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

김은영; 김혜민; 송정은; 이현수; 주천기; 강길선; Eun Young Kim; Hye Min Kim; Jeong Eun Song; Hyun Soo Lee; Choun-Ki Joo; Gilson Khang

Polymer(Korea), Vol.38, No.6, 702-707, November, 2014

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

김은영, 김혜민, 송정은 , 이현수¹, 주천기 ¹, 강길선 ^†

전북대학교 BIN 융합공학과, 고분자나노공학과, 고분자BIN융합연구소
¹가톨릭대학교 의과대학 서울성모병원 안과

Eun Young Kim, Hye Min Kim, Jeong Eun Song , Hyun Soo Lee¹, Choun-Ki Joo¹, and Gilson Khang^†

Dept. of BIN Fusion Tech., Polymer Fusion Res Center & Dept. of PolymerNano Sci. Tech., Chonbuk National University, 567 Baekje-daero, Jeonju 561-756, Korea
¹Department of Ophthalmology and Visual Science, College of Medicine, Gangnam St. Mary Hospital, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul 137-701, Korea

E-mail:

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+-ATPase의 발현을 확인하였으며, RT-PCR을 통해 각막 내피세포의 인자들을 확인하였다. 실험 결과, 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 RTPCR 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.

Keywords:corneal endothelium (CE);PLGA;extracellular matrix (ECM).

[References]

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Joyce NC, Exp. Eye Res., 81, 629 (2005)
Joyce NC, Prog. Retin. Eye Res., 22, 359 (2003)
Hollingsworth J, Perez-Gomez I, Mutalib HA, Efron N, Optom. Vis. Sci., 78, 706 (2001)
Sabater AL, Guarnieri A, Espana EM, Li W, Prosper F, Moreno-Montanes J, Regen. Med., 8, 183 (2013)
Basky G, JAMC, 162, 558 (2000)
Fagerholm P, Lagali1 NS, Merrett K, Jackson WB, Munger R, Liu Y, Polarek JW, Soderqvist M, Griffith M, Sci. Transl. Med., 2, 46 (2010)
Liang Y, Liu W, Han B, Yang C, Ma Q, Zhao W, Rong M, Li H, J. Mater. Sci. Mater. Med., 22, 175 (2010)
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Ishino Y, Sano Y, Nakamura T, Connon CJ, Rigby H, Fullwood NJ, Kinoshita S, Invest. Ophthalmol. Vis. Sci., 45, 800 (2004)
Choi JS, Williams JK, Greven M, Walter KA, Laber PW, Khang G, Soker S, Biomaterials, 31, 6738 (2010)
Joo CK, Green WR, Pepose JS, Fleming TP, Graefes. Arch. Clin. Exp. Ophthalmol., 238, 174 (2000)
Amano S, Cornea, 22, 66 (2003)
Yamagami S, Mimura T, Yokoo S, Takato T, Amano S, Cornea, 25, 90 (2006)
Egami M, Haraguchi Y, Shimizu T, Yamato M, Okano T, Arch. Pharm. Res., 37, 96 (2014)
Kim EY, Song JE, Park CH, Joo CK, Khang G, Inflamm. Regen., 34, 4 (2014)
Hirenkumar KM, Siegel SJ, Polymers, 3, 1377 (2011)
Lee JH, Kim EY, Lee CJ, Joo CK, Khang G, Int. J. Tissue Regen., 4, 53 (2013)
Engler C, Kelliher C, Speck CL, Jun AS, Cornea, 28, 1050 (2009)
Choi JS, Kim EY, Kim MJ, Giegengack M, Khan FA, Khang G, Soker S, Biomed. Mater., 8, 014108 (2013)
Yamaguchi M, Ebihara N, Shima N, Kimoto M, Funaki T, Yokoo S, Murakami A, Yamagami S, Invest. Ophthalmol. Vis. Sci., 52, 679 (2011)
Yamamoto A, Tabata Y, Int. J. Tissue. Regen., 4, 36 (2013)
Armitage WJ, Dick AD, Bourne WM, Invest. Ophthalmol. Vis. Sci., 44, 3326 (2003)
Yamamoto M, Tabata Y, Int. J. Tissue Regen., 4, 29 (2013)
Suzuki M, Morita T, Iwamoto T, Cell. Mol. Life Sci., 63, 12 (2006)
Sampson MJ, Lovell RS, Davison DB, Craigen WJ, Genomics, 36, 192 (1996)
McCarthy KM, Skare IB, Stankewich MC, Furuse M, Tsukita S, Rogers RA, Lynch RD, Schneeberger EE, J. Cell. Sci., 109, 2287 (1996)
Joyce NC, Harris DL, Zieske JD, Invest. Ophthalmol. Vis. Sci., 39, 2572 (1998)

[Referenced By]

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[1] Tuft SJ, Coster DJ, Eye, 4, 389 (1990)

[2] Joyce NC, Exp. Eye Res., 81, 629 (2005)

[3] Joyce NC, Prog. Retin. Eye Res., 22, 359 (2003)

[4] Hollingsworth J, Perez-Gomez I, Mutalib HA, Efron N, Optom. Vis. Sci., 78, 706 (2001)

[5] Sabater AL, Guarnieri A, Espana EM, Li W, Prosper F, Moreno-Montanes J, Regen. Med., 8, 183 (2013)

[6] Basky G, JAMC, 162, 558 (2000)

[7] Fagerholm P, Lagali1 NS, Merrett K, Jackson WB, Munger R, Liu Y, Polarek JW, Soderqvist M, Griffith M, Sci. Transl. Med., 2, 46 (2010)

[8] Liang Y, Liu W, Han B, Yang C, Ma Q, Zhao W, Rong M, Li H, J. Mater. Sci. Mater. Med., 22, 175 (2010)

[9] Madden PW, Lai JN, George KA, Giovenco T, Harkin DG, Chirila TV, Biomaterials, 32, 4076 (2011)

[10] Ishino Y, Sano Y, Nakamura T, Connon CJ, Rigby H, Fullwood NJ, Kinoshita S, Invest. Ophthalmol. Vis. Sci., 45, 800 (2004)

[11] Choi JS, Williams JK, Greven M, Walter KA, Laber PW, Khang G, Soker S, Biomaterials, 31, 6738 (2010)

[12] Joo CK, Green WR, Pepose JS, Fleming TP, Graefes. Arch. Clin. Exp. Ophthalmol., 238, 174 (2000)

[13] Amano S, Cornea, 22, 66 (2003)

[14] Yamagami S, Mimura T, Yokoo S, Takato T, Amano S, Cornea, 25, 90 (2006)

[15] Egami M, Haraguchi Y, Shimizu T, Yamato M, Okano T, Arch. Pharm. Res., 37, 96 (2014)

[16] Kim EY, Song JE, Park CH, Joo CK, Khang G, Inflamm. Regen., 34, 4 (2014)

[17] Hirenkumar KM, Siegel SJ, Polymers, 3, 1377 (2011)

[18] Lee JH, Kim EY, Lee CJ, Joo CK, Khang G, Int. J. Tissue Regen., 4, 53 (2013)

[19] Engler C, Kelliher C, Speck CL, Jun AS, Cornea, 28, 1050 (2009)

[20] Choi JS, Kim EY, Kim MJ, Giegengack M, Khan FA, Khang G, Soker S, Biomed. Mater., 8, 014108 (2013)

[21] Yamaguchi M, Ebihara N, Shima N, Kimoto M, Funaki T, Yokoo S, Murakami A, Yamagami S, Invest. Ophthalmol. Vis. Sci., 52, 679 (2011)

[22] Yamamoto A, Tabata Y, Int. J. Tissue. Regen., 4, 36 (2013)

[23] Armitage WJ, Dick AD, Bourne WM, Invest. Ophthalmol. Vis. Sci., 44, 3326 (2003)

[24] Yamamoto M, Tabata Y, Int. J. Tissue Regen., 4, 29 (2013)

[25] Suzuki M, Morita T, Iwamoto T, Cell. Mol. Life Sci., 63, 12 (2006)

[26] Sampson MJ, Lovell RS, Davison DB, Craigen WJ, Genomics, 36, 192 (1996)

[27] McCarthy KM, Skare IB, Stankewich MC, Furuse M, Tsukita S, Rogers RA, Lynch RD, Schneeberger EE, J. Cell. Sci., 109, 2287 (1996)

[28] Joyce NC, Harris DL, Zieske JD, Invest. Ophthalmol. Vis. Sci., 39, 2572 (1998)