Interfacial phenomena: an in vitro study of the effect of calcium phosphate (Ca-P) ceramic on bone formation.

In previous studies we developed a RF magnetron sputter technique for the production of thin Ca-P coatings. With this technique coatings can be produced that vary in Ca/P ratio as well as in structural appearance. The aim of this investigation was to obtain more understanding of the biological behavior of these coatings by way of in vitro experiments. The effect of noncoated titanium (Ti) and three different Ca-P-sputtered surfaces on the proliferation and differentiation (morphology and matrix production) of osteoblast-like cells was studied. Proliferation was determined using counting procedures; morphology was studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Fluorescent markers and energy-dispersive X-ray microanalysis (EDX) were used to obtain quantitative and compositional information about the resultant calcified extracellular matrix (ECM). Results demonstrated that proliferation of the osteoblast-like cells was significantly (p < 0.05) higher on noncoated than on Ca-P-coated samples. On the other hand, more mineralized ECM was formed on the coated surfaces. In addition, TEM confirmed that the cells on the coated substrates were surrounded by ECM with collagen fibers embedded in crystallized, needle-shaped structures. On the basis of these findings, we concluded that: (1) the investigated Ca-P sputter coatings possess the capacity to activate the differentiation and expression of osteogenic cells, and (2) bone formation proceeds faster on Ca-P surfaces than on Ti substrates. Further, this bone-inductive effect appeared to be dependent on the Ca-P ratio of the deposited coatings.

Mechanical and histologic evaluation of Ca-P plasma-spray and magnetron sputter-coated implants in trabecular bone of the goat.

To aim of this study was to investigate the bone response to calcium phosphate (Ca-P) plasma-spray and radiofrequency magnetron sputter-coated implants with comparable roughness. Therefore, tapered conical screw designed implants were installed in the trabecular bone of the femurs of nine goats. They were provided with two types of coatings, a plasma-spray dual coating of fluorapatite and hydroxyapatite (FA/HA-PS) and a titanium plasma-spray coating, covered with an amorphous Ca-P magnetron sputtercoating (TPS/Ca-P-a). These implants were evaluated histologically and mechanically after 3 months of implantation. A well-controlled method to apply and measure a torsional force to load the screw-type implants to the point of failure was introduced. All implants healed uneventful and were well fixed. No significant difference (Student t test, p > 0.05) for the torsional failure force was measured for both type of coatings. Nevertheless, SEM revealed differently situated fracture planes. Light microscopy showed intimate bone-implanted contact for both types of coatings; original drill margins were still visible. A lamellar type of bone with some remodeling lacunae was shown. Histomorphometry revealed a higher percentage of bone contact for the FA/HA-PS-coated implants (students t test, p < 0.05). Measurement of the amount of bone revealed more bone mass around TPS/Ca-P-a-coated implants (analysis of variance and Turkey multiple comparison, p < 0.05).

Initial interfacial healing events around calcium phosphate (Ca-P) coated oral implants.

The bone response to different calcium phosphate (Ca-P) coated and non-coated titanium implants was evaluated in a goat animal model. Two types of Ca-P coatings have been investigated: an experimental plasma-spray bilayered Ca-P coating (FA-HA) and an amorphous RF magnetron sputter coating (Ca-P-a). Fifty-four conical screw shaped implants were inserted in the lateral and medial femoral condyles of 18 Saanen goats. After implantation periods of 3, 12 and 24 days, the bone-implant interface was evaluated histologically and histomorphometrically. Light microscopical evaluation revealed that bone formation on the Ca-P coated implants proceeded faster. At 24 days higher percentages of bone contact were measured for both Ca-P coated implants than for non-coated implants. However, this difference was only significant for the FA-HA coated implants. On basis of these findings, we concluded that Ca-P coatings show improved bone response due to an initial difference in bone cell response.

Evaluation of plasma-spray and magnetron-sputter Ca-P-coated implants: an in vivo experiment using rabbits.

The bone response to different plasma-spray and magnetron-sputter calcium phosphate (Ca-P)-coated implants was evaluated in a rabbit animal model. Four types of Ca-P coatings have been investigated: a plasma-spray Ca-P coating (HA-PS), a heat-treated plasma-spray Ca-P coating (HA-PS/ht), an amorphous magnetron-sputter coating (Ca-P-a), and a crystalline magnetron-sputter coating (CA-P-c). Seventy-two specially designed cylindrical implants were inserted in the lateral and medial femoral condyles of 18 New Zealand White rabbits. The four differently coated implants were positioned in one animal according to a split-plot design. After implantation periods of 3, 6, and 9 weeks, the bone-implant interface was evaluated histologically. Besides descriptive light microscopical evaluation, quantitative histomorphometrical measurements were done to determine bone contact and the amount of bone surrounding the implant-bone interface. Light microscopical examination revealed that all types of coatings followed the same process of bone healing. Measurements of bone contact at 6 and 9 weeks did not reveal significant differences between the various coatings. For the amount of bone, in a circular region at a certain distance from the implant, the Ca-P-c-coated implants showed a significantly greater amount of bone after 6 weeks of implantation than did the other three Ca-P coatings. At 9 weeks this difference could no longer be measured. On the basis of these findings we concluded that magnetron-sputtered Ca-P coatings show the same process of bone healing as the plasma-sprayed Ca-P coatings when inserted into the trabecular femoral bone of rabbits.

Biological evaluation of the effect of magnetron sputtered Ca/P coatings on osteoblast-like cells in vitro.

A rat bone marrow cell culture was used to evaluate the osteogenic potential of amorphous and crystalline thin calcium phosphate (Ca/P) coatings. The coatings were deposited on titanium discs using a radiofrequency magnetron sputter procedure. Amorphous and crystalline plasma spray Ca/P coated and noncoated titanium discs served as reference material. The cellular behavior was analyzed with quantitative (attachment and proliferation rates) and qualitative (scanning electron microscopy) techniques. No significant differences were found in cell attachment and proliferation rates between the various materials. Scanning electron microscopy showed extracellular matrix formation after 18 days of culture on amorphous plasma-sprayed and the two types of magnetron sputtered coatings. Furthermore, no severe degradation of the magnetron sputtered coatings was observed. They even appeared to induce apatite formation. On basis of the results, we conclude that magnetron sputtering appears to be a promising method to manufacture bioactive ceramic coatings.