학 서울성모병원 안과) ;
  • 주천기 (가톨릭대학교 의과대학 서울성모병원 안과) ;
  • 강길선 (전북대학교 BIN 융합공학과, 고분자나노공학과, 고분자BIN융합연구소)
  • Kim, Eun Young (Dept. of BIN Fusion Tech., Polymer Fusion Res. Center & Dept. of PolymerNano Sci. Tech., Chonbuk National University) ;
  • Kim, Hye Min (Dept. of BIN Fusion Tech., Polymer Fusion Res. Center & Dept. of PolymerNano Sci. Tech., Chonbuk National University) ;
  • Song, Jeong Eun (Dept. of BIN Fusion Tech., Polymer Fusion Res. Center & Dept. of PolymerNano Sci. Tech., Chonbuk National University) ;
  • Lee, Hyun Soo (Department of Ophthalmology and Visual Science, College of Medicine, The Catholic University of Korea, Gangnam St. Mary Hospital) ;
  • Joo, Choun-Ki (Department of Ophthalmology and Visual Science, College of Medicine, The Catholic University of Korea, Gangnam St. Mary Hospital) ;
  • Khang, Gilson (Dept. of BIN Fusion Tech., Polymer Fusion Res. Center & Dept. of PolymerNano Sci. Tech., Chonbuk National University)
  • 투고 : 2014.02.11
  • 심사 : 2014.04.25
  • 발행 : 2014.11.25

초록

각막 내피세포는 각막 가장 안쪽의 단일 세포층이며, 데스메막 위에 놓여있다. 데스메막은 피브로넥틴, 콜라겐, 라미닌, 단백당 등의 포함하는 세포외 기질이라 불리는 다양한 단백질들로 구성되어 있다. 본 연구에서, 조직공학에서 널리 이용되고 있는 락타이드 글리콜라이드 공중합체를 이용하여 투명한 필름을 제작하였으며, 표면에 다양한 부착 분자들(피브로넥틴, 콜라겐 타입 I, IV, 라미닌, FNC 코팅 믹스)을 코팅한 후, 세포 형태 관찰, 세포 증식 및 부착, ZO-1과 $Na^+/K^+-ATPase$의 발현을 확인하였으며, RT-PCR을 통해 각막 내피세포의 인자들을 확인하였다. 실험결과, in vitro 상에서 PLGA 필름은 각막내피세포 전달체로서 역할을 하며 코팅된 세포외 기질들은 각막 내피세포의 거동에 긍정적인 영향을 미침을 확인하였다.

Corneal endothelium is mono-inner cell layer of cornea and lay on Descmet's membrane which comprised of various proteins called extracellular matrix such as fibronectin, collagen, laminin, and proteoglycan, etc. In this study, we fabricated transparent poly(lactic-co-glycolic acid) (PLGA) film because PLGA is widely used for tissue engineering based on their properties. We investigated the behaviors of rabbit corneal endothelial cells (rCEnCs) on PLGA film surfaces coated with various cell-adhesive molecules like fibronectin, laminin, collagen type I and IV and FNC coating mix. The morphologic images, proliferation and adhesion assay, immunofluorescence for ZO-1 and $Na^+/K^+-ATPase$ and RT-PCR for expression of specific markers were conducted. These results showed that PLGA film plays a role as CEnC carriers in vitro and the cell-adhesive molecules give positive effects on the behaviors of rCEnC.

키워드

과제정보

연구 과제 주관 기관 : 한국연구재단

참고문헌

  1. S. J. Tuft and D. J. Coster, Eye, 4, 389 (1990). https://doi.org/10.1038/eye.1990.53
  2. N. C. Joyce, Exp. Eye Res., 81, 629 (2005). https://doi.org/10.1016/j.exer.2005.06.012
  3. N. C. Joyce, Prog. Retin. Eye Res., 22, 359 (2003). https://doi.org/10.1016/S1350-9462(02)00065-4
  4. J. Hollingsworth, I. Perez-Gomez, H. A. Mutalib, and N. Efron, Optom. Vis. Sci., 78, 706 (2001). https://doi.org/10.1097/00006324-200110000-00010
  5. A. L. Sabater, A. Guarnieri, E. M. Espana, W. Li, F. Prosper, and J. Moreno-Montanes, Regen. Med., 8, 183 (2013). https://doi.org/10.2217/rme.13.11
  6. G. Basky, JAMC, 162, 558 (2000).
  7. P. Fagerholm, N. S. Lagali1, K. Merrett, W. B. Jackson, R. Munger, Y. Liu, J. W. Polarek, M. Soderqvist, and M. Griffith, Sci. Transl. Med., 2, 46 (2010).
  8. Y. Liang, W. Liu, B. Han, C. Yang, Q. Ma, W. Zhao, M. Rong, and H. Li, J. Mater. Sci. Mater. Med., 22, 175 (2010).
  9. P. W. Madden, J. N. Lai, K. A. George, T. Giovenco, D. G. Harkin, and T. V. Chirila, Biomaterials, 32, 4076 (2011). https://doi.org/10.1016/j.biomaterials.2010.12.034
  10. Y. Ishino, Y. Sano, T. Nakamura, C. J. Connon, H. Rigby, N. J. Fullwood, and S. Kinoshita, Invest. Ophthalmol. Vis. Sci., 45, 800 (2004). https://doi.org/10.1167/iovs.03-0016
  11. J. S. Choi, J. K. Williams, M. Greven, K. A. Walter, P. W. Laber, G. Khang, and S. Soker, Biomaterials, 31, 6738 (2010). https://doi.org/10.1016/j.biomaterials.2010.05.020
  12. C. K. Joo, W. R. Green, J. S. Pepose, and T. P. Fleming, Graefes. Arch. Clin. Exp. Ophthalmol., 238, 174 (2000). https://doi.org/10.1007/s004170050029
  13. S. Amano, Cornea, 22, S66-74 (2003). https://doi.org/10.1097/00003226-200310001-00010
  14. S. Yamagami, T. Mimura, S. Yokoo, T .Takato, and S. Amano, Cornea, 25, S90-92 (2006). https://doi.org/10.1097/01.ico.0000247221.95424.d7
  15. M. Egami, Y. Haraguchi, T. Shimizu, M. Yamato, and T. Okano, Arch. Pharm. Res., 37, 96 (2014). https://doi.org/10.1007/s12272-013-0299-8
  16. E. Y. Kim, J. E. Song, C. H. Park, C. K. Joo, and G. Khang, Inflamm. Regen., 34, 4 (2014). https://doi.org/10.2492/inflammregen.34.004
  17. K. M. Hirenkumar and S. J. Siegel, Polymers, 3, 1377 (2011). https://doi.org/10.3390/polym3031377
  18. J. H. Lee, E. Y. Kim, C. J. Lee, C. K. Joo, and G. Khang, Int. J. Tissue Regen., 4, 53 (2013).
  19. C. Engler, C. Kelliher, C. L. Speck, and A. S. Jun, Cornea, 28, 1050 (2009). https://doi.org/10.1097/ICO.0b013e3181a165a3
  20. J. S. Choi, E. Y. Kim, M. J. Kim, M. Giegengack, F. A. Khan, G. Khang, and S. Soker, Biomed. Mater., 8, 014108 (2013). https://doi.org/10.1088/1748-6041/8/1/014108
  21. M. Yamaguchi, N. Ebihara, N. Shima, M. Kimoto, T. Funaki, S. Yokoo, A. Murakami, and S. Yamagami, Invest. Ophthalmol. Vis. Sci., 52, 679 (2011). https://doi.org/10.1167/iovs.10-5555
  22. A. Yamamoto and Y. Tabata, Int. J. Tissue. Regen., 4, 36 (2013).
  23. W. J. Armitage, A. D. Dick, and W. M. Bourne, Invest. Ophthalmol. Vis. Sci., 44, 3326 (2003). https://doi.org/10.1167/iovs.02-1255
  24. M. Yamamoto and Y. Tabata, Int. J. Tissue Regen., 4, 29 (2013).
  25. M. Suzuki, T. Morita, and T. Iwamoto, Cell. Mol. Life Sci., 63, 12 (2006). https://doi.org/10.1007/s00018-005-5336-4
  26. M. J. Sampson, R. S. Lovell, D. B Davison, and W. J. Craigen, Genomics, 36, 192 (1996). https://doi.org/10.1006/geno.1996.0445
  27. K. M. McCarthy, I. B. Skare, M. C. Stankewich, M. Furuse, S. Tsukita, R. A. Rogers, R. D. Lynch, and E. E. Schneeberger, J. Cell. Sci., 109, 2287 (1996).
  28. N. C. Joyce, D. L. Harris, and J. D. Zieske, Invest. Ophthalmol. Vis. Sci., 39, 2572 (1998).