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Biomedical subjects

H J Busscher

Publications and source records attributed to H J Busscher.

At least 37 records · Page 2Linked to original sources

Influence of dentifrices and dietary components in saliva on wettability of pellicle-coated enamel in vitro and in vivo.

In vitro salivary pellicles were found to be less hydrophobic by water contact angles than clinically formed pellicles. In this study, water contact angles were measured on enamel coated with pellicles adsorbed from reconstituted human whole saliva (RHWS) and after exposure to dentifrices or dietary components. In addition, adhesion of Streptococcus oralis J22 to pellicles formed from RHWS with minor amounts of milk added and after exposure to dentifrices was studied. Exposure of RHWS-pellicles to milk or salad oil yielded an increase in the hydrophobicity of in vitro pellicles to values observed in vivo, but a decrease was seen after exposure to a sugar solution. Pellicles formed from saliva with 0.4% milk added attracted less S. oralis cells than pellicles formed in the absence of milk components. Exposure of pellicles formed from saliva with milk added to various dentifrices had a variable effect on bacterial adhesion: markedly lower numbers of adhering S. oralis were found for a dentifrice with NaF, but exposure to dentifrices containing SnF2 or hexametaphosphate showed slightly increased adhesion. In summary, dietary components have influence on the hydrophobicity of enamel pellicles, while combinations of dietary components and dentifrices certainly influence the adhesiveness of the pellicles. The effects of dietary components on pellicle conditioning film should be taken into consideration in research on the development of ingredients to control intraoral surface chemistry and microbiology, as well as in the development of oral products.

Animals↗

Functional variation of the antigen I/II surface protein in Streptococcus mutans and Streptococcus intermedius.

Although Streptococcus intermedius and Streptococcus mutans are regarded as members of the commensal microflora of the body, S. intermedius is often associated with deep-seated purulent infections, whereas S. mutans is frequently associated with dental caries. In this study, we investigated the roles of the S. mutans and S. intermedius antigen I/II proteins in adhesion and modulation of cell surface characteristics. By using isogenic mutants, we show that the antigen I/II in S. mutans, but not in S. intermedius, was involved in adhesion to a salivary film under flowing conditions, as well as in binding to rat collagen type I. Binding to human fibronectin was a common function associated with the S. mutans and S. intermedius antigen I/II. Adhesion of S. mutans or S. intermedius to human collagen types I or IV was negligible. Hydrophobicity, as measured by water contact angles, and zeta potentials were unaltered in the S. intermedius mutant. The S. mutans isogenic mutants, on the other hand, exhibited more positive zeta potentials at physiological pH values than did the wild type. The results indicate common and species-specific roles for the antigen I/II in mediating the attachment of S. mutans and S. intermedius to host components and in determining cell surface properties.

Adhesins, Bacterial↗

Adhesion of Enterococcus faecalis 1131 grown under subinhibitory concentrations of ampicillin and vancomycin to a hydrophilic and a hydrophobic substratum.

The effect of two subinhibitory antibiotic concentrations of ampicillin and vancomycin during growth on the adhesion of Enterococcus faecalis 1131 to glass and silicone rubber was studied in a parallel plate flow chamber. Initial deposition rates and numbers of adhering bacteria after 4 h were higher on hydrophilic glass than on hydrophobic silicone rubber, regardless of growth conditions. The presence of 1/4 minimal inhibitory concentration (MIC) of ampicillin during growth reduced enterococcal adhesion to both substrata, but growth in the presence of 1/4 MIC vancomycin did not affect the adhesion of E. faecalis. Moreover, enterococcal adhesion increased after growth in the presence of 1/8 MIC vancomycin. The increased adhesion after growth in the presence of subinhibitory concentrations of vancomycin may have strong implications for patients living with implanted biomaterials, as they may suffer adverse effects from use of this antibiotic, especially since bacteria once adhered are less sensitive to antibiotics.

Ampicillin↗

Charge transfer during staphylococcal adhesion to TiNOX coatings with different specific resistivity.

Adhesion of the bacterial strain Staphylococcus epidermidis 3399 to titanium-oxy-nitride (TiNOX) substrata with different specific resistivities was studied in a parallel plate flow chamber, while simultaneously measuring the electric potential of the substrata. During adhesion, bacteria either donated or accepted electrons to the substrata depending on the specific resistivity of the substratum and bacteria that had donated electrons to the substratum adhered more strongly than bacteria that had accepted electrons from the substratum. These results demonstrate that electron transfer plays a role in bacterial adhesion to conducting surfaces, which has hitherto been neglected.

Algorithms↗

Electric field induced desorption of bacteria from a conditioning film covered substratum.

Desorption of three oral bacterial strains from a salivary conditioning film on an indium tin oxide electrode during application of a positive (bacterial adhesion to the anode) or a negative electric current was studied in a parallel plate flow chamber. Bacterial adhesion was from a flowing suspension of high ionic strength, after which the bacterial suspension was replaced by a low ionic strength solution without bacteria and currents ranging from -800 to +800 microA were applied. Streptococcus oralis J22 desorbed during application of a positive and negative electric current with a desorption probability that increased with increasing electric current. Two actinomyces strains, however, could not be stimulated to desorb by the electric currents applied. The desorption forces acting on adhering bacteria are electroosmotic in origin and working parallel to the electrode surface in case of a positive current, whereas they are electrophoretic and electrostatic in origin and working perpendicular to the surface in case of a negative current. By comparison of the effect of positive and negative electric currents, it can be concluded that parallel forces are more effective in stimulating bacterial desorption than perpendicular forces. The results of this study point to a new pathway of cleaning industrial and biomedical surfaces without the use of detergents or biocides.

Actinomyces↗

Air-flow resistances of silicone rubber voice prostheses after formation of bacterial and fungal biofilms.

Laryngectomized patients use silicone rubber voice prostheses to rehabilitate their voice. However, biofilm formation limits the lifetime of voice prostheses by causing leakage or an increased air-flow resistance and the prosthesis has to be replaced. To determine which bacterial or yeast strains, isolated from explanted voice prostheses, contribute most to increases in air-flow resistance of silicone rubber voice prostheses, biofilms consisting of either a bacterial or a yeast strain were grown on voice prostheses in the artificial throat model. The effects of these biofilms on air-flow resistances were determined by calculating the difference in air-flow resistance of the individual voice prosthesis as covered with a 7-day-old biofilm with the situation prior to biofilm formation. Conspicuously, voice prosthetic biofilms formed by the bacterial strains Staphylococcus aureus GB 2/1 and Rothia dentocariosa GBJ 41/25B and their excreted organic matter showed larger increases in air-flow resistance (more then 30 cm H(2)O.s/L) than biofilms formed by Candida species. This is contrary to the literature, where there seems to be agreement that Candida species are mainly responsible for clinical failure of silicone rubber voice prostheses.

Bacteria↗

Biofilm formation and design features of indwelling silicone rubber tracheoesophageal voice prostheses--an electron microscopical study.

After total laryngectomy, voice can be restored with a silicone rubber tracheoesophageal voice prosthesis. However, biofilm formation and subsequent deterioration of the silicone material of the prosthesis will limit device life by impairing valve function. To simulate the natural process of biofilm development under dynamic nutrient conditions, a modified Robbins device was used to evaluate the biofilm-related valve dysfunction of the Groningen, Provox2, Blom-Singer indwelling, and VoiceMaster voice prostheses. Obstruction of the semicircular slit-valved Groningen prosthesis leading to increased airway resistance was caused not only by a buildup of deposits on the esophageal flange and valve hat, but also by accumulation of deposits on the semicircular valve seating. The hinged flap valved Provox2 and indwelling Blom-Singer prostheses failed to close sufficiently because of biofilm formation on the valve seating. The esophageal flange of the VoiceMaster prosthesis was affected, but the tripod structure of the ball valve was fully colonized up to the titanium sleeve, which interfered with proper valve opening and closure. These findings on biofilm formation could be used for the further development and modification of critical design features of voice prostheses to facilitate tracheoesophageal speech.

Bacterial Adhesion↗

Staphylococcus aureus biofilm formation on different gentamicin-loaded polymethylmethacrylate bone cements.

In this in vitro study, the formation of a Staphylococcus aureus biofilm on six gentamicin-loaded bone cements (CMW1, CMW3, CMW Endurance, CMW2000, Palacos, and Palamed) was determined in a modified Robbins device over a 3 days time span and related with previously (Van de Belt et al., Biomaterials 21 (2000) 1981) measured kinetics of antibiotic release by these cement brands. The influence of gentamicin release on biofilm formation was quantified by expressing the number of colony-forming units on gentamicin-loaded cement relative to the number of viable organisms on unloaded cement of the same brand. Biofilms formed on all gentamicin-loaded cements, despite the release of antibiotics, followed a consistent pattern in time with a maximum number of colony-forming units per unit cement area found between 24 and 30 h after inoculation. None of the gentamicin-loaded cements showed a reduction in biofilm formation relative to unloaded cements within 6 h after inoculation, whereas only gentamicin-loaded CMW1 and Palacos reduced biofilm formation 24 h after inoculation. Alternatively, CMW Endurance, CMW2000, and Palamed did not exhibit any initial reductions in biofilm formation, but effects started after 72, 48, and 72 h, respectively. Biofilm reduction by gentamicin-loaded CMW3 lasted the longest from 24 to 72 h. Interestingly, each cement seemed to have a different "window-of-effectiveness" with regard to reduction in biofilm formation that did not relate with the gentamicin-release kinetics. Summarising, this study demonstrates that although gentamicin loading of bone cements yields reductions in biofilm formation, S. aureus is able to grow on gentamicin-loaded bone cements.

Biocompatible Materials↗

Bacterial adhesion to surface hydrophilic and hydrophobic contact lenses.

The aim of this paper was to determine the adhesion of two physico-chemically characterized bacterial strains to a surface hydrophilic (CL A, water contact angle 57 degrees) and hydrophobic (CL B, water contact angle 106 degrees) hydrogel contact lens (CL) with and without an adsorbed tear film in a parallel plate flow chamber. Hydrophobicity (by water contact angles), charge (by particulate microelectrophoresis) and elemental composition (by XPS) of the surfaces of seven bacterial strains were characterized, after which two strains were selected for further studies. On CL surfaces, hydrophobicity, elemental composition, and mean surface roughness (by AFM) were determined, as well as the protein composition of tear films adsorbed on these lenses (by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE)). Bacterial cell surfaces were relatively uncharged and water contact angles on lawns of different strains ranged from hydrophobic to hydrophilic. After adsorption of tear film components, N/C elemental surface concentrations increased on CL A and CL B and differences in water contact angles between both lenses reduced to range from 57 degrees (CL A) to 69 degrees (CL B). However, different protein compositions were inferred. The surface roughness of CL A increased from 4 to 13 nm. while it remained 16 nm for CL B. Adhesion of hydrophobic Pseudomonas aeruginosa #3 was more extensive than of hydrophilic Staphylococcus aureus 799, with no differences between both lenses. The hydrophobicity of P. aeruginosa #3 after cell surface damage decreased and its adhesion was reduced on CL A and strongly on CL B. In addition, passage of an air-liquid interface yielded more detachment of S. aureus 799 than of P. aeruginosa #3 from the CL surfaces. In conclusion, the hydrophobicity of CL surfaces dictates the composition of the adsorbed tear film and therewith plays an important role in bacterial adhesion to lenses. Adhesion of hydrophobic P. aeruginosa #3 was more tenacious than of hydrophilic S. aureus 799.

Bacterial Adhesion↗

Effects of cell surface damage on surface properties and adhesion of Pseudomonas aeruginosa.

Bacterial cell surfaces play a crucial role in their adhesion to surfaces. In the present study, physico-chemical cell surface properties of Pseudomonas aeruginosa, isolated from a case of contact lens associated keratitis, are determined for mid-exponential and early stationary phase cells and for cells after exposure to a lens care solution or after mechanical damage by sonication. Exposure to a lens care solution and mechanical cell surface damage reduced the cell surface hydrophobicity and water contact angles decreased from 129 degrees to 96 degrees and 83 degrees, respectively. Zeta potentials in saline (-9 mV) were hardly affected after mechanical damage, but tri-modal zeta potential distributions, with subpopulation zeta potentials at -11, -28 and -41 mV, were observed after exposure of bacteria to a lens care solution. X-ray photoelectron spectroscopy indicated changes in the amounts of oxygen-, nitrogen- and phosphorus-rich cell surface components. Mid-exponential phase cells had more nitrogen-rich cell surface components than early stationary phase cells, but water contact angles and zeta potentials were not very different. In addition, mid-exponential phase cells adhered better than early stationary phase cells to hydrophobic and hydrophilic substrata in a parallel plate flow chamber. The capacity of P. aeruginosa to adhere was decreased after inflicting cell surface damage. Exposure to a lens care solution yielded a larger reduction in adhesion capacity than sonication, likely because sonication left most of the cells in a viable state, in contrast to exposure to a lens care solution. It is argued that for clinically relevant experiments, it may be preferable to work with surface damaged cells rather than with gently harvested organisms.

Bacterial Adhesion↗

Probing molecular interactions and mechanical properties of microbial cell surfaces by atomic force microscopy.

Knowledge of the surface properties of microbial cells is a key to gain a detailed understanding of their functions in the natural environment and to efficiently exploit them in biotechnological processes. In this paper, we present force-distance curves recorded, by atomic force microscopy (AFM) in aqueous solutions, on various microbial samples: reconstituted S-layers, whole fungal spores and several bacterial strains. The approach and retraction curves exhibited important differences--depending on the type of microorganism, on the physiological state (dormancy versus germination) and on the environmental conditions (ionic strength)--which were shown to reflect differences in long-range surface forces, adhesion forces and mechanical properties. These data illustrate the great potential of AFM force measurements to elucidate the physical properties of microbial cells and to understand, at the molecular level, biointerfacial phenomena such as cell adhesion and cell aggregation.

Cell Adhesion↗

Adhesive interactions between voice prosthetic yeast and bacteria on silicone rubber in the absence and presence of saliva.

Biofilms on silicone rubber voice prostheses are the major cause for frequent failure and replacement of these devices. The presence of both bacterial strains and yeast has been suggested to be crucial for the development of voice prosthetic biofilms. Adhesive interactions between Candida albicans, Candida krusei, and Candida tropicalis with 14 bacterial strains, all isolated from explanted voice prostheses were investigated in a parallel plate flow chamber. Bacteria were first allowed to adhere to silicone rubber, after which the flow chamber was perfused with yeast, suspended either in saliva or buffer. Generally, when yeast were adhering from buffer and saliva, the presence of adhering bacteria suppressed adhesion of yeast. In saliva, Rothia dentocariosa and Staphylococcus aureus enhanced adhesion of yeast, especially of C. albicans. This study shows that bacterial adhesion mostly reduces subsequent adhesion of yeast, while only a few bacterial strains stimulate adhesion of yeast, provided salivary adhesion mediators are present. Interestingly, different clinical studies have identified R. dentocariosa and S. aureus in biofilms on explanted prostheses of patients needing most frequent replacement, while C. albicans is one of the yeast generally held responsible for silicone rubber deterioration.

Bacteria↗

Co-adhesion and removal of adhering bacteria from salivary pellicles by three different modes of brushing.

This study compares removal of pairs of co-adhering and non-co-adhering oral actinomyces and streptococci by hand, electric and sonic brushing from salivary pellicles. In addition, re-deposition of a co-adhering and non-co-adhering streptococcal strain to brushed pellicles was studied. First, actinomyces were allowed to adhere to a pellicle in a flow chamber, after which streptococci suspended in saliva were perfused through the chamber at 33 C. Pellicles with adhering bacterial pairs were brushed and the number of bacteria remaining determined. Whereas sonic brushing removed nearly all adhering bacteria, greater numbers of larger aggregates of the co-adhering pair, involving Streptococcus oralis J22, were left behind after hand and electric brushing than of the non-co-adhering pair with Streptococcus sanguis PK1889. Re-deposition of streptococci to electrically and sonically brushed pellicles, however, was two-fold higher for the co-adhering pair than for the non-co-adhering pair. This demonstrates a role for co-adhesion in de novo plaque formation. Removal by the three different modes of brushing was not affected by the presence of fluoride, indicating that fluoride is not able to disrupt calcium bridges between co-adhering pairs. In contrast, in the presence of lactose only small aggregates of co-adhering pair were left behind.

Actinomyces↗

Infection of orthopedic implants and the use of antibiotic-loaded bone cements. A review.

Infections by bacteria are a serious complication following orthopedic implant surgery, that can usually only be cured by removing the implant, since the biofilm mode of growth of infecting bacteria on an implant surface protects the organisms from the host immune system and antibiotic therapy. Over the past few decades, attempts have been made to prevent and cure orthopedic implant infections by incorporating antibiotics in polymethylmethacrylate bone cements, in primary and revision surgery. However, the clinical efficacy of antibiotic-releasing bone cements is not accepted by all and the long-term exposure to low doses from antibiotic-releasing bone cements in patients is strongly related to the emerging threat of antibiotic resistance in medicine today. In this article, we start by reviewing the mechanisms governing the formation of an infectious biofilm on orthopedic implant materials, the release mechanisms and properties of clinically-used, antibiotic-loaded bone cements. The clinical efficacy of antibiotic-loaded bone cements is evaluated analyzing separatedly the prophylactic and therapeutic uses of these products.

Antibiotic Prophylaxis↗

Biomaterial-associated infection of gentamicin-loaded PMMA beads in orthopaedic revision surgery.

In two-stage orthopaedic revision surgery, high local levels of antibiotics are achieved after removal of an infected prosthesis through temporary implantation of gentamicin-loaded beads. However, despite their antibiotic release, these beads act as a biomaterial surface to which bacteria preferentially adhere, grow and potentially develop antibiotic resistance. Gentamicin-loaded beads were retrieved from 20 patients with prosthesis-related infections. Excised tissue samples were taken for routine culture, while beads were analysed in an extensive laboratory procedure. Extensive culture procedures indicated the presence of bacteria on gentamicin-loaded beads in 18 of the 20 patients involved, while 12 of these 18 patients were considered free of infection by routine culture. Nineteen of 28 bacterial strains isolated were gentamicin resistant and cultures from three patients yielded highly gentamicin-resistant sub-populations. It is concluded that routine culture of excised tissues in orthopaedic revision surgery is inadequate to ascertain full eradication of infection, especially as infecting, antibiotic-resistant bacteria preferentially adhere to and grow on gentamicin-loaded beads. Extensive examination of the bead surfaces is proposed as a more reliable indication that infection has been eradicated.

Anti-Bacterial Agents↗

Antimicrobial effects of positively charged surfaces on adhering Gram-positive and Gram-negative bacteria.

The infection of biomaterials is determined by an interplay of adhesion and surface growth of the infecting organisms. In this study, the antimicrobial effects on adhering bacteria of a positively charged poly(methacrylate) surface (xi potential +12 mV) were compared with those of negatively charged poly(methyl methacrylate) (-12 mV) and a highly negatively charged poly(methacrylate) (-18 mV) surface. Initial adhesion of Staphylococcus aureus ATCC 12600, Staphylococcus epidermidis HBH(2) 102, Escherichia coli O2K2 and Pseudomonas aeruginosa AK1 to these surfaces was measured in a parallel plate flow chamber in phosphate-buffered saline. Adhering bacteria were allowed to multiply by perfusing the flow chamber with growth medium. All bacteria adhered most rapidly to the positively charged surface, but there was no subsequent surface growth of the Gram-negative strains. On the negatively charged surfaces, despite a slower initial adhesion, surface growth of the adhering bacteria was exponential for both Gram-positive and Gram-negative strains. These results suggest that positively charged biomaterial surfaces exert an antimicrobial effect on adhering Gram-negative bacteria, but not on Gram-positive ones.

Bacterial Adhesion↗