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J Funct Biomater
2018 Nov 23;94:. doi: 10.3390/jfb9040067.
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Development of Phosphatized Calcium Carbonate Biominerals as Bioactive Bone Graft Substitute Materials, Part II: Functionalization with Antibacterial Silver Ions.
Sethmann I
,
Völkel S
,
Pfeifer F
,
Kleebe HJ
.
Abstract
Porous calcium phosphate (CaP) materials as bone graft substitutes can be prepared from Ca carbonate biomineral structures by hydrothermal conversion into pseudomorphic CaP scaffolds. The present study aims at furnishing such phosphatized Ca carbonate biomineral (PCCB) materials with antibacterial Ag ions in order to avoid perisurgical wound infections. Prior to this study, PCCB materials with Mg and/or Sr ions incorporated for stimulating bone formation were prepared from coral skeletons and sea urchin spines as starting materials. The porous PCCB materials were treated with aqueous solutions of Ag nitrate with concentrations of 10 or 100 mmol/L, resulting in the formation of Ag phosphate nanoparticles on the sample surfaces through a replacement reaction. The materials were characterized using scanning electron microscopy (SEM) energy-dispersive X-ray spectroscopy (EDS) and X-ray diffractometry (XRD). In contact with Ringer`s solution, the Ag phosphate nanoparticles dissolved and released Ag ions with concentrations up to 0.51 mg/L, as shown by atomic absorption spectroscopy (AAS) analyses. In tests against Pseudomonas aeruginosa and Staphylococcus aureus on agar plates, antibacterial properties were similar for both types of Ag-modified PCCB materials. Concerning the antibacterial performance, the treatment with AgNO₃ solutions with 10 mmol/L was almost as effective as with 100 mmol/L.
Figure 1. Phosphatized and Sr-modified coral skeleton with additional Ag functionalization (sample C Ag 100). (a) Porous structure of the material (environmental scanning electron microscopy-backscatter electron (ESEM-BSE)); (b) a fractured trabecula (fracture edge indicated) showing the internal microstructure of the material on the right hand side (darker shading; center of trabecula indicated) and the material surface (indicated) with a layer of small crystals composed of heavier elements (lighter shading) on the left hand side and dispersed nanocrystals composed of heavy elements (white dots; see inset) (ESEM-BSE). Energy-dispersive X-ray spectroscopy analyses show the chemical composition at (c) the surface and of the interior of the material at depths of (d) 10 µm, (e) 20 µm, and (f) 30 µm. (g) Crystalline phases contained in the material, identified by XRD (aragonite PDF 00-041-1475, hydroxyapatite PDF 00-009-0432, Sr-β-TCP, c.f. [59], Ag phosphate PDF 00-006-0505); XRD raw data S1: C33.
Figure 2. Phosphatized and Sr-modified sea urchin spine with additional Ag functionalization (sample SU Ag 100). (a) Porous structure of the material (ESEM-BSE); (b) a fractured trabecula showing the internal microstructure of the material and rose-shaped aggregates of crystals grown on the surface with additional microcrystals composed of heavy elements dispersed on the surface (white dots; ESEM-BSE). Energy-dispersive X-ray spectroscopy analyses show the chemical composition of (c) the surface material and (d) the bulk material. (e) Cubic microcrystals on the material’s surface. (f) chemical composition of these microcrystals (EDS). (g) Mineral phases contained in the material, identified by XRD (Mg-calcite PDF 00-043-0697, merrillite PDF 01-076-8368, hydroxyapatite PDF 00-009-0432, and Ag phosphate PDF 00-006-0505); XRD raw data S2: SU25.
Figure 3. Antibacterial effects of phosphatized and Sr-modified materials with and without additional Ag functionalization on bacterial strains of Pseudomonas aeruginosa (Gram-negative) and Staphylococcus aureus (Gram-positive). Pseudomonas aeruginosa-seeded agar plates inoculated with (a) coral-derived materials and (b) sea urchin-derived materials and (c) measurements (with standard deviations) of the lateral bacterial inhibition areas for the different materials. Staphylococcus aureus-seeded agar plates inoculated with (d) coral-derived materials and (e) sea urchin-derived materials and (f) measurements (with standard deviations) of the lateral bacterial inhibition areas for the different materials. (C Ag 0 and SU Ag 0: no treatment with Ag ions; C Ag 10 and SU Ag 10: treated with 10 mM Ag solution; C Ag 100 and SU Ag 100: treated with 100 mM Ag solution.)
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