Nanoceramics composite biomaterials in the calcium carbonate hydroxyapatite system Margarita Goldberg 1,a, Valeriy Smirnov 1,b, Sergey

Kucev1,a, Natalia Sergeeva2,c, Irina Sviridova2,c, Valentina Kirsanova2,c, Syraia Akhmedova 2,c, Sergey Barinov1,a
1

Institution of Russian Academy of Sciences A.A. Baikov Institute of Metallurgy and Material Science RAS, Leninskii pr. 49, Moscow, Russia
2

Moscow Herzen Research Oncological Institute, 2-i Botkinskii pr. 3, Moscow, Russia
a

naiv.syper@gmail.com, bsmirnov2007@mail.ru, cprognoz.06@mail.ru

Keywords: Bio-materials, Hydroxyapatite, Calcium carbonate

Abstract Composite powders in the calcium carbonate (CC) hydroxyapatite (HA) system with carbonate content up to 50 wt. % were synthesized by the precipitation from aqueous solution and characterized in detail. Specific area of the powders decreases with an increase in carbonate content due to appearance of large elongated CC crystals of about 0,5 micrometer size. The use of highly active to sintering nanodisperse powders resulted in lowering of sintering temperature below 720 0C, preventing thermal decomposition. Ceramics of about 20 wt.% carbonate content revealed the highest strength of about 77 MPa, having the grain size of about 100 nm. According to in vitro testing, the materials are non toxic for human fibroblast cells. Introduction New medical engineering processes for repair of human bone tissue damaged upon diseases or injury are based on implanting porous scaffolds made of biocompatible materials with cultivated osteoforming cells into the defect site [13]. With time, this structure is replaced by bone tissue formed de novo. Apart from biocompatibility with the body, the requirements to the scaffold material also include time coherence between the resorption and the formation of a new bone tissue. Materials based on calcium phosphates, especially hydroxyapatite (HA), which is analogous in the chemical and phase composition to the mineral component of bone tissue, are best suited for this purpose. However, HA is the calcium phosphate most resistant to resorption. In order to achieve the optimal characteristics of resorption, it was suggested to create composite materials based on the hydroxyapatite with the second more soluble phase a calcium carbonate. Experimental and Results According to reaction (1), materials containing 20, 40, 50, 70 and 80 wt. % of a calcium carbonate, were synthesized by the precipitation from aqueous solution.

(10y+x)Ca(O)2+6y(NH4)2PO4+x(NH4)CO3yCa10(PO4)6(OH)2+xCaCO3+ (12y+x)NH4OH+(6y+x)H2O

(1)

According to powder X-ray diffraction data (Shimadzu XRD-6000), the dried powders had a low degree of crystallization and mainly contained several phases: CC and HA. As the content of carbonate groups increased, the degree of crystallization increased. (Fig. 1, a). After powders calcinating at temperature 600 for an hour in air, the main crystallized phases were CC and HA (Fig. 1, b). Powders 3, 4 and 5 contain a minor amount of calcium oxide (O in Fig. 1b, up to 5 wt %) formed by thermal decomposition by heating in air.

Fig.1 XRD pattern of as synthesized (a) and calcinated (b) powders, A hydroxyapatite, C CaCO3, O CaO. Fourier transform IR spectroscopy was carried out on Avatar, as the samples there were powders in KBr. The IR spectra of all powders exhibit absorption bands for PO43-, OH- and CO32- groups (Fig.2) [4,5]. Compared to the intensity of the bands of phosphate groups, the intensity of the bands of CO32- close to 1300-1650 (mode 3) and 873-877 (2) m-1, that present in spectra and especially the band at 714 m-1 of CC increased [5]. Fig.2. Fo ur ier t r ans fo r m infr ar ed spect r a of as synthesized powders,: * -, ^ , # 43-, 32- of a3, + 32- carbonated hydroxyapatite.

For definition of materials thermal stability, differential scanning calorimetry (DSC), thermogravimetry (STA 409 Luxx) and mass-spectroscopy (quadrupole mass spectrometer Alos) have been made. According to mass-spectroscopy data, extraction by thermal decomposition in air

gas had molecular weight 44 g/mol that corresponds 2. According to DSC and thermogravimetry data the mass loss for materials with low content of CC started at 684-697 C (materials 1 and 2) and there were 6,89-7,27 wt. %. With the increase of CC content, the peak shifted to the higher temperature and mass loss for material 3 started at 724 C 18,62 wt. %, for materials 4 and 5 the mass loss was 29,76 and 35,85 wt.% began at 756,1 and 766C respectively. The quantity of carbon removed from carbonate hydroxyapatite and calcium carbonate by calcination with air present has been measured by Multiphase Carbon/Hydrogen/Moisture Determinator Leco RC-412. Also, this method allowed determining the content of CO32-groups in phases. The thermal decomposition in oxidizing atmosphere showed 3 peaks of carbon extraction. Supposedly, appearance of first peak was connected with adsorption of carbon dioxide from the environment. Two other peaks belong to decomposition of CaCO3 (temperature range about 680720 C) and carbonate hydroxyapatite (CHA) (temperature range about 800-880 C) which was formed as solid solution between CC and HA. This analysis allows measuring of total content of 32- and it was shown that it was close to design value (table 1). Table.1 Composition and properties of powders Parameter Specified content of carbonate groups [wt %] Content of carbone, Ctotal [ wt %] (RC-412) Content of carbonate groups,CO3 [wt %] Content of carbone in CaCO3 [wt %] Content of carbone in CHA [wt %] Specific surface area [m2/g]
2-

Powder 1 12 2,5 12,5 2,1 0,4 149 2 24 3,1 15,5 2,2 0,7 146 3 30 5,8 29,0 5,1 0,5 110 4 42 8,2 41,0 8,1 0,1 33 5 48 10,1 50,5 9,5 0,4 32,8

According to scanning electron microscopy data (LEO 1420), the noncalcinated powders were composed of aggregates with ~3-5 m size formed by particles with average sizes 30-50 nm. Round shape particles predominated in materials 1, 2 and 3, and large rod-like crystals with about 500 nm size of C appeared in materials 4 and 5. With increase of CC content, specific surface area of powders decreased from 149 to 32,8 m2/g (Table 1). The sintering of composite materials CC-HA was carried out with a part of liquid phase, which was formed by melting of additives containing carbonate ions. Also, these additives stabilized phase composition in the sintering process by the increase of carbonate-ions concentration in liquid phase, what prevented decomposition of CC at temperature up to 600 0. With the increase of sintering temperature, the strength of sintered samples rises, the highest bending strength (Instron 5581) was reached in material 1 and amounted to 77 MPa at 7200 due to decrease of open pores content (dense ceramics with porosity lower than 1% were obtained) and forming of uniform microstructure with average grain diameter ~100-200 nm. According to SEM data with increase of CC content in composite materials the HA grain shape changed from hexagonal to round, size of CC prismatic crystals increased. Average grain diameter

for material 1 was ~100-200 nm and increased to 500 nm for material 5. Solubility was measured at temperature 370 by calcium ion-selective electrode (Econix-Expert Expert-001) in deionized water and isotonic solution for injection (0,9 wt. % NaCl solution) on composites, sintered at 680 0. The Ca2+ ratio in deionized water changed from 8 to 12 mg/l with increase of CC content and in isotonic solution from 10 to 12 mg/l respectively. The growing of solubility in isotonic solution was corresponding to [6]. According to in vitro tests, all analyzed samples were non toxic for cells and support their adhesion, spreading, and proliferation of human fibroblasts. Conclusions Composite powders in the calcium carbonate hydroxyapatite system, containing 12 50 wt. % of CO32- groups were obtained by precipitation from aqueous solution. The specific surface area was reached 150 m2/g. Due to nanodispersion of powders and low-melting additives the sintering temperature was decreased to 7200 and intensive thermal decomposition of composite materials was prevented. There were obtained nanoceramics with grain sizes from 100 to 500 nm and bending strength value of 77 MPa depending on the composite composition. The developed composite materials are perspective for bone tissue engineering due to them solubility and absence of toxic reaction. Acknowledgment This work was supported by the Russian Foundation for Basic Research (project no. 0808 00224a). References [1] [2] Barinov S.M., Komlev V.S. Bioceramics based on calcium phosphates. M: Science. 2005. LeGeros R., Lin S., Rohanizaden R., Mijares E.D., LeGeros J.P. Biphase xcalcium phosphate bioceramics: Preparation, properties and applications // J. Mater. Sci. Mater. Med. 2003, v. 14, p. 201-209. A.G. Veresov, V.I. Putlyaev, Y.D. Tret'yakov Advances in Calcium Phosphate Biomaterials Ross. Khim. Zh, 2000 Rehman I., Bonfield W. Characterisation of hydroxyapatite and carbonated apatite by photo acoustic FTIR spectroscopy // J. Mater. Sci. Mater. Med. 1997, v. 8, p. 1-4.

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