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Orthopaedic Proceedings
Vol. 98-B, Issue SUPP_2 | Pages 68 - 68
1 Jan 2016
Iwakiri K Kobayashi A Takaoka K Ando Y Hoshino M Tsujio T Seki M Nakamura H
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[Introduction]

Surgical-site-infections (SSI) prolong hospital stay, and they are leading nosocomial cause of morbidity and a source of excess cost. Recently, a waterless hand-rubbing protocol containing aqueous 1% chlorhexidine gluconate was developed before surgery, but there is no literature in orthopaedic surgery. The aim was to compare the SSI rates between waterless hand-rubbing and traditional hand-scrubbing protocol.

[Materials and Methods]

STUDY 1: A total of 996 consecutive patients who underwent orthopaedic surgery between August 1, 2012 and January 31, 2014, were screened for SSI within 30 days after surgery. 500 patients from August 1, 2012 to April 1, 2013 were used by traditional hand-scrubbing, and 496 patients from June 1, 2013 to January 1, 2014 were by waterless hand-rubbing.

STUDY 2: The twelve operating room staff members were randomly recruited, and the participants were assigned equally to use either a traditional hand-scrubbing protocol or a waterless hand-rubbing on 2 separate days. Washing times were recorded and microorganisms on hands were sampled on bacterial culture plates. Two days after sampling, the grown colonies were counted.


Orthopaedic Proceedings
Vol. 95-B, Issue SUPP_15 | Pages 352 - 352
1 Mar 2013
Tsukamoto M Ando Y Noda I Akiyama T Eto S Yonekura Y Kawano S Sonohata M Miyamoto H Mawatari M
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Problems

Biofilm infections are increasingly associated with orthopedic implants. Bacteria form biofilms on the surfaces of orthopedic devices. The biofilm is considered to be a common cause of persistent infections at a surgical site. The growth and the maturation of biofilm are enhanced by the flow of broth in culture environment. In order to reduce the incidence of implant-associated infections, we developed a novel coating technology of hydroxyapatite (HA) containing silver (Ag). We previously reported that the Ag-HA coating inhibits biofilm formation under flow condition of Trypto Soy Broth + 0.25% glucose for 7 days. In this study, we evaluated whether the Ag-HA coating continuously inhibits the biofilm formation on its surface under flow condition of fetal bovine serum, which contains many in vivo substrates such as proteins.

Materials and Method

The commercial pure titanium disks were used as substrates. Ag-HA or HA powder was sprayed onto the substrates using a flame spraying system. The HA coating disks were used as negative control. The biofilm-forming methicillin resistant Staphylococcus aureus (BF-MRSA; UOEH6) strain was used. The bacterial suspension (about 105 colony forming units) was inoculated into 24-well sterile polystyrene tissue culture plates. The Ag-HA and HA coating disks were aseptically placed in the wells. After cultivation at 37°C for 1 hour, the disks were rinsed twice with 500 μL sterile PBS (−) to eliminate the non-adherent bacteria. After rinsing, the disks were transferred into petri-dish containing heat-inactivated FBS with a stirring bar on the magnetic stirrer and they were cultured at 37 °C for 24 hours, 7 and 14 days. In the meantime, the stirring bar was spun at 60 rounds per minute. Then, the disks were immersed in a fluorescent reagent to stain the biofilm. Finally, the biofilm on each disk was observed by a fluorescence microscope and the biofilm-covered rate (BCR) on the surfaces of them was calculated using the NIH image software.


Orthopaedic Proceedings
Vol. 94-B, Issue SUPP_XXV | Pages 4 - 4
1 Jun 2012
Ando Y Noda I Miyamoto H Akiyama T Shimazaki T Yonekura Y Miyazaki M Mawatari M Hotokebuchi T
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Bacterial infection related to prosthetic replacement is one of the serious types of complications. Recently, there has been a greater interest in antibacterial biomaterials. In order to reduce the incidence of replacement-associated infections, we developed a novel coating technology of Hydroxyapatite (HA) containing silver (Ag). We reported the Ag-HA coating showed high antibacterial activity against E. coli, S. aureus and methicillin-resistant S. aureus (MRSA) under static condition. However, human bodies have a circulating body fluid, which is not a static condition. And the growth and the maturation of biofilm, which is said that a common course of persistent infections at a surgical site, are enhanced by the flow of broth in culture environment. Therefore, we evaluated whether the Ag-HA coating inhibits the biofilm formation on its surface or not by a biofilm-forming test under flow condition in this study.

Ag-HA or HA powder was sprayed onto the commercial pure titanium disks using a flame spraying system. The HA coating disks were used as negative control. The biofilm-forming methicillin sensitive S. aureus (BF-MSSA; Seattle 1945) strain and the BF-MRSA (UOEH6) strain were used. The pre-culture bacterial suspension (about 105 colony forming units; CFU) was inoculated onto the Ag-HA and HA coating disks. After cultivation at 37 °C for 1 h, the disks were rinsed twice with 500 μL sterile PBS (-) to eliminate the non-adherent bacteria. The number of the adherent bacteria on these disks was counted using culture method. After rinsing, the disks were transferred into petri-dish containing Trypto–Soy Broth (TSB) + 0.25% glucose with a stirring bar on the magnetic stirrer and they were cultured at 37°C for 7 days. In the meantime, the stirring bar was spun at 60 rounds per minute. Then, the disks were immersed in a fluorescent reagent to stain the biofilm. Finally, the biofilm on each disk was observed by a fluorescence microscope and the biofilm-covered rate on the surfaces of them was calculated using the NIH image software.

The number of the bacteria on these disks was not so different between Ag-HA and HA coating after rinsing. After biofilm-forming test, the coverage of the biofilm of BF-MSSA was 2.1% and 81.0% on the Ag-HA and HA coatings, respectively. Similarly, in the case of BF-MRSA, it was 7.7% and 72.0% on the Ag-HA and HA coatings, respectively. Though bacteria slightly adhered, biofilm was hardly observed on the Ag-HA coating. The biofilm on the HA coating was extensive and mature. The inhibition effect of biofilm formation on the Ag-HA coating might be ascribed to the antibacterial effect by Ag ions released from the coating. Because Ag ions have a broad spectrum of antibacterial activity against pathogens, including biofilm forming bacteria, they inhibited the biofilm formation on the Ag-HA coating by killing adherent bacteria. Even in a flow condition, it was suggested that the AgHA shows the antibacterial activity, though the conditions in this work are different from those in living body.