Nitric acid passivation of Ti6Al4V reduces thickness of surface oxide layer and increases trace element release.

Passivation of Ti6Al4V and cpTi implants using methods based on the ASTM-F86 nitric acid protocol are used with the intention of reducing their surface reactivity, and consequently the corrosion potential, in the highly corrosive biologic milieu. The ASTM-F86 passivation protocol was originally developed for surgical implants made of stainless steel and chrome cobalt alloy. Using X-ray photoelectron spectroscopy (XPS) to examine the effect of nitric acid passivation on the surface oxide layer of mill-annealed Ti6Al4V and cpTi, we have found that such treatment actually reduced the oxide thickness on the alloy while having no significant effect on the pure metal. These results correlated with observations obtained using graphite furnace atomic absorption spectrophotometry (GFAAS) to detect trace element release from solid, mill-annealed, Ti6Al4V and cpTi into serum-containing culture medium. We detected significantly greater levels of Ti, Al, and V in the presence of passivated compared to nonpassivated Ti6Al4V. In contrast, nitric acid passivation did not influence Ti release from mill-annealed cpTi. These results, derived from two mill-annealed Ti-based metals, would indicate that re-examination of ASTM-F86-based passivation protocols with respect to Ti6Al4V should be considered in view of the widespread use of this alloy for biomedical devices.

A quantitative analysis of the migration, attachment, and orientation of human gingival fibroblasts to human dental root surfaces in vitro.

The purpose of this study was to assess the migration, attachment, and orientation of human gingival fibroblasts to human dental roots over a period of 21 days in vitro. The fibroblasts were incubated with a total of 120 periodontally diseased and non-diseased root slices (200 microns thickness) which had been treated in the following manner: 1) Root planed diseased root (DT); 2) Root planed and citric acid demineralized diseased root (DTD); 3) Non-treated diseased root (DNT); 4) Citric acid demineralized diseased root (DNTD); 5) Non-diseased control (ND); 6) Citric acid demineralized non-diseased root (CA); 7) Citric acid and collagenase digested non-diseased root (CAC); 8) EDTA demineralized non-diseased root (E); and 9) EDTA-demineralized and collagenase-digested non-diseased root (EC). The results showed that that most active phase of cell attachment and orientation occurred during the first 10 days of the experiment. Statistical differences were observed between the variables, and, in terms of cell attachment and orientation to the root slices, it was concluded that: 1) Root planing improves diseased roots; 2) Acid demineralization subsequent to root planing improved diseased roots to such an extent as to render them comparable to non-diseased roots; 3) Citric acid demineralization alone improved diseased roots to the same extent as root planed diseased roots; 4) The exposure of collagen fibrils resulting from acid demineralization of the tools is not the sole reason for the improvement of the root surface, but rather a combination of the exposed collagen fibrils with the creation of a more hospitable environment was found to be responsible.

Cell attachment of human gingival fibroblasts in vitro to porous-surfaced titanium alloy discs coated with collagen and platelet-derived growth factor.

The influence of biological coating, with or without the incorporation of growth factor, on the migration, attachment and orientation of human gingival fibroblasts in relation to porous-surfaced titanium alloy (Ti6AI4V) discs, was measured. Comparison was made between coating the discs with collagen and with collagen incorporating platelet-derived growth factor (PDGF); controls comprised porous-surfaced discs coated with agar or collagen containing bovine serum albumin (used as a carrier for the PDGF), uncoated porous-surfaced Ti6AI4V discs (with or without additional protein additives) exhibited significantly higher attachment indices (AI) and orientation indices (OI) compared with naked control discs (p less than 0.01); OI was also significantly higher than that of surface-demineralized root slices (p less than 0.001) on days 1, 2 and 3. Addition of PDGF to the collagen resulted in a further enhancement in OI on days 1 and 2 (p less than 0.01) over that shown by discs coated with collagen incorporating the bovine serum albumin vehicle. There was no cell attachment and consequently, no cell orientation, in relation to Ti alloy discs that had been coated with agar. These data suggest that attachment and orientation of cells following migration in relation to porous-surfaced Ti6AI4V discs can be modified by the application of biological molecules to the surface of the disc. This may have a useful application in clinical implantology.

Migration, attachment, and orientation of human gingival fibroblasts to root slices, naked and porous-surfaced titanium alloy discs, and Zircalloy 2 discs in vitro.

Cell migration, attachment, and orientation of cultured human gingival fibroblasts (HGF) were measured in relation to four types of specimens: Ti-6Al-4V alloy discs with either ground surfaces or porous structured surfaces, and Zr alloy discs (Zircalloy 2) with either ground surfaces or surfaces modified by being heated to form a thicker oxide cover. Surface-demineralized and non-demineralized root slices were used as controls. Initial cell attachment was measured by means of a 51Cr assay, and cell attachment and orientation following migration by an assay that measures development of the resulting refractile material in relation to the specimen. No significant difference between initial cell attachment to Ti alloy and control demineralized root slices could be detected after one hr using the 51Cr assay. However, with the second assay, cell attachment and orientation at three, seven, 10, and 14 days were significantly higher in relation to surface-demineralized root slices than to all of the metal discs and the non-demineralized root slices. While cell attachment to the surface-ground Ti alloy discs appeared to be higher than that to the porous-surfaced Ti alloy discs at three days and seven days, cell orientation to the porous-surfaced Ti alloy discs and the non-demineralized root slices at days 7, 10, and 14 was greater than that to all of the other metal discs. These data suggest that surface geometry could affect attachment and orientation of cells in vitro.

Initial attachment of fibroblast-like cells to periodontally-diseased root surfaces in vitro.

The precise factors mediating both initial cell attachment and longer term connective tissue reattachment after tissue destruction due to periodontal disease are not known. An in vitro model was used to assess initial attachment of fibroblast-like cells to periodontally-diseased root surfaces. Root fragments were obtained from freshly extracted teeth from 6 different patients. Individual roots were prepared such that a comparison could be made of initial attachment to non-instrumented diseased root surface, curetted diseased root surface and the non-diseased, non-instrumented portion of the same root. The amount of hard tissue removed by instrumentation was quantitated and kept constant between 0.9-1.0 mm. The unsterilized fragments were incubated with human gingival fibroblasts (HGF) for 1 h at 37 degrees, after which the roots were first washed to remove non-adherent cells and then photographed. The number of attached cells per unit area was quantitated from the photographs using a grid system. No significant differences could be detected between the numbers of cells attached to the 3 types of root surfaces studied on the individual roots or between any of the roots studied. Thus, initial attachment of HGF to diseased root surfaces is not inhibited by the presence of plaque or endotoxins.