Abstract
In this study, the effect of addition of magnesium as a stabilizer of amorphous phase on the structure and bioactivity of amorphous calcium phosphate for biomedical applications was investigated. Therefore, amorphous calcium phosphate was synthesized by a precipitation and freeze drying method with different molar ratios of magnesium to calcium (Mg/Ca). Then the samples were incubated in simulated body fluid (SBF) for 3, 7 and 14 days to evaluate their bioactivity and the investigation of their phase conversion. X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy were employed to characterize the prepared samples. According to X-ray diffraction patterns, the most stable amorphous phase was obtained at a magnesium to calcium (Mg/Ca) ratio of 0.3 and the magnesium acted as a stabilizer of the amorphous phase. After incubation in SBF solution, the hydroxyapatite phase was detected in XRD patterns of the samples, but with the addition of the magnesium, the crystallinity and formation of the hydroxyapatite phase was decreased. Also the FTIR analysis showed that the intensity of the peaks related to phosphate and hydroxyl groups decreased with the addition of magnesium, which indicated a more stable amorphous calcium phosphate phase. Finally, in-vitro experiments like MTT assay and alkaline phosphatase activity with human osteosarcoma cell lines (G-292) on the prepared samples were evaluated and discussed. The results indicated that amorphous calcium phosphate with a calcium to magnesium ratio of 0.3 revealed optimal cell proliferation and ALP activity. Eventually, this material is non-toxic and compatible to be used as a bone substitute material.
Similar content being viewed by others
References
Maciejewski M, Brunner TJ, Loher SF, Stark WJ, Baiker A (2008) Phase transitions in amorphous calcium phosphates with different Ca/P ratios. Thermochim Acta 468(1–2):75–80. https://doi.org/10.1016/j.tca.2007.11.022
Medvecky L, Sopcak T, Girman V, Briancin J (2013) Amorphous calcium phosphates synthesized by precipitation from calcium D-gluconate solutions. Colloids Surf A: Physicochem Eng Asp 417:191–200. https://doi.org/10.1016/j.colsurfa.2012.11.015
Ding G-J, Zhu Y-J, Qi C, Lu B-Q, Wu J, Chen F (2015) Porous microspheres of amorphous calcium phosphate: Block copolymer templated microwave-assisted hydrothermal synthesis and application in drug delivery. J Colloid Interface Sci 443:72–79. https://doi.org/10.1016/j.jcis.2014.12.004
Cuneyt Tas A (2014) Submicron spheres of amorphous calcium phosphate forming in a stirred SBF solution at 55 C. J Non-Cryst Solids 400:27–32. https://doi.org/10.1016/j.jnoncrysol.2014.04.031
Gross KA, Young CJ, Beck MA, Keebaugh EW, Bronts TJ, Saber-Samandari S, Riley DP (2011) Characterization and dissolution of functionalized amorphous calcium phosphate biolayers using single-splat technology. Acta Biomater 7(5):2270–2275. https://doi.org/10.1016/j.actbio.2011.01.020
Oyane A, Araki H, Nakamura M, Shimizu Y, Shubhra QTH, Ito A, Tsurushima H (2016) Controlled superficial assembly of DNA–amorphous calcium phosphate nanocomposite spheres for surface-mediated gene delivery. Colloids Surf B: Biointerfaces 141:519–527. https://doi.org/10.1016/j.colsurfb.2016.02.010
Schumacher GE, Antonucci JM, O’Donnell JNR, Skrtic D (2007) The use of amorphous calcium phosphate composites as bioactive basing materials: Their effect on the strength of the composite/adhesive/dentin bond. J Amer Dent Assoc 138(11):1476–1484. https://doi.org/10.14219/jada.archive.2007.0084
Zyman Z, Rokhmistrov D, Glushko V (2012) Structural changes in precipitates and cell model for the conversion of amorphous calcium phosphate to hydroxyapatite during the initial stage of precipitation. J Cryst Growth 353(1):5–11. https://doi.org/10.1016/j.jcrysgro.2012.04.041
Raz M, Moztarzadeh F, Kordestani SS (2016) Synthesis, Characterization and in-vitro Study of Chitosan/Gelatin/Calcium Phosphate Hybrid Scaffolds Fabricated Via Ion Diffusion Mechanism for Bone Tissue Engineering. Silicon:1–10. https://doi.org/10.1007/s12633-016-9439-3
Zhao J, Liu Y, W-b Sun, Yang X (2012) First detection, characterization, and application of amorphous calcium phosphate in dentistry. J Dent Sci 7(4):316–323. https://doi.org/10.1016/j.jds.2012.09.001
Melo MAS, Cheng L, Zhang K, Weir MD, Rodrigues LKA, Xu HHK (2013) Novel dental adhesives containing nanoparticles of silver and amorphous calcium phosphate. Dent Mater 29(2):199–210. https://doi.org/10.1016/j.dental.2012.10.005
Julien M, Khairoun I, LeGeros RZ, Delplace S, Pilet P, Weiss P, Daculsi G, Bouler JM, Guicheux J (2007) Physico-chemical–mechanical and in vitro biological properties of calcium phosphate cements with doped amorphous calcium phosphates. Biomaterials 28(6):956–965. https://doi.org/10.1016/j.biomaterials.2006.10.018
Xu HHK, Moreau JL, Sun L, Chow LC (2011) Nanocomposite containing amorphous calcium phosphate nanoparticles for caries inhibition. Dent Mater 27(8):762–769. https://doi.org//10.1016/j.dental.2011.03.016
Zhou R, Xu W, Chen F, Qi C, Lu B-Q, Zhang H, Wu J, Qian Q-R, Zhu Y-J (2014) Amorphous calcium phosphate nanospheres/polylactide composite coated tantalum scaffold: Facile preparation, fast biomineralization and subchondral bone defect repair application. Colloids Surf B: Biointerfaces 123:236–245. https://doi.org/10.1016/j.colsurfb.2014.09.021
Niu X, Wang L, Tian F, Wang L, Li P, Feng Q, Fan Y (2016) Shear-mediated crystallization from amorphous calcium phosphate to bone apatite. J Mech Behav Biomed Mater 54:131–140. https://doi.org/10.1016/j.jmbbm.2015.09.024
Zhang H, Fu Q-W, Sun T-W, Chen F, Qi C, Wu J, Cai Z-Y, Qian Q-R, Zhu Y-J (2015) Amorphous calcium phosphate, hydroxyapatite and poly(d,l-lactic acid) composite nanofibers: Electrospinning preparation, mineralization and in vivo bone defect repair. Colloids Surf B: Biointerfaces 136:27–36. https://doi.org/10.1016/j.colsurfb.2015.08.015
Van den Vreken NMF, Pieters IY, Declercq HA, Cornelissen MJ, Verbeeck RMH (2010) Characterization of calcium phosphate cements modified by addition of amorphous calcium phosphate. Acta Biomater 6(2):617–625. https://doi.org/10.1016/j.actbio.2009.07.038
Seyedmomeni SS, Naeimi M, Raz M, Mohandesi JA, Moztarzadeh F, Baghbani F, Tahriri M (2016) Synthesis, characterization and biological evaluation of a new Sol-Gel Derived B and Zn-Containing bioactive glass: In Vitro study. Silicon:1–7. https://doi.org/10.1007/s12633-016-9414-z
Cobourne G, Mountjoy G, Rodriguez-Blanco JD, Benning LG, Hannon AC, Plaisier JR (2014) Neutron and X-ray diffraction and empirical potential structure refinement modelling of magnesium stabilised amorphous calcium carbonate. J Non-Cryst Solids 401:154–158. https://doi.org/10.1016/j.jnoncrysol.2013.12.023
Dong G, Zheng Y, He L, Wu G, Deng C (2016) The effect of silicon doping on the transformation of amorphous calcium phosphate to silicon-substituted α-tricalcium phosphate by heat treatment. Ceram Int 42(1, Part A):883–890. https://doi.org/10.1016/j.ceramint.2015.09.013
Cheng L, Weir MD, Xu HHK, Antonucci JM, Kraigsley AM, Lin NJ, Lin-Gibson S, Zhou X (2012) Antibacterial amorphous calcium phosphate nanocomposites with a quaternary ammonium dimethacrylate and silver nanoparticles. Dent Mater 28(5):561–572. https://doi.org/10.1016/j.dental.2012.01.005
Lee D, Kumta PN (2010) Chemical synthesis and characterization of magnesium substituted amorphous calcium phosphate (MG-ACP). Mater Sci Eng: C 30(8):1313–1317. https://doi.org/10.1016/j.msec.sec2010.05.009
Torres PMC, Abrantes JCC, Kaushal A, Pina S, Döbelin N, Bohner M, Ferreira JMF (2016) Influence of Mg-doping, calcium pyrophosphate impurities and cooling rate on the allotropic \(\leftrightarrow \)-tricalcium phosphate phase transformations. J Eur Ceram Soc 36(3):817–827. https://doi.org/10.1016/j.jeurceramsoc.2015.09.037
Qian J, Ma J, Su J, Yan Y, Li H, Shin J-W, Wei J, Zhao L (2016) PHBV-based ternary composite by intermixing of magnesium calcium phosphate nanoparticles and zein: In vitro bioactivity, degradability and cytocompatibility. Eur Polym J 75:291–302. https://doi.org/10.1016/j.eurpolymj.2015.12.026
Khan NI, Ijaz K, Zahid M, Khan AS, Abdul Kadir MR, Hussain R, Anis ur R, Darr JA, Ihtesham ur R, Chaudhry AA (2015) Microwave assisted synthesis and characterization of magnesium substituted calcium phosphate bioceramics. Mater Sci Eng: C 56:286–293. https://doi.org/10.1016/j.msec.2015.05.025
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Shahrezaee, M., Raz, M., Shishehbor, S. et al. Synthesis of Magnesium Doped Amorphous Calcium Phosphate as a Bioceramic for Biomedical Application: In Vitro Study. Silicon 10, 1171–1179 (2018). https://doi.org/10.1007/s12633-017-9589-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12633-017-9589-y