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Title Retention of protein repulsive character and antimicrobial activity of PEO brush layers following nisin entrapment
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Date Available
Date Accessioned
Degree MS
Discipline/Department Chemical Engineering
Degree Level masters
University/Publisher Oregon State University
Abstract Nisin, an amphiphilic, antimicrobial peptide, has been shown to integrate into the hydrophobic inner region of poly(ethylene oxide) (PEO) brush layers; however, the presence of integrated nisin may compromise the protein repulsive character of the PEO layer. In particular, the introduction of fibrinogen to nisin-loaded brush layers has been observed to cause changes consistent with partial elution of nisin and/or location of fibrinogen at the interface. Questions surrounding the possibility of fibrinogen adsorption warrant further investigation, as the location of procoagulant proteins at a peptide-loaded PEO layer would significantly reduce the viability of a medical device coating based on such an approach. In this work, the preferential location of fibrinogen at PEO brush layers was investigated by: detection of FITC-labeled fibrinogen after sequential introduction of nisin and labeled fibrinogen; measurement of changes in the zeta potential of PEO coated and uncoated surfaces following nisin, fibrinogen, and/or buffer challenges; and evaluation of adsorption and elution kinetics in label-free, sequential adsorption experiments using optical waveguide lightmode spectroscopy (OWLS). PEO layers were constructed through radiolytic grafting of Pluronic® F108 or F68 onto silanized silica surfaces producing long-chain or short-chain PEO layers, respectively. Adsorption results indicated that sequential introduction of nisin and fibrinogen to PEO brush layers, based on F108, does not result in fibrinogen adsorption beyond that expected for a nisin-free PEO layer. No evidence of nisin entrapment in fibrinogen-repellent F68 layers was recorded. Low-level fibrinogen adsorption observed at F68 layers following the introduction of nisin was determined to be a result of nisin adsorption at (uncoated) defect regions on the surface. In conclusion, retention of PEO layer capacity for protein repulsion after nisin entrapment is owing to a steric repulsive barrier provided by PEO segments extending beyond the level of entrapped nisin. It was then hypothesized that the immobilized, pendant PEO chains will inhibit exchange of entrapped nisin by competing proteins, and therefore prolong nisin activity retention. In order to evaluate nisin function following its entrapment, the antimicrobial activity of nisin-loaded, F108-coated silica surfaces was evaluated against the Gram-positive indicator strain, Pediococcus pentosaceous. The retained biological activity of these nisin-loaded layers was evaluated after incubation in the presence of bovine serum albumin (BSA), for contact periods up to one week. Surfaces were withdrawn at selected times and placed on plates inoculated with P. pentosaceous to measure kill zone radius in order to quantify nisin activity. In the presence of BSA, F108-coated surfaces retained more antimicrobial activity than the uncoated, hydrophobic surfaces. These results strongly suggest that PEO brush layers may serve as a viable drug storage platform due to the retained non-fouling character after bioactive…
Subjects/Keywords nisin adsorption; Nisin  – Absorption and adsorption
Contributors McGuire, Joseph (advisor); Schilke, Karl F. (committee member)
Language en
Country of Publication us
Record ID handle:1957/35800
Repository oregonstate
Date Indexed 2017-03-17
Grantor Oregon State University
Issued Date 2012-11-30 00:00:00
Note [] Graduation date: 2013; [peerreview] no;

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…56 B.5 Nisin activity against P. pentosaceus and S. epidermidis ............ 58 Appendix C Appendix D Nisin adsorption to polyethylene oxide layers and its resistance to elution in the presence of fibrinogen .................. 66 Quantifying…

nisin adsorption behavior at pendant PEO layers 87 LIST OF FIGURES Figure Page 2.1 Mechanism of nisin entrapment within PEO brush ...................................... 6 2.2 Schematic of layer creation and experimental methods…

…12 2.5a Comparison of nisin adsorption onto uncoated, F108, and F68 coated surfaces quantified with OWLS ..................................................................... 14 2.5b Comparison of nisin adsorption onto uncoated, F108, and F68…

…coated surfaces quantified with OWLS ..................................................................... 14 2.6a Relative nisin and fibrinogen adsorption onto F68 surfaces with and without nisin, quantified with OWLS…

…16 2.6b Relative nisin and fibrinogen adsorption onto F108 surfaces with and without nisin, quantified with OWLS ............................................................ 16 3.1 3.2 3.3 3.4 Proposed mechanism of activity retention of…

…entrapped nisin versus surface adsorbed nisin ................................................................................... 26 Diluted plasma adsorption to uncoated, F108 coated, and F108 coated with nisin surfaces, quantified with OWLS…

…78 Zeta potential measurements of nisin and fibrinogen adsorption to TCVS silanized silica microspheres ......................................................................... 79 Zeta potential measurements of nisin and fibrinogen adsorption to…

…F108 coated and TCVS silanized surfaces ............................................................. 80 Zeta potential detection of nisin and fibrinogen adsorption to EGAP-NTA coated and TCVS silanized surfaces…

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