Synthesis, characterization and biocompatibility evaluation of hydroxyapatite - gelatin polyLactic acid ternary nanocomposite

Document Type: Research Paper

Authors

1 Faculty of Metallurgical and Materials Engineering, Semnan University, Semnan, Iran

2 Faculty of New Science and Technology, Semnan University, Semnan, Iran

10.7508/nmj.2016.02.006

Abstract

Objective(s): The current study reports the production and biocompatibility evaluation of a ternary nanocomposite consisting of HA, PLA, and gelatin for biomedical application.
Materials and Methods: Hydroxyapatite nanopowder (HA: Ca10(PO4)6(OH)2) was produced by burning the bovine cortical bone within the temperature range of 350-450 oC followed by heating in an oven at 800. Synthesis of the ternary nanocomposite was carried out in two steps: synthesis of gelatin-hydroxyapatite binary nanocomposite and addition of poly lactic acid with different percentages to the resulting composition. The crystal structure was determined by X-ray diffraction (XRD), while major elements and impurities of hydroxyapatite were identified by elemental analysis of X-ray fluorescence (XRF). Functional groups were determined by Fourier transform infrared spectroscopy (FTIR). Morphology and size of the nanocomposites were evaluated using field emission scanning electron microscope (FE-SEM).Biocompatibility of nanocomposites was investigated by MTT assay.
Results: XRD patterns verified the ideal crystal structure of the hydroxyapatite, which indicated an appropriate synthesis process and absence of disturbing phases. Results of FTIR analysis determined the polymers’ functional groups, specified formation of the polymers on the hydroxyapatite surface, and verified synthesis of nHA/PLA/Gel composite. FESEM images also indicated the homogeneous structure of the composite in the range of 50 nanometers. MTT assay results confirmed the biocompatibility of nanocomposite samples.
Conclusion: This study suggested that the ternary nanocomposite of nHA/PLA/Gel can be a good candidate for biomedical application such as drug delivery systems, but for evaluation of its potential in hard tissue replacement, mechanical tests should be performed.

Keywords


[1]  Bose S, Tarafder S. Calcium phosphate ceramic systems in growth factor and drug delivery for bone tissue engineering: a review. Acta Biomater. 2012; 8(4): 1401-1421.

[2]  Verron E, Khairoun I, Guicheux J, Bouler JM. Calcium phosphate biomaterials as bone drug delivery systems: a review. Drug DiscovToday. 2010; 15(13): 547-552.

[3]  Meseguer Olmo L, Ros Nicolás MJ, Vicente Ortega V, Alcaraz Baños M, Clavel Sainz M, Arcos D, Ragel CV, Vallet Regí M, Meseguer Ortiz C. A bioactive sol gel glass implant for in vivo gentamicin release. Experimental model in Rabbit. J Orthopaed Res. 2006; 24(3): 454-460.

[4]  Zakaria SM, Sharif Zein SH, Othman MR, Yang F, Jansen JA. Nanophase hydroxyapatite as a biomaterial in advanced hard tissue engineering: a review. Tissue Eng Part B: Reviews. 2013; 19(5): 431-441.

[5]  Dion A, Langman M, Hall G, Filiaggi M. Vancomycin release behaviour from amorphous calcium polyphosphate matrices intended for osteomyelitis treatment. Biomaterials. 2005; 26(35): 7276-7285.

[6]  Rhee SH. Synthesis of hydroxyapatite via mechanoch- emical treatment. Biomaterials. 2002; 23(4): 1147-1152.

[7]  Liu DM, Troczynski T, Hakimi D. Effect of hydrolysis on the phase evolution of water-based sol–gel hydroxyapatite and its application to bioactive coatings. J Mater Sci  Mater Med. 2002; 13(7): 657-665.

[8]  Gopi D, Indira J, Kavitha L, Sekar M, Mudali UK. Synthesis of hydroxyapatite nanoparticles by a novel ultrasonic assisted with mixed hollow sphere template method. Spectrochim Acta Part A. 2012; 93: 131-134.

[9]  Guo H, Khor KA, Boey YC, Miao X. Laminated and fun- ctionally graded hydroxyapatite/yttria stabilized tetragonal zirconia composites fabricated by spark plasma sintering. Biomaterials. 2003; 24(4): 667-675.

[10] Ho WF, Hsu HC, Hsu SK, Hung CW, Wu SC. Calcium phosphate bioceramics synthesized from eggshell powders through a solid state reaction. Ceram Int. 2013; 39(6): 6467-6473.

[11] Bahrololoom ME, Javidi M, Javadpour S, Ma J. Charac- terisation of natural hydroxyapatite extracted from bovine cortical bone ash. J Ceram Process. Res. 2009; 10: 129-138.

[12] Russias J, Saiz E, Nalla RK, Gryn K, Ritchie RO, Tomsia AP. Fabrication and mechanical properties of PLA/HA composites: a study of in vitro degradation. Mate Sci Eng C. 2006; 26(8): 1289-1295.

[13] Mohamadi Y, Mirzadeh H, Moztarzadeh F, Soleimani M, Jabbari E. Design and Fabrication of Biodegadable Porous Chitosan/Gelatin/Tricalcium Phosphate Hybrid Scaffolds for Tissue Engineering. Iran J Polym Sci Technol. (In Persion). 2007; 20: 297-308.

[14] Hsu FY, Tsai SW, Lan CW, Wang YJ. An in vivo study of a bone grafting material consisting of hydroxyapatite and reconstituted collagen. J Mater Sci  Mater Med. 2005; 16(4): 341-345.

[15] Zandi M, Mirzadeh H, Mayer C, Urch H, Eslaminejad MB, Bagheri F, Mivehchi H. Biocompatibility evaluation of nano rod hydroxyapatite/gelatin coated with nano HAp as a novel scaffold using mesenchymal stem cells. J  Biomed. Mater Res. Part A. 2010; 92(4): 1244-1255.

[16] Xu HH, Smith DT, Simon CG. Strong and bioactive composites containing nano-silica-fused whiskers for bone repair. Biomaterials. 2004; 25(19): 4615-4626.

[17] Khan Y, Yaszemski MJ, Mikos AG, Laurencin CT. Tissue engineering of bone: material and matrix considerations. J  Bone  Joint Surg. 2008; 90(Supplement 1): 36-42.

[18] Vincent J. Structural biomaterials. Third edition, Princeton University Press, USA. 2012; 84-86.

[19] LeGeros RZ, Lin S, Rohanizadeh R, Mijares D, LeGeros JP. Biphasic calcium phosphate bioceramics: preparation, properties and applications. J Mater Sci Mater Med. 2003; 14(3): 201-209.

[20] Murugan R, Ramakrishna S. Development of nano- composites for bone grafting. Compos  Sci Technol. 2005; 65(15): 2385-406.

[21] Chen L, Mccrate JM, Lee JC, Li H. The role of surface charge on the uptake and biocompatibility of hydroxyapatite nanoparticles with osteoblast cells. Nanotechnology. 2011; 22(10): 105708.

[22] Tanner KE. Bioactive ceramic-reinforced composites for bone augmentation. J R Soc Interface. 2010; 7(Suppl 5): S541-57.

[23] Wang M. Developing bioactive composite materials for tissue replacement. Biomaterials. 2003; 24(13): 2133-2151.

[24] Liu B, Yang C, Yan X, Wang J, Lv Y. Interaction of avelox with bovine serum albumin and effect of the coexistent drugs on the reaction. Int J Anal Chem. 2012; 2012.

 [25] Jamshidian M, Tehrany EA, Imran M, Jacquot M, Desobry S. Poly Lactic Acid: production, applications, nanocomposites, and release studies. Compr Rev Food Sci Food Saf. 2010; 9(5): 552-571.

[26] Rose JB, Pacelli S, Haj AJ, Dua HS, Hopkinson A, White LJ, Rose FR. Gelatin-based materials in ocular tissue engineering. Materials. 2014; 7(4): 3106-3135.

[27] Mohammadi Y, Mirzadeh H, Moztarzadeh F, Soleimani  M, Jabbari E. Osteogenic Differentiation of Mesenchymal Stem Cells on Novel Three-Dimensional Poly (L-Lactic Acid) /Chitosan/ Gelatin/ Beta- Tricalcium Phosphate Hybrid Scaffolds. Iran Polym J. 2007; 16(1): 57.

[28] Kumaraswamy G. Crystallization of polymers from stressed melts. J Macromol Sci Pol R. 2005; 45(4): 375-397.

[29] Chang MC, Ko CC, Douglas WH. Preparation of hydroxyapatite-gelatin nanocomposite. Biomaterials. 2003; 24(17): 2853-2862.

[30] Pradeesh TS, Sunny MC, Varma HK, Ramesh P. Preparation of microstructured hydroxyapatite microspheres using oil in water emulsions. Bull Mater Sci. 2005; 28(5): 383-390.

[31] Tsai SW, Hsu FY, Chen PL. Beads of collagen–nano- hydroxyapatite composites prepared by a biomimetic process and the effects of their surface texture on cellular behavior in MG63 osteoblast-like cells. Acta Biomater. 2008; 4(5): 1332-1341.

[32] Chang SH, Hsu YM, Wang YJ, Tsao YP, Tung KY, Wang TY. Fabrication of pre-determined shape of bone segment with collagen-hydroxyapatite scaffold and autogenous platelet-rich plasma. J Mater Sci  Mater Med. 2009; 20(1): 23-31.

[33] Kim HW, Yoon BH, Kim HE. Microsphere of apatite-gelatin nanocomposite as bone regenerative filler. J Mater Sci Mater Med. 2005; 16(12): 1105-1109.

[34] Sivakumar M, Manjubala I, Rao KP. Preparation, charact- erization and in-vitro release of gentamicin from coralline hydroxyapatite–chitosan composite microspheres. Carbohydr Polym. 2002; 49(3): 281-288.

[35] Sivakumar M, Rao KP. Preparation, characterization and in vitro release of gentamicin from coralline hydroxyapatite–gelatin composite microspheres. Biomaterials. 2002; 23(15): 3175-3181.

[36] Wei HJ, Yang HH, Chen CH, Lin WW, Chen SC, Lai PH, Chang Y, Sung HW. Gelatin microspheres encapsulated with a nonpeptide angiogenic agent, ginsenoside Rg 1, for intramyocardial injection in a rat model with infarcted myocardium. J Controlled Release. 2007; 120(1): 27-34.

[37] Bahrololoom ME, Javidi M, Javadpour S, Ma J. Charac- terisation of natural hydroxyapatite extracted from bovine cortical bone ash. J Ceram Process Res. 2009; 10: 129-138.

[38] Chang MC, Ko CC, Douglas WH. Preparation of hydrox- yapatite-gelatin nanocomposite. Biomaterials. 2003; 24(17): 2853-2862.

[39] Haberko K, Buæko MM, Brzeziñska-Miecznik J, Haberko M, Mozgawa W, Panz T, Pyda A, Zarêbski J. Natural hydroxyapatite its behaviour during heat treatment. J Eur Ceram Soc. 2006; 26(4): 537-542.