Antimicrobial and osteoconductive properties of two different types of titanium silver coating.

In prosthetic joint surgery, Ag coating of implant areas in direct contact with bone has been met with hesitation for fear of compromising osseointegration. The physicochemical, antibacterial and osteoconductive properties of three different Ti samples were studied: Ti6Al4V alloy that was grit-blasted (GB), Ti6Al4V alloy with an experimental Ti-Ag-nitride layer (SN) applied by physical vapour deposition (PVD) and commercially available PVD-coated Ti6Al4V alloy with a base Ag layer and a surface Ti-Ag-nitride layer (SSN, clinically known as PorAg®). Ag content on the surface of experimental SN and SSN discs was 27.7 %wt and 68.5 % wt, respectively. At 28 d, Ag release was 4 ppm from SN and 26.9 ppm from SSN substrates. Colonisation of discs by Staphylococcus aureus was the highest on GB [944 (± 91) × 10 4 CFU/mL], distinctly lower on experimental SN discs [414 (± 117) × 104 CFU/mL] and the lowest on SSN discs [307 (± 126) × 10 4 CFU/mL]. Primary human osteoblasts were abundant 28 d after seeding on GB discs but their adhesion and differentiation, measured by alkaline-phosphatase production, was suppressed by 73 % on SN and by 96 % on SSN discs, in comparison to GB discs. Thus, the PVD-applied Ag coatings differed considerably in their antibacterial effects and osteoconductivity. The experimental SN coating had similar antibacterial effects to the commercially available SSN coating while providing slightly improved osteoconductivity. Balancing the Ag content of Ti implants will be vital for future developments of implants designed for cementless fixation into bone.

Resting microglial cells in vitro: analysis of morphology and adhesion molecule expression in organotypic hippocampal slice cultures.

Neurons in organotypic hippocampal slice cultures (OHSCs) are known to preserve morphological and physiological features of the in vivo situation; however, little is known about the properties of microglial cells under these in vitro conditions. In this study, we addressed the question whether microglial cells in OHSCs are initially activated following explantation but return to a resting state during in vitro cultivation. Thus, we analyzed a) microglial cell morphology, b) microglial cell distribution, and c) expression of integrin adhesion molecules as putative markers of microglial activation. Hippocampal slices fixed immediately following explantation showed only resting microglial cells, mainly located in the paraventricular regions. After 3 days in vitro (div) OHSC surfaces were covered by activated microglia, whereas intermediate layers contained fewer microglial cells, giving the slices a sandwich-like appearance with the intact hippocampal formation being surrounded by glial tissue. After 3 div, microglial cells in intermediate layers of OHSCs showed activated morphology with ovaloid cytoplasm and no or merely few cytoplasmic processes; after 6 div, however, an increasing degree of ramification could be observed. After 9 div, microglia in intermediate layers had almost regained the morphological appearance of resting cells with filigrane cytoplasmic processes extended in all directions. The integrin adhesion molecules LFA-1 (alpha and beta chains) and VLA-4 were expressed on most microglial cells with activated morphology, as verified by co-localization with double immunofluorescence labeling for LFA-1 or VLA-4 and Griffonia simplicifolia isolectin B4 (GFS-B4). In contrast, only low levels of integrin adhesion molecule expression were also found on reactive astrocytes along slice surfaces. However, LFA-1 or VLA-4 were never found on ramified microglial cells, and double immunofluorescence labeling of LFA-1 or VLA-4 with ramified GFS-B4+ microglia never occurred. We conclude that a) originally resting microglial cells activated in an early phase of in vitro culture but regain a resting status after at least 6 div; and b) integrin adhesion molecules LFA1 and VLA-4 are potential markers of microglial activation, as they were found on activated but never on resting microglial cells. This enables further investigations on immunological and electrophysiological features of resting and activated microglial cells under in vitro conditions.