Synthetic Optimization of Gelatin-Oleic Conjugate and Aqueous-Based Formation of Self-Assembled Nanoparticles without Cross-Linkers

Van Hong Nguyen; Beom-Jin Lee

Macromolecular Research, Vol.25, No.5, 466-473, May, 2017

DOI10.1007/s13233-017-5056-2 Export Citation

Synthetic Optimization of Gelatin-Oleic Conjugate and Aqueous-Based Formation of Self-Assembled Nanoparticles without Cross-Linkers

Van Hong Nguyen, and Beom-Jin Lee^†

Bioavailability Control Laboratory, College of Pharmacy, Ajou University, Suwon, Gyeonggi 16499, Korea

E-mail:

Amphiphilically-modified gelatin has the ability to form self-assembled NPs which own hydrophobic inner core for water-insoluble drugs loading and gelatin layer for circulation extension and biocompatibility. Gelatin-oleic acid conjugates (GOC) was synthesized from oleic acid (OA) and gelatin, using 1-ethyl-3 (3 dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) in 60% ethanol. The effect of various gelatin concentrations, oleic acid-gelatin molar ratio and reaction time on the substitution of GOC was evaluated. Fourier transform infrared (FT-IR) spectroscopy and proton nuclear magnetic resonance (1H NMR) revealed the successful synthesis of GOC. The ratio of oleic acid-reacted amino groups within gelatin was above 50% as quantified using the 2,4,6-trinitrobenzene sulfonic acid (TNBS) method. Based on the physicochemical changes of GOC upon the modification of various synthesis factors, the optimized process for the GOC synthesis was well established. Self-assembled gelatin-oleic acid nanoparticles (GON) were fabricated simply by the desolvation method using dialysis membrane, without adding a cross-linker. GON was formed at a lower critical micelle concentration (CMC) in water, 0.3 mg/mL. GON has a homogenous size of 100-300 nm, which was showed in the electrophoretic light scattering (ELS) results, scanning electron microscope (SEM) and transmission electron microscope (TEM) images. The zeta potential of GON indicated a slightly negative surface charge (-6 mV). The GON showed no toxicity in human embryonic kidney cells (HEK 293) and significantly improved coumarin loading in adenocarcinomic human alveolar basal epithelial cells (A549). The current GOC and GON could provide versatile strategies to deliver cargo into the body by acting as a safe and biocompatible nanoparticulate carrier.

Keywords:gelatin-oleic acid conjugate;aqueous-based self-assembled nanoparticles;physicochemical properties;synthetic optimization;cellular uptake

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[2] Dash RN, Mohammed H, Humaira T, J. Pharm. Invest., 46, 153 (2016)

[3] Naseripour M, Abrishami M, Sedaghat A, Abrishami M, Kanavi MR, Mosallaei N, Falavarjani KG, Pourmatin R, Safarian O, Malaekeh-Nikouei B, J. Pharm. Invest., 46, 575 (2016)

[4] Du YZ, Weng Q, Yuan H, Hu FQ, ACS Nano, 4, 6894 (2010)

[5] Nesporova K, Sogorkova J, Smejkalova D, Kulhanek J, Huerta-Angeles G, Kubala L, Velebny V, Int. J. Pharm., 511, 638 (2016)

[6] Kirtane AR, Narayan P, Liu G, Panyam J, J. Pharm. Invest., 47, 65 (2017)

[7] Rao KM, Rao KSYK, Ramanjaneyulu G, Ha CS, Int. J. Pharm., 478, 788 (2015)

[8] Nunes PS, Rabelo AS, Souza JC, Santana BV, da Silva TM, Serafini MR, Menezes PDP, Lima BDS, Cardoso JC, Alves JC, Frank LA, Guterres SS, Pohlmann AR, Pinheiro MS, de Albuquerque RL, Araujo AA, Int. J. Pharm., 513(1-2), 473 (2016)

[9] Park C, Vo CLN, Kang T, Oh E, Lee BJ, Eur. J. Pharm. Biopharm., 89, 365 (2015)

[10] Tran PHL, Tran TTD, Vo TV, Vo CLN, Lee BJ, J. Biomed. Nanotechnol., 9, 1416 (2013)

[11] Won YW, Yoon SM, Sonn CH, Lee KM, Kim YH, ACS Nano, 5, 3839 (2011)

[12] Kim KJ, Byun Y, Biotechnol. Bioprocess Eng., 4, 210 (1999)

[13] Tanigo T, Takaoka R, Tabata Y, J. Control. Release, 143, 201 (2010)

[14] Seo J, Lee J, Na K, Macromol. Res., 25(1), 63 (2017)

[15] Hermanson GT, Bioconjugate Techniques, 3rd ed., Academic Press, London, 2013.

[16] Gao FP, Zhang HZ, Liu LR, Wang YS, Jiang Q, Yang XD, Zhang QQ, Carbohydr. Polym., 71, 606 (2008)

[17] Tran TH, Bae BC, Lee YK, Na K, Huh KM, Carbohydr. Polym., 92, 1615 (2013)

[18] Muller RH, Ruhl D, Luck M, Paulke BR, Pharm. Res., 14, 18 (1997)

[19] Jain A, Gulbake A, Jain A, Shilpi S, Hurkat P, Jain A, Jain SK, J. Microencapsul., 29(1), 95 (2012)

[20] Schrieber R, Gareis H, Gelatine Handbook: Theory and Industrial Practice, John Wiley & Sons, Weinheim, 2007.

[21] Cavalieri F, Postma A, Lee L, Caruso F, ACS Nano, 3, 234 (2009)

[22] Zhou HF, Yu WT, Guo X, Liu XD, Li N, Zhang Y, Ma XJ, Biomacromolecules, 11(12), 3480 (2010)

[23] Yinsong W, Lingrong L, Jian W, Zhang Q, Carbohydr. Polym., 69, 597 (2007)

[24] Jiang GB, Quan D, Liao K, Wang H, Carbohydr. Polym., 66, 514 (2006)

[25] Fuguet E, Rafols C, Roses M, Bosch E, Anal. Chim. Acta, 548, 95 (2005)

[26] Liu WG, De Yao K, Wang GC, Li HX, Polymer, 41(20), 7589 (2000)

[27] Won YW, Kim YH, Macromol. Res., 17(7), 464 (2009)