헤스페리딘을 함유한 락타이드 글리콜라이드 공중합체 지지체에서 섬유륜 세포의 증식과 거동 평가

장나금; 고현아; 차세롬; 이선의; 정현기; 김수영; 신재훈; 김은영; 송정은; 강길선; Na Keum Jang; Hyun Ah Ko; Se Rom Cha; Seon Eui Lee; Hyun Ki Jeong; Su Young Kim; Jae Hun Shin; Eun Young Kim; Jeong Eun Song; Gilson Khang

Polymer(Korea), Vol.39, No.5, 782-787, September, 2015

DOI10.7317/pk.2015.39.5.782 Export Citation

헤스페리딘을 함유한 락타이드 글리콜라이드 공중합체 지지체에서 섬유륜 세포의 증식과 거동 평가

Proliferation and Growth Behavior of Annulus Fibrosus Cells on Hesperidin Loaded Poly(lactide-co-glycolic acid) Scaffold

장나금, 고현아, 차세롬, 이선의, 정현기, 김수영, 신재훈, 김은영, 송정은 , 강길선 ^†

전북대학교 BIN융합공학과, 고분자나노공학과, 고분자융합소재연구소

Na Keum Jang, Hyun Ah Ko, Se Rom Cha, Seon Eui Lee, Hyun Ki Jeong, Su Young Kim, Jae Hun Shin, Eun Young Kim, Jeong Eun Song , and Gilson Khang^†

Dept. of BIN Fusion Tech & Dept. of PolymerNanoSci Tech, Chonbuk National University, 567 Baekje-daero, Jeonju 561-756, Korea

E-mail:

초록

항산화와 항염증으로 잘 알려진 헤스페리딘은 디스크 재생을 위해 효과적인 지지체로 제작하기 위해 사용하였다. 지지체는 poly(lactide-co-glycolic acid) (PLGA)에 헤스페리딘(0, 3, 5, 10%)을 첨가하여 염 침출법으로 제작하였고, 섬유륜 세포(AF)를 파종한 후 세포의 변화를 연구하였다. SEM, WST 그리고 RT-PCR 분석을 통해 세포의 부착과 증식, 세포의 표현형 분석을 실시하였다. 또한 면역조직화학염색을 통하여 AF세포의 생체 내 거동을 확인하였다. 그 결과, 5% 헤스페리딘이 함유된 PLGA 지지체가 가장 좋은 세포의 형태와 생체 적합성을 보여, 디스크 재생을 위한 지지체로서의 활용 가능성을 확인하였다.

Hesperidin, a well-known antioxidant and anti-inflammatory agent, was smartly used for designing efficient scaffolds for disc regeneration. The scaffolds were synthesized via a salt-leaching method by adding hesperidin (0, 3, 5, & 10%) to poly(lactide-co-glycolic acid) (PLGA) followed by a cellular behaviors study by seeding annulus fibrosus (AF) cells. The cellular adhesion, proliferation and phenotypes were further evaluated by SEM, WST and RT-PCR, respectively. Additionally, AF cell behavior was studied by immunohisto-chemical staining in vivo. Results showed that PLGA scaffold with 5% hesperidin are best suited for disc regeneration, showing good cellular morphology and biocompatibility.

Keywords:hesperidin;PLGA;annulus fibrosus;scaffold

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[1] Buckwalter JA, Spine, 20, 1307 (1995)

[2] Bibby SR, Jones DA, Lee RB, Yu J, Urban JPG, Joint Bone Spine, 68, 537 (2001)

[3] Hadjipaviou AG, Simmons JW, Pope MH, Necessary JT, Goel VK, Am. J. Orthop., 28, 561 (1999)

[4] Kim SH, Yun SJ, Jang JW, Kim MS, Khang G, Lee HB, Polym.(Korea), 30(1), 14 (2006)

[5] Hedbom E, Heinegard D, J. Biol. Chem., 264, 6898 (1989)

[6] Yeung AT, Yeung CA, Orthop. Clin. North Am., 38, 363 (1989)

[7] Carragee EJ, Han MY, Suen PW, Kim D, J. Bone Joint Surg. Am., 85, 102 (2003)

[8] Choy DS, J. Clin. Laser Med. Surg., 18, 29 (2000)

[9] Hegewald AA, Ringe J, Sittinger M, Thome. C. Front. Biosci., 13, 1507 (2008)

[10] Atlas SJ, Keller RB, Wu YA, Deyo RA, Spine, 30, 927 (2005)

[11] Masato S, Takashi A, Masayuki I, Miya I, Toshiyuki K, Makoto K, Kyosuke F, Spine, 28, 548 (2003)

[12] Yang S, Leong KF, Du Z, Chua CK, Tissue Eng., 7, 679 (2001)

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

[14] Nishida K, Hirano T, Cancer Sci., 94, 1029 (2003)

[15] Karageorgiou V, Kaplan D, Biomaterials, 26, 5474 (2005)

[16] Whitaker MJ, Quirk RA, Howdle SM, Shakesheff KM, J. Pharm. Pharmacol., 53, 1427 (2001)

[17] Lowes KN, Croager EJ, Olynyk JK, Abraham J, Yeoh GCT, J. Gastroent. Hepat., 18, 4 (2003)

[18] Cha SR, Jang NK, Kuk H, Kim EY, Song JE, Park CH, Khang G, Int. J. Tissue Regen., 5, 84 (2014)

[19] Zheng X, Yang F, Wang S, Lu S, Zhang W, Liu S, Huang J, Wang A, Yin B, Ma N, Zhang L, Xu W, Guo Q, J. Mater. Sci. -Mater. Med., 22, 693 (2011)

[20] Makadia HK, Siegel SJ, Polymer, 3, 1377 (2011)

[21] Taylor MS, Daniels AU, Andriano KP, Heller J, J. Appl. Biomater., 5, 151 (1994)

[22] Athanasiou K, Niederauer GG, Agrawal CM, Athanasiou K, Niederauer GG, Agrawal CM, Biomaterials, 17, 93 (1996)

[23] Kim HY, Lee SJ, Kim YL, Park CH, Kwon SY, Chung JW, Lee D, Khang G, Inter. J. Tissue Regen., 4, 95 (2013)

[24] Verheyen CC, Wijn JR, Blitterswijk CA, J. Biomed. Mater. Res., 27, 1115 (1993)

[25] Anderson JM, Shive MS, Adv. Drug. Deliv. Rev., 28, 5 (1997)

[26] Rouseff RL, J. Assoc. Off. Anal. Chem., 71, 798 (1988)

[27] Son HS, Kim HS, Kwon TB, Ju JS, J. Korea. Soc. Food Nutr., 21, 136 (1992)

[28] Isabelle M, Gerard L, Pascale C, Odile S, Nicole P, Pierre B, Pierre C, Josiane C, Biochem. Pharmacol., 45, 13 (1993)

[29] Guardia T, Rotelli AE, Juarez AO, Pelzer LE, Il Farmaco, 56, 683 (2001)

[30] Kaur G, Tirkey N, Chopra K, Toxicology, 226, 152 (2006)

[31] Song JE, Shim CR, Lee Y, Ko HA, Yoon H, Lee D, Khang G, Polym.(Korea), 37(3), 323 (2013)

[32] Cho SA, Cha SR, Park SM, Kim KH, Lee HG, Kim EY, Lee D, Khang G, J. Biomater. Sci.-Polym. Ed., 25, 625 (2014)