A gentamicin-releasing coating for cementless hip prostheses-Longitudinal evaluation of efficacy using in vitro bio-optical imaging and its wide-spectrum antibacterial efficacy.

Cementless prostheses are increasingly popular in total hip arthroplasties. Therewith, common prophylactic measures to reduce the risk of postoperative infection like the use of antibiotic-loaded bone cements, will no longer be available. Alternative prophylactic measures may include the use of antibiotic-releasing coatings. Previously, we developed a gentamicin-releasing coating for cementless titanium hip prostheses and derived an appropriate dosing of this coating by adjusting the amount of gentamicin in the coating to match the antibacterial efficacy of clinically employed gentamicin-loaded bone cement. In this manuscript, we investigated two important issues regarding the prophylactic use of this 1 mg cm(-2) bioactive gentamicin-releasing coating in cementless total hip arthroplasty: (1) its ability to prevent bacterial growth in a geometrically relevant set-up and (2) its antibacterial spectrum. A geometrically relevant set-up was developed in which miniature titanium stems were surrounded by agar, contaminated with bioluminescent Staphylococcus aureus. Novel, bio-optical imaging was performed allowing noninvasive, longitudinal monitoring of staphylococcal growth around miniature stems with and without the gentamicin-releasing coating. Furthermore, the antibacterial efficacy of the gentamicin-releasing coating was determined against a wide variety of clinical isolates, including bioluminescent Staphylococcus aureus strains, using traditional zone of inhibition measurements. The gentamicin-releasing coating demonstrated a wide-spectrum of antibacterial efficacy and successfully prevented growth of bioluminescent staphylococci around a miniature stem mounted in bacterially contaminated agar for at least 60 h. This implies that the gentamicin-releasing coating has potential to contribute to the improvement of infection prophylaxis in cementless total hip arthroplasty.

Antibacterial efficacy of a new gentamicin-coating for cementless prostheses compared to gentamicin-loaded bone cement.

Cementless prostheses are increasingly popular but require alternative prophylactic measures than the use of antibiotic-loaded bone cements. Here, we determine the 24-h growth inhibition of gentamicin-releasing coatings from grit-blasted and porous-coated titanium alloys, and compare their antibacterial efficacies and gentamicin release-profiles to those of a commercially available gentamicin-loaded bone cement. Antibacterial efficacy increased with increasing doses of gentamicin in the coating and loading with 1.0 and 0.1 mg gentamicin/cm(2) on both grit-blasted and porous-coated samples yielded comparable efficacy to gentamicin-loaded bone cement. The coating had a higher burst release than bone cement, and also inhibited growth of gentamicin-resistant strains. Antibacterial efficacy of the gentamicin coatings disappeared after 4 days, while gentamicin-loaded bone cement exhibited efficacy over at least 7 days. Shut-down after 4 days of gentamicin-release from coatings is advantageous over the low-dosage tail-release from bone cements, as it minimizing risk of inducing antibiotic-resistant strains. Both gentamicin-loaded cement discs and gentamicin-coated titanium coupons were able to kill gentamicin-sensitive and -resistant bacteria in a simulated prothesis-related interfacial gap. In conclusion, the gentamicin coating provided similar antibacterial properties to those seen by gentamicin-loaded bone cement, implying protection of a prosthesis from being colonized by peri-operatively introduced bacteria in cementless total joint arthroplasty.

Spectroscopic investigations of intermediates in the reaction of cytochrome P450(BM3)-F87G with surrogate oxygen atom donors.

Rapid mixing of substrate-free ferric cytochrome P450(BM3)-F87G with m-chloroperoxybenzoic acid (mCPBA) resulted in the sequential formation of two high-valent intermediates. The first was spectrally similar to compound I species reported previously for P450(CAM) and CYP 119 using mCPBA as an oxidant, and it featured a low intensity Soret absorption band characterized by shoulder at 370nm. This is the first direct observation of a P450 compound I intermediate in a type II P450 enzyme. The second intermediate, which was much more stable at pH values below 7.0, was characterized by an intense Soret absorption peak at 406nm, similar to that seen with P450(CAM) [T. Spolitak, J.H. Dawson, D.P. Ballou, J. Biol. Chem. 280 (2005) 20300-20309]. Double mixing experiments in which NADPH was added to the transient 406nm-absorbing intermediate resulted in rapid regeneration of the resting ferric state, with the flavins of the flavoprotein domain in their reduced state. EPR results were consistent with this stable intermediate species being a cytochrome c peroxidase compound ES-like species containing a protein-based radical, likely localized on a nearby Trp or Tyr residue in the active site. Iodosobenzene, peracetic acid, and sodium m-periodate also generated the intermediate at 406nm, but not the 370nm intermediate, indicating a probable kinetic barrier to accumulating compound I in reactions with these oxidants. The P450 ES intermediate has not been previously reported using iodosobenzene or m-periodate as the oxygen donor.