Biologically Inspired Collagen/Apatite Composite Biomaterials for Potential Use in Bone Tissue Regeneration-A Review.

Type I collagen and nanocrystalline-substituted hydroxyapatite are the major components of a natural composite-bone tissue. Both of these materials also play a significant role in orthopedic surgery and implantology; however, their separate uses are limited; apatite is quite fragile, while collagen's mechanical strength is very poor. Therefore, in biomaterial engineering, a combination of collagen and hydroxyapatite is used, which provides good mechanical properties with high biocompatibility and osteoinduction. In addition, the porous structure of the composites enables their use not only as bone defect fillers, but also as a drug release system providing controlled release of drugs directly to the bone. This feature makes biomimetic collagen-apatite composites a subject of research in many scientific centers. The review focuses on summarizing studies on biological activity, tested in vitro and in vivo.

The solid-state proton NMR study of bone using a dipolar filter: apatite hydroxyl content versus animal age.

The hydroxyl content of bone apatite mineral has been measured using proton solid-state NMR performed with a multiple-pulse dipolar filter under slow magic angle spinning (MAS). This new method succeeded in resolving and relatively enhancing the main hydroxyl peak at ca. 0 ppm from whole bone, making it amenable to rigorous quantitative analysis. The proposed methodology, involving line fitting, the measurement of the apatite concentration in the studied material and adequate calibration, was proved to be convenient and suitable for monitoring bone mineral hydroxylation in different species and over the lifetime of the animal. It was found that the hydroxyl content in the cranial bone mineral of pig and rats remained in the 5-10% range, with reference to stoichiometric hydroxyapatite. In rats, the hydroxyl content showed a non-monotonic increase with age, which was governed by biological processes rather than by chemical, thermodynamically driven apatite maturation.

Kinetics of 1H --> 31P NMR cross-polarization in bone apatite and its mineral standards.

Kinetics of NMR cross-polarization (CP) from protons to phosphorus-31 nuclei was studied in the following samples: mineral of whole human bone, apatite prepared from bone, natural brushite, synthetic hydroxyapatite (hydrated and calcined), and synthetic carbonatoapatite of type B with 9 wt% of CO(3) (2-). In order to avoid an effect of magic angle spinning on CP and relaxation, the experiments were carried out on static samples. Parameters of the CP kinetics were discussed for trabecular and cortical bone tissue from adult subjects in comparison to the synthetic mineral standards. It was found that carbonatoapatite shows similar CP behavior to the bone mineral. Both materials undergo two-component CP kinetics. The fast-relaxing classical component is from the surface of apatite crystals and the slow-relaxing nonclassical component comes from the crystal interior. The components have been unambiguously assigned using inverse CP from phosphorus-31 to protons. The study provides information on a structured water layer, which covers crystal surface of carbonato- and bone apatite. The layer encompasses ca 40% of apatite phosphorus and its thickness is more than ca 2 nm.

Substitution of strontium and boron into hydroxyapatite crystals: Effect on physicochemical properties and biocompatibility with human Wharton-Jelly stem cells.

Hydroxyapatite (HA) enriched with strontium and boron ions was synthesized using two different methods: the precipitation method (Sr,B-HAw) and the dry method (Sr,B-HAd). Additionally, for the sake of comparison, the "pure" unsubstituted HA was prepared together with HAs substituted only with one type of a foreign ion. The obtained materials were subjected to physicochemical analysis with the use of various analytical methods, such as powder X-ray diffraction (PXRD), transmission electron microscopy (TEM), inductively coupled plasma optical emission spectroscopy (ICP-OES), Fourier transform infrared spectroscopy (FT-IR) and solid-state proton nuclear magnetic resonance (1H ssNMR). All the obtained materials were also biologically tested for their potential cytotoxicity. The obtained materials (Sr,B-HAw and Sr,B-HAd) were homogeneous and respectively showed nano- and microcrystal apatitic structures. The simultaneous introduction of Sr2+ and BO33- ions turned out to be more effective in respect of the dry method. Of importance, doped materials obtained using both synthesis routes have been demonstrated to be biocompatible, opening the way for medical applications.

Phosphorus-31 spin-lattice NMR relaxation in bone apatite and its mineral standards.

Phosphorus-31 spin-lattice relaxation, both in the laboratory (B(0)=4.7 T) and rotating frame (B(1)=2.2 mT), was studied in the following samples: mineral of whole human bone (samples B1-B6), apatite prepared from bone (BHA), natural brushite (BRU), synthetic hydroxyapatite hydrated (HAh) and calcined (HAc), and synthetic carbonatoapatite of type B (CHA-B) with 9 wt% of CO(3)(2-). The T(1)(P) relaxation time was determined directly using the saturation recovery technique, while the T(1 rho)(P) relaxation time was measured via (1)H-->(31)P CP by incrementing the (31)P spin-lock. In order to avoid an effect of magic-angle spinning (MAS) on CP and relaxation, the experiments were carried out on static samples. The (31)P spin-lattice relaxation was discussed for trabecular and cortical bone tissue from adult subjects in comparison to the synthetic mineral standards. None of the reference materials has matched accurately the relaxation behaviour of the bone mineral. The most striking differences between the examined substances were observed for T(1)(P), which for human bone was sample dependent and appeared in the range 55-100 s, while for HAh, HAc, and CHA-B was 7.2, 10.0, and 25.8 s, respectively. Possible reasons of so large relaxation diversity were discussed. It has been suggested that T(1)(P) of apatites is to some extent dependent on the concentration of the structural hydroxyl groups, and this in turn is controlled by the material crystallinity. It was also found that T(1)(P) decreased on hydration by ca. 30%. For T(1rho)(P), both its magnitude and dependence on the CP contact time gave useful structural information. The dehydrated samples (HAc and BHA) had long T(1 rho)(P) over 250 ms. Those, which contained water, either structural (BRU) or adsorbed on the crystal surface (HAh, CHA-B, and B1-B6), had shorter T(1 rho)(P) below 120 ms. It was concluded that the effect of water on T(1 rho)(P) is much more pronounced than on T(1)(P). The interpretation has involved P-OH groups and adsorbed water, which cover the apatite crystal surface.

Efficiency of 1H --> 31P NMR cross-polarization in bone apatite and its mineral standards.

Human bone mineral was studied using solid-state 31P NMR with cross-polarization (CP) from protons. The CP efficiency was determined for trabecular and cortical bone tissue from human adults and compared with synthetic mineral standards. The study shows the similarity between carbonatoapatite of type B and bone mineral as shown by their CP behaviour. The method can be used for the characterization of synthetic apatite-based implant materials.