헤스페리딘/PLGA 필름에서 망막색소상피세포의 부착과 증식거동

이소진; 강수지; 김혜윤; 이정환; 김은영; 권순용; 정진화; 주천기; 강길선; So Jin Lee; Su Ji Kang; Hye Yun Kim; Jung Hwan Lee; Eun Young Kim; Soon Yong Kwon; Jin Wha Chung; Choun-Ki Joo; Gilson Khang

Polymer(Korea), Vol.38, No.1, 24-30, January, 2014

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헤스페리딘/PLGA 필름에서 망막색소상피세포의 부착과 증식거동

Adhesion and Proliferation Behavior of Retinal Pigment Epithelial Cells on Hesperidin/PLGA Films

이소진, 강수지, 김혜윤, 이정환, 김은영, 권순용 ¹, 정진화 ^{1, †}, 주천기 ², 강길선 ^†

전북대학교 BIN 융합공학과 고분자나노공학과
¹가톨릭대학교 의과대학 여의도성모병원 정형외과
²가톨릭대학교 의과대학 여의도성모병원 안과

So Jin Lee, Su Ji Kang, Hye Yun Kim, Jung Hwan Lee, Eun Young Kim, Soon Yong Kwon¹, Jin Wha Chung^{1, †}, Choun-Ki Joo², 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
¹Dept. of Orthopedic Surgery, Yeouido St. Mary's Hospital, Catholic University of Korea, 62 Yeouido-dong, Yeongdeungpo-gu, Seoul 150-896, Korea
²Dept. of Ophthalmology and Visual Science, Seoul St. Mary’s Hospital, College of Medicine, Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul 137-040, Korea

E-mail:,

초록

망막색소상피(retinal pigment epithelium, RPE)는 시기능을 유지하는데 중요한 역할을 하여 RPE의 퇴화는 여러 망막변성질병을 유발한다. 현재 이에 대한 효과적인 치료법이 부족하여 세포 이식에 적합한 지지체를 제작하기 위해, 생분해성 고분자인 PLGA와 항염증, 항산화 작용 등의 기능이 있는 헤스페리딘을 이용하여 하이브리드 필름을 제조하였다. ARPE-19를 파종한 후, MTT 분석법을 이용하여 세포 증식률을 확인하고, 세포의 부착 및 세포 형태를 SEM을 통하여 확인하였다. 또한 RPE 세포의 특이적 유전자 발현정도를 확인하기 위하여 RT-PCR을 수행하였고, RPE65의 발현을 확인하기 위해 AEC 면역화학적 염색을 실시하였다. 그 결과, 헤스페리딘/PLGA 필름은 PLGA보다 RPE 세포의 부착, 증식 및 표현형 유지가 우수함을 확인하였고, 이를 통해 헤스페리딘/PLGA 필름의 망막재생을 위한 조직공학적 담체로써 응용 가능성을 확인할 수 있었다.

Retinal pigment epithelium (RPE) plays an important role in maintaining the visual function and the degeneration of the RPE causes several retinal degeneration disease. In order to fabricate the suitable carrier for RPE transplantation, the hybrid poly(lactide-co-glycolide) (PLGA) film with hesperidin was prepared. Hesperidin has an antiinflammatory and antioxidant characteristics. ARPE-19 was seeded on hesperidin/PLGA film and then, cell proliferation was determined by the MTT assay, and cell adhesion and cell morphology were confirmed by SEM. Also, RT-PCR was performed to confirm the expression of the specific genes, and AEC immunohistochemical staining was performed to determine the expression of RPE65. As a result, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells were more excellent on hesperidin/PLGA film than PLGA film, thereby we were able to confirm the potential applications of hesperidin/PLGA film as tissue engineering carrier for regeneration of retina.

Keywords:retinal pigment epithelial cells;hesperidin;PLGA;hesperidin/PLGA hybrid film.

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[1] Beatty S, Koh H, Phil M, Henson D, Boulton M, Surv. Ophthalmol., 45, 115 (2000)

[2] Ambati J, Ambati BK, Yoo SH, Ianchulev S, Adamis AP, Surv. Ophthalmol., 48, 257 (2003)

[3] Organisciak DT, Vaughan DK, Prog. Retin. Eye Res., 29, 113 (2010)

[4] Aisenbrey S, Lafaut BA, Szurman P, Grisanti S, Luke C, Krott R, Thumann G, Fricke J, Neugebauer A, Hilgers RD, Esser P, Walter P, Bartz-Schmidt KU, Arch. Ophthalmol., 120, 451 (2002)

[5] Strauss O, Physiol. Rev., 85, 845 (2005)

[6] Binder S, Stanzel BV, Krebs I, Glittenberg C, Prog. Retin. Eye Res., 26, 516 (2007)

[7] van Leeuwen R, Klaver CC, Vingerling JR, Hofman A, de Jong PT, Eur. J. Epidemiol., 18, 845 (2003)

[8] Klein R, Chou CF, Klein BE, Zhang X, Meuer SM, Saaddine JB, Arch. Ophthalmol., 129, 75 (2011)

[9] Shah CA, Indian. J. Med. Sci., 62, 500 (2008)

[10] Kubota A, Nishida K, Yamato M, Yang J, Kikuchi A, Okano T, Tano Y, Biomaterials, 27, 3639 (2006)

[11] Yaji N, Yamato M, Yang J, Okano T, Hori S, Biomaterials, 30, 797 (2009)

[12] Lee JH, Park SJ, Chun HJ, Kim CH, Inter. J. Tissue Regen., 1, 1 (2010)

[13] Lee CT, Huang CP, Lee YD, Biomol. Eng., 24, 131 (2007)

[14] Yoshioka T, Kawazoe N, Tateishi T, Chen G, Biomaterials, 29(24-25), 3438 (2008)

[15] Gu BK, Kim MS, Park SJ, Kim CH, Inter. J. Tissue Regen., 2, 83 (2011)

[16] Jo EH, Kim SJ, Cho SJ, Lee GY, Kim OY, Lee EY, Cho WH, Lee DW, Khang G, Polym.(Korea), 35(4), 289 (2011)

[17] Garg A, Garg S, Zaneveld LJ, Singla AK, Phytother. Res., 15, 655 (2001)

[18] Gil-Izquierdo A, Gil MI, Ferreres F, Tomas-Barberan FA, J. Agric. Food Chem., 49, 1035 (2001)

[19] Ahmad ST, Arjumand W, Nafees S, Seth A, Ali N, Rashid S, Sultana S, Toxicol. Lett., 208, 149 (2012)

[20] Shi X, Liao S, Mi H, Guo C, Qi D, Li F, Zhang C, Yang Z, Molecules, 17, 12868 (2012)

[21] Thumann G, Viethen A, Gaebler A, Walter P, Kaempf S, Johnen S, Salz AK, Biomaterials, 30, 287 (2009)

[22] Udono T, Takahashi K, Nakayama M, Murakami O, Durlu YK, Tamai M, Shibahara S, Invest. Ophthalmol. Vis. Sci., 41, 1962 (2000)

[23] Oh YA, Kim SH, Lee SJ, Yoo JJ, van Dyke M, Rhee JM, Khang G, Polym.(Korea), 32(5), 403 (2008)

[24] Tirkey N, Pilkhwal S, Kuhad A, Chopra K, BMC Pharmacol., 5, 2 (2005)

[25] Kuntic V, Filipovic I, Vujic Z, Molecules, 16, 1378 (2011)

[26] Song YS, Kwon JI, Kim BK, Jeon YY, Khang G, Lee W, J. Biomed. Mater. Res. A, 98, 517 (2011)

[27] Rao YK, Fang SH, Tzeng YM, Phytother. Res., 22, 957 (2008)

[28] Xiaoting L, Xiangyun Z, Shumei L, Minghua D, Liang X, Adv. Exp. Med. Biol., 664, 193 (2010)