Hydroxyapatite nanocomposites for biological application J. Kumar 1, a

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1 Materials Science Forum Online: ISSN: , Vol. 760, pp doi: / Trans Tech Publications, Switzerland Hydroxyapatite nanocomposites for biological application J. Kumar 1, a 1 Crystal Growth Centre, Anna University, Chennai, India . a marsjk@gmail.com Keywords: Hydroxyapatite, Bioceramics, Drug delivery, Ciprofloxacin, Sodium alginate, Abstract: - Hydroxyapatite is a bioceramic which has a wide range of medical application for bone diseases. To enhance its usage, we have prepared ciprofloxacin loaded nano hydroxyapatite (HA) composite with a natural polymer, alginate, using wet chemical method at low temperature. The prepared composites were analyzed by various physicochemical methods. The results show that the nano HA crystallites are well intact with the alginate macromolecules. For the composite system FT-IR study is done. The drug is pre-adsorbed onto the ceramic particle before the formation of composite. The thermal behavior of composite has been studied using thermo gravimetric analysis (TGA). This paper communicated the Influence of Polymer concentration on the Controlled drug release from hydroxyapatite Nanocomposites Introduction: Drug delivery is an interesting interdisciplinary area which involves pharmaceutical, chemical engineering, biological, biomaterials and medical sciences. Calcium phosphate is a good biocompatible ceramic material which is used as a material for drug delivery for bone disorders. Hydroxyapatite can be simultaneously used as a local drug delivery system for various classes of drugs, growth factors and proteins. Nano hydroxyapatite is a bioceramic which has a wide range of medical application for bone diseases. To enhance its usage, we have prepared ciprofloxacin loaded Hydroxyapatite (HA) composite with a natural polymer, alginate, using wet chemical method at low temperature [1-4]. Ciprofloxacin (CPX), 1-cyclopropyl 6-fluro-1, 4-dihydro-4-oxo-7-(1- piperazinyl)-3-quinoline carbolitic acid is a broad spectrum fluroquinolone antibacterial agent used in the treatment of both Gram positive and Gram negative microorganisms. 11 It stops cell division by inhibiting DNA gyrase, a type II topoisomerase and topoisomerase IV. The minimum inhibition concentration (MIC) of CPX is µg/ml for pathogens like Staphylococcus aureus and Pseudomonas aeruginosa which cause osteomyelitis [5]. Biopolymer/ inorganic material composites are used as drug carrier for controlled drug delivery. They have unique structure and properties. Inorganic materials including clays, hydroxyapatite (HA), silica and carbon nanotubes are the different inorganic materials which have been used for the preparation of the composites. Biopolymer and inorganic material have a strong interfacial interaction between them via electrostatic interaction and hydrogen bonding. The mechanical properties, swelling behavior, drug loading efficiency and controlled release behavior are enhanced because of this synergistic effect [6]. In our previous paper we have reported the synthesis and charecterization of hydroxyapatite nanoparticles and ciprofloxacin loaded hydroxyapatite nanoparticles. In this paper we report the Influence of Polymer concentration on the Controlled drug release from hydroxyapatite Nanocomposites Experimental procedure: a) Synthesis of Ciprofloxacin loaded Hydroxyapatite-Alginate composite: Ciprofloxacin loaded Hydroxyapatite powder was dispersed in Na-alginate solution under gentle stirring until a homogeneous paste was obtained. The Ciprofloxacin loaded Hydroxyapatite powder was mixed with 1 to 3.0% w/v of sodium alginate solution at a weight ratio of All rights reserved. No part of contents of this paper may be reproduced or transmitted in any form or by any means without the written permission of Trans Tech Publications, (ID: , Pennsylvania State University, University Park, USA-06/03/16,23:53:20)

2 40 Functional Nanomaterials for Energy and Environmental Applications hydroxyapatite/alginate 40 wt%. The pastes were extruded drop wise into a 2 M CaCl2 cross linking solution, where spherical-shaped particles instantaneously formed and were allowed to harden for 30 min. At completion of the gelling period the microspheres were recovered and rinsed in water in order to remove the excess CaCl2. Finally, they were dried overnight in a vacuum oven at 30 C [7]. b) Drug release- in vitro study: In order to determine the drug release profile, 100 mg of the drug loaded hydroxyapatite and drug loaded hydroxyapatite with alginate coating were introduced into a screw capped glass bottle containing 50 ml of phosphate buffered saline (PBS) medium at 37 C and ph 7.4 under sterile condition. The drug release study was done for a period of 600 hours. 5 ml samples were withdrawn by a pipette and replaced immediately with 5 ml of fresh PBS medium, which was accounted for when calculating the amount released. Ciprofloxacin concentration in the collected samples was measured spectrophotometrically at a wavelength of 274 nm. Mean, standard deviation and one way analysis of variance (Anova) is carried out. Results and Discussions Drug loaded Polymer-Ceramic mixtures of different composition were prepared and drop wise extruded into a cross linking solution containing Ca 2+. The formation of spherical particles is due to the rapid establishment of calcium-mediated associations between polyguluronic acid sequences on the polymer backbone. Fig 1 shows the FT-IR spectrum of Ciprofloxacin-hydroxyapatite-alginate particles. The alginate did not induce subsequent modification in the hydroxyapatite structure. Bands corresponding to the presence of calcium/sodium alginate, namely at 2925 cm_1, 2169 cm _1, 1316 cm _1, 901 cm _ 1, 820 cm _1, 3445 cm _1 (υoh), 1629 and 1418 cm _1 (υcoo_), identified in the literature as the combination of three possible vibrational modes (τco+δcco+δcch) are observed. The broadening of the υ3po4 region ( cm _1 ) denotes the presence of the polymer. The presence of ciprofloxacin is confirmed by the appearance of bands corresponding to ciprofloxacin CH in plane bending at 1272, CN stretching at 869 cm_1, 1491 cm _1, 716 cm _1, 740 cm _1, 782 cm _1, 894 cm _1, 869 cm _1, 568 cm _1 [8]. Fig 1. FTIR spectrum of HAp-Cipro-Alg

3 Materials Science Forum Vol TGA curve of hydroxyapatite and with various percentage of polymer is shown in Fig 2. Naalginate decomposition starts at about 60 C with loss of water then the decomposition implies two weight losses: at 240 C and at 600 C. The ciprofloxacin loaded hydroxyapatite/alginate composite shows a different profile characterized by broad transitions, but the relative weight losses for each step are the same as those of pure alginate. In the ciprofloxacin loaded hydroxyapatite no distinct thermal decomposition can be observed at temperatures above ~100 C. The weight of ciprofloxacin loaded hydroxyapatite/alginate composite sample decreased gradually from room temperature to 200 C; in the range of C weight declined sharply and then decreased slightly from 300 to 600 C; after 600 C, the sample lost the weight rapidly. The weight loss below 200 C could be ascribed to the water evaporation and decomposition of oligosaccharide, while the rapid weight loss upon 200 C involved the complex process of degradation of polysaccharides. As the polymer concentration increases the attachment of alginate to Hap nanoparticles increases. Fig 2. TGA analysis of HAp-Cipro-Alg The drug release profile is given in Fig 3. The percentage of drug released in 600 hours were estimated to be 85 %, 88, 92% from 3 %, 2 %, 1 % alginate coated samples and and % from Hap-drug sample respectively. There is a sudden outburst of drug in the initial few hours and then the drug is released in a controlled manner. The drug release percentage decreases with increase in the alginate concentration from 1% to 3%. The drug release reduces with increase in the polymer concentration. 100 Drug release(%) % alginate 2% alginate 1% alginate H A p-c iprofloxacin3% alginate T im300 e (hours) Fig 3. Drug Release Profile

4 42 Functional Nanomaterials for Energy and Environmental Applications Conclusion: The synthesis and characterization of nano hydroxyapatite, to be used as drug delivery system as bone replacement material was done in this present work. The drug is coated on HAp nanoparticles before alginate encapsulation. FTIR analysis shows the appearance of bands corresponding to HAP, ciprofloxacin and alginate. TGA analysis shows the increase in alginate attachment with increase in alginate concentration. The increase in the polymer concentration delays the release of ciprofloxacin. Reference: [1] J. Guicheux, G. Grimandi, M. Trecant, A. Faivre, S. Takahashi and G. Daculsi, Apatite as carrier for growth hormone: in-vitro characterization of loading and release. J. Biomed Mater Res.34, (1997), pp 165. [2] P.K. Bajpai and H.A. Benghuzzi, Ceramic systems for long-term delivery of chemicals and biologicals. J. Biomed Mater Res. 22, (1988), pp [3] Donncha Harverty, Syed A.M Tofail, T. Kenneth, Stanton and James B. McMonagle, Structure and Stability of Hydroxyapatite: Density functional calculation and Rietveld analysis. Physical Review. 71, (2005),pp 1-9. [4] M. Bohner, Physical and Chemical aspects of calcium phosphates used in spinal surgery. Eur Spine J. 10, (2001), pp114. [5] S.A Patel, N.M Patel and M.M Patel, Simultaneous spectrophotometric estimation of ciprofloxacin and ornidazole in tablets. Indian journal of Pharmaceutical Sciences. 68, (2006), pp 665. [6] J. Zhang, Q. Wanga and A. Wang, In situ generation of sodium alginate/hydroxyapatite nanocomposite beads as drug-controlled release matrices. Acta Biomaterialia.6, (2010), pp.445. [7] T. Yoshioka, T. Ikoma, A. Monkawa, S. Yunoki, T. Abe, M. Sakane and J. Tanaka, Preparation of hydroxyapatite-alginate gels as a carrier for controlled release of Paclitaxel. Key engineering materials. 330, (2007), pp1053. [8] B. Dupuy, A. Arien and A.P. Minnot, FT-IR membranes made with alginate/polylysin complexes. Variations with mannuronic or guluronic content of the polysaccharides. Art Cells, Blood Subs. Immob Biotech. 20 (1994) pp,71.

5 Functional Nanomaterials for Energy and Environmental Applications / Hydroxyapatite Nanocomposites for Biological Application /

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