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Biomedical subjects

Brian J Nablo

Publications and source records attributed to Brian J Nablo.

6 recordsLinked to original sources

Miniaturized glucose biosensor modified with a nitric oxide-releasing xerogel microarray.

An enzyme-based glucose biosensor modified to release nitric oxide (NO) via a xerogel microarray is reported. The biosensor design is as follows: (1) glucose oxidase (GOx) is immobilized in a methyltrimethoxysilane (MTMOS) xerogel layer; (2) a blended polyurethane/hydrophilic polyurethane coating prevents enzyme leaching and imparts selectivity for glucose; and (3) micropatterned xerogel lines (5 microm wide) separated by distances of 5 or 20 microm provide NO-release capability. This configuration allows for increased glucose sensitivity relative to sensors modified with NO-releasing xerogel films since significant portions of the sensor surface remain unmodified. Glucose diffusion to the GOx layer is thus less inhibited. The micropatterned NO-releasing biosensors generate sufficient NO levels to reduce both Pseudomonas aeruginosa and platelet adhesion without significantly compromising the enzymatic activity of GOx. The glucose response, linearity and stability of the NO-releasing micropatterned sensors are reported.

Absorption↗

Nitric oxide-releasing sol-gels as antibacterial coatings for orthopedic implants.

To assess the benefits of nitric oxide (NO)-releasing sol-gels as potential antibacterial coatings for orthopedic devices, medical-grade stainless steel is coated with a sol-gel film of 40% N-aminohexyl-N-aminopropyltrimethoxysilane and 60% isobutyltrimethoxysilane. Upon converting the diamine groups in these films to diazeniumdiolate NO donors, the NO release from the sol-gel-coated stainless steel is evaluated at both ambient and physiological temperature. Sol-gel films incubated at 25 degrees C have a lower NO flux over the first 24 h compared to those at 37 degrees C, but release more than five times longer. The bacterial adhesion resistance of NO-releasing coatings is evaluated in vitro by exposing bare steel, sol-gel, and NO-releasing sol-gel-coated steel to cell suspensions of Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis at 25 degrees C and 37 degrees C. Cell adhesion to bare and sol-gel-coated steel is similar, while NO-releasing surfaces have significantly less bacterial adhesion for all species and temperatures investigated.

Anti-Bacterial Agents↗

In vitro cytotoxicity of nitric oxide-releasing sol-gel derived materials.

The cytotoxicity of bare and PU-coated nitric oxide (NO)-releasing sol-gel derived materials (sol-gels) was investigated using L929 mouse fibroblasts in both direct and indirect contact models to differentiate between the biological impact of the sol-gel matrix and NO release. The flux of NO was varied up to 150 pmol cm(-2) s(-1) using N-(6-aminohexyl)-aminopropyltrimethoxysilane (balance iso-butyltrimethoxysilane) diazeniumdiolate (NO donor)-modified sol-gels. The addition of a polyurethane (PU) outer membrane greatly improved the stability of the sol-gel matrix without significantly suppressing the NO flux. Direct contact studies demonstrated a cytotoxic effect that was dependent on the aminosilane content of the sol-gel. The use of the thin PU overcoat eliminated this effect. A direct cytotoxicity dependence of NO release for L929 fibroblasts was discovered from indirect contact studies, where 24 h exposure to NO fluxes in excess of 50 pmol cm(-2) s(-1) was cytotoxic.

Animals↗

Inhibition of implant-associated infections via nitric oxide release.

The in vivo antibacterial activity of nitric oxide (NO)-releasing xerogel coatings was evaluated against an aggressive subcutaneous Staphylococcus aureus infection in a rat model. The NO-releasing implants were created by coating a medical-grade silicone elastomer with a sol-gel-derived (xerogel) film capable of storing NO. Four of the bare or xerogel-coated silicone materials were subcutaneously implanted into male rats. Ten rats were administered 10 microl of a 10(8) cfuml(-1)S. aureus colony directly into the subcutaneous pocket with the implant prior to wound closure. Infection was quantitatively and qualitatively evaluated after 8d of implantation with microbiological and histological methods, respectively. A 82% reduction in the number of infected implants was achieved with the NO-releasing coating. Histology revealed that the capsule formation around infected bare silicone rubber controls was immunoactive and that a biofilm may have formed. Capsule formation in response to NO-releasing implants had greater vascularity in comparison with uninoculated or untreated controls. These results suggest that NO-releasing coatings may dramatically reduce the incidence of biomaterial-associated infection.

Animals↗

Antibacterial properties of nitric oxide-releasing sol-gels.

The antibacterial characteristics of nitric oxide (NO)-releasing sol-gel coatings are described. The NO release from these surfaces is steady over short periods (approximately 1 h) and measurable over several days. The ability of NO to prevent bacterial adhesion is evaluated by exposing controls and NO-releasing sol-gels to approximately 10(8) colony-forming units (cfu)/mL saline suspensions of Pseudomonas aeruginosa. Pseudomonas aeruginosa adhesion to sol-gel controls varies depending on the sol-gel formulation. Sol-gel surfaces capable of NO release decrease bacterial adhesion by 30% to 95% relative to controls. The contact angle measurements of control and NO-releasing surfaces are similar, supporting NO's action as an antibacterial agent against bacterial adhesion.

Anti-Bacterial Agents↗

Poly(vinyl chloride)-coated sol-gels for studying the effects of nitric oxide release on bacterial adhesion.

Nitric oxide (NO) releasing sol-gel materials coated with poly(vinyl chloride) (PVC) films exhibit increased stability at ambient and physiological temperatures. The polymer overcoat, however, reduces the NO fluxes by 5-35% over the initial week of release. The variation in NO fluxes between unmodified and PVC-coated sol-gels is negligible after 7 days. The PVC polymeric layer provides controlled surface chemistry for systematic studies of the effects of NO release on bacterial adhesion. As an example, the adhesion of Pseudomonas aeruginosa and Proteus mirabilis at PVC-coated NO-releasing sol-gels is investigated. A direct NO dependence on the reduction of P. aeruginosa adhesion is observed for NO fluxes up to 20 pmol cm(-2) s(-1). Although decreased by 50% in the presence of NO release, P. mirabilis adhesion does not appear to correlate to the flux of NO release. PVC-coated NO-releasing sol-gels may prove useful for studying the effects of localized NO release on other biological and chemical systems.

Bacterial Adhesion↗