Bioactive composite bone cement based on a-tricalcium phosphate/ tricalcium silicate

Loreley Morejon-Alonso,1 Oscar Jacinto Bareiro Ferreira,1 Raul Garcia Carrodeguas,2 Luis Alberto dos Santos1
1 2

Escola de Engenharia, Departamento de Materiais, Universidade Federal do Rio Grande do Sul, Porto Alegre (RS), Brasil Departamento de Ceramica, Instituto de Ceramica y Vidrio - CSIC, Madrid, Spain

Received 7 January 2011; revised 13 May 2011; accepted 25 May 2011 Published online 17 October 2011 in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/jbm.b.31926 Abstract: Silicon compounds are known as bioactive materials that are able to bond to the living bone tissue by inducing an osteogenic response through the stimulation and activation of osteoblasts. To improve the bioactive and mechanical properties of an a-Ca3PO4-based cement, the effects of the addition of Ca3SiO5 (C3S) on physical, chemical, mechanical, and biological properties after soaking in simulated body uid (SBF) were studied. The morphological and structural changes of the material during immersion were analyzed by X-ray diffraction and scanning electron microscopy. The results showed that it is possible to increase the compressive strength of the cement by adding 5% of C3S. Higher C3S contents enhance bioactivity and biocompatibility by the formation of a dense and homogeneous hydroxyapatite layer within 7 days; however, compressive strength decreases drastically as a consequence of delayed hydrolysis of a-Ca3(PO4)2. An increment in setting times and degradation rate of composites containing C3S was C also observed. V 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part
B: Appl Biomater 100B: 94102, 2012.

Key Words: calcium phosphates cements, hydroxyapatite, tricalcium silicate, in vitro

How to cite this article: Morejon-Alonso L, Bareiro Ferreira OJ, Garca Carrodeguas R, Santos LA. 2012. Bioactive composite bone cement based on a-tricalcium phosphate/tricalcium silicate. J Biomed Mater Res Part B 2012:100B:94102.

INTRODUCTION

Calcium phosphate cements (CPCs) represent a clinical alternative to the use of traditional bioceramics as they can be easily manipulated and shaped, they provide intimate adaptation to the contours of defect surfaces, and set in situ in the bone cavity to form a solid restoration.1 They also have attracted great interest since they were developed in the mid 80s owing to their chemical similarity to the mineral phase of bone tissue and good osteoconductivity.2 One of the most important formulations is that based on a-tricalcium phosphate [a-Ca3(PO4)2; a-TCP] which sets in situ and forms a calcium decient hydroxyapatite [Ca9(HPO4)(PO4)5(OH); CDHA] when hydrated.3 However, it is only strong enough under compression4 and has low mechanical strength when compared with cortical bone5; consequently, its application is restricted to places that require low mechanical loads.6 Being aware of the excellent bioresorbability of CDHA, researchers are concentrating their efforts in trying to overcome the mechanical weakness of calcium phosphates cements by using different llers, bers, and reinforcing additives that lead to the formation of various multiphase composites based on the idea that a ller in the matrix might stop crack propagation.7 Nevertheless, adding llers

affects the capacity to allow bone ingrowths into pores and produces a denser cement with a slower resorption rate and hence a slower bone substitution.8 Therefore, it is difcult to increase the strength of cements without having a negative impact on the other properties. Taking into account that silicon plays an important role in bone formation9-11 and its presence improves the bioactivity of materials by enhancing mesenchymal cell differentiation and increasing osteoblasts activity12,13; the addition of calcium silicates could be an effective way to improve the biocompatibility and bioactivity of calcium phosphates cements. On the other hand, since silicates have spontaneous development of strength towards water14 (spontaneous consolidation), the introduction of these additives in traditional formulations, could also enhance the mechanical properties of these materials. Due to the rapid development of strength during the rst stages of hydration and the higher content of silicon with respect to Ca2SiO4, dicalcium silicate (C2S), Ca3SiO5 (C3S) was chosen as additive. Different formulations of a-TCP/Ca3SiO5 were prepared and the effects of C3S addition on setting times, chemical composition, bioactivity, and mechanical properties before, during, and after ageing in simulated body uid (SBF) were studied. Cytotocixity and degradability were also determined.

Correspondence to: L. Morejon-Alonso; e-mail: loreley.morejon@ufrgs.br Contract grant sponsors: Coordenac o de Aperfeic a oamento de Pessoal de Nvel Superior (CAPES), Brasil

94

C V 2011 WILEY PERIODICALS, INC.

ORIGINAL RESEARCH REPORT

MATERIALS AND METHODS

Materials To prepare the a-TCP/C3S cement, all chemicals of analytical grade were used. a-TCP was prepared as described by Monma et al.15 using c-Ca2P2O7 and CaCO3 as raw materials in stoichiometric amounts. After calcination, the product was wet milled for 4 h in a polyethylene jar with alumina balls using an alcoholic medium (anhydrous ethanol). Tricalcium silicate powders were synthesized by solgel route, using Ca(NO3)2.4H2O and Si(OC2H5)4 (TEOS).16 Repeated grindings and calcinations at 1400 C were necessary to reach the product as monophasic as possible. To obtain powders with similar particle size distributions, the same milled treatment was made that in the case of a-TCP. Preparation of composite samples Synthesized C3S (7.11 lm, average diameter) was mixed with a-TCP (10.71 lm, average diameter) in powder ratios of 0, 5.0, and 10.0 mass%. The liquid phase was a sodium phosphate buffer prepared from NaH2PO4 and Na2HPO4.12H2O, and the liquid/powder ratios (L/P) was dependent of the content of C3S added ranging from 0.4 to 0.44 mL/g. Each powder sample was carefully weighed and mixed with the liquid phase in appropriate liquid-to-powder ratio, packed into silicon molds and aged at 36.5 C with controlled humidity for 24 h. Setting time measurement Setting time of samples was measured according to ASTM C266-89 using a Gillmore Needles method.17 Three specimens for each formulation were performed and standard deviation was used as a measure of the standard uncertainty. Initial setting time was determined as the end of moldability without serious damage to the cement structure, and the nal setting as the time beyond which it is possible to touch the cement without causing serious damage.18 In vitro tests To assess in vitro bioactivity, the 24 h set pastes were soaking in SBF at 36.5 C19,20 for 14 days and afterward, gently rinsed with deionized water followed by ethanol dehydration and drying in atmospheric temperature. For degradation tests, the disks were accurately weighed before and after immersion in SBF. The weight loss (WL) was calculated according to Eq. (1), being W0 the initial weight of the specimen and Wd the weight of the specimen dried after different degradation times (7, 14, and 21 days). Each measurement was performed three times and the average value was calculated. WL% W0 Wd =W0 100 (1)

FIGURE 1. Initial setting and nal setting time of the paste samples with various contents of C3S [L/P) 0.4 mL/g].

and negative controls and the number of viable cells was quantitatively assessed by MTT test. Experimental values were analyzed via one-way ANOVA test follow by Tukeys multiple comparison test. Characterization techniques Phase composition of the samples was determined by X-ray diffraction (XRD) in a PHILLIPSV diffractometer (XPert MPD) and Cu-target. Diffractograms were recorded using Ni-ltered radiation (k 1.5406 ) with a step size of 0.05 and a time/step ratio of 1 s. Morphological differences before and after soaking in SBF were characterized by Scanning Electron Microscopy (SEM) using a JEOL microscope (JSM-6060) on gold-coated samples. Compressive strength (CS) was measured in servohydraulic Universal Testing Machine (MTS 810) with a load measuring cell of 10 kN and a loading rate of 1 mm/min. The number of replicas was n 10 and student multiple comparison test was used to compare mean values. pH measures were carried out during soaking in SBF and lectures were made in an mPA-210 pH meter at 36.5 C.
R

RESULTS

Effect of C3S on the setting time of a-TCP cement Figure 1 shows the initial and nal setting times of composites containing 0% (a-TCP), 5% (C3S) and 10% (10C3S). The initial and nal setting times of a-TCP were higher than those reported in the literature for similar materials22 and the addition of C3S produced a remarkable increase in setting times. No signicant differences were found when the amount of C3S added was raised. Characterization of CPC/C3S composites cement Figures 24 show the powder XRD patterns of composites after setting (24 h setting) and after soaking in SBF for 7 and 14 days. After 24 h setting (Figure 1), for a-TCP-based cement, mainly peaks of CDHA (JCPDS 46-0905) were

Cytotoxicity test for cements The cell viability assay was performed by direct contact test according to ISO 10993-5 using peripheral blood mononuclear cells (PBMCs) and a procedure described elsewhere.21 Latex (1 cm2) and culture medium were used as positive

JOURNAL OF BIOMEDICAL MATERIALS RESEARCH B: APPLIED BIOMATERIALS | JAN 2012 VOL 100B, ISSUE 1

95

FIGURE 2. XRD patterns of cements 24h set. n a-Ca3(PO4)2; ~ b-Ca3(PO4)2;

Ca9(HPO4)(PO4)5(OH).

observed. With the addition of C3S, the diffraction peaks of CHDA appear less intense and unreacted peaks of a-TCP were also detected. The greater the amount of C3S added, the greater the intensity of the peaks of unreacted a-TCP and the lower the CDHA formed. After 7 days of soaking (Figure 2) the intensity of a-TCP lines decreased in relation to set cements and CDHA lines appeared stronger. Within

14 days of SBF soaking, the hydration reaction seemed to be complete for a-TCP and 5C3S, whereas great amount of unreacted a-TCP in addition to CDHA can be observed for 10C3S (Figure 3). For all times and all formulations, the characteristic peaks of b-TCP (JCPDS 09-0169), which appeared as a secondary phase in a-TCP powder and is not involved in the hydration process, were present.

FIGURE 3. XRD patterns of cements after 7 days in SBF. n a-Ca3(PO4)2; ~ b-Ca3(PO4)2;

Ca9(HPO4)(PO4)5(OH).

96

MOREJON-ALONSO ET AL.

a-TRICALCIUM PHOSPHATE/TRICALCIUM SILICATE COMPOSITE CEMENT

ORIGINAL RESEARCH REPORT

FIGURE 4. XRD patterns of cements after 14 days in SBF. n a-Ca3(PO4)2; ~ b-Ca3(PO4)2;

Ca9(HPO4)(PO4)5(OH).

Furthermore, no calcium-silicate-hydrate (C-S-H) or calcium hydroxide [Ca(OH)2] resulting from hydration of C3S were detected by this method. The bone-like apatite formation: Microstructural changes during soaking Since the main hydration product of a-TCP-based cement systems is CDHA, it is difcult to account for the ability of the materials to induce the deposition of apatite in SBF; however, the observation by SEM is an effective way to estimate the bioactivity of materials as the apatite grains and layers that are formed have characteristic features. Figure 5 shows the SEM surface images of a-TCP, 5C3S, and 10C3S at different times of immersion. For a-TCP-based cement [Figure 5(a)] a ne layer of CDHA crystals deposited onto the a-TCP grains was observed before immersion, while a network of entangled plate-like apatite crystals was detected for subsequent immersion times. Fourteen days after soaking, a layer of bone-like apatite, precipitated under physiological conditions, was clearly observed. With the addition of 5C3S, the formation of a gelatinous coating covering the surface of unreacted grains at early times of hydration was detected [Figure 5(b)]. After soaking in SBF, the morphology of 5C3S surface varied slightly and a needle microstructure appeared the same as type I C-S-H23; whereas a homogeneous layer of CDHA with globular shape morphology, typical of bioactive materials after immersion in SBF was detected within 14 days.24 With larger additions of C3S [Figure 5(c)], the surface before immersion appeared covered or speckled with small distinct features that could be identied as small crystals of dry C-S-H. After immersion for 7 days, a well dened bone-like apatite layer is formed and transformed

into a more dense and homogeneous layer with increasing immersion time. Although measures were taken to maintain aseptic conditions, bacterial contamination by Bacillus and Cocci colonies was observed at the surface of samples.25 Different microstructural features were found in the fracture surface of different composites (Figure 6). Since early stages, a-TCP showed the typical petal-like plates covering the biggest grains and the growth of these plates with time, at the expense of smaller grains, within the interstices of fracture surface [Figure 6(a)]. With the addition of C3S neither petal or needle-like crystals distinctive of the beginning of setting and hardening of the CPC, nor the microstructure of rod-like characteristics of more aged systems,22 were observed. With low contents of C3S [Figure 6(b)], the deposition of small CDHA crystals on the top of the larger a-TCP particles could be seen. These crystals evolve and grow into acicular crystals encasing some individual grains of b-TCP. The presence of some characteristic hollow-shells (Hadley grains),26 showing an empty shell of hydration products inside which an a-TCP or C3S had fully reacted, could be clearly identied in both the aTCP and the 5C3S cement. For greater additions of C3S [Figure 6(c)] no signicant differences with soaking time were found and the most common morphology observed was a mixed result of brillar and amorphous type I C-S-H which are characteristic of the early stages of C3S hydration. Mechanical strength Figure 7 shows the compressive strength of a-TCP and aTCP/C3S composites before and after soaking in SBF for 7 and 14 days. The results indicate that the compressive strength of a-TCP decreased slightly with soaking, whereas

JOURNAL OF BIOMEDICAL MATERIALS RESEARCH B: APPLIED BIOMATERIALS | JAN 2012 VOL 100B, ISSUE 1

97

FIGURE 5. SEM micrographs of surface after soaking in SBF (A) a-TCP, (B) 5C3S, and (C) 10C3S.

the compressive strength of 5C3S increased with ageing time, reaching values similar to those of a-TCP after hardening (16.86 MPa). For 10C3S, values of compressive strength were very low and no signicant changes were observed during immersion. pH analysis on in vitro immersion Figure 8 shows the changes in pH values of SBF caused by the cement pastes. The results showed that for a-TCP, the pH value decreased slightly in the rst 24 h after immersion and then ranges from 7.3 to 7.5. With the addition of C3S the pH increased rapidly, reaching a maximum of 10.4 for a 10% addition, and then decreased gradually and stabilized after 72 h. In vitro degradation Figure 9 shows the degradation of the a-TCP and a-TCP/C3S composite pastes with different contents of C3S after soaking in SBF solution for various time periods. It was found that the weight of a-TCP increased with time, whereas the weight of composites containing C3S experienced a mass
MOREJON-ALONSO ET AL.

loss during the rst 7 days and then began to gain mass. It was also found that the degradation rate of the composites containing C3S was higher than that of a-TCP, while the degradation rate raised proportionate to the increase in the amount of C3S added. Cytotocixity test Figure 10 shows the result of a direct cytotoxicity test for composites against the PBMCs after 24 and 48 h of incubation. It was found that the cytotoxicity of the composites containing C3S was signicantly lower than the positive control (p < 0.05) at 24 h of incubation, and biocompatibility was higher than in traditional a-TCP. It was also found that the viability of PBMCs decreased in all compositions after incubating for 48 h.
DISCUSSION

Setting times for CPCs usually ranged from 5 to 8 min when a neutral phosphate such as disodium hydrogen phosphate (Na2HPO4) or sodium dihydrogen phosphate (NaH2PO4) was
a-TRICALCIUM PHOSPHATE/TRICALCIUM SILICATE COMPOSITE CEMENT

98

ORIGINAL RESEARCH REPORT

FIGURE 6. SEM micrographs of fracture surface after soaking in SBF (A) a-TCP, (B) 5C3S, and (C) 10C3S.

FIGURE 7. Compressive strength of the a-TCP a-TCP/C3S composites after soaking in SBF for various times.

FIGURE 8. Changes in pH value of SBF soaked with a-TCP and a-TCP/ C3S composite.

JOURNAL OF BIOMEDICAL MATERIALS RESEARCH B: APPLIED BIOMATERIALS | JAN 2012 VOL 100B, ISSUE 1

99

FIGURE 9. Weight loss of the a-TCP and a-TCP/C3S composites after soaking in SBF for various times.

added to the liquid phase.22 When b-TCP is present, setting times are often delayed due to the non-participation of same in the hydration reaction [Eq. (2)]; and in this case, a large quantity of b-TCP (18%) was present in the original powder as a result of the doping of Mg2 raw materials (0.28% as MgO).21 With the addition of C3S, the solubility of a-TCP decreases due to the formation of Ca(OH)2 during Ca3SiO5 hydrolysis [Eq. (3)], which results in alkaline circumstances and delayed dissolution of a-TCP. Furthermore, the formation of a dense calcium silicate hydrate gel (C-S-H) on the surface of a-TCP particles [Figure 5(b)] also retards dissolution of a-TCP grains and the precipitation of CDHA. 3a Ca3 PO4 2s H2 O Ca9 HPO4 PO4 5 OHs Ca3 SiO5s 3H2 O CaO:SiO2 :H2 Ogel 2CaOH2s (2) (3)

As in the case with C3S hydrolysis,27 the hydration mechanism of a-TCP-based cements is diffusion dependent of the reactive species through the layer of formed product. Therefore, the degree of extension of the a-TCP hydration, usually 80% at 24 h,28 is lower as evidenced by the presence of intense unreacted peaks in the diffraction patterns of composites containing C3S (Figure 2). Even when the presence of silicate ions formed from C3S promotes apatite nucleation,29 the main effect of adding C3S at early stages seems to be the delay in the dissolution of a-TCP and the precipitation of CDHA. While initially retarded by the presence of C3S, hydration reaction occurs when soaking time is increased and the unreacted peaks of a-TCP disappear resulting in more intense peaks of CDHA. For formulations with higher content of C3S, a-TCP peaks can be found within 14 days and the CDHA peaks appear more intense, probably due to its precipitation from SBF solution (Figures 3 and 4). When compared with bioactive bone substitution materials such as A-W glass ceramic and Bioglass , the CPCs appear to be less capable of inducing an homogeneous bone-like apatite layer on the surface to form chemical
R V

bond to bone tissue at early stages of implantation.30 The results of SEM analyses (Figure 5) showed that a-TCP/C3S composite pastes induce homogeneous apatite deposition on the paste surface due to the presence of C3S, and the formation time of same was dependent on the content of C3S. As is generally accepted, the deposition of apatite on the surface of calcium phosphate cement is largely dependent on supersaturation of Ca2 and PO43 ions and indeed such process proceeds with low rate resulting in the absence of homogeneous apatite layer formation at early stages of implantation.24 In the presence of C3S, the HSiO3 ions are released during hydration of the composite paste, acting as sites for nucleation of apatite crystals and hence accelerating the deposition of apatite on the surface. The apatite layer in a-TCP-based cement was formed within 14 days. This is the time which is reported as required to the deposition of new bone on the surface of the implant during in vivo experiments,31 which shows the superior bioactivity of composites. A critical problem that limits wider clinical application of CPCs is their mechanical properties. Since C3S had a negative impact on setting times and the hydration rate of aTCP, the mechanical strength of composites at early stages was not improved. Another factor that could explain this behavior is related to the crystal growth of hydroxyapatite. The SEM results show (Figure 5) that a-TCP set by entanglement of plate-like crystals, while composites containing C3S had a gelatinous coating of C-S-H on the surface covering the a-TCP particles, also showed an incipient precipitation of CDHA crystals on the fracture surface in the rst 24 h, which caused the fall of the mechanical properties since the precipitation of CHDA is responsible for the adherence and interlocking of the crystalline grains, which results in hardening. With the consumption of C3S and the growth of CHDA needles-like crystals on composites, the mechanical properties begin to increase reaching values similar to those of aTCP-based cement after setting. Adding more C3S produces more C-S-H, which delays the solubilization of the a-TCP grains, and consequently, the precipitation of CDHA. In this case, the C-S-H does not act as ller does not bond with the

FIGURE 10. Cell viability of PBMCs on the different pastes after culturing for 24 and 48 h. *p < 0.05 compared with () control group.

100

MOREJON-ALONSO ET AL.

a-TRICALCIUM PHOSPHATE/TRICALCIUM SILICATE COMPOSITE CEMENT

ORIGINAL RESEARCH REPORT

apatite formed during the setting, thus the compressive strength is not enhanced. The dissolution of calcium ions into body uids plays an important role in the nucleation and growth of hydroxyapatite layers on a materials surface. When immersed in SBF, the pH of the a-TCP-based cement decreases to values close to 7.0 probably due to the formation of some H3PO4 during hydrolyzation of Ca3PO4 into Ca10(PO4)6(OH)232 and after this time values remain almost unchanged. With the addition of C3S, OH ions are released to SBF solution raising the pH of SBF solution in the rst 72 h, after which the pH stabilizes implying that OH ions are no longer released to the medium. Conventional CPCs usually have a slow degradability and often experience a mass gain after long soaking periods as a result of hydrolysis of a-TCP or due to the formation of apatite on the surface of the samples.33 Considering the fact that the degradability is primarily governed by the chemical composition and the physical characteristics of the material, the reason for the higher degradation rate of the a-TCP/C3S composites could be the higher solubility of the C-S-H as compared with CDHA. A mixture of two crystallized calcium silicate hydrated identied by DRX as 1.5CaO.SiO2.H2O (JCPDS 33-0306) and 2CaO.SiO2.0.5H2O (JCPDS 12-0199)34 were found in the supernatant solution during the rst 72 h, indicating the solubilization of the silicate hydrate at that time. After 7 days of soaking, a reversal behavior can be seen and the amount of apatite deposited on the cement surface and formed due to the hydrolysis of a-TCP was larger than dissolution of C-S-H and/or other phases, so the weight of samples increased. It is generally accepted that the in vitro cellmaterial interaction is a useful criterion in the evaluation of new biomaterials. The results of cell culture showed that composites containing C3S are less cytotoxic and more compatible than pure a-TCP-based cement and the positive effect in cytotoxicity could be attributed to the dissolution of silicate ions present in a-TCP/C3S pastes that stimulate cell proliferation.35,36 The slight increase in cytotoxicity 48 h after culturing may be explained due to the occurrence of some chemical transformations of the material in culture medium since the pastes continue hydrating after 7 days in aqueous media. Nevertheless, the study indicated that the composite cement was not only biocompatible but also non-cytotoxic.
CONCLUSIONS

and were not only more biocompatible but also more noncytotoxic as showed by the in vitro cytotoxic assay. The results showed that it is only possible to incorporate up to 5% with an improvement of the bioactivity and biocompatibility without compromising signicantly the mechanical strength of materials. The Brazilian Government entity dedicated to the training of human resources.
REFERENCES
1. Brown WE, Chow LC. A new calcium phosphate water-setting cement. In: Brown WE, editor. Cements Research Progress. Westerville, OH: American Ceramic Society; 1986. pp352379. 2. LeGeros RZ, Cohayeb A, Shulman A. Apatitic calcium phosphates: Possible dental restorative materials. J Dent Res 1982;61:343. 3. Monma H. The hydration of alpha-tricalcium phosphate. Yogo Kyokai Shi 1976;84:209213. 4. Ginebra MP, Boltong MG, Fernandez E, Planell JA, Driessens FCM. Effect of various additives and temperature on some properties of an apatitic calcium phosphate cement. J Mater Sci Mater Med 1995;6:612616. 5. Santos LA, Cristina de Oliveira L, Cristina da Silva E, Garcia R, Ortega A, Arruda A. Fiber reinforced calcium phosphate. Artif Organs 2000;24:212216. 6. Yamamoto H, Niwa S, Hori M, Hattori T, Sawai K, Aoki S, Hirano M, Takeuchi H. Mechanical strength of calcium phosphate cement in vivo and in vitro. Biomaterials 1998;19:15871591. 7. Dorozhkin S. Calcium orthophosphate cements and concretes. Materials 2009;2:221291. 8. Ishikawa K, Asaoka K. Estimation of ideal mechanical strength and critical porosity of calcium phosphate cement. J Biomed Mater Res 1995;29:15371543. 9. Carlisle EM. Silicon: An essential element for the chick. Science 1972;178:619621. 10. Carlisle EM. A silicon requirement for normal skull formation in chicks. J Nutr 1980;1:352359. 11. Schwarz K, Milne DB. Growth-promoting effects of silicon in rats. Nature 1972;239:333334. 12. Hench LL. Bioceramics: From concept to clinic. J Am Ceram Soc 1991;74:14871410. 13. Zhao W, Chang J. Preparation and characterization of novel tricalcium silicate bioceramics. J Biomed Res A 2005;73:8689. 14. Lea FM. The Chemistry of Cement and Concrete. London: Edward Arnold Ltd.; 1970. 15. Monma H, Goto M, Kohmura T. Effect of additives on hydration and hardness of tricalcium phosphate. Gypsum Lime 1984;188: 1116. 16. Zhao W, Chang J. Solgel synthesis and in vitro bioactivity of tricalcium silicate powders. Mater Lett 2004;58:23502353. 17. ASTM. Standard Test Method for Time of Setting of HydraulicCement Paste by Gillmore Needles, ASTM. C-266-89 A. ASTM; 1995. Available at: http://www.astm.org/Standards/C266.htm 18. Driessens FMC, Planell JA, Gil J. Calcium phosphates bone cements. In: Wise D, Trantolo D, Altobelli D, Yaszernski M, Gresser J, Schwartz E, editors. Encyclopedic Handbook of Biomaterials and Bioengineering, Part B: Applications; New York: Marcel Dekker; 1995. pp855871. 19. Kim HM, Miyazaki T, Kokubo T, Nakamura T. Revised simulated body uid. Key Eng Mater 2001;192195:4750. 20. ISO/FDIS. Implants for Surgery. In Vitro Evaluation for ApatiteForming Ability of Implant Materials, ISO/FDIS 23317. ISO; 2007. Available at: http://www.iso.org/iso/iso_catalogue/catalogue_tc/ catalogue_detail.htm?csnumber=54163 21. Morejon-Alonso L, Ferrari MB, Camassola M, Garcia Carrodeguas R, Santos LA. In vitro cytotoxicity of a calcium phosphate-silicate composite bone cement. Presented at the Sixth Latinoam Congress of Biomaterials and Articial Organs, Gramado, RS, August 1720, 2010. 22. Ambard A, Mueninghoff L. Calcium phosphate cement: Review of mechanical and biological properties. J Prosthodon 2006;15: 321328.

In this article, novel bioactive composite cements were prepared by adding Ca3SiO5 into the traditional a-Ca3(PO4)2based cement. The composites thus obtained showed an increase in setting times, and a considerable delay in the rate of a-TCP hydration, which caused the loss of initial mechanical properties together with the microstructural differences found due to the formation of a C-S-H gel. Furthermore, the presence of high contents of C3S increases the degradability of cements and raises the pH of medium at early stages, compromising the precipitation of CDHA. However, the a-TCP/C3S composites possessed excellent bioactivity, as indicated by the formation of bone-like apatite in SBF

JOURNAL OF BIOMEDICAL MATERIALS RESEARCH B: APPLIED BIOMATERIALS | JAN 2012 VOL 100B, ISSUE 1

101

23. Fonseca PC, Jennings HM. The effect of drying on early-age morphology of CSH as observed in environmental SEM. Cement Concr Res 2010;40:16731680. 24. Kokubo T, Takadama H. How useful is SBF in predicting in vivo bone bioactivity? Biomaterials 2006;27:29072915. 25. Gil J, Padros A, Manero JM, Aparicio C, Nilsson M, Planell JA. Growth of bioactive surfaces on titanium and its alloys for orthopaedic and dental implants. Mater Sci Eng C 2002;22:5360. 26. Hadley DH, Dolch WL, Diamond S. On the occurrence of hollowshell hydration grains in hydrated cement paste. Cement Concr Res 2000;30:16. 27. Greenberg SA, Chang TN. The hydration of tricalcium silicate. Port Cement Assoc 1965;69:553561. 28. Ginebra MP, Fernandez E, Driessens FCM, Planell JA. Modeling of the hydrolysis of a-tricalcium phosphate. J Am Ceram Soc 1999; 82:28082812. 29. Kokubo T. Bioactive glass-ceramics: Properties and applications. Ceram Soc Japan 1991;99:965. 30. Kobayashi M, Nakamura T, Okada Y, Fukumoto A, Furukawa T, Kato H, Kokubo T, Kikutani T. Bioactive bone cement: Comparison

31.

32.

33.

34.

35. 36.

of apatite and wollastonite containing glass-ceramic, hydroxyapatite, and b-tricalcium phosphate llers on bone-bonding strength. Biomed Mater Res 1998;42:223. Duan YR, Zhang ZR, Chen CY, Zhang XD. Dynamic study of calcium phosphate formation on porous HA/TCP ceramics. J Mater Sci Mater Med 2005;16:795801. Santos LA, Carrodeguas RG, Rogero SO, Higa OZ, Boschi AO, de Arruda ACF. a-Tricalcium phosphate cement: In vitro cytotoxicity. Biomaterials 2002;23:20352042. Ishikawa K, Takagi S, Chow LC, Ishikawa Y, Eanes ED, Asaoka K. Behavior of a calcium phosphate cement in simulated blood plasma in vitro. Dent Mater 1994;10:2632. Morejon-Alonso L. Avaliac o de cimentos osseos de Fosfatos de a Calcio com adic es de Aluminato e Silicato de Calcio. PhD Tese, o Universidade Federal do Rio Grande do Sul, Porto Alegre, 2011. Hench LL, West JK. Biological applications of bioactive glasses. Life Chem Rep 1996;13:187241. Zhao WY, Wang JY, Zhai WY, Wang Z, Chang J. The self-setting properties and in vitro bioactivity of tricalcium silicate. Biomaterials 2005;26:61136121.

102

MOREJON-ALONSO ET AL.

a-TRICALCIUM PHOSPHATE/TRICALCIUM SILICATE COMPOSITE CEMENT