Bone regeneration: molecular and cellular interactions with calcium phosphate ceramics.

Calcium phosphate bioceramics are widely used in orthopedic and dental applications and porous scaffolds made of them are serious candidates in the field of bone tissue engineering. They have superior properties for the stimulation of bone formation and bone bonding, both related to the specific interactions of their surface with the extracellular fluids and cells, ie, ionic exchanges, superficial molecular rearrangement and cellular activity.

Bone tissue engineering on amorphous carbonated apatite and crystalline octacalcium phosphate-coated titanium discs.

Poor fixation of bone replacement implants, e.g. the artificial hip, in implantation sites with inferior bone quality and quantity may be overcome by the use of implants coated with a cultured living bone equivalent. In this study, we tested, respectively, amorphous carbonated apatite (CA)- and crystalline octacalcium phosphate (OCP)-coated discs for their use in bone tissue engineering. Subcultured rat bone marrow cells were seeded on the substrates and after 7 days of culture, the implants were subcutaneously implanted in nude mice for 4 weeks. After 7 days of culture, the cells had formed a continuous multi-layer that covered the entire surface of the substrates. The amount of cells was visually higher on the crystalline OCP-coated discs compared to the amorphous CA-coated discs. Furthermore, the amorphous CA-coated discs exhibited a visually higher amount of mineralized extracellular matrix compared to the crystalline OCP-coated discs. After 4 weeks of implantation, clear de novo bone formation was observed on all discs with cultured cells. The newly formed bone on the crystalline OCP-coated discs was more organized and revealed a significantly higher volume compared to the amorphous CA-coated discs. The percentage of bone contact with the discs was also significantly higher on the OCP-coated discs. Overall, the results suggest that a crystalline OCP coating is more suitable for bone tissue engineering than an amorphous CA coating.

Nano-scale study of the nucleation and growth of calcium phosphate coating on titanium implants.

The nucleation and growth of a calcium phosphate (Ca-P) coating deposited on titanium implants from simulated body fluid was investigated by using atomic force microscopy (AFM) and environmental scanning electron microscopy (ESEM). Forty titanium alloy plates were assigned into two groups. One group with a smooth surface having a maximum roughness R(max) < 0.10 microm (s-Ti6Al4V) and a group with a rough surface with an R(max) < 0.25 microm (r-Ti6Al4V) were used. Titanium samples were immersed in SBF concentrated by five (SBF x 5) from 10 min to 5 h and examined by AFM and ESEM. Scattered Ca-P deposits of approximately 15 nm in diameter appeared after only 10 min of immersion in SBF x 5. These Ca-P deposits grew up to 60-100 nm after 4 h on both s- and r-Ti6Al4V substrates. With increasing immersion time, the packing of Ca-P deposits with size of tens of nanometers in diameter formed larger globules and then a continuous Ca-P film on titanium substrates. A direct contact between the Ca-P coating and the Ti6Al4V surface was observed. The Ca-P coating was composed of nanosized deposits and of an interfacial glassy matrix. This interfacial glassy matrix might ensure the adhesion between the Ca-P coating and the Ti6Al4V substrate. In the case of s-Ti6Al4V substrate, failures within this interfacial glassy matrix were observed overtime. Part of the glassy matrix remained on s-Ti6Al4V while part detached with the Ca-P film. The Ca-P coating detached from the smooth substrate, whereas the Ca-P film extended onto the whole rough titanium surface over time. In the case of r-Ti6Al4V, the Ca-P coating covered evenly the substrate after immersion in SBF x 5 for 5 h. The present study suggested that the heterogeneous nucleation of Ca-P on titanium was immediate and did not depend on the Ti6Al4V surface topography. The further growth and mechanical attachment of the final Ca-P coating strongly depended on the surface, for which a rough topography was beneficial.

Osteogenecity of octacalcium phosphate coatings applied on porous metal implants.

The biomimetic route allows the homogeneous deposition of calcium phosphate (Ca-P) coatings on porous implants by immersion in simulated physiologic solution. In addition, various Ca-P phases, such as octacalcium phosphate (OCP) or bone-like carbonated apatite (BCA), which are stable only at low temperatures, can be deposited. In this pilot study, experiments were designed with a twofold-purpose: (1) to investigate the osteoinduction of OCP-coated and noncoated porous tantalum cylinders and of dense titanium alloy cylinders (5 mm in diameter and 10 mm in length) in the back muscle of goats at 12 and 24 weeks (n = 4); and (2) to compare the osteogenic potentials of BCA-coated, OCP-coated, and bare porous tantalum cylinders in a gap of 1 mm created in the femoral condyle of a goat at 12 weeks (n = 2). In the goat muscle, after 12 weeks the OCP-coated porous cylinder had induced ectopic bone as well as bone within the cavity of the OCP-coated dense titanium cylinder. In the femoral condyle, bone did not fill the gap in any of the porous implants. In contrast with the two other groups, OCP-coated porous cylinders exhibited bone formation in the center of the implant. The nature of the Ca-P coating, via its microstructure, its dissolution rate, and its specific interactions with body fluids, may influence the osteogenecity of the Ca-P biomaterial.

Calcium phosphate interactions with titanium oxide and alumina substrates: an XPS study.

Besides the excellent mechanical properties of titanium and alumina (Al(2)O(3)) in the case of load bearing applications, their bone-bonding properties are very different. In osseous environment, Al(2)O(3) ceramic is encapsulated by fibrous tissues, whereas bone can bind directly to titanium, via its natural titanium dioxide (TiO(2)) passivation layer. So far, this calcification dissimilarity between TiO(2) and Al(2)O(3) was attributed to respectively their negative and positive surface charge under physiological conditions. The present study aims at studying the chemical interactions between TiO(2) and Al(2)O(3) (phase alpha) with the diverse ions contained in simulated body fluids (SBFs) buffered with trishydroxymethyl aminomethane (TRIS) at pH=6.0 and pH=7.4. After 1 h of immersion, TiO(2) and alpha-Al(2)O(3) powders were analyzed by X-ray photoelectron spectroscopy (XPS). The results indicated that Ca and HPO(4) groups were present on TiO(2) surface. In addition, HPO(4) groups were found to be in a higher amount than Ca on TiO(2), which does not comply with the surface charge theory. With regard to Al(2)O(3), little HPO(4) but no Ca was detected on its surface, and TRIS bound to Al(2)O(3) substrate in all of the immersion experiments. The fact that both Ca and HPO(4) were present at the vicinity of TiO(2) might be at the origin of its calcification ability. On the other hand, Al(2)O(3) did not show any affinity towards Ca and HPO(4) ions. This might explain the inability of Al(2)O(3) substrate to calcify.