Journal

J. Am. Ceram. Soc., 92 [7] 15921595 (2009) DOI: 10.1111/j.1551-2916.2009.03093.x r 2009 The American Ceramic Society

Synthesis and Structure Refinement of Zinc-Doped b-Tricalcium Phosphate Powders

Sanjeevi Kannan,z Friedlinde Goetz-Neunhoeffer,y Jurgen Neubauer,y and Jose M.F. Ferreiraw,z
z

Department of Ceramics and Glass Engineering, University of Aveiro, CICECO, 3810-193 Aveiro, Portugal
y

GeoZentrum Nordbayern, Mineralogy, University of Erlangen-Nuremberg, 91054 Erlangen, Germany

Zinc substituted b-tricalcium phosphate [b-Ca3(PO4)2] was formed by substituting a zinc precursor in calcium-decient apatite through aqueous precipitation technique. Heat treatment at 10001C led to the formation of well crystalline b-Ca3(PO4). Renement technique was used to determine the inuence of incorporated zinc in the b-Ca3(PO4) structure. The structural data for all the four different zinc substituted b-Ca3(PO4) ranging from 09 mol% of zinc investigated in the present study conrmed the rhombohedral structure of b-Ca3(PO4) in the hexagonal setting (space group R3c). The incorporation of lower sized Zn21 (0.745 A for sixfold coordination with O) at the higher sized Ca21 (1.00 A for sixfold coordination with O) site in the bCa3(PO4) structure led to the contraction of unit cell parameters. The added zinc prefers to occupy the Ca(5) site of bCa3(PO4) structure.

I. Introduction
b-TRICALCIUM phosphate [b-TCP, b-Ca3(PO4)2] is a well known potential bone substitute due to its favorable features like biocompatibility, osteoinductivity and its safety in living tissues.1,2 In recent years, ionic substitutions in b-Ca3(PO4)2 ceramics both in the form of cationic (Na1, Mg21, Sr21, Zn21) and anionic (SiO4) have been the subject of immense interest owing to the 4 critical roles of these ions in the biological process upon implantation.36 Zinc as an essential trace element has been detected in all the human tissues, with bone containing the major part of total body zinc.7 Zn content ranges from 0.0126 to 0.0217 wt% in human bone and is relatively higher than the average Zn content of whole fat-free adult tissues (0.0030 wt%) and that of plasma (1217 Zn mM).811 The role of zinc has been well known as it actively participates in the: (i) bone regeneration process11; (ii) improvement of osteoblasts activity, playing an essential role12; (iii) enzymatic activity13; (iv) nucleic acid metabolism14; and (v) protein synthesis.15 Owing to these favorable features, studies involving the development of zinc substituted bioceramics have been the subject of extensive investigation for the past 2 years as suggested by recent literature reports.1619 Among zinc-doped bioceramics, zinc containing b-TCP is considered a promising bone repair material due to the expected signicant role of this trace element in enhancing the osteoblast activity and promoting bone generation. The synthesis of zinc containing b-TCP could be obtained both through solid-state reaction and wet chemical synthesis. However, the literature review has revealed the lack of sufcient structural information on the inuence of zinc in b-Ca3(PO4)2. A recent
T. Troczynskicontributing editor

study on the neutron diffraction studies of zinc and silicon cosubstituted b-Ca3(PO4)2 was reported.20 Given the trace level of zinc presence in human bone, a more reliable information about the inuence of zinc on the structure of b-Ca3(PO4)2 could be obtained while dealing with the studies of zinc-doped b-Ca3(PO4)2 powders in a wider Zn concentration range. The present study is an attempt on the synthesis of zinc-doped b-Ca3(PO4)2 through wet precipitation technique and to gain structural details on the zinc inuence in b-Ca3(PO4)2 through Rietveld renement technique. It has to be mentioned that Rietveld method is a standard technique for the detailed analysis of crystallographic structures. The synthesis procedure to form divalent ion substituted b-Ca3(PO4)2 through aqueous precipitation technique has been documented by the authors from their previous work that involves the formation of Mg-doped b-Ca3(PO4)221 and Sr-doped b-Ca3(PO4)2.22 A similar procedure has been adopted in the present study to form Zn-doped b-Ca3(PO4)2 by introducing a zinc precursor in the calcium decient apatite and thereby heating the ionic-doped calcium decient apatite at temperatures ! 8001C to obtain zinc-doped b-Ca3(PO4)2. Four different compositions of zinc substituted b-Ca3(PO4)2, with the zinc content ranging from 09 mol% were synthesized and taken for the renement study. The results were compared with single phase b-Ca3(PO4)2 obtained from the same method.

II. Experimental Procedure (1) Synthesis Four different zinc substituted b-Ca3(PO4)2 compositions were prepared through aqueous precipitation technique. In brief, calcium decient apatites were synthesized via a wet chemical route according to predetermined Ca/P molar ratios reported in Table I by the slow addition of (NH4)2HPO4 solution to the continuously stirred (1000 rpm) solution of Ca(NO3)2 4H2O. Table I also details the concentrations of the precursors used for the synthesis and also the elemental analysis of zinc contents in the heat-treated powders determined through inductively coupled plasma (ICP) technique. A predetermined concentration of Zn(NO3)2 6H2O was added slowly to the above solution containing calcium and phosphorous ions. The pH of the mixed solution/suspension was then increased to 7.4 and maintained at this value by adding the required amounts of 8M ammonium hydroxide (NH4OH) solution. After the completion of addition, the reaction was performed at 901C for 2 h under constant stirring conditions (1000 rpm). In parallel, a calcium decient apatite of Ca/P molar ratio of 1.5 was prepared without added zinc precursor under the same conditions to compare the results. The precipitated suspensions were poured out from the reactor and the precipitates were separated through vacuum ltration and dried at 801C overnight. The dried cakes were ground to ne powders, sieved through a mesh size of 30 mm and used for Rietveld renement studies. Infrared spectra for the powders were obtained using an infrared Fourier Spectrometer (FT-IR, model 1592

Manuscript No. 25587. Received December 3, 2008; approved March 17, 2009. This work was supported by the Portuguese Foundation for Science and Technology from the project POCTI/CTM/60207/2004 and nancially supported by GRICES-DAAD. w Author to whom correspondence should be addressed. e-mail: jmf@ua.pt

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Synthesis and Structure Renement of Zinc-Doped b-TCP

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Table I. Molar Concentrations of the Precursors Ca, P, and Zn Used in the Synthesis and Results from ICP Analysis
Molar concentration of the precursors Sample code Ca P Zn Ca/P ratio (Ca1Zn)/P ratio Mol% of zinc Experimentally planned From ICP analysis

TCP Zn-TCP1 Zn-TCP2 Zn-TCP3 Zn-TCP4

0.900 0.880 0.860 0.840 0.820

0.600 0.600 0.600 0.600 0.600

0.000 0.020 0.040 0.060 0.080

1.500 1.466 1.433 1.400 1.366

1.50 1.50 1.50 1.50 1.50

0.00 2.22 4.44 6.66 8.88 III. Results and Discussion

2.43 4.53 6.59 8.78

Mattson Galaxy S-7000). For this purpose each powder was mixed with KBr in the proportion of 1/150 (by weight) for 15 min and pressed into a pellet using a hand press.

(2) Structure Renement For renement studies, all the powders were heat treated in a Thermolab furnace (Pt30%Rh/Pt6%Rh-thermocouple) with a heating rate of 51/min to achieve a predetermined temperature of 10001C, followed by a dwelling time of 2 h, and then cooled to room temperature at the rate of 51/min. Although low crystalline b-Ca3(PO4)2 forms at about 8001C,21,22 a higher heat treatment temperature of 10001C was required to obtain the desired well-crystallized phase for Rietveld renement. Collection of Xray powder diffraction (XRPD) pattern data was performed with a Siemens D5000 (Karlsruhe, Germany) equipped with a diffracted beam graphite monochromator. The software TOPAS 3 (Bruker AXS, Karlsruhe, Germany) with the fundamental parameters approach was used for Rietveld renements. All powders were prepared and measured three times independently. Rietveld renement was performed using the structural model of ICSD card number of # 619123 for b-TCP. Rened parameters were scale factor, specimen displacement, background as Chebyshev polynomial of fth grade, crystallite size, microstrain, lattice parameters, and occupancy factors. As the scattering factor for P51 is not implemented in the TOPAS 3 Rietveld software, the P sites were rened with a combination of Si41 and P for pure b-Ca3(PO4)2 phase. This leads to an improved renement of Ca21 occupancy and of the zinc substitution at Ca21 sites. Zinc content in the heat-treated powders was analyzed by the ICP technique.

All the investigated compositions have led to the formation of well-dened crystalline b-Ca3(PO4)2 phase after heat treatment at 10001C. No signicant differences in the phase behavior could be noticed in the X-ray diffraction patterns. The formation of zinc substituted b-Ca3(PO4)2 has been evidenced according to Ca9x Znx HPO4 PO4 5 OH 1000 C 3 Ca3x=3 Znx=3 PO4 2 H2 O ! (1)

These results were found to be consistent with the previous work on the Mg-doped b-Ca3(PO4)221 and Sr-doped bCa3(PO4)222 bioceramics reported by the authors. The elemental analysis data for zinc presented in Table I conrmed to have good coincidence with the experimental planned values, which enables to infer that the zinc added during the synthesis has been incorporated in the structural lattice of b-Ca3(PO4)2 bioceramic. The renement of XRPD data for b-Ca3(PO4)2 without zinc substitution was performed assuming the space group R3c. The unit cell and structural parameters reported by Dickens et al.23 were used as starting values in the renement. Rened data for b-Ca3(PO4)2 conrmed that crystalline b-TCP has been formed with unit cell parameters a 5 b 5 10.4380(4) A, c 5 37.4016(17) A, a 5 b 5 901, and g 5 1201 in the hexagonal setting (space group R3c). The rened lattice parameter values had close similarities with the values reported in the literature.23,24 Using the rened occupancy factors, the Ca/P ratio was determined to be 1.51(1) in agreement with the value of 1.5 calculated from the chemical formula of b-Ca3(PO4)2. In accordance with the previous reports,23 the rened occupancy factor for the Ca(4) site

1.000 900 800 700 600 Counts 500 400 300 200 100 0 100 5 10 15 20 25 30 35 40 2 Theta Degree Cu Kalpha 45 50

Whitlockite 100.00 %

55

60

65

Fig. 1. Rietveld analysis pattern of powder diffraction data of ZnTCP1 (2.43 mol% zinc). The solid lines are calculated intensities and the marked ones are observed intensities. The difference between the observed and calculated intensities is plotted below the prole.

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Journal of the American Ceramic SocietyKannan et al.

3540 3525 Unit Cell Volume( ) 3510 3495 3480 3465 3450 3435 0 2 4 6 Mole % of Zinc

Vol. 92, No. 7

showed partial occupancy of 0.55(4), due to larger Ca(4)O distances with weak bonding between oxygen and Ca(4) atoms along c-axis. The rened values of pure b-Ca3(PO4)2 obtained from the present investigation were taken as the starting values to rene zinc containing b-Ca3(PO4)2. The rened XRPD pattern for Zn-TCP1 presented in Fig. 1 conrms the single-phase b-Ca3(PO4)2 with no other additional phases detected in the pattern. In fact, all the zinc containing b-Ca3(PO4)2 compositions investigated in the present study have conrmed the formation of crystalline b-Ca3(PO4)2 phase. The structural data obtained after renement for the different zinc containing b-TCP powders are presented in Figs. 2 and 3. The structural data for ZnTCP1 Ca2.93Zn0.07 (PO4)2 after renement showed b-Ca3(PO4)2-structure with unit cell parameters a 5 b 5 10.415 (3) A, c 5 37.3517 (13) A, a 5 b 5 901, and g 5 1201 in the hexagonal setting (space group R3c). All the Zn-substituted bCa3(PO4)2 investigated in the present study were also conrmed to retain the rhombohedral structure. It can be easily noticed from the rened parameters of zinc substituted b-Ca3(PO4)2 that both a and c unit cell parameters (Fig. 2) have indicated a decreasing trend thereby leading to the observed contraction in the unit cell volume (Fig. 3) when compared with the pure bCa3(PO4)2. This decreasing trend in the lattice data can be ascribed to the incorporation of lower ionic radii of Zn21 (0.745 A for sixfold coordination with O) at Ca21 (1.00 A for sixfold coordination with O) sites in the b-TCP structure. The decreasing trend of both lattice parameters a and c for zinc containing b-Ca3(PO4)2, has been found consistent with the previously reported results on Mg-containing b-Ca3(PO4)2 ceramics, which have also shown a decreasing trend in the c-axis parameter till a similar concentration limit of Mg.25 The preferential distribution of zinc on the ve crystallographical independent Ca21 sites of b-Ca3(PO4)2 was analyzed by rening the occupancy sites of all Ca21 positions. All the ve Ca21 sites were rened simultaneously by including Zn21 ion at each Ca21 site. Renement of the ve different Ca21 sites has conrmed that no zinc was detected on Ca(1), Ca(2), Ca(3), and Ca(4) sites of b-Ca3(PO4)2 structure. The rened occupancy values show that the accommodation of Zn21 in the sixfold coordinated Ca(5) site of b-Ca3(PO4)2 structure is favored. The rened occupancy factors for the Ca(5) site of four different zinc containing b-Ca3(PO4)2 have been represented graphically in Fig. 4. A gradual increase in the occupancy factors with the increasing concentration of zinc could be well seen from Fig. 4. The reason for the preferred occupancy of Zn21 at the Ca(5) site could be explained as follows. The accommodation of a foreign ion into the atomic site purely depends on the prevailing conditions in the crystallographic site and also on the nature and

Unit Cell Volume

10

Fig. 3. Inuence of unit cell volume with respect to the different contents of zinc in b-tricalcium phosphate structure.

size of the ion. In the present investigation, given the concentrations of investigated Zn21, which possesses lower ionic radii when compared with the Ca21 ion, it has been found that Zn21 preferentially occupies at the Ca(5) site. This is based on the fact that the shortest mean distances (2.238 and 2.287 A) in the structure were determined23 for the Ca(5)O bonds and hence this Ca(5) site then can easily accommodate the Zn21 ions. FTIR spectra for all the zinc-doped b-Ca3(PO4)2 presented in Fig. 5 have conrmed the formation of crystalline phase. No broad bands corresponding to the amorphous content in the samples have been identied. The infra red absorption bands located at 1132, 1030, 990, 960, 610, and 560 cm1 contribute for the conrmation of PO4 tetrahedra of b-Ca3(PO4)2 structure. However, with the increasing zinc concentration in solid solution, the degeneracy of the absorption bands increases as it is evident from Fig. 5 and this fact also echoes with the view of Bigi et al.5 Additionally, no bands for the presence of hydroxyl group (OH) could be witnessed in the FT-IR spectrum reveals that the transformation of calcium decient apatite to b-Ca3(PO4)2 has been completed.

4. Conclusions
A simple aqueous precipitation technique has been used to form zinc substituted b-Ca3(PO4)2 by heating calcium decient apatite. The transformation mechanism that occurred at 8001C to form low crystallinity b-Ca3(PO4)2 with substituted zinc is similar to that of calcium decient apatite of Ca/P molar ratio 5 1.5 to form pure b-Ca3(PO4)2 without any substituted zinc. Wellcrystalline b-Ca3(PO4)2 phase has been detected at 10001C for all the compositions investigated in the present study. Elemental
1.00

10.455 10.440 a-axis Lattice Parameter a () 10.425 10.410 10.395 10.380 10.365 10.350 10.335 0 2 4 6 Mole % of Zinc 8 10 c-axis

37.425 37.400 37.375 Lattice Parameter c () 37.350 37.325 37.300 37.275 37.250 37.225 37.200 37.175 37.150 37.125

Occupancy at Ca(5) site

0.80 0.60 0.40 0.20 0.00 0.20 0 2 4 6 Mole % of Zinc 8 10

Fig. 2. Inuence of lattice a- and c-axis parameters with respect to the different contents of zinc in b-tricalcium phosphate structure.

Fig. 4. Graphical representation of the zinc occupancy at the Ca(5) site of b-tricalcium phosphate structure.

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Synthesis and Structure Renement of Zinc-Doped b-TCP

5

1595

400

800

1200

1600

Wave number (cm1)

Fig. 5. Fourier transmitted infrared spectra for the different zinc-doped b-tricalcium phosphate (b-TCP) after heat treatment at 10001C.

analyses of zinc made for all the powders after heat treatment at 10001C have shown a good coincidence with the experimental values. Rietveld renement of the XRPD data have conrmed linear decrease in the lattice parameters a and c with the increased concentration of zinc substitution in the b-Ca3(PO4)2 structure. Renement of ve different Ca21 sites of b-Ca3(PO4)2, has proven that the substituted zinc prefers to occupy the Ca(5) site. Thus the quantitative data on zinc substituted b-Ca3(PO4)2 from the present study will be of immense value in terms of low zinc contents detected in the natural bone.811

Acknowledgments
Thanks are due to the fellowship grants of S. Kannan (SFRH/BPD/18737/ 2004). The authors thank Daniele Lutz for performing the ICP analysis.

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Transmittance (a.u)