여러 가지 천연고분자를 함유한 PLGA 하이브리드 지지체에서 토끼 골수줄기세포의 골분화 능력 평가

전하얀; 김수민; 전유신; 국연지; 송정은; 강길선; Ha Yan Jeon; Soo Min Kim; Yoo Shin Jeon; Yeon Ji Kook; Jeong Eun Song; Gilson Khang

Polymer(Korea), Vol.41, No.5, 867-873, September, 2017

DOI10.7317/pk.2017.41.5.867 Export Citation

여러 가지 천연고분자를 함유한 PLGA 하이브리드 지지체에서 토끼 골수줄기세포의 골분화 능력 평가

Osteogenic Differentiation of Rabbit Bone Marrow Mesenchymal Stem Cell in Several Natural Source Biomaterials/PLGA Hybrid Scaffolds

전하얀, 김수민, 전유신, 국연지, 송정은 , 강길선 ^†

전북대학교 BIN 융합공학과, 고분자나노공학과, 고분자 BIN융합 연구소

Ha Yan Jeon, Soo Min Kim, Yoo Shin Jeon, Yeon Ji Kook, Jeong Eun Song , and Gilson Khang^†

Department of BIN Convergence Tech, Polymer BIN Fusion Res Center & Department of Polymer·Nano Sci Tech, Chonbuk National University, 567 Deokjin, Jeonju, Chonbuk 54898, Korea

E-mail:

초록

합성고분자인 poly(lactic-co-glycolic acid)(PLGA)는 우수한 생분해능을 가진 생체재료로서 널리 이용되고 있으며, 천연유래 고분자인 실크와 소장 점막하 조직, 탈 미네랄화 골분 그리고 오리발 유래의 콜라겐은 생체 적합한특성을 가지고 있다. 본 논문에서는 여러 가지 천연유래 고분자를 함유한 PLGA를 제작한 후, 지지체의 물성 및 표면을 확인하였으며 토끼 골수유래줄기세포를 이용하여 지지체에 파종 후, 세포의 생존율과 세포분화 정도를 측정하였고 in vivo 실험에서 면역조직학적 염색인 H&E와 Von Kossa을 실시하여 골분화 정도를 확인하였다. 결과적으로 오리발 유래 콜라겐과 소장 점막하 조직을 함유한 지지체에서 빠른 골 분화의 모습을 확인할 수 있었다.

Poly(lactic-co-glycolic acid) (PLGA) has been most widely used owing to its advantages such as good biodegradability and biocompatibility. Scaffolds made of natural source bio-materials like small intestinal submucosa (SIS), demineralized bone particles (DBP), silk fibroin (SF), and collagen-derived duck’s feet collagen (DC) have been proved to be biocompatible. We manufactured composite scaffolds of all these natural biomaterials/PLGA. After making hybrid scaffolds, we measured mechanical strength of scaffolds and observed a porosity and surface morphology of the scaffolds by scanning electronic microscopy. Rabbit bone marrow mesenchymal stem cells (rBMSCs) were cultured, expanded and seeded on scaffolds. Inflammatory responses were checked at 1, 4 and 8 weeks using H&E & Von Kossa staining. DC and SIS group were found to be faster in bone regeneration than other groups. Finally, we have observed that scaffolds using biomaterials are effective in cell attachment, stem cell differentiation, and mineralized tissue formation.

Keywords:collagen;silk;PLGA;scaffold;bone marrow mesenchymal stem cells

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[1] Levenberg S, Langer R, Curr. Top. Dev. Biol., 61, 113 (2004)

[2] Shin H, Jo S, Mikos AG, Biomaterials, 24, 4353 (2003)

[3] Black CRM, Goriainov V, Gibbs D, Kanczler J, Tare RS, Oreffo ROC, Curr. Mol. Bio. Rep., 1, 132 (2015)

[4] Santin M, Motta A, Freddi G, Cannas M, J. Biomed. Mater. Res., 46, 382 (1999)

[5] Ko HA, Kim HM, Ha HJ, Kim AR, Song JE, Lee KH, Sang JE, Yoon H, Lee OJ, Lee D, Park CH, Khang G, Int. J. Tissue Regen., 3, 103 (2012)

[6] Jin HJ, Chen J, Karageorgiou V, Altman GH, Kaplan DL, Biomaterials, 25, 1039 (2004)

[7] Meinel L, Karageorgiou V, Hofmann S, Fajardo R, Snyder B, Li C, Zichner L, Langer R, Vunjak-Novakov G, Kaplan DL, J. Biomed. Mater. Res., 71, 25 (2004)

[8] Miao X, Tan DM, Li J, Xiao Y, Crawford R, Acta Biomater., 4, 638 (2008)

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

[10] Nair LS, Laurencin CT, Prog. Polym. Sci, 32, 762 (2007)

[11] Lu L, Peter SJ, Lyman MD, Lai HL, Leite SM, Tamada JA, Uyama S, Vacanti JP, Langer R, Mikos AG, Biomaterials, 21, 1837 (2000)

[12] Oh SH, Kang SG, Kim ES, Cho SH, Lee JH, Biomaterials, 24, 4011 (2003)

[13] Liu C, Xia Z, Czernuszka JT, Chem. Eng. Res. Des., 85(A7), 1051 (2007)

[14] Caputo I, Lepretti M, Scarabino C, Esposito C, Proto A, Anal. Biochem., 421, 92 (2012)

[15] Awidi A, Ababneh N, Alkilani H, Salah B, Nazzal S, Zoghool M, Shomaf M, Mater. Med., 26, 239 (2015)

[16] Vepari C, Kaplan DL, Prog. Polym. Sci, 32, 991 (2007)

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

[18] Yoshimoto H, Shin YM, Terai H, Biomaterials, 24, 2077 (2003)

[19] Fan H, Hu Y, Zhang C, Li X, Lu R, Qin L, Zhn R, Biomaterials, 27, 4573 (2006)

[20] Wu YC, Shaw SY, Lin HR, Lee TM, Yang CY, Biomaterials, 27, 896 (2006)

[21] Goh SK, Bertera S, Olsen P, Candielio JE, Halfter W, Uechi G, Balasubramani M, Johnson SA, Sicari BM, Kollar E, Badylak SF, Banerjee I, Biomaterials, 34, 6760 (2013)

[22] Long K, Liu Y, Li W, Wang L, Liu S, Wang Y, Wang Z, J. Biomed. Mater. Res., 103, 1159 (2015)

[23] Lee OJ, Lee JM, Kim JH, Kim J, Kweon HY, Jo YY, Park CH, J. Biomed. Mater. Res., 100, 2018 (2012)

[24] Kim SH, Park HS, Lee OJ, Chao JR, Park HJ, Lee JM, Ju HW, Moon BM, Park YR, Song JE, Khang G, Park CH, Int. J. Biol. Macromol., 85, 442 (2016)

[25] Kim KS, Lee JY, Kang YM, Kim ES, Kim GH, Rhee SD, Cheon HG, Kim JH, Min BH, Lee HB, Kim MS, Biomaterials, 31, 1104 (2010)

[26] Voytik-Harbin SL, Brightman AO, Kraine MR, Waisner B, Badylak SF, J. Cell Biochem., 67, 478 (1997)

[27] Hodde JP, Badylak SF, Brightman AO, Voytikharbin SL, Tissue Eng., 2, 209 (1996)

[28] McPherson TB, Badylak SF, Tissue Eng., 4, 75 (1998)

[29] Thitiset T, Damrongsakkul S, Bunaprasert T, Leeanansaksiri W, Honsawek S, Int. J. Mol. Sci., 14(1), 2056 (2013)

[30] Silvipriya KS, Kumar KK, Bhat AR, Kumar BD, Anish J, J. Appl. Pharmaceuti. Sci., 5, 123 (2015)

[31] Zhang L, Zhao J, Duan M, Zhang H, Jiang J, Yu R, Anal. Chem., 85, 3797 (2013)