DBP 스폰지와 DBP/PLGA 지지체에서의 인간 디스크세포 거동분석 비교

이선경; 홍희경; 김수진; 김용기; 송이슬; 하윤; 이동원; 강길선; Seon Kyoung Lee; Hee Kyung Hong; Su Jin Kim; Yong Ki Kim; Yi Seul Song; Yoon Ha; Dongwon Lee; Gilson Khang

Polymer(Korea), Vol.34, No.5, 398-404, September, 2010

The Comparison of Sponges and PLGA Scaffolds Impregnated with DBP on Growth Behaviors of Human Intervertebral Disc Cells

이선경, 홍희경, 김수진, 김용기, 송이슬, 하윤¹, 이동원 , 강길선 ^†

전북대학교 BIN 융합공학과, 고분자 나노공학과
¹연세대학교 의과대학 신경외과

Seon Kyoung Lee, Hee Kyung Hong, Su Jin Kim, Yong Ki Kim, Yi Seul Song, Yoon Ha¹, Dongwon Lee , and Gilson Khang^†

Department of BIN Fusion Technology & Department of PolymerㆍNano Science & Technology, Chonbuk National University, 664-14, Dukjin Ku, Jeonju 561-756, Korea
¹Department of Neurosurgery, Spine & Spinal Cord Institute, College of Medicine, Yonsei University, Shinchon-dong, Seodaemon-ku, Seoul 120-750, Korea

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초록

본 연구팀은 DBP를 함침시킨 물성이 서로 다른 스폰지와 PLGA 지지체를 제작한 후 세포 부착, 증식 및 형태 유지를 알아보기 위한 실험을 수행하였다. WST 분석법과 SEM 관찰을 통하여 스폰지에 비해서 PLGA 지지체에서의 세포의 증식이 활발한 것을 확인하였고, RT-PCR을 통해 디스크세포에서 특이적으로 발현하는 제 2형 콜라겐과 어그리칸의 발현을 확인하였다. WST 결과, 세포 증식률은 DBP/PLGA 지지체가 DBP를 함침시킨 스폰지보다 세포 증식률이 높음을 확인하였다. 본 연구팀은 스폰지보다 PLGA 지지체가 인간디스크의 표현형 유지 및 증식에 있어서 긍정적인 영향을 미치는 것을 확인하였다.

We fabricated sponge and poly(lactide-co-glycolide)(PLGA) scaffolds impregnated demineralized bone particle(DBP)(DBP/PLGA) and investigated proper condition to proliferation and phenotype maintenance of intervertebral disc(IVD) cells by comparison between DBP/PLGA scaffold and DBP sponge. DBP/PLGA scaffolds were prepared by solvent casting/salt leaching. Human IVD cells were seeded in scaffolds of two types. Cell viability and proliferation according to scaffolds were analyzed by WST assay and SEM. RT-PCR was assessed to measure mRNA expression of aggrecan and type II collagen of human IVD cells. In WST assay results, cell viability in scaffolds impregnated DBP/PLGA scaffold were higher than DBP sponge. We could observe that disc cell mRNA expressed better in DBP/PLGA scaffold than DBP sponge. We concluded that the using of DBP/PLGA in terms of scaffold fabrication for bio-disc with human IVD cells is helpful growth of disc cells maintenance of phenotypes.

Keywords:DBP;PLGA;scaffold;sponge;H-disc

[References]

Hochschuler RF, Rashbaum RG, Johnson D, Ohnmeiss V, Moo H, Vanharanta BL, Sachs MA, Spivey RD, Guyer SH, Spine, 12, 295 (1987)
Gruber HE, Johnson TL, Leslie K, Ingram JA, Martin D, Hoelscher G, Banks D, Phieffer L, Coldham G, Hanley EN, Spine, 27, 1626 (2002)
Buckwalter JA, Spine, 20, 1307 (1995)
Hedbom E, Heinegard D, J. Biol. Chem., 264, 6898 (1989)
Hedbom E, Heinegard D, J. Biol. Chem., 268, 27307 (1993)
Nishida K, Kang JD, Gilbertson LG, Moon SH, Suh JK, Vogt MT, Robbins PD, Evans CH, Spine, 24, 2419 (1999)
Mochida J, Nishimura K, Nomura T, Toh E, Chiba M, Spine, 21, 1556 (1996)
Lee CK, Spine, 12, 357 (1988)
Schlegel JD, Smith JA, Schleusener RL, Spine, 21, 970 (1996)
Mizuno H, Roy AK, Vacanti CA, Kojima K, Ueda M, Bonassar LJ, Spine, 29, 1299 (2004)
Seguin CA, Grynpas MD, Pillar RM, Walden SD, Kandal RA, Spine, 29, 1299 (2004)
Tsai WB, Chen CH, Chen JF, Chang KY, J. Mater. Sci. Med., 17, 337 (2006)
Atala A, J. Endourol., 14, 49 (2000)
Zhang N, Yan H, Wen X, Brain Res. Rev., 49, 48 (2005)
Urist MR, Peltier LF, Science, 150, 893 (1965)
Glowacki J, Kaban LB, Murray JE, Folkman J, Mulliken JB, Lancet, 1, 959 (1981)
Khang G, Kim MS, Cho SH, Lee I, Rhee JM, Lee HB, Tissue Eng. Regen. Me., 1, 9 (2004)
Goegan P, Johnson G, Vincent R, Toxic in Vitro, 9, 257 (1995)
Kim KS, Cho MH, Ahn HH, Song SB, Seo SJ, Kim MS, Lee B, Khang G, Lee HB, Tissue Eng. Regen. Med., 4, 168 (2007)
Huang W, Carlsen B, Wulur I, Rudkina G, Ishidaa K, Wuc B, Yamaguchia DT, Miller TA, Experimental Cell Res., 299, 325 (2004)
Sun HG, Wu C, Dai K, Changc J, Tang T, Biomaterials, 27, 5651 (2006)
Kim SH, Yoon SJ, Choi BS, Ha HJ, Rhee JM, Kim MS, Yang YS, Lee HB, Khang G, Adv. Exp. Med. Biol., 585, 167 (2006)
Choi BS, Kim SH, Yoon SJ, Ha HJ, Kim MS, Yang YI, Son Y, Khang G, Rhee JM, Lee HB, Tissue Eng. Regen. Med., 3, 295 (2006)
Jang JW, Baek MO, Kim SH, Choi JH, Yang JC, Hong HH, Hong HK, Rhee JM, Min BH, Khang GS, Polym.(Korea), 33(2), 104 (2009)

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[1] Hochschuler RF, Rashbaum RG, Johnson D, Ohnmeiss V, Moo H, Vanharanta BL, Sachs MA, Spivey RD, Guyer SH, Spine, 12, 295 (1987)

[2] Gruber HE, Johnson TL, Leslie K, Ingram JA, Martin D, Hoelscher G, Banks D, Phieffer L, Coldham G, Hanley EN, Spine, 27, 1626 (2002)

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

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

[5] Hedbom E, Heinegard D, J. Biol. Chem., 268, 27307 (1993)

[6] Nishida K, Kang JD, Gilbertson LG, Moon SH, Suh JK, Vogt MT, Robbins PD, Evans CH, Spine, 24, 2419 (1999)

[7] Mochida J, Nishimura K, Nomura T, Toh E, Chiba M, Spine, 21, 1556 (1996)

[8] Lee CK, Spine, 12, 357 (1988)

[9] Schlegel JD, Smith JA, Schleusener RL, Spine, 21, 970 (1996)

[10] Mizuno H, Roy AK, Vacanti CA, Kojima K, Ueda M, Bonassar LJ, Spine, 29, 1299 (2004)

[11] Seguin CA, Grynpas MD, Pillar RM, Walden SD, Kandal RA, Spine, 29, 1299 (2004)

[12] Tsai WB, Chen CH, Chen JF, Chang KY, J. Mater. Sci. Med., 17, 337 (2006)

[13] Atala A, J. Endourol., 14, 49 (2000)

[14] Zhang N, Yan H, Wen X, Brain Res. Rev., 49, 48 (2005)

[15] Urist MR, Peltier LF, Science, 150, 893 (1965)

[16] Glowacki J, Kaban LB, Murray JE, Folkman J, Mulliken JB, Lancet, 1, 959 (1981)

[17] Khang G, Kim MS, Cho SH, Lee I, Rhee JM, Lee HB, Tissue Eng. Regen. Me., 1, 9 (2004)

[18] Goegan P, Johnson G, Vincent R, Toxic in Vitro, 9, 257 (1995)

[19] Kim KS, Cho MH, Ahn HH, Song SB, Seo SJ, Kim MS, Lee B, Khang G, Lee HB, Tissue Eng. Regen. Med., 4, 168 (2007)

[20] Huang W, Carlsen B, Wulur I, Rudkina G, Ishidaa K, Wuc B, Yamaguchia DT, Miller TA, Experimental Cell Res., 299, 325 (2004)

[21] Sun HG, Wu C, Dai K, Changc J, Tang T, Biomaterials, 27, 5651 (2006)

[22] Kim SH, Yoon SJ, Choi BS, Ha HJ, Rhee JM, Kim MS, Yang YS, Lee HB, Khang G, Adv. Exp. Med. Biol., 585, 167 (2006)

[23] Choi BS, Kim SH, Yoon SJ, Ha HJ, Kim MS, Yang YI, Son Y, Khang G, Rhee JM, Lee HB, Tissue Eng. Regen. Med., 3, 295 (2006)

[24] Jang JW, Baek MO, Kim SH, Choi JH, Yang JC, Hong HH, Hong HK, Rhee JM, Min BH, Khang GS, Polym.(Korea), 33(2), 104 (2009)