RESUMO
Dissecting the mechanism of action of surface-tethered antimicrobial and immunomodulatory peptides is critical to the design of optimized anti-infection coatings on biomedical devices. To address this, we compared the biomembrane interactions of host defense peptide IDR-1010cys (1) in free form, (2) as a soluble polymer conjugate, and (3) with one end tethered to a solid support with model bacterial and mammalian lipid membranes. Our results show that IDR-1010cys in all three distinct forms interacted with bacterial and mammalian lipid vesicles, but the extent of the interactions as monitored by the induction of secondary structure varied. The enhanced interaction of surface-tethered peptides is well correlated with their very good antimicrobial activities. Our results demonstrate that there may be a difference in the mechanism of action of surface-tethered versus free IDR-1010cys.
Assuntos
Bicamadas Lipídicas/metabolismo , Peptídeos/química , Peptídeos Catiônicos Antimicrobianos/síntese química , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/metabolismo , Proteínas Imobilizadas/síntese química , Proteínas Imobilizadas/química , Proteínas Imobilizadas/metabolismo , Bicamadas Lipídicas/química , Modelos Biológicos , Peptídeos/síntese química , Peptídeos/metabolismo , Polímeros/química , Estrutura Secundária de Proteína , Quartzo/química , Propriedades de SuperfícieRESUMO
Primary amine containing copolymer, poly(N,N-dimethylacrylamide-co-N-(3-aminopropyl)methacrylamide hydrochloride) (poly(DMA-co-APMA)), brushes were synthesized on Ti surface by surface-initiated atom transfer radical polymerization (SI-ATRP) in aqueous conditions. A series of poly(DMA-co-APMA) copolymer brushes on titanium (Ti) surface with different molecular weights, thicknesses, compositions, and graft densities were synthesized by changing the SI-ATRP reaction conditions. Cysteine-functionalized cationic antimicrobial peptide Tet213 (KRWWKWWRRC) was conjugated to the copolymers brushes using a maleimide-thiol addition reaction after initial modification of the grafted chains using 3-maleimidopropionic acid N-hydroxysuccinimide ester. The modified surfaces were characterized by X-ray photoelectron spectroscopy (XPS), water contact angle measurements, attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, atomic force microscopy (AFM), and ellipsometry analysis. The conjugation of the Tet213 onto brushes strongly depended on graft density of the brushes at different copolymer brush compositions. The peptide density (peptides/nm(2)) on the surface varied with the initial composition of the copolymer brushes. Higher graft density of the brushes generated high peptide density (pepetide/nm(2)) and lower number of peptides/polymer chain and vice versa. The peptide density and graft density of the chains on surface greatly influenced the antimicrobial activity of peptide grafted polymer brushes against Pseudomonas aeruginosa.
Assuntos
Acrilamidas/química , Antibacterianos/síntese química , Peptídeos Catiônicos Antimicrobianos/síntese química , Materiais Revestidos Biocompatíveis/síntese química , Proteínas Imobilizadas/síntese química , Infecções por Pseudomonas/prevenção & controle , Pseudomonas aeruginosa/efeitos dos fármacos , Antibacterianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Materiais Revestidos Biocompatíveis/farmacologia , Humanos , Proteínas Imobilizadas/farmacologia , Testes de Sensibilidade Microbiana , Microscopia de Força Atômica , Peso Molecular , Espectroscopia Fotoeletrônica , Polimerização , Polímeros/química , Próteses e Implantes/microbiologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/crescimento & desenvolvimento , Espectroscopia de Infravermelho com Transformada de Fourier , Relação Estrutura-Atividade , Succinimidas/química , Propriedades de SuperfícieRESUMO
Bacterial colonization on implant surfaces and subsequent infections are one of the most common reasons for the failure of many indwelling devices. Several approaches including antimicrobial and antibiotic-eluting coatings on implants have been attempted; however, none of these approaches succeed in vivo. Here we report a polymer brush based implant coating that is non-toxic, antimicrobial and biofilm resistant. These coating consists of covalently grafted hydrophilic polymer chains conjugated with an optimized series of antimicrobial peptides (AMPs). These tethered AMPs maintained excellent broad spectrum antimicrobial activity in vitro and in vivo. We found that this specially structured robust coating was extremely effective in resisting biofilm formation, and that the biofilm resistance depended on the nature of conjugated peptides. The coating had no toxicity to osteoblast-like cells and showed insignificant platelet activation and adhesion, and complement activation in human blood. Since such coatings can be applied to most currently used implant surfaces, our approach has significant potential for the development of infection-resistant implants.
Assuntos
Antibacterianos/química , Biofilmes/crescimento & desenvolvimento , Materiais Revestidos Biocompatíveis/efeitos adversos , Próteses e Implantes/efeitos adversos , Próteses e Implantes/microbiologia , Animais , Antibacterianos/farmacologia , Linhagem Celular Tumoral , Dicroísmo Circular , Feminino , Humanos , Microscopia de Força Atômica , Peptídeos/efeitos adversos , Peptídeos/química , Peptídeos/farmacologia , Ativação Plaquetária/efeitos dos fármacos , Polímeros/efeitos adversos , Polímeros/química , Polímeros/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
N-Propargylamide-terminated peptide-based macromonomers with a degree of polymerization ranging from 4 to 40 were synthesized by the polymerization of gamma-benzyl and gamma-stearyl-L-glutamate-N-carboxy anhydrides initiated with propargylamine. The macromonomers took an alpha-helical structure, which was confirmed by signals at 208 and 220 nm in CD spectra. The macromonomers were subjected to polymerization and copolymerization with an alanine-derived N-propargylamide [N-(tert-butoxycarbonyl)-L-alanine-N'-propargylamide] catalyzed with (2,5-norbornadiene)Rh+[eta6-C6H5B- (C6H5)3]. It was confirmed through a CD spectroscopic study that the copolymer obtained from the copolymerization of the gamma-stearyl-L-glutamate-based macromonomer with the alanine-derived N-propargylamide had a helical polyacetylene main chain and helical polypeptide side chains.