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Biological systems have a remarkable capability to produce perfect fine structures such as seashells, pearls, bones, teeth and corals. These structures are composites of interacting inorganic (calcium phosphate or carbonate minerals) and organic counterparts. It is difficult to say with certainty which part has the primary role. For example, the growth of molluscan shell crystals is thought to be initiated from a solution by the extracellular organic matrix (ECM). According to this theory, the matrix induces nucleation of calcium containing crystals. Recently, an alternative theory has been put forward, stating that a class of granulocytic hemocytes would be directly involved in shell crystal production in oysters. In the work presented here the surface of AISI 316 stainless steel was modified by deposition of ECM proteins. The ability of the modified substrates to induce nucleation and growth of hydroxyapatite (HA) from simulated body fluid (SBF) was examined by a kinetic study using two methods: (1) a simple soaking process in SBF and (2) a laser-liquid-solid interaction (LLSI) process which allows interaction between a scanning laser beam and a solid substrate immersed in SBF. The deposited HA layers were investigated by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). It was found that a coating of stainless steel surface with native ECM proteins induced nucleation and growth of HA and facilitated its crystallization. By the process of simple soaking of the samples, irrespective of their horizontal or vertical position in the solution, HA layers were grown due to the reactive ECM-coated stainless steel surface. It was shown that the process occurring in the first stages of the growth was not only a result of the force of gravity. The application of the LLSI process strongly influenced HA formation on the ECM-modified substrates by promoting and enhancing the HA nucleation and growth through a synergistic effect of a few stimuli, i.e., the modified solid surface, the laser beam and the aqueous solution.

Serratia sp. NCIMB 40259 is a non-pathogenic Gram-negative bacterium that is able to produce hydroxyapatite by a mechanism involving enzymic cleavage of organic phosphates. Serratia bacteria can attach and form a biofilm on glass, plastics, ceramics and metals and the method can be used to form three dimensional porous scaffolds and for coating 3D structures with hydroxyapatite. The production of calcium phosphate is driven by an acid phosphatase enzyme located in the bacterial cell wall, on fimbriae and within the bacterial extracellular polymeric matrix. Calcium phosphate ceramic may be obtained by two methods: In the first, crystals of calcium phosphate are formed extracellularly within the pre-formed bacterial biofilm grown on the substrata. In the second method, planktonic bacteria catalyse the formation of CaP in suspension and on solid substrata placed in the same container. Composite thin layer of silicon-based polymer and detonated nanodiamond (DND) particles was used as substrate for the process of biomineralization by a species of Serratia. The plasma polymerization (PP) method was chosen to obtain composites of silicon-based polymer, in which DND particles were incorporated. Over the past decades carbon-based nanostructures have been the focus of intense research due to their unique chemical and physical properties. Recently it was shown that the incorporation of the DND particles in a polymer matrix (an organosilicon polymer) changes their physico-chemical properties. The composite films are homogeneous, chemically resistant, thermally and mechanically stable, thus allowing a large amount of biological components to be loaded onto their surface and to be used in tissue engineering, regenerative medicine, implants, stents, biosensors and other medical and biological devices. The aim of this study was to investigate the process of biomineralisation by Serratia bacteria on various composites of silicon-based polymer and detonated nanodiamond particles, and to determine the effect of composite surface on the process of biomineralisation. [ABSTRACT FROM AUTHOR]

Recently, nanodiamonds (NDs) have attracted great interest due to their unique physical and chemical properties that could be used in various biological applications. However, depending on the origin, NDs often contain different impurities which may affect cellular functions and viability. Therefore, before their biomedical application, the cytotoxicity of newly produced NDs should be assessed. In the present study, we have evaluated cytotoxicity of four types of ND particles with two cell models: a human osteosarcoma cell line, MG-63, and primary rat mesenchymal stem cells (rMSCs). Detonation-generated nanodiamond (DND) particles were purified with different acid oxidizers and impurities' content was determined by elemental analysis. The particles size distribution was measured revealing that the DND particles have an average size in the range of 51-233 nm. Cytotoxicity was assessed by optical microscopy and proliferation assay after 72 hours exposure of the cells to nanoparticles. We observed cell-specific and material-specific toxicity for all tested particles. Primary stem cells demonstrated higher sensitivity to DND particles than osteosarcoma cells. The most toxic were the DND particles with the smallest grain size and slight content of non-diamond carbon, while DNDs with higher grain size and free from impurities had no significant influence on cell proliferation and morphology. In addition, the smaller DND particles were found to form large aggregates mainly during incubation with rMSCs. These results demonstrate the role of the purification method on the properties of DND particles and their cytotoxicity as well as the importance of cell types used for evaluation of the nanomaterials.