Development of Phosphatized Calcium Carbonate Biominerals as Bioactive Bone Graft Substitute Materials, Part I: Incorporation of Magnesium and Strontium Ions.

Synthetic materials based on calcium phosphate (CaP) are frequently used as bone graft substitutes when natural bone grafts are not available or not suitable. Chemical similarity to bone guarantees the biocompatibility of synthetic CaP materials, whereas macroporosity enables their integration into the natural bone tissue. To restore optimum mechanical performance after the grafting procedure, gradual resorption of CaP implants and simultaneous replacement by natural bone is desirable. Mg and Sr ions released from implants support osteointegration by stimulating bone formation. Furthermore, Sr ions counteract osteoporotic bone loss and reduce the probability of related fractures. The present study aimed at developing porous Ca carbonate biominerals into novel CaP-based, bioactive bone implant materials. Macroporous Ca carbonate biominerals, specifically skeletons of corals (aragonite) and sea urchins (Mg-substituted calcite), were hydrothermally converted into pseudomorphic CaP materials with their natural porosity preserved. Sr ions were introduced to the mineral replacement reactions by temporarily stabilizing them in the hydrothermal phosphate solutions as Sr-EDTA complexes. In this reaction system, Na, Mg, and Sr ions favored the formation of correspondingly substituted ß-tricalcium phosphate over hydroxyapatite. Upon dissolution, the incorporated functional ions became released, endowing these CaP materials with bioactive and potentially osteoporotic properties.

Development of Phosphatized Calcium Carbonate Biominerals as Bioactive Bone Graft Substitute Materials, Part II: Functionalization with Antibacterial Silver Ions.

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.

Microstructures of Randall's plaques and their interfaces with calcium oxalate monohydrate kidney stones reflect underlying mineral precipitation mechanisms.

Randall's plaques (RP) are preferred sites for the formation of calcium oxalate monohydrate (COM) kidney stones. However, although processes of interstitial calcium phosphate (CaP) plaque formation are not well understood, the potential of plaque microstructures as indicators of CaP precipitation conditions received only limited attention. We investigated RP-associated COM stones for structural details of the calcified tissues and microstructural features of plaque-stone interfaces as indicators of the initial processes of stone formation. Significantly increased CaP supersaturation can be expected for interstitial fluid, if reabsorbed ions from the tubular system continuously diffuse into the collagenous connective tissue. Densely packed, fine-grained CaP particles were found in dense textures of basement membranes while larger, laminated particles were scattered in coarse-meshed interstitial tissue, which we propose to be due to differential spatial confinements and restrictions of ion diffusion. Particle morphologies suggest an initial precipitation as metastable amorphous calcium phosphate (ACP). Morphologies and arrangements of first COM crystals at the RP-stone interface ranged from stacked euhedral platelets to skeletal morphologies and even porous, dendritic structures, indicating, in this order, increasing levels of COM supersaturation. Furthermore, these first COM crystals were often coated with CaP. On this basis, we propose that ions from CaP-supersaturated interstitial fluid may diffuse through porous RP into the urine, where a resulting local increase in COM supersaturation could trigger crystal nucleation and, hence, initiate stone formation. Ion-depleted fluid in persistent pores of initial COM layers may get replenished from interstitial fluid, leading to CaP precipitation in porous COM.

Structure and composition of calcareous sponge spicules: a review and comparison to structurally related biominerals.

Since the early 19th century, the skeletons of calcareous sponges (Porifera: Calcarea) with their mineralized spicules have been investigated for their morphologies, structures, and mineralogical and organic compositions. These biomineral spicules, up to about 10mm in size, with one to four rays called actines, have various specific shapes and consist mainly of magnesium-calcite: in only one case has an additional phase of stabilized amorphous CaCO3 (ACC) been discovered. The spicules are invariably covered by a thin organic sheath and display a number of intriguing properties. Despite their complex morphologies and rounded surfaces without flat crystal faces they behave largely as single crystal individuals of calcite, and to some degree crystallographic orientation is related to morphology. Despite their single-crystalline nature, most spicules show nearly isotropic fracture behaviour, not typical for calcite crystals, indicating enhanced fracture resistance. These unusual morphological and mechanical properties are the result of their mechanism of growth. Each spicule is formed by specialized cells (sclerocytes) that supply mineral ions or particles associated by organic macromolecules to extracellular cavities, where assembly and crystallization in alignment with an initial seed crystal (nucleus) takes place. As a result of discontinuous mineral deposition, cross-sections of larger spicules display concentric layering that mantles a central calcitic rod. On a smaller scale, the entire spicule displays a 'nano-cluster' structure with crystallographically aligned and putatively semicoherent crystal domains as well as a dispersed organic matrix intercalated between domain boundaries. This ultrastructure dissipates mechanical stress and deflects propagating fractures. Additionally, this nano-cluster construction, probably induced by intercalated organic substances, enables the formation of complex crystal morphologies independent of crystal faces. In this review, the current knowledge about the structure, composition, and formation of calcareous sponge spicules is summarised and discussed. Comparisons of calcareous sponge spicules with the amorphous silica spicules of sponges of the classes Hexactinellida and Demospongiae, as well as with calcitic skeletal elements of echinoderms are drawn. Despite the variety of poriferan spicule mineralogy and the distant phylogenetic relationship between sponges and echinoderms, all of these biominerals share similarities regarding their nano-scale construction. Furthermore, echinoderm skeletal elements resemble calcareous sponge spicules in that they represent magnesium-bearing calcite single-crystals with extremely complex morphologies.

Nano-cluster composite structure of calcitic sponge spicules--a case study of basic characteristics of biominerals.

Spicules of calcareous sponges are elaborately shaped skeletal elements that nonetheless show characteristics of calcite single-crystals. Our atomic force microscopic and transmission electron microscopic investigation of the triradiate spicules of the sponge Pericharax heteroraphis reveals a nano-cluster structure with mostly well-aligned small crystal domains and pockets with accumulated domain misalignments. Combined high-resolution and energy-filtering transmission electron microscopy revealed carbon enrichments located in between crystal domain boundaries, which strongly suggests an intercalated network-like proteinaceous organic matrix. This matrix is proposed to be involved in the nano-clustered calcite precipitation via a transient phase that may enable a 'brick-by-brick' formation of composite and yet single-crystalline spicules with elaborate morphologies. This composite cluster structure reduces the brittleness of the material by dissipating strain energy and deflecting crack propagation from the calcite cleavage planes, but the lattice symmetry and anisotropic growth properties of calcite still play a major role in the morphogenesis of these unusual calcite single-crystals. Our structural, crystallographic, textural, and chemical analysis of sponge spicules corroborates the view that nano-clustered crystal growth, induced by organic matrices, is a basic characteristic of biomineralisation that enables the production of composite materials with elaborate morphologies.