Fabrication of Porous PLGA Microspheres with BMP-2 Releasing Polyphosphate-Functionalized Nano-Hydroxyapatite for Enhanced Bone Regeneration

Byeong Jin Jeon; Seo Young Jeong; Ahn Na Koo; Byung-Chul Kim; Yu-Shik Hwang; Sang Cheon Lee

Macromolecular Research, Vol.20, No.7, 715-724, July, 2012

DOI10.1007/s13233-012-0103-5 Export Citation

Fabrication of Porous PLGA Microspheres with BMP-2 Releasing Polyphosphate-Functionalized Nano-Hydroxyapatite for Enhanced Bone Regeneration

Byeong Jin Jeon, Seo Young Jeong, Ahn Na Koo¹, Byung-Chul Kim¹, Yu-Shik Hwang¹, and Sang Cheon Lee^{1, †}

Department of Life and Nanopharmaceutical Science, Kyung Hee University, Seoul, 130-701, Korea
¹Department of Maxillofacial Biomedical Engineering & Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul, 130-701, Korea

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This paper introduces a novel bone-regenerative scaffold that is based on the systematic combination of porous polymer microspheres, nano-hydroxyapatite, and bone morphogenetic protein-2 (BMP-2), where each component was rationally incorporated to express its intrinsic activity in bone tissue formation. Poly(lactide-co-glycolide) (PLGA) microspheres, with interconnected pore structures, were fabricated by a gas-forming method in a water-in-oil-in-water double emulsion and solvent evaporation process. Polyphosphate-functionalized nano-hydroxyapatite (PP-n-HAp) was employed as a main component and was immobilized on the pore surface of the PLGA microspheres to controllably incorporate and release BMP-2. The surface polyphosphate functionalities of PP-n-HAp enabled the stable chemical immobilization of nano-hydroxyapatite (n-HAp) on the amine-treated pore surface of the PLGA microspheres. Field-emission scanning electron microscopy (FE-SEM) and X-ray photoelectron spectroscopy (XPS) confirmed the nano-level exposure of n-HAp on the pore surface of the PLGA microspheres. BMP-2 with a positive charge was bound at a high efficiency onto the anionic phosphates of surface-immobilized PP-n-HAp and was controllably released for approximately 1 month. The release rate was manipulated by adjusting the amount of loaded BMP-2. The osteogenic differentiation and proliferation of human adipose-derived stem cells (hADSCs) within the n-HAp/BMP-2-incorporated microspheres were monitored in a dynamic 3D cell culture system. Histological, immunohistochemical, and quantitative real-time polymerase chain reaction (PCR) analyses showed that the PP-n-HAp-immobilized surface promoted cell adhesion/proliferation and osteoconduction. With its osteoinductive property, the sustained release of BMP-2 further enhanced the bone tissue regenerative activity of the porous microspheres.

Keywords:nano-hydroxyapatite;polyphosphate;bone morphogenetic protein-2;microsphere;surface immobilization.

[References]

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Ribeiro N, Sousa SR, Monteiro FJ, J. Colloid Interface Sci., 351(2), 398 (2010)
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Kothapalli CR, Shaw MT, Wei M, Acta Biomater., 1, 653 (2005)
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Chen FM, Chen R, Wang XJ, Sun HH, Wu ZF, Biomaterials., 30, 5215 (2009)
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[Referenced By]

[1] Liu Y, Lu Y, Tian X, Cui G, Zhao Y, Yang Q, Yu S, Xing G, Zhang B, Biomaterials., 30, 6276 (2009)

[2] Kim SS, Gwak SJ, Kim BS, J. Biomed. Mater. Res., 87A, 245 (2008)

[3] Kim K, Dean D, Lu A, Mikos AG, Fisher JP, Acta Biomater., 7, 1249 (2011)

[4] Shen H, Hu X, Yang F, Bei J, Wang S, Acta Biomater., 6, 455 (2010)

[5] Wei G, Ma PX, Biomaterials., 25, 4749 (2004)

[6] Ribeiro N, Sousa SR, Monteiro FJ, J. Colloid Interface Sci., 351(2), 398 (2010)

[7] Kim SS, Park MS, Jeon O, Choi CY, Kim BS, Biomaterials., 27, 1399 (2006)

[8] Kothapalli CR, Shaw MT, Wei M, Acta Biomater., 1, 653 (2005)

[9] Kokubo T, Acta Mater., 46, 2519 (1998)

[10] Landis WJ, Silver FH, Freeman JW, J. Mater. Chem., 16, 1495 (2006)

[11] Lee J, Choi WI, Tae G, Kim YH, Kang SS, Kim SE, Kim SH, Jung Y, Kim SH, Acta Biomater., 7, 244 (2011)

[12] Li C, Vepari C, Jin HJ, Kim HJ, Kaplan DL, Biomaterials., 27, 3115 (2006)

[13] Song I, Kim BS, Kim CS, Im GI, Biochem. Biophys. Res. Commun., 408(1), 126 (2011)

[14] Lee SC, Choi HW, Lee HJ, Kim KJ, Chang JH, Kim SY, Choi J, Oh KS, Jeong YK, J. Mater. Chem., 17, 174 (2007)

[15] Chung HJ, Kim IK, Kim TG, Park TG, Tissue Eng.Part A,, 14, 607 (2008)

[16] Park K, Park JS, Woo DG, Yang HN, Chung HM, Park KH, Biomaterials., 29, 2490 (2008)

[17] Kim SE, Choi HW, Lee HJ, Chang JH, Choi J, Kim KJ, Lim HJ, Jun YJ, Lee C, J. Mater. Chem., 18, 4994 (2008)

[18] Lee HJ, Choi HW, Kim KJ, Lee SC, Chem. Mater., 18, 5111 (2006)

[19] Ciapetti G, Ambrosio L, Savarino L, Granchi D, Cenni,E, Baldini N, Pagani S, Guizzardi S, Causa F, Giunti A, Biomaterials., 24, 3815 (2003)

[20] Choi HW, Lee HJ, Kim KJ, Kim HM, Lee SC, J. Colloid Interface Sci., 304(1), 277 (2006)

[21] Chen FM, Chen R, Wang XJ, Sun HH, Wu ZF, Biomaterials., 30, 5215 (2009)

[22] Liu Y, Lu Y, Tian X, Cui G, Zhao Y, Yang Q, Yu S, Xing G, Biomaterials., 30, 6276 (2009)

[23] Chung HJ, Park TG, Tissue Eng. Part A., 15, 1391 (2008)

[24] Shi ZL, Neoh KG, Kang ET, Poh CK, Wang W, Biomacromolecules, 10(6), 1603 (2009)

[25] Balasundaram G, Yao C, Webster TJ, J. Biomed. Mater.Res., 84A, 447 (2008)

[26] Kretlow JD, Mikos AG, Tissue Eng., 13, 927 (2007)

[27] Stancu IC, Filmon R, Cincu C, Marculescu B, Zaharia C,Tourmen Y, Basle MF, Chappard D, Biomaterials., 25, 205 (2004)

[28] Yin YJ, Luo XY, Cui JF, Wang CY, Guo XM, Yao KD, Macromol. Biosci., 4, 971 (2004)

[29] Murphy WL, Mooney DJ, J. Am. Chem. Soc., 124(9), 1910 (2002)

[30] Tan J, Gemeinhart RA, Ma M, Saltzman WM, Biomaterials., 26, 3663 (2005)

[31] Gemeinhart RA, Bare CM, Haasch RT, Gemeinhart EJ, J. Biomed. Mater. Res. A., 78, 433 (2006)

[32] Jadlowiec J, Koch H, Zhang X, Campbel PGl, Seyedain M, Sfeir C, J. Biol. Chem., 279, 53323 (2004)