Immobilization effect of bone morphogenetic protein-2 on collagen membrane via photoreactive gelatin derivatives: Biocompatibility and preservability of osteoinductive activity

Yun Heo; Eun-Hye Kim; Hyung-Jae Lee; Jin-Hong Jeong; Do-Gi Pyun; Eui-Chan Jang; Gang-Un Kim; Sung-Min Rhee; Tae-Il Son; Kwang-Sup Song

Macromolecular Research, Vol.23, No.6, 525-530, June, 2015

DOI10.1007/s13233-015-3068-3 Export Citation

Immobilization effect of bone morphogenetic protein-2 on collagen membrane via photoreactive gelatin derivatives: Biocompatibility and preservability of osteoinductive activity

Yun Heo, Eun-Hye Kim, Hyung-Jae Lee, Jin-Hong Jeong, Do-Gi Pyun¹, Eui-Chan Jang², Gang-Un Kim², Sung-Min Rhee², Tae-Il Son, and Kwang-Sup Song^{2, †}

Department of Systems Biotechnology and Bio-Environmental Technology (BET) Research Institute, Chung-Ang University, Anseong, Gyeonggi-do, 456-756, Korea
¹Department of Polymer Science and Engineering, Sungkyunkwan University, Suwon, Gyeonggi-do, 440-746, Korea
²Department of Orthopedic Surgery, College of Medicine, Chung-Ang University, 224-1, Heukseok-dong, Dongjak-gu, Seoul, 156-755, Korea

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To photo-immobilize bone morphogenetic protein-2 (BMP-2), UV light-reactive gelatin (azidophenyl gelatin, Az-gel), which is introduced with an N-(4-azidobenzoyloxy) succinimide-containing an azide group, was prepared as a carrier of BMP-2. Characterization of the photo-reactivity of Az-gel was evaluated by the curing ratio and it was micropatterned using a photomask. The cross-linking ratio and an enzyme-linked immunosorbent assay, enzymelinked immunospecific assay (ELISA)-based BMP-2-release test were used to determine the optimal density of Azgel (1, 3, and 5%) and UV treatment time (10, 30, 60 s) of the BMP-2 carrier in collagen matrix. The results showed that 5% Az-gel with 60 s of UV treatment was the most efficient condition for cross-linking. Cytotoxicity assays using the C2C12 mouse myoblast cell line showed that there were no cytotoxic effects of Az-gel. Alkaline phosphatase activity and calcium ion detection assays to evaluate the osteoinductive activities were conducted in assumed optimal condition of Az-gel (5% and 60 s UV treatment) 14 days post-treatment. In our study, the osteoinductive activities were the highest in cells treated with BMP-2 and the photo-reactive Az-gel carrier compared to BMP-2 alone or untreated control cells in collagen matrix at all time points (3, 7, and 14 days).

Keywords:azidophenyl gelatin;photo-immobilization;collagen membrane;sustained protein release;bone morphogenetic protein-2 (BMP-2)

[References]

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[2] Petite H, Viateau V, Bensaid W, Meunier A, Pollak CD, Bourguignon M, Oudina K, Sedel L, Guillemin G, Nat. Biotechnol., 18, 959 (2000)

[3] Yuan H, Fernandes H, Habibovic P, de Boer J, Barradas AM, de Ruiter A, Walsh WR, van Blitterswijk CA, de Bruijn JD, Proc. Natl. Acad. Sci., 107, 13614 (2010)

[4] Nique T, Fonseca RJ, Upton LG, Scott R, J. Oral Maxillofac. Surg., 45, 386 (1987)

[5] Powles R, Clink H, Spence D, Morgenstern G, Watson J, Selby P, Woods M, Barrett A, Jameson B, Sloane J, Lancet, 315, 327 (1980)

[6] Jin Q, Anusaksathien O, Webb S, Rutherford R, Giannobile W, J. Periodontol., 74, 202 (2003)

[7] Blanc KL, Rasmusson I, Sundberg B, Gotherstrom C, Hassan M, Uzunel M, Ringden O, Lancet, 363, 1439 (2004)

[8] Boden SD, Titus L, Hair G, Liu Y, Viggeswarapu M, Nanes MS, Baranowski C, Spine, 23, 2486 (1998)

[9] Park J, Ries J, Gelse K, Kloss F, Mark KVD, Wiltfang J, Neukam F, Schneider H, Gene Ther., 10, 1089 (2003)

[10] Kempen DH, Lu L, Heijink A, Hefferan TE, Creemers LB, Maran A, Yaszemski MJ, Dhert WJ, Biomaterials, 30, 2816 (2009)

[11] Mason JM, Breitbart AS, Barcia M, Porti D, Pergolizzi RG, Grande DA, Clin. Orthop. Rel. Res, 379, S171 (2000)

[12] Franceschi R, J. Dent. Res., 84, 1093 (2005)

[13] Dai K, Xu X, Tang T, Zhu Z, Yu C, Lou J, Zhang X, Calcif. Tissue Int., 77, 55 (2005)

[14] Wozney JM, Rosen V, Clin. Orthop. Rel. Res, 346, 26 (1998)

[15] Sellers RS, Zhang R, Glasson SS, Kim HD, Peluso D, D’augusta DA, Beckwith K, Morris EA, J. Bone Joint Surg., 82, 151 (2000)

[16] Giannoudis PV, Einhorn TA, Schmidmaier G, Marsh D, Injury, 39, S5 (2008)

[17] Ozaki J, Fujimoto S, Masuhara K, Tamai S, Yoshimoto S, Clin. Orthop. Rel. Res, 202, 173 (1986)

[18] Ye L, Mosbach K, Chem. Mater., 20, 859 (2008)

[19] Rowley JA, Madlambayan G, Mooney DJ, Biomaterials, 20, 45 (1999)

[20] Lu K, Lu J, Mater. Sci. Eng. A-Struct. Mater. Prop. Microstruct. Process., 375, 38 (2004)

[21] Johnson WL, Prog. Mater. Sci., 30, 81 (1986)

[22] Hubbell JA, Nat. Biotechnol., 13, 565 (1995)

[23] Na HN, Kim KI, Han JH, Lee JG, Son TI, Han DK, Ito Y, Song KS, Jang EC, Macromol. Res., 18(10), 1001 (2010)

[24] Kim KI, Na HN, Ito Y, Son TI, Macromol. Res., 19(3), 216 (2011)

[25] Park S, Seo S, Na H, Kim K, Lee J, Woo H, Lee J, Seok H, Lee J, Chung S, Chung KH, Han DK, Ito Y, Jang E, Son T, Macromol. Res., 19(9), 921 (2011)

[26] Park SH, Seo SY, Lee HJ, Na HN, Lee JW, Woo HD, Son TI, Macromol. Res., 20(8), 842 (2012)

[27] Ito Y, Hasuda H, Yamauchi T, Komatsu N, Ikebuchi K, Biomaterials, 25, 2293 (2004)

[28] Ito Y, Mater. Sci. Eng. C-Biomimetic Supramol. Syst., 6, 267 (1998)

[29] Son TI, Sakuragi M, Takahashi S, Obuse S, Kang J, Fujishiro M, Matsushita H, Gong J, Shimizu S, Tajima Y, Acta Biomater., 6, 4005 (2010)

[30] Heo Y, Lee HJ, Kim EH, Kim MK, Ito Y, Son TI, J. Appl. Polym. Sci., 131 (2014)

[31] Park SH, Seo SY, Kang JH, Ito Y, Son TI, J. Appl. Polym. Sci., 127(1), 154 (2013)

[32] Matsuda T, Sugawara T, Langmuir, 11(6), 2272 (1995)

[33] Ito Y, Hasuda H, Terai H, Kitajima T, J. Biomed. Mater. Res. A, 74, 659 (2005)