scholarly journals Identification and Morphological Characterization of Biofilms Formed by Strains Causing Infection in Orthopedic Implants

Pathogens ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 649
Author(s):  
Débora C. Coraça-Huber ◽  
Lisa Kreidl ◽  
Stephan Steixner ◽  
Maximilian Hinz ◽  
Dietmar Dammerer ◽  
...  

Objectives: For a better understanding of the mechanisms involved in biofilm formation, we performed a broad identification and characterization of the strains affecting implants by evaluating the morphology of biofilms formed in vitro in correlation with tests of the strains’ antibiotic susceptibility in planktonic form. The ability of the strains to form biofilms in vitro was evaluated by means of colony forming units counting, metabolic activity tests of biofilm cells, and scanning electron microscopy. Methods: A total of 140 strains were isolated from patients with orthopedic implant-related infections during the period of 2015 to 2018. The identification of the isolates was carried out through microbiological cultures and confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Antibiotic susceptibility rates of the isolates were accessed according to EUCAST (European Committee on Antimicrobial Susceptibility Testing). The ability of all isolates to form biofilms in vitro was evaluated by counting the colony forming units, by measuring the metabolic activity of biofilm cells, and by analyzing the morphology of the formed biofilms using scanning electron microscopy. Results: From all the isolates, 41.84% (62 strains) were Staphylococcus epidermidis and 15.60% (22 strains) were Staphylococcus aureus. A significant difference in the capacity of biofilm formation was observed among the isolates. When correlating the biofilm forming capacity of the isolates to their antibiotic susceptibility rates, we observed that not all strains that were classified as resistant were biofilm producers in vitro. In other words, bacteria that are not good biofilm formers can show increased tolerance to multiple antibiotic substances. Conclusion: From 2015 until 2018, Staphylococcus epidermidis was the strain that caused most of the orthopedic implant-related infections in our hospital. Not all strains causing infection in orthopedic implants are able to form biofilms under in vitro conditions. Differences were observed in the number of cells and morphology of the biofilms. In addition, antibiotic resistance is not directly related to the capacity of the strains to form biofilms in vitro. Further studies should consider the use of in vitro culture conditions that better reproduce the joint environment and the growth of biofilms in humans.

2002 ◽  
Vol 81 (9) ◽  
pp. 659-661 ◽  
Author(s):  
William A. Jarrett ◽  
Julie Ribes ◽  
Jose M. Manaligod

An increased awareness of biofilms and their mechanisms has led to a better understanding of bacterial infections that occur following the placement of tracheostomy tubes and other implanted devices and prostheses. One aspect of biofilm formation that is still subject to debate is whether the specific material that is used to manufacture a tube has any bearing in the incidence of infection. We conducted a test of four different tube materials—polyvinyl chloride, silicone, stainless steel, and sterling silver—to ascertain how bacterial biofilms form on tracheostomy tubes and to determine if there is a material-dependent difference in biofilm formation. Scanning electron microscopy demonstrated that Pseudomonas aeruginosa and Staphylococcus epidermidis both formed bacterial biofilms on tracheostomy tubes in vitro. We also found that there was no difference in susceptibility to biofilm formation among the four tube materials tested.


2019 ◽  
Vol 98 (2) ◽  
pp. 209-219 ◽  
Author(s):  
Sascha Beck ◽  
Carolin Sehl ◽  
Sylvia Voortmann ◽  
Hedda Luise Verhasselt ◽  
Michael J. Edwards ◽  
...  

Abstract Periprosthetic infection (PPI) is a devastating complication in joint replacement surgery. On the background of an aging population, the number of joint replacements and associated complications is expected to increase. The capability for biofilm formation and the increasing resistance of different microbes to antibiotics have complicated the treatment of PPI, requiring the need for the development of alternative treatment options. The bactericidal effect of the naturally occurring amino alcohol sphingosine has already been reported. In our study, we demonstrate the antimicrobial efficacy of sphingosine on three different strains of biofilm producing Staphylococcus epidermidis, representing one of the most frequent microbes involved in PPI. In an in vitro analysis, sphingosine’s capability for prevention and treatment of biofilm-contamination on different common orthopedic implant surfaces was tested. Coating titanium implant samples with sphingosine not only prevented implant contamination but also revealed a significant reduction of biofilm formation on the implant surfaces by 99.942%. When testing the antimicrobial efficacy of sphingosine on sessile biofilm-grown Staphylococcus epidermidis, sphingosine solution was capable to eliminate 99.999% of the bacteria on the different implant surfaces, i.e., titanium, steel, and polymethylmethacrylate. This study provides evidence on the antimicrobial efficacy of sphingosine for both planktonic and sessile biofilm-grown Staphylococcus epidermidis on contaminated orthopedic implants. Sphingosine may provide an effective and cheap treatment option for prevention and reduction of infections in joint replacement surgery. Key messages • Here we established a novel technology for prevention of implant colonization by sphingosine-coating of orthopedic implant materials. • Sphingosine-coating of orthopedic implants prevented bacterial colonization and significantly reduced biofilm formation on implant surfaces by 99.942%. • Moreover, sphingosine solution was capable to eliminate 99.999% of sessile biofilm-grown Staphylococcus epidermidis on different orthopedic implant surfaces.


Author(s):  
William J. Lamoreaux ◽  
David L. Smalley ◽  
Larry M. Baddour ◽  
Alfred P. Kraus

Infections associated with the use of intravascular devices have been documented and have been reported to be related to duration of catheter usage. Recently, Eaton et al. reported that Staphylococcus epidermidis may attach to silastic catheters used in continuous ambulatory peritoneal dialysis (CAPD) treatment. The following study presents findings using scanning electron microscopy (SEM) of S. epidermidis adherence to silastic catheters in an in vitro model. In addition, sections of polyvinyl chloride (PVC) dialysis bags were also evaluated by SEM.The S. epidermidis strain RP62A which had been obtained in a previous outbreak of coagulase-negative staphylococcal sepsis at local hospitals was used in these experiments. The strain produced surface slime on exposure to glucose, whereas a nonadherent variant RP62A-NA, which was also used in these studies, failed to produce slime. Strains were grown overnight on blood agar plates at 37°C, harvested from the surface and resuspended in sterile saline (0.85%), centrifuged (3,000 rpm for 10 minutes) and then washed twice in 0.1 M phosphate-buffered saline at pH 7.0. Organisms were resuspended at a concentration of ca. 106 CFU/ml in: a) sterile unused dianeal at 4.25% dextrose, b) sterile unused dianeal at 1.5% dextrose, c) sterile used dialysate previously containing 4.25% dextrose taken from a CAPD patient, and d) sterile used dialysate previously containing 1.5% dextrose taken from a CAPD patient.


2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Lucas Pereira Borges ◽  
Julio Cesar Campos Ferreira-Filho ◽  
Julia Medeiros Martins ◽  
Caroline Vieira Alves ◽  
Bianca Marques Santiago ◽  
...  

The purpose of this work was to verifyin vitroadherence ofE. corrodensandS. oralisto the surface of tongue piercings made of surgical steel, titanium, Bioplast, and Teflon. For this, 160 piercings were used for the count of Colony Forming Units (CFU) and 32 piercings for analysis under scanning electron microscopy. Of these, 96 (24 of each type) were individually incubated in 5 mL of BHI broth and 50 μL of inoculum at 37°C/24 h. The other 96 piercings formed the control group and were individually incubated in 5 mL of BHI broth at 37°C/24 h. Plates were incubated at 37°C/48 h for counting of CFU/mL and data were submitted to statistical analysis (pvalue<0.05). ForE. corrodens, difference among types of material was observed (p<0.001) and titanium and surgical steel showed lower bacterial adherence. The adherence ofS. oralisdiffered among piercings, showing lower colonization (p<0.007) in titanium and surgical steel piercings. The four types of piercings were susceptible to colonization byE. corrodensandS. oralis, and bacterial adhesion was more significant in those made of Bioplast and Teflon. The piercings presented bacterial colonies on their surface, being higher in plastic piercings probably due to their uneven and rough surface.


2016 ◽  
Vol 29 (04) ◽  
pp. 347-350 ◽  
Author(s):  
Mohammed Azab ◽  
Matthew Allen ◽  
Joshua Daniels

Summary Objectives: To evaluate the in vitro anti -bacterial activity of a silver-impregnated coating against a biofilm-forming strain of methicillin-resistant Staphylococcus pseud -intermedius (MRSP). Methods: A clinical MRSP isolate sourced from a failed canine knee implant was evaluated for biofilm production and used in the present study. Using a standard test method and a clinically approved titanium substrate, the antimicrobial activity of a novel silver plasma coating was determined at two times: five minutes after inoculation of the specimens (T0) and after 24 hours of incubation (T24). Scanning electron microscopy was used to evaluate the biofilm formation on specimens. Results: The tested clinical MRSP isolate was classified as a strong biofilm producer. The silver coating significantly reduced the MRSP growth more than four log steps compared to the non-coated specimens and showed more than 99.98% reduction in the number of colony forming units after 24 hours. Scanning electron microscopy images revealed that silver-coated surfaces did not manifest detectable biofilm, while biofilm formation was readily observed on the control specimens. Clinical significance: The silver coating exhibited excellent activity against the multi-drug resistant biofilm-forming MRSP isolate. The next stage of this work will involve testing in an animal model of orthopaedic infection. Positive results from animal studies would support the introduction of the silver plasma coating as a new strategy for preventing implant contamination, biofilm formation, and surgical infection in dogs undergoing orthopaedic surgery.Supplementary Material for this paper is available online at: http://dx.doi.org/10.3415/VCOT-15-08-0134


F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 267
Author(s):  
Phat L. Tran ◽  
Keaton Luth ◽  
James Wang ◽  
Coby Ray ◽  
Anselm de Souza ◽  
...  

Background:It is necessary to develop new strategies to protect against bacteria such as Streptococcus mutans, Streptococcus sanguis, andStreptococcus salivarius, which contribute to tooth decay and plaque formation. Our current study investigated the efficacy of a colloidal silver gel in inhibiting biofilm formation by these principal oral bacteria, in vitro. The aim of this study was to assess the efficacy of a colloidal silver gel formulation for inhibiting bacterial biofilm formation (Ag-gel) by the principal bacteria that cause plaque formation and tooth decay.Methods:The effect of Ag-gel on viability ofS. mutans,S. sanguis,and S. salivariuswas assessed by quantifying their colony forming units (CFU) in presence or absence of the test gel. The effect of this formulation on biofilm-forming ability of these bacteria was studied through scanning electron microscopy.Results:Using the CFU assays, over 6 logs of inhibition (100%) were found forS. mutans,S. sanguis, andS. salivariusfor the Ag-gel-treated bacteria when compared with the control gel. In addition, the Ag-gel also inhibited biofilm formation by these three bacteria mixed together. These results were confirmed by scanning electron microscopy.Conclusions:The Ag-gel was effective in preventing biofilm formation byS. mutans, S. sanguis, and S. salivarius. This Ag-gel should be tested for the ability to block plaque formation in the mouth, through its use as a tooth paste.


2012 ◽  
Vol 727-728 ◽  
pp. 1113-1118 ◽  
Author(s):  
A.G.S. Galdino ◽  
Cecília A.C. Zavaglia

Along decades, bioceramics have been used as materials for bone reconstruction, where hydroxyapatite is one of the most used bioceramics. But hydroxyapatite mechanical strength is not so high when compared with another bioceramics. This research aimed to characterize hydroxyapatite-titanium oxide scaffolds with different compositions. Samples were made using a polyurethane sponge with compositions of 70%-30% wt., 60%-40% wt. and 50%-50% wt. of hydroxyapatite-titanium oxide, calcined at 550°C for burning the polymeric sponge and sintered at 1250°C, 1300°C and 1350°C. Samples were characterized by scanning electron microscopy (SEM), x-ray diffraction (XRD) and energy dispersive x-ray spectroscopy (EDS). Results showed that all compositions maintained the polymeric sponge pores structure without any residual traces of the polymeric sponge. Those results are in accordance with technical literature and it is indicated to do in vitro essays to study the scaffolds biocompatibility for using as bone reconstruction materials.


2016 ◽  
Vol 43 (1) ◽  
pp. 5-13
Author(s):  
N. Stamenov ◽  
G. Tomov ◽  
Z. Denkova ◽  
I. Dobrev

SummaryOral/perioral piercing may provide an ideal environment for adhesion and colonization of microorganisms. The aim of this study is to perform an “in vitro” research on the capabilities of adhesion of Candida albicans on oral piercings made of plastic and metal. Acrylic and metal piercings were incubated with Candida albicans and then were observed using scanning electron microscopy under different magnifications. A lot of irregularities and roughness were observed on the surface of the plastic piercing unlike the surface of the metal one, which is not so rough. Nevertheless, the number of Candida albicans colonies was considerably larger on the scanned metal surface in comparison to the plastic surface. In vitro the metal surface of the piercing creates better environment for the adhesion and colonization of microorganisms than the acrylic. This could be attributed to the electrostatic forces that most likely attract Candida albicans to the metal piercing in the early stages of biofilm formation.


Author(s):  
Bianca Dorana de Oliveira Souza ◽  
Janneth Josefina Escobar Arcos ◽  
Celso Vataru Nakamura ◽  
Luciene Airy Nagashima ◽  
Ayako Sano ◽  
...  

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