Antimicrobial activity and biologic potential of silver-substituted calcium phosphate constructs produced with self-propagating high-temperature synthesis

Abstract

There is significant demand for synthetic bone substitute materials that can decrease the incidence of implant-based bacterial infections. The intent of this research was to evaluate the antimicrobial activity and biologic potential of calcium phosphate (CaP) constructs substituted with silver (Ag) that were produced via self-propagating high-temperature synthesis (SHS). SHS is a combustion synthesis technique that has successfully generated porous CaP bioceramics intended for use in bone repair. SHS reactions are highly versatile; dopants can be added to the reactant powders to alter product chemistry and morphology. In this research, Ag powder was added to the reactants generating porous CaP constructs containing 0.5, 1, or 2 wt% Ag. Antibacterial performance of the constructs was assessed against Escherichia coli, a representative model for Gram-negative bacteria. Liquid solutions (1 μg/mL) of CaP–Ag particles to phosphate buffered saline were incubated with 105 cells/mL. After 24 h, 10 μL of solution were spread on an LB agar plate and cultured for 24 h at 37 °C. Samples cultured with CaP–Ag showed complete bacterial inhibition while the controls (E. coli only and CaP without Ag) exhibited significant colony formation. The effects of Ag concentration on cytotoxicity and biocompatibility were tested in vitro. At 7 days, osteoblasts uniformly enveloped the CaP–Ag particles and displayed a healthy flattened morphology suggesting the concentrations of Ag incorporated into constructs were not cytotoxic. CaP–Ag constructs produced via SHS represent a source of synthetic bone substitute materials that could potentially inhibit, or reduce the incidence of post-operative bacterial infections.

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References

  1. 1.

    Barrere F, van Blitterswijk C, de Groot K. Bone regeneration molecular and cellular interactions with calcium phosphate ceramics. Int J Nanomed. 2006;3:317–32.

    Google Scholar 

  2. 2.

    Rezwan K, Chen Q, Blaker J, Boccaccini A. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials. 2006;27(18):3413–31. doi:10.1016/j.biomaterials.2006.01.039.

    Article  Google Scholar 

  3. 3.

    Laurencin CT. Bone graft substitute materials. eMedicine from Web MD. 2006.

  4. 4.

    Dorozhkin S. Medical application of calcium orthophosphate bioceramics. Bio. 2011;1(1):1–51. doi:10.5618/bio.2011.v1.n1.1.

    Article  Google Scholar 

  5. 5.

    Ewald A, Hösel D, Patel S, Grover LM, Barralet JE, Gbureck U. Silver-doped calcium phosphate cements with antimicrobial activity. Acta Biomater. 2011;7(11):4064–70. doi:10.1016/j.actbio.2011.06.049.

    Article  Google Scholar 

  6. 6.

    Do Y, Horiguchi T, Moriwaki Y, Kitago H, Kajimoto T, Iwayama Y. Formation of apatite collagen complexes. J Biomed Mater Res. 1996;31:43–9.

    Article  Google Scholar 

  7. 7.

    Du C, Cui F, Feng Q, Zhu X, De Groot K. Tissue response to nanohydroxyapatite collagen composite implants in marrow cavity. J Biomed Mater Res. 1998;42:540–8.

    Article  Google Scholar 

  8. 8.

    LeGeros R. Biodegradation and bioresorption of calcium phosphate ceramics. Clin Mater. 1993;14:65–88.

    Article  Google Scholar 

  9. 9.

    Albrektsson T, Johansson C. Osteoinduction, osteoconduction and osseointegration. Eur Spine J. 2001;10:96–101. doi:10.1007/s005860100282.

    Article  Google Scholar 

  10. 10.

    Declercq H, Verbeeck R, Deridder L, Schacht E, Cornelissen M. Calcification as an indicator of osteoinductive capacity of biomaterials in osteoblastic cell cultures. Biomaterials. 2005;26(24):4964–74. doi:10.1016/j.biomaterials.2005.01.025.

    Article  Google Scholar 

  11. 11.

    Bagambisa F, Joos U. Preliminary studies on the phenomonological behavior of osteoblasts cultured on HA ceramics. Biomaterials. 1990;11:50–6.

    Article  Google Scholar 

  12. 12.

    Hench L, Paschall H. Direct chemical bonding between bioactive glassceramic materials and bone. J Biomed Mater Res Symp. 1973;4:25–42.

    Article  Google Scholar 

  13. 13.

    Dorozhkin SV. Calcium orthophosphates. J Mater Sci. 2007;42(4):1061–95. doi:10.1007/s10853-006-1467-8.

    Article  Google Scholar 

  14. 14.

    Kim T, Feng Q, Kim J, Wu J, Wang H, Chen G, et al. Antimicrobial effects of metal ions (Ag+, Cu2+, Zn2+) in hydroxyapatite. J Mater Sci Mater Med. 1998;9:129–34.

    Article  Google Scholar 

  15. 15.

    Hendriks JGE, van Horn JR, van der Mei HC, Busscher HJ. Backgrounds of antibiotic-loaded bone cement and prosthesis-related infection. Biomaterials. 2004;25(3):545–56. doi:10.1016/S0142-9612(03)00554-4.

    Article  Google Scholar 

  16. 16.

    LeGeros R, Lin S, Rohanizadeh R, Mijares D, LeGeros P. Biphasic calcium phosphate bioceramics preparation, properties and applications. J Mater Sci Mater Med. 2003;14:201–9.

    Article  Google Scholar 

  17. 17.

    Dorozhkin S. Biphasic, triphasic and multiphasic calcium orthophosphates. Acta Biomater. 2011;8(3):963–77.

    Article  Google Scholar 

  18. 18.

    Alt V, Bechert T, Steinrücke P, Wagener M, Seidel P, Dingeldein E, et al. An in vitro assessment of the antibacterial properties and cytotoxicity of nanoparticulate silver bone cement. Biomaterials. 2004;25(18):4383–91. doi:10.1016/j.biomaterials.2003.10.078.

    Article  Google Scholar 

  19. 19.

    Sondi I, Salopek-Sondi B. Silver nanoparticles as antimicrobial agent: a case study on E. coli as a model for Gram-negative bacteria. J Colloid Interface Sci. 2004;275(1):177–82. doi:10.1016/j.jcis.2004.02.012.

    Article  Google Scholar 

  20. 20.

    Raffi M, Hussain F, Bhatti T, Akhter J, Hameed A, Hasan M. Antibacterial characterization of silver nanoparticles against E. coli ATCC-15224. J Mater Sci Technol. 2008;24(2):192–6.

    Google Scholar 

  21. 21.

    Hwang K-S, Hwangbo S, Kim J-T. Silver-doped calcium phosphate nanopowders prepared by electrostatic spraying. J Nanopart Res. 2008;10(8):1337–41. doi:10.1007/s11051-008-9404-1.

    Article  Google Scholar 

  22. 22.

    Chen W, Oh S, Ong AP, Oh N, Liu Y, Courtney HS, et al. Antibacterial and osteogenic properties of silver-containing hydroxyapatite coatings produced using a sol gel process. J Biomed Mater Res, Part A. 2007;82A(4):899–906. doi:10.1002/jbm.a.31197.

    Article  Google Scholar 

  23. 23.

    Munir Z, Naselmi-Tamburini U. Self-propagating exothermic reaction: the synthesis of high temperature materials by combustion. Mater Sci Rep. 1989;3:277–365.

    Article  Google Scholar 

  24. 24.

    Ayers R, Hannigan N, Vollmer N, Unuvar C. Combustion synthesis of heterogeneous calcium phosphate bioceramics from calcium oxide and phosphate precursors. Int J Self Propag High Temp Synth. 2010;20:6–14.

    Article  Google Scholar 

  25. 25.

    Ayers RA, Burkes DE, Gottoli G, Yi H-C, Zhim F, Yahia LH, et al. Combustion synthesis of porous biomaterials. J Biomed Mater Res, Part A. 2007;81A(3):634–43. doi:10.1002/jbm.a.31017.

    Article  Google Scholar 

  26. 26.

    Castillo M. Combustion synthesis of porous tricalcium phosphate, titanium-carbide, and nonstoichiometric titanium-carbide. Golden: Colorado School of Mines; 2004.

    Google Scholar 

  27. 27.

    Gibson IR, Bonfield W. Novel synthesis and characterization of an AB-type carbonate-substituted hydroxyapatite. J Biomed Mater Res. 2002;59(4):697–708.

    Article  Google Scholar 

  28. 28.

    Barralet J, Best S, Bonfield W. Carbonate substitution in precipitated hydroxyapatite An investigation into the effects of reaction temperature and bicarbonate ion concentration. J Biomed Mater Res. 1998;41:79–86.

    Article  Google Scholar 

  29. 29.

    Koutsopoulos S. Synthesis and characterization of hydroxyapatite crystals: a review study on the analytical methods. J Biomed Mater Res. 2002;62:600–12.

    Article  Google Scholar 

  30. 30.

    Chen L, McCrate JM, Lee JCM, Li H. The role of surface charge on the uptake and biocompatibility of hydroxyapatite nanoparticles with osteoblast cells. Nanotechnology. 2011;22(10):105708. doi:10.1088/0957-4484/22/10/105708.

    Article  Google Scholar 

  31. 31.

    Ottani V, Raspanti M, Ruggeri A. Collagen structure and functional implications. Micron. 2001;32:251–60.

    Article  Google Scholar 

  32. 32.

    Harris S, Enger R, Riggs LB, Spelsberg T. Development and characterization of a conditionally immortalized human osteoblast cell line. J Bone Miner Res. 1995;10(2):178–86.

    Article  Google Scholar 

  33. 33.

    Baxter L, Frauchiger V, Textoe M, Ap Gwynn I, Richards R. Fibroblast and osteoblast adhesion and morphology on calcium phosphate surfaces. Eur Cells Mater. 2002;4:1–17.

    Google Scholar 

  34. 34.

    Moore J, Feng H. Combustion synthesis of advanced materials: part II. Classification, applications and modelling. Prog Mater Sci. 1995;39:275–316. doi:10.1016/0079-6425(94)00012-3.

    Article  Google Scholar 

  35. 35.

    Eslamloo-Grami M, Munir ZA. Effect of nitrogen pressure and diluent content on the combustion synthesis of titanium nitride. J Am Ceram Soc. 1990;73(8):2222–7.

    Article  Google Scholar 

  36. 36.

    Descamps M, Hornez J, Leriche A. Effects of powder stoichiometry on the sintering of β-tricalcium phosphate. J Eur Ceram Soc. 2007;27(6):2401–6. doi:10.1016/j.jeurceramsoc.2006.09.005.

    Article  Google Scholar 

  37. 37.

    Zhou J, Zhang X, Chen J. High temperature characteristics of synthetic HA. J Mater Sci Mater Med. 1993;4:83–5.

    Article  Google Scholar 

  38. 38.

    Nuechterlein J. Production of ceramic nanoparticles through self-propagating high-temperature synthesis (SHS) and their introduction into a metallic matrix to form metal matrix composites (MMC). Golden: Colorado School of Mines; 2013.

    Google Scholar 

  39. 39.

    Faust R. Formal toxicity summary for SILVER. 1992; 2013(1-24-13). https://rais.ornl.gov/tox/profiles/silver_f_V1.html.

  40. 40.

    Yamada S, Heymann D, Bouler J, Daculsi G. Osteoclastic resorption of calcium phospahte ceramics with different HA-bTCP ratios. Biomaterials. 1997;18(15):1037–41. doi:10.1016/S0142-9612(97)00036-7.

    Article  Google Scholar 

  41. 41.

    Moore J, Feng H. The combustion synthesis of advanced materials: part I. Reaction parameters. Prog Mater Sci. 1995;39(4–5):243–73. doi:10.1016/0079-6425(94)00011-5.

    Article  Google Scholar 

  42. 42.

    Wang X, Nyman J, Reyes M, Dong X, Leng H, Athanasiou K. Fundamental biomechanics in bone tissue engineering synthesis lectures on tissue engineering. San Rafael: Morgan and Claypool Publishers; 2010. doi:10.2200/S00246ED1V01Y200912TIS004.

    Google Scholar 

  43. 43.

    Le Huec J, Schaeverbeke T, Clement D, Faber J, Le Rebeller A. Influence of porosity on the mechanical resistance of HA ceramics under compressive stress. Biomaterials. 1995;16:113–6.

    Article  Google Scholar 

  44. 44.

    Meille S, Lombardi M, Chevalier J, Montanaro L. Mechanical properties of porous ceramics in compression: on the transition between elastic, brittle, and cellular behavior. J Eur Ceram Soc. 2012;32(15):3959–67. doi:10.1016/j.jeurceramsoc.2012.05.006.

    Article  Google Scholar 

  45. 45.

    Barsoum M. Fundamentals of ceramics. McGraw-Hill series in materials science and engineering. New York: The McGraw-Hill Companies Inc.; 1997.

    Google Scholar 

  46. 46.

    Daculsi G, Passuti N. Effect of the macroporosity for osseous substitution of calcium phosphate ceramics. Biomaterials. 1990;11:86–7.

    Google Scholar 

  47. 47.

    Vincent J. Structural biomaterials. Princeton: Princeton University Press; 1990.

    Google Scholar 

  48. 48.

    Jackson S, Cartwright A, Lewis D. The morphology of bone mineral crystals. Calcif Tissue Int. 1978;25:217–22.

    Article  Google Scholar 

  49. 49.

    Carter DR, Spengler DM. Mechanical properties and composition of cortical bone. Clin Orthop Relat Res. 1978;135:192–217.

    Google Scholar 

  50. 50.

    de Bruijn J, Klein C, de Groot K, van Blitterswijk C. The ultrastructure of the bone hydroxyapatite interface invitro. J Biomed Mater Res. 1992;26:1365–82.

    Article  Google Scholar 

  51. 51.

    Glimcher M. The nature of the mineral component of bone and the mechanism of calcification. Instr Course Lect. 1987;36:49–69.

    Google Scholar 

  52. 52.

    LeGeros RZ. Effect of carbonate on the lattice parameters of apatite. Nature. 1965;206:403–4. doi:10.1038/206403a0.

    Article  Google Scholar 

  53. 53.

    Sudarsanan K, Young R. Structure of strontium hydroxide phosphate, Srs(PO4)3OH. Acta Crystallogr A. 1972;B28:3668–70.

    Article  Google Scholar 

  54. 54.

    Elliott JC. Structure and chemistry of the apatites and other calcium orthophosphates. Amsterdam: Elsevier; 1994.

    Google Scholar 

  55. 55.

    Kreidler E, Hummel F. Phase relationships in the system SrO–P2O5 and the influence of water vapor on the formation of Sr4P2O5. J Inorg Chem. 1967;6(5):884–91.

    Article  Google Scholar 

  56. 56.

    Dorozhkin S. Amorphous calcium (ortho)phosphates. Acta Biomater. 2010;6:4457–73.

    Article  Google Scholar 

  57. 57.

    Walsh W. Bone composite behaviour: effects of mineral organic bonding. J Mater Sci - Mater Med. 1994;5:72–9.

    Article  Google Scholar 

  58. 58.

    Gloria A, Ronca D, Russo T, D’Amora U, Chierchia M, De Santis R, et al. Technical features and criteria in designing fiber-reinforced composite materials: from the aerospace and aeronautical field to biomedical applications. J Appl Biomater Biomech: JABB. 2011;9(2):151–63. doi:10.5301/jabb.2011.8569.

    Google Scholar 

  59. 59.

    Rice R. Mechanisms of toughening in ceramic matrix composites. In: Proceedings of the 5th annual conference on composites and advanced ceramic materials: ceramic engineering and science proceedings. 1981;2(7/8):661–98. doi:10.1002/9780470291092.ch20.

  60. 60.

    Power L, Itier S, Hawton M, Schraft H. Time lapse confocal microscopy studies of bacterial adhesion to self assembled monolayers. Langmuir. 2007;23:5622–9.

    Article  Google Scholar 

  61. 61.

    Nishizawa K, Toriyama M, Suzuki T, Kawamoto Y, Yokogawa Y, Nagae H. Effects of the surface wettability and zeta potential of bioceramics. J Ferment Bioeng. 1993;75(6):435–7.

    Article  Google Scholar 

  62. 62.

    Tamada Y, Ikada Y. Fibroblast growth on polymer surfaces and biosynthesis of collagen. J Biomed Mater Res. 1994;28:783–9.

    Article  Google Scholar 

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Acknowledgments

The authors wish to acknowledge Dr. John Chandler and Gary Zito for their assistance during materials characterization. Additionally, the authors would like to thank Ryan Hort for his aseptic E. coli training, and Josh Cruz, Casey Davis, and Iris Vollmer for their assistance conducting this research. This work was accomplished under the Foundation for the National Institutes of Health Grant 1R15AR060011-01, and the Colorado Bioscience Discovery Evaluation Grant Program Grant 11BGF-48.

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Correspondence to N. L. Vollmer.

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Vollmer, N.L., Spear, J.R. & Ayers, R.A. Antimicrobial activity and biologic potential of silver-substituted calcium phosphate constructs produced with self-propagating high-temperature synthesis. J Mater Sci: Mater Med 27, 104 (2016). https://doi.org/10.1007/s10856-016-5715-4

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Keywords

  • Contact Angle
  • Bone Replacement Material
  • Combustion Synthesis Technique
  • Bone Cell Attachment
  • Human Fetal Osteoblast Cell Line