Abstract
Synthesis of unsubstituted and multi-element (magnesium, zinc and cobalt) substituted hydroxyapatites (HAP) with varying stoichiometric compositions and evaluation of their morphological and structural characteristics, degree of crystallinity, bioactivity, cytotoxicity and antibacterial activity are addressed. The morphological features are not altered much following the substitution of Mg2+, Zn2+, and Co2+ in the HAP lattice. Nevertheless, their substitution exerts a strong influence on the structural characteristics HAP. Rietveld refinement analysis of the X-ray diffraction patterns indicates a decrease in crystallinity and mineralogical composition of HAP phase, which is accompanied with an increase of β-tricalcium phosphate (β-TCP) phase along with Co3O4 phase. Broadening of the PO43− peaks and a decrease in intensity of the OH− peak are observed by Fourier-transform infrared spectra. A decrease in intensity, broadening and a slight shift in Raman band (at 961 cm−1 for HAP) towards the lower side suggest the incorporation of Mg, Zn, and Co, disordering of the crystal structure of HAP and formation of β-TCP as additional phase besides HAP. The MgZnCo-HAP’s exhibits a better bioactivity, cell viability and anti-bacterial activity than the unsubstituted HAP. However, a decrease in cell viability and anti-bacterial activity are observed when the stoichiometric ratio of the substituent elements is relatively higher.
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References
Mucalo M (ed) (2015) Hydroxyapatite (HAp) for biomedical applications, woodhead publishing series in biomaterials. Elsevier-Woodhead Publishers, Cambridge
Dorozhkin SV (2010) Bioceramics of calcium orthophosphates Biomaterials 31:1465–1485
Dorozhkin SV (2012) Calcium orthophosphates: applications in Nature, Biology, and Medicine. Pan Stanford Publishing, Singapore
Koutsopoulos S (2002) Synthesis and characterization of hydroxyapatite crystals: A review study on the analytical methods. J Biomed Mater Res 62:600–612
Gómez-Morales J, Iafisco M, Delgado-López JM, Sarda S, Drouet C (2013) Progress on the preparation of nanocrystalline apatites and surface characterization: overview of fundamental and applied aspects. Prog Cryst Growth Charact Mater 59:1–46
Feng W, Mu-sen L, Yu-peng L, Yong-xin Q (2005) A simple sol–gel technique for preparing hydroxyapatite nanopowders. Mater Lett 59:916–919
Fathi MH, Hanifi A (2007) Evaluation and characterization of nanostructure hydroxyapatite powder prepared by simple sol–gel method. Mater Lett 61:3978–3983
Shepherd JH, Shepherd DV, Best SM (2012) Substituted hydroxyapatites for bone repair. J Mater Sci Mater Med 23:2335–2347
Boanini E, Gazzano M, Bigi A (2010) Ionic substitutions in calcium phosphates synthesized at low temperature. Acta Biomater 6:1882–1894
Cacciotti I (2015) Cationic and anionic substitutions in hydroxyapatite. In: Antoniac IV (ed) Handbook of bioceramics and biocomposites. Springer International Publishing, Switzerland, p 1–68
Šupová M (2015) Substituted hydroxyapatites for biomedical applications: A review. Ceram Int 41:9203–9231
Percival M (1999) Bone health and osteoporosis. Appl Nutr Sci Rep 5:1–6
Moonga BS, Dempster DW (1995) Zinc is a potent inhibitor of osteoclastic bone resorption in vitro. J Bone Miner Res 10:453–457
Yamaguchi M (1998) Role of zinc in bone formation and bone resorption. J Trace Elem Exp Med 11:119–135
Aina V, Lusvardi G, Annaz B, Gibson IR, Imrie FE, Malavasi G, Menabue L, Cerrato G, Martra G (2012) Magnesium and strontium-co-substituted hydroxyapatite: the effects of doped-ions on the structure and chemico-physical properties. J Mater Sci Mater Med 23:2867–2879
Cox SC, Jamshidi P, Grover LM, Mallick KK (2014) Preparation and characterisation of nanophase Sr, Mg, and Zn substituted hydroxyapatite by aqueous precipitation. Mater Sci Eng C35:106–114
Bigi A, Falini G, Foresti E, Gazzano M, Ripamonti A, Roveri N (1993) Magnesium influence on hydroxyapatite crystallization. J Inorg Biochem 49:69–78
Landi E, Logroscino G, Proietti L, Tampieri A, Sandri M, Sprio S (2008) Biomimetic Mg substituted hydroxyapatite: from synthesis to in vivo behaviour. J Mater Sci Mater Med 19:239–247
Batra U, Kapoor S, Sharma S (2013) Influence of magnesium ion substitution on structural and thermal behavior of nanodimensional hydroxyapatite. J Mater Eng Perform 22:1798–1806
Cacciotti I, Bianco A, Lombardi M, Montanaro L (2009) Mg-substituted hydroxyapatite nanopowders: synthesis, thermal stability and sintering behaviour. J Eur Ceram Soc 29:2969–2978
Thian ES, Konishi T, Kawanobe Y, Lim PN, Choong C, Ho B, Aizawa M (2013) Zinc-substituted hydroxyapatite: a biomaterial with enhanced bioactivity and antibacterial properties. J Mater Sci Mater Med 24:437–445
Bigi A, Foresti E, Gandolfi M, Gazzano M, Roveri N (1995) Inhibiting effect of zinc on hydroxylapatite crystallization. J Inorg Biochem 58:49–58
Guerra-Lopez JR, Echeverria GA, Guida JA, Vina R, Punte G (2015) Synthetic hydroxyapatite doped with Zn(II) studied by X-Ray diffraction, infrared, Raman and thermal analysis. J Phys Chem Solids 81:57–65
Tang Y, Chappell HF, Dove MT, Reeder RJ, Lee YJ (2009) Zinc incorporation into hydroxylapatite. Biomaterials 30:2864–2872
Stanic′ V, Dimitrijevic′ S, Antic′-Stankovic′ J, Mitric′ M, Jokic′ B, Plec′aš I, Raičevic′ S (2010) Synthesis, characterization and antimicrobial activity of copper and zinc-doped hydroxyapatite nanopowders. Appl Surf Sci 256:6083–6089
Miyaji F, Kono Y, Suyama Y (2005) Formation and structure of zinc-substituted calcium hydroxyapatite. Mater Res Bull 40:209–220
Ren F, Xin R, Ge X, Leng Y (2009) Characterization and structural analysis of zinc-substituted hydroxyapatites. Acta Biomater 5:3141–3149
Tank KP, Chudasama KS, Thaker VS, Joshi MJ (2013) Cobalt-doped nano hydroxyapatite: synthesis, characterization, antimicrobial and hemolytic studies. J Nanopart Res 15:1644
Stojanovic Z, Veselinovic L, Markovic S, Ignjatovic N, Uskokovic D (2009) Hydrothermal synthesis of nanosized pure and cobalt-exchanged hydroxyapatite. Mater Manuf Process 24:1096–1103
Kramer E, Itzkowitz E, Wei M (2014) Synthesis and characterization of cobalt-substituted hydroxyapatite powders. Ceram Int 40:13471–13480
Kulanthaivel S, Roy B, Agarwal T, Giri S, Pramanik K, Pal K, Ray SS, Maiti TK, Banerjee I (2016) Cobalt doped proangiogenic hydroxyapatite for bone tissue engineering application. Mater Sci Eng C58:648–658
Ignjatović N, Ajduković Z, Savić V, Najman S, Mihailović D, Vasiljević P, Stojanović Z, Uskoković V, Uskoković D (2013) Nanoparticles of cobalt-substituted hydroxyapatite in regeneration of mandibular osteoporotic bones. J Mater Sci Mater Med 24:343–354
Moreira MP, Soares GDA, Dentzer J, Anselme K, Sena LÁ, Kuznetsov A, Santos EA (2016) Synthesis of magnesium and manganese-doped hydroxyapatite structures assisted by the simultaneous incorporation of strontium. Mater Sci Eng C61:736–743
Iqbal N, Kadir MRA, Mahmood NH, Salim N, Froemming GRA, Balaji HR, Kamarul T (2014) Characterization, antibacterial and in vitro compatibility of zinc–silver doped hydroxyapatite nanoparticles prepared through microwave synthesis. Ceram Int 40:4507–4513
Lowry N, Han Y, Meenan BJ, Boyd AR (2017) Strontium and zinc co-substituted nanophase hydroxyapatite. Ceram Int 43:12070–12078
Robinson L, Salma-Ancane K, Stipniece L, Meenan BJ, Boyd AR (2017) The deposition of strontium and zinc co-substituted hydroxyapatite coatings. J Mater Sci Mater Med 29:51
Kulanthaivel S, Mishra U, Agarwal T, Giri S, Pal K, Pramanik K, Banerjee I (2015) Improving the osteogenic and angiogenic properties of synthetic hydroxyapatite by dual doping of bivalent cobalt and magnesium ion. Ceram Int 41:11323–11333
Kolmas J, Jaklewicz A, Zima A, Buc′ko M, Paszkiewicz Z, Lis J, Lósarczyk S ́ (2011) Incorporation of carbonate and magnesium ions into synthetic hydroxyapatite: the effect on physicochemical properties J Mol Struct 987:40–50
Suresh Kumar G, Thamizhavel A, Yokogawa Y, Narayana Kalkura S, Girija EK (2012) Synthesis, characterization and in vitro studies of zinc and carbonate co-substituted nano-hydroxyapatite for biomedical applications. Mater Chem Phys 134:1127–1135
Gomes S, Vichery C, Descamps S, Martinez H, Kaur A, Jacobs A, Nedelec JM, Renaudin G (2018) Copper doping of calcium phosphate bioceramics from mechanism to the control of cytotoxicity. Acta Biommaterialia 65:465–474
Kokubo T, Kushitani H, Sakka S, Kitsugi T, Yamamuro T (1990) Solutions able to reproduce in vivo surface-structure changes in bioactive glass ceramic A-W3. J Biomed Mater Res 24:721–734
Esakkirajan M, Prabhu NM, Arulvasu C, Beulaja M, Manikandan R, Thiagarajan R, Govindarajan K, Prabhu D, Dinesh D, Babu G, Dhanasekaran G (2014) Anti-proliferative effect of a compound isolated from Cassia auriculate against human colon cancer cell line HCT 15. Spectrochim Acta Part A 120:462–466
Elkabouss K, Kacimi M, Ziad M, Ammar S, Bozon-Veduraz F (2004) Cobalt-exchanged hydroxyapatite catalysts: magnetic studies, spectroscopic investigations, performance in 2-butanol and ethane oxidative dehydrogenations. J Catal 226:16–24
Jarcho M, Bolen CH, Thomas MB, Bobick J, Kay JF, Doremus RH (1976) Hydroxylapatite synthesis and characterization in dense polycrystalline form. J Mater Sci 11:2027–2035
Li MO, Xiao XF, Liu RF, Chen CY, Huang LZ (2008) Structural characterization of zinc-substituted hydroxyapatite prepared by hydrothermal method. J Mater Sci Mater Med 19:797–803
Shepherd D, Best SM (2013) Production of zinc substituted hydroxyapatite using various precipitation routes. Biomed Mater 8:025003
Lorenzo LMR, Regi MV (2000) Controlled crystallization of calcium phosphate apatites. Chem Mater 12:2460–2465
Gomes S, Renaudin G, Jallot E, Nedelec JM (2009) Structural characterization and biological fluid interaction of sol-gel derived Mg substituted biphasic phosphate ceramic. App Mater Interface 1(2):505–513
Gomes S, Karur A, Nedelec JM, Renaudin G (2014) X-Ray absorption spectroscopy shining (synchrotron) light onto the insertion of Zn2+ in calcium phosphate ceramics and its influence on their behavior under biological conditions. J Mater Chem 2:536–545
Gomes S, Karur A, Greneche JM, Nedelec JM, Renaudin G (2017) Atomic scale modeling of iron-doped biphasic calcium phosphate bioceramics. Acta Biomater 50:78–88
Renaudin G, Gomes S, Nedelec JM (2017) First-row transition metal doping in calcium phosphate bioceramics: a detailed crystallographic study Mater 10(1):92
Gomes S, Nedelec JM, Jallot E, Sheptyakov D, Renaudin G (2011) Unexpected mechanism of Zn2+ insertion in calcium phosphate bioceramics. Chem Mater 23:3072–3085
Antonakos A, Liarokapis E, Leventouri T (2007) Micro-Raman and FTIR studies of synthetic and natural apatites. Biomaterials 28:3043–3054
Yang Y, Perez-Amodio S, Barre‘ re-de Groot FYF, Everts V, van Blitterswijk CA, Habibovic P (2010) The effects of inorganic additives to calcium phosphate on in vitro behaviour of osteoblasts and osteoclasts. Biomaterials 31:2976–2989
Zilm ME, Chen L, Sharma V, McDannald A, Jain M, Ramprasad R, Wei M (2016) Hydroxyapatite substituted by transition metals: experiment and theory. Phys Chem Chem Phys 18:16457–16465
Penel G, Leroy G, Rey C, Bres E (1998) Micro Raman spectral study of the PO4 and CO3 vibrational modes in synthetic and biological apatites. Calcif Tissue Int 63:475–481
Awonusi A, Morris M, Tecklenburg M (2007) Carbonate assignment and calibration in the Raman spectrum of apatite. Calcif Tissue Int 81:46–52
Pasteris JD, Wopenka B, Freeman JJ, Rogers K, Valsami-Jones E, van der Houwen JAM, Silva MJ (2004) Lack of OH in nanocrystalline apatite as a function of degree of atomic order: implications for bone and biomaterials. Biomaterials 25:229–238
Darimont GL, Gilbert B, Cloots R, Non-destructive R (2003) evaluation of crystallinity and chemical composition by Raman spectroscopy in hydroxyapatite-coated implants. Mater Lett 58:71–73
Aminzadeh A, Shahabi S, Walsh LJ (1999) Raman spectroscopic studies of CO2 laser-irradiated human dental enamel. Spectrochim Acta Part A 55:1303–1308
Kim HM, Himeno T, Kokubo T, Nakamura T (2005) Process and kinetics of bonelike apatite formation on sintered hydroxyapatite in a simulated body fluid. Biomaterials 26:4366–4373
LeGeros RZ (1993) Biodegradation and bioresorption of calcium phosphate ceramics. Clin Mater 14:65–88
Xue W, Liu X, Zheng X, Ding C (2005) Effect of hydroxyapatite coating crystallinity on dissolution and osseointegration in vivo. J Biomed Mater Res A 74:553–561
Ding Q, Zhang X, Huang Y, Yan Y, Pang X (2015) In vitro cytocompatibility and corrosion resistance of zinc-doped hydroxyapatite coatings on a titanium substrate. J Mater Sci 50:189–202
Wang X, Ito A, Sogo Y, Li X, Oyane A (2010) Zinc-containing apatite layers on external fixation rods promoting cell activity. Acta Biomater 6:962–968
Ito A, Ojima K, Naito H, Ichinose N, Tateishi T (2000) Preparation, solubility, and cytocompatibility of zinc-releasing calcium phosphate ceramics. J Biomed Mater Res 50:178–183
Lu J, Wei J, Yan Y, Li H, Jia J, Wei S, Guo H, Xiao T, Liu C (2011) Preparation and preliminary cytocompatibility of magnesium doped apatite cement with degradability for bone regeneration. J Mater Sci Mater Med 22:607–615
de Lima IR, Alves GG, Soriano CA, Campaneli AP, Gasparoto TH, Ramos Jr ES, de Sena LA, Rossi AM, Granjeiro JM (2011) Understanding the impact of divalent cation substitution on hydroxyapatite: An in vitro multiparametric study on biocompatibility. J Biomed Mater Res Part A 98A:351–358
Tin OM, Gopalakrishna V, Samsuddin AR, Al-Salihi KA, Shamsuria O (2007) Antibacterial property of locally produced hydroxyapatite. Arch Orofac Sci 2:41–44
Sahithi K, Swetha M, Prabaharan M, Moorthi A, Saranya N, Ramasamy K, Srinivasan N, Partridge NC, Selvamurugan N (2010) Synthesis and characterization of nanoscale-hydroxyapatite-copper for antimicrobial activity towards bone tissue engineering applications. J Biomed Nanotechnol 6:333–339
Kolmas J, Groszyk E, Kwiatkowska-Rozycka D (2014) Substituted hydroxyapatites with antibacterial properties BioMed, Res Int 123:1–15Article ID 178
Chung R, Hsieh M, Huang C, Perng L, Wen H, Chin T (2006) Antimicrobial effect and human gingival biocompatibility of hydroxyapatite sol-gel coatings. J Biomed Mater Res B 76B:169–178
Swetha M, Sahithi K, Moorthi A, Saranya N, Saravanan S, Ramasamy K, Srinivasan N, Selvamurugan N (2012) Synthesis, characterization, and antimicrobial activity of nano-hydroxyapatite-zinc for bone tissue engineering applications. J Nanosci Nanotechnol 12:167–172
Udhayakumar G, Muthukumarasamy N, Velauthapillai D, Santhosh SB, Asokan V (2016) Magnesium incorporated hydroxyapatite nanoparticles: preparation, characterization, antibacterial and larvicidal activity. Arabian J Chem. https://doi.org/10.1016/j.arabjc.2016.05.010
Yang Y-C, Chen C-C, Wang J-B, Wang Y-C, Lin F-H (2017) Flame sprayed zinc doped hydroxyapatite coating with antibacterial and biocompatible properties. Ceram Int. https://doi.org/10.1016/j.ceramint.2017.05.318
Acknowledgements
The authors thank the Director, National Centre for Nanoscience and Nanotechnology (NCNSNT) for extending characterization facilities such as SEM and EDS, Department of Nuclear Physics, University of Madras for providing X-ray diffraction facilities and Dr. C. Arulvasu, Department of Zoology, University of Madras for his kind help and valuable suggestions in performing cell growth studies. The authors also thank Dr. S. Kannan, Assistant professor, Centre for Nanoscience and Technology, Pondicherry University, Puducherry, for his guidance and help in performing the Rietveld refinement analysis.
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Highlights
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The HAP and MgZnCo-HAP’s was synthesized by sol-gel method
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The MgZnCo-HAP1 exhibits a better bioactivity, cell viability and good anti-bacterial activity than the un-HAP
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MgZnCo-HAP1 is considered as a promising material for biomedical applications.
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Rajendran, A., Balakrishnan, S., Kulandaivelu, R. et al. Multi-element substituted hydroxyapatites: synthesis, structural characteristics and evaluation of their bioactivity, cell viability, and antibacterial activity. J Sol-Gel Sci Technol 86, 441–458 (2018). https://doi.org/10.1007/s10971-018-4634-x
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DOI: https://doi.org/10.1007/s10971-018-4634-x