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1.
Molecules ; 19(6): 8610-28, 2014 Jun 24.
Artículo en Inglés | MEDLINE | ID: mdl-24962395

RESUMEN

Poly(ethylene glycol) (PEG)-decorated polystyrene (PS) nanoparticles with mean hydrodynamic diameter (D) and zeta-potential (ζ) of (286 ± 15) nm and (-50 ± 5) mV, respectively, were modified by the adsorption of Congo red (CR). The PS/PEG/CR particles presented D and ζ values of (290 ± 19) nm and (-36 ± 5) mV, respectively. The adsorption of lipase onto PS/PEG or PS/PEG/CR particles at (24 ± 1) °C and pH 7 changed the mean D value to (380 ± 20) and (405 ± 11) nm, respectively, and ζ value to (-32 ± 4) mV and (-25 ± 2) mV, respectively. The kinetic parameters of the hydrolysis of p-nitrophenyl butyrate were determined for free lipase, lipase immobilized onto PS/PEG and PS/PEG/CR particles. Lipase on PS/PEG/CR presented the largest Michaelis-Menten constant (KM), but also the highest Vmax and kcat values. Moreover, it could be recycled seven times, losing a maximum 10% or 30% of the original enzymatic activity at 40 °C or 25 °C, respectively. Although lipases immobilized onto PS/PEG particles presented the smallest KM values, the reactions were comparatively the slowest and recycling was not possible. Hydrolysis reactions performed in the temperature range of 25 °C to 60 °C with free lipases and lipases immobilized onto PS/PEG/CR particles presented an optimal temperature at 40 °C. At 60 °C free lipases and lipases immobilized onto PS/PEG/CR presented ~80% and ~50% of the activity measured at 40 °C, indicating good thermal stability. Bioconjugation effects between CR and lipase were evidenced by circular dichroism spectroscopy and spectrophotometry. CR molecules mediate the open state conformation of the lipase lid and favor the substrate approaching.


Asunto(s)
Butiratos/metabolismo , Enzimas Inmovilizadas/farmacocinética , Lipasa/metabolismo , Lipasa/farmacocinética , Nanopartículas/metabolismo , Adsorción , Butiratos/química , Candida/enzimología , Dominio Catalítico , Dicroismo Circular , Rojo Congo/química , Enzimas Inmovilizadas/química , Interacciones Hidrofóbicas e Hidrofílicas , Lipasa/química , Nanopartículas/química , Polietilenglicoles/química , Poliestirenos/química , Espectrofotometría
2.
Biochim Biophys Acta ; 1818(12): 3064-71, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22960286

RESUMEN

The interaction between the antimicrobial peptide gramicidin (Gr) and dipalmitoylphosphatidylcholine (DPPC)/dioctadecyldimethylammonium bromide (DODAB) 1:1 large unilamellar vesicles (LVs) or bilayer fragments (BFs) was evaluated by means of several techniques. The major methods were: 1) Gr intrinsic fluorescence and circular dichroism (CD) spectroscopy; 2) dynamic light scattering for sizing and zeta-potential analysis; 3) determination of the bilayer phase transition from extrinsic fluorescence of bilayer probes; 4) pictures of the dispersions for evaluation of coloidal stability over a range of time and NaCl concentration. For Gr in LVs, the Gr dimeric channel conformation is suggested from: 1) CD and intrinsic fluorescence spectra similar to those in trifluoroethanol (TFE); 2) KCl or glucose permeation through the LVs/Gr bilayer. For Gr in BFs, the intertwined dimeric, non-channel Gr conformation is evidenced by CD and intrinsic fluorescence spectra similar to those in ethanol. Both LVs and BFs shield Gr tryptophans against quenching by acrylamide but the Stern-Volmer quenching constant was slightly higher for Gr in BFs confirming that the peptide is more exposed to the water phase in BFs than in LVs. The DPPC/DODAB/Gr supramolecular assemblies may predict the behavior of other antimicrobial peptides in assemblies with lipids.


Asunto(s)
1,2-Dipalmitoilfosfatidilcolina/metabolismo , Gramicidina/metabolismo , Membrana Dobles de Lípidos/metabolismo , Compuestos de Amonio Cuaternario/metabolismo , 1,2-Dipalmitoilfosfatidilcolina/química , Gramicidina/química , Membrana Dobles de Lípidos/química , Lípidos/química , Conformación Molecular , Transición de Fase , Compuestos de Amonio Cuaternario/química
3.
Langmuir ; 29(31): 9677-84, 2013 Aug 06.
Artículo en Inglés | MEDLINE | ID: mdl-23841487

RESUMEN

The purpose of this Article is to characterize polymeric particles of poly(methylmethacrylate) (PMMA) synthesized in the presence of one of two different quaternary ammonium surfactants (QACs): cetyltrimethylammonium bromide (CTAB) or dioctadecyldimethylammonium bromide (DODAB). The methods used are dynamic light scattering for sizing, polydispersity and zeta potential analysis, scanning electron microscopy (SEM) for morphology visualization, and plating plus colony-forming unities (CFU) counting for the determination of antimicrobial activity. The results point out the high QAC concentration required to obtain cationic and bioactive antimicrobial particles with good colloidal stability and a permanent load of the polymeric network with QACs. Over a range of micromolar QAC concentrations, there is remarkable antimicrobial activity of PMMA/CTAB or PMMA/DODAB particles, which is much higher than those determined for the QACs by themselves. Loading the biocompatible polyacrylate particles with QACs is a facile, fast, low-cost approach to obtaining highly efficient antimicrobial nanoparticles.


Asunto(s)
Antibacterianos/farmacología , Polimetil Metacrilato/farmacología , Pseudomonas aeruginosa/efectos de los fármacos , Compuestos de Amonio Cuaternario/química , Staphylococcus aureus/efectos de los fármacos , Tensoactivos/química , Antibacterianos/síntesis química , Antibacterianos/química , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Emulsiones/síntesis química , Emulsiones/química , Emulsiones/farmacología , Pruebas de Sensibilidad Microbiana , Tamaño de la Partícula , Polimerizacion , Polimetil Metacrilato/síntesis química , Polimetil Metacrilato/química , Pseudomonas aeruginosa/citología , Staphylococcus aureus/citología , Relación Estructura-Actividad , Propiedades de Superficie
4.
Pharmaceuticals (Basel) ; 16(6)2023 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-37375764

RESUMEN

Cationic and hydrophilic coatings based on casting and drying water dispersions of two different nanoparticles (NPs) onto glass are here described and evaluated for antimicrobial activity. Discoid cationic bilayer fragments (BF) surrounded by carboxy-methylcellulose (CMC) and poly (diallyl dimethyl ammonium) chloride (PDDA) NPs and spherical gramicidin D (Gr) NPs dispersed in water solution were cast onto glass coverslips and dried, forming a coating quantitatively evaluated against Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. From plating and colony forming units (CFU) counting, all strains interacting for 1 h with the coatings lost viability from 105 to 106, to zero CFU, at two sets of Gr and PDDA doses: 4.6 and 25 µg, respectively, or, 0.94 and 5 µg, respectively. Combinations produced broad spectrum, antimicrobial coatings; PDDA electrostatically attached to the microbes damaging cell walls, allowing Gr NPs interaction with the cell membrane. This concerted action promoted optimal activity at low Gr and PDDA doses. Further washing and drying of the deposited dried coatings showed that they were washed out so that antimicrobial activity was no longer present on the glass surface. Significant applications in biomedical materials can be foreseen for these transient coatings.

5.
Biochim Biophys Acta ; 1808(3): 649-55, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21147062

RESUMEN

The interaction between cationic bilayer fragments and a model oligonucleotide was investigated by differential scanning calorimetry, turbidimetry, determination of excimer to monomer ratio of 2-(10-(1-pyrene)-decanoyl)-phosphatidyl-choline in bilayer fragment dispersions and dynamic light scattering for sizing and zeta-potential analysis. Salt (Na2HPO4), mononucleotide (2'-deoxyadenosine-5'-monophosphate) or poly (dA) oligonucleotide (3'-AAA AAA AAA A-5') affected structure and stability of dioctadecyldimethylammonium bromide bilayer fragments. Oligonucleotide and salt increased bilayer packing due to bilayer fragment fusion. Mononucleotide did not reduce colloid stability or did not cause bilayer fragment fusion. Charge neutralization of bilayer fragments by poly (dA) at 1:10 poly (dA):dioctadecyldimethylammonium bromide molar ratio caused extensive aggregation, maximal size and zero of zeta-potential for the assemblies. Above charge neutralization, assemblies recovered colloid stability due to charge overcompensation. For bilayer fragments/poly (dA), the nonmonotonic behavior of colloid stability as a function of poly (dA) concentration was unique for the oligonucleotide and was not observed for Na2HPO4 or 2'-deoxyadenosine-5'-monophosphate. For the first time, such interactions between cationic bilayer fragments and mono- or oligonucleotide were described in the literature. Bilayer fragments/oligonucleotide assemblies may find interesting applications in drug delivery.


Asunto(s)
Cationes/química , Cationes/metabolismo , Membrana Dobles de Lípidos/química , Membrana Dobles de Lípidos/metabolismo , Compuestos de Amonio Cuaternario/química , Compuestos de Amonio Cuaternario/metabolismo , Rastreo Diferencial de Calorimetría , Lípidos/química , Nefelometría y Turbidimetría
6.
BMC Biotechnol ; 11: 40, 2011 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-21496246

RESUMEN

BACKGROUND: Cationic bilayers based on the inexpensive synthetic lipid dioctadecyldimethylammonium bromide (DODAB) have been useful as carriers for drug delivery, immunoadjuvants for vaccines and active antimicrobial agents. METHODS: Rifampicin (RIF) or isoniazid (ISO) interacted with DODAB bilayer fragments (BF) or large vesicles (LV). Dispersions were evaluated by dynamic light-scattering for zeta-average diameter (Dz) and zeta-potential (ζ) analysis; dialysis for determination of drug entrapment efficiency; plating and CFU counting for determination of cell viability of Mycobacterium smegmatis or tuberculosis, minimal bactericidal concentration (MBC) and synergism index for DODAB/drug combinations. RESULTS: DODAB alone killed micobacteria over a range of micromolar concentrations. RIF aggregates in water solution were solubilised by DODAB BF. RIF was incorporated in DODAB bilayers at high percentiles in contrast to the leaky behavior of ISO. Combination DODAB/RIF yielded MBCs of 2/2 and 4/0.007 µg/mL against Mycobacterium smegmatis or Mycobacterium tuberculosis, respectively. Synergism indexes equal to 0.5 or 1.0, indicated synergism against the former and independent action, against the latter species. CONCLUSIONS: In vitro, DODAB acted effectively both as micobactericidal agent and carrier for rifampicin. The novel assemblies at reduced doses may become valuable against tuberculosis.


Asunto(s)
Antibacterianos/farmacología , Viabilidad Microbiana/efectos de los fármacos , Compuestos de Amonio Cuaternario/farmacología , Rifampin/farmacología , Antibacterianos/química , Antiinfecciosos/química , Antiinfecciosos/farmacología , Antibióticos Antituberculosos/química , Antibióticos Antituberculosos/farmacología , Portadores de Fármacos , Composición de Medicamentos/métodos , Sinergismo Farmacológico , Membrana Dobles de Lípidos/química , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Mycobacterium smegmatis/efectos de los fármacos , Mycobacterium smegmatis/crecimiento & desarrollo , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/crecimiento & desarrollo , Compuestos de Amonio Cuaternario/química , Rifampin/química , Tecnología Farmacéutica/métodos
7.
Langmuir ; 26(14): 12300-6, 2010 Jul 20.
Artículo en Inglés | MEDLINE | ID: mdl-20578678

RESUMEN

Hybrid nanoparticles from cationic lipid and polymers were prepared and characterized regarding physical properties and antimicrobial activity. Carboxymethylcellulose (CMC) and polydiallyldimethylammonium chloride (PDDA) were sequentially added to cationic bilayer fragments (BF) prepared from ultrasonic dispersion in water of the synthetic and cationic lipid dioctadecyldimethylammonium bromide (DODAB). Particles thus obtained were characterized by dynamic light-scattering for determination of z-average diameter (Dz) and zeta-potential (zeta). Antimicrobial activity of the DODAB BF/CMC/PDDA particles against Pseudomonas aeruginosa or Staphylococcus aureus was determined by plating and CFU counting over a range of particle compositions. DODAB BF/CMC/PDDA particles exhibited sizes and zeta-potentials strictly dependent on DODAB, CMC, and PDDA concentrations. At 0.1 mM DODAB, 0.1 mg/mL CMC, and 0.1 mg/mL PDDA, small cationic particles with Dz = 100 nm and zeta = 30 mV were obtained. At 0.5 mM DODAB, 0.5 mg/mL CMC and 0.5 mg/mL PDDA, large cationic particles with Dz = 470 nm and zeta = 50 mV were obtained. Both particulates were highly reproducible regarding physical properties and yielded 0% of P. aeruginosa viability (10(7) CFU/mL) at 1 or 2 microg/mL PDDA dissolved in solution or in form of particles, respectively. 99% of S. aureus cells died at 10 microg/mL PDDA alone or in small or large DODAB BF/CMC/PDDA particles. The antimicrobial effect was dependent on the amount of positive charge on particles and independent of particle size. A high microbicide potency for PDDA over a range of nanomolar concentrations was disclosed. P. aeruginosa was more sensitive to all cationic assemblies than S. aureus.


Asunto(s)
Antiinfecciosos/química , Antiinfecciosos/farmacología , Electrólitos/química , Lípidos/química , Nanopartículas/química , Polímeros/química , Carboximetilcelulosa de Sodio/química , Supervivencia Celular/efectos de los fármacos , Modelos Moleculares , Conformación Molecular , Polietilenos/química , Pseudomonas aeruginosa/citología , Pseudomonas aeruginosa/efectos de los fármacos , Compuestos de Amonio Cuaternario/química , Staphylococcus aureus/citología , Staphylococcus aureus/efectos de los fármacos
8.
BMC Biotechnol ; 9: 5, 2009 Jan 19.
Artículo en Inglés | MEDLINE | ID: mdl-19152701

RESUMEN

BACKGROUND: Silica particles cationized by dioctadecyldimethylammonium bromide (DODAB) bilayer were previously described. This work shows the efficiency of these particulates for antigen adsorption and presentation to the immune system and proves the concept that silica-based cationic bilayers exhibit better performance than alum regarding colloid stability and cellular immune responses for vaccine design. RESULTS: Firstly, the silica/DODAB assembly was characterized at 1 mM NaCl, pH 6.3 or 5 mM Tris.HCl, pH 7.4 and 0.1 mg/ml silica over a range of DODAB concentrations (0.001-1 mM) by means of dynamic light scattering for particle sizing and zeta-potential analysis. 0.05 mM DODAB is enough to produce cationic bilayer-covered particles with good colloid stability. Secondly, conditions for maximal adsorption of bovine serum albumin (BSA) or a recombinant, heat-shock protein from Mycobacterium leprae (18 kDa-hsp) onto DODAB-covered or onto bare silica were determined. At maximal antigen adsorption, cellular immune responses in vivo from delayed-type hypersensitivity reactions determined by foot-pad swelling tests (DTH) and cytokines analysis evidenced the superior performance of the silica/DODAB adjuvant as compared to alum or antigens alone whereas humoral response from IgG in serum was equal to the one elicited by alum as adjuvant. CONCLUSION: Cationized silica is a biocompatible, inexpensive, easily prepared and possibly general immunoadjuvant for antigen presentation which displays higher colloid stability than alum, better performance regarding cellular immune responses and employs very low, micromolar doses of cationic and toxic synthetic lipid.


Asunto(s)
Adyuvantes Inmunológicos/farmacología , Presentación de Antígeno , Compuestos de Amonio Cuaternario/inmunología , Dióxido de Silicio/inmunología , Adyuvantes Inmunológicos/química , Adsorción , Animales , Formación de Anticuerpos , Cationes , Células Cultivadas , Citocinas/análisis , Ensayo de Inmunoadsorción Enzimática , Femenino , Hipersensibilidad Tardía/inmunología , Membrana Dobles de Lípidos/química , Membrana Dobles de Lípidos/inmunología , Ratones , Ratones Endogámicos BALB C , Compuestos de Amonio Cuaternario/química , Albúmina Sérica Bovina/metabolismo , Dióxido de Silicio/química
9.
J Phys Chem B ; 112(51): 16422-30, 2008 Dec 25.
Artículo en Inglés | MEDLINE | ID: mdl-19367939

RESUMEN

The interaction between giant bacteriophage DNA and cationic biomimetic particles was characterized from sizing by dynamic light-scattering, zeta-potential analysis, turbidimetry, determination of colloid stability, visualization from atomic force microscopy (AFM), and determination of cytotoxicity against E. coli from colony forming unities counting. First, polystyrene sulfate (PSS) particles with different sizes were covered by a dioctadecyldimethylammonium bromide (DODAB) bilayer yielding the so-called cationic biomimetic particles (PSS/DODAB). These cationic particles are highly organized, present a narrow size distribution and were obtained over a range of particle sizes. Thereafter, upon adding lambda, T5 or T2-DNA to PSS/DODAB particles, supramolecular assemblies PSS/DODAB/DNA were obtained and characterized over a range of DNA concentrations and particle sizes (80-700 nm). Over the low DNA concentration range, PSS/DODAB/ DNA assemblies were cationic, colloidally stable with moderate polydispersity and highly cytotoxic against E. coli. From DNA concentration corresponding to charge neutralization, neutral or anionic supramolecular assemblies PSS/DODAB/DNA exhibited low colloid stability, high polydispersity and moderate cytotoxicity. Some nucleosome mimetic assemblies were observed by AFM at charge neutralization (zeta-potential equal to zero).


Asunto(s)
Bacteriófagos/genética , ADN Viral/metabolismo , Imitación Molecular , Cationes
10.
J Phys Chem B ; 112(31): 9301-10, 2008 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-18630858

RESUMEN

Intermolecular associations between a cationic lipid and two model polymers were evaluated from preparation and characterization of hybrid thin films cast on silicon wafers. The novel materials were prepared by spin-coating of a chloroformic solution of lipid and polymer on silicon wafer. Polymers tested for miscibility with the cationic lipid dioctadecyldimethylammonium bromide (DODAB) were polystyrene (PS) and poly(methyl methacrylate) (PMMA). The films thus obtained were characterized by ellipsometry, wettability, optical and atomic force microscopy, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and activity against Escherichia coli. Whereas intermolecular ion-dipole interactions were available for the PMMA-DODAB interacting pair producing smooth PMMA-DODAB films, the absence of such interactions for PS-DODAB films caused lipid segregation, poor film stability (detachment from the silicon wafer) and large rugosity. In addition, the well-established but still remarkable antimicrobial DODAB properties were transferred to the novel hybrid PMMA/DODAB coating, which is demonstrated to be highly effective against E. coli.


Asunto(s)
Cationes/química , Lípidos/química , Polimetil Metacrilato/química , Poliestirenos/química , Rastreo Diferencial de Calorimetría , Escherichia coli/citología , Viabilidad Microbiana , Microscopía de Fuerza Atómica , Compuestos de Amonio Cuaternario/química , Dióxido de Silicio/química , Agua/química
11.
Chem Phys Lipids ; 152(1): 38-45, 2008 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-18258186

RESUMEN

Dioctadecyldimethylammonium bromide (DODAB)/dipalmitoylphosphatidylcholine (DPPC) large and cationic vesicles obtained by vortexing a lipid film in aqueous solution and above the mean phase transition temperature (T(m)) are characterized by means of determination of phase behaviour, size distribution, zeta-potential analysis and colloid stability. The effect of increasing % DODAB over the 0-100% range was a nonmonotonic phase behaviour. At 50% DODAB, the mean phase transition temperature and the colloid stability were at maximum. There is an intimate relationship between stability of the bilayer structure and colloid stability. In 1, 50 and 150mM NaCl, the colloid stability for pure DPPC or pure DODAB vesicles was very low as observed by sedimentation or flocculation, respectively. In contrast, at 50% DODAB, remarkable colloid stability was achieved in 1, 50 or 150mM NaCl for the DODAB/DPPC composite vesicles. Vesicle size decreased but the zeta-potential remained constant with % DODAB, due to a decrease of counterion binding with vesicle size. This might be important for several biotechnological applications currently being attempted with cationic bilayer systems.


Asunto(s)
1,2-Dipalmitoilfosfatidilcolina/química , Membrana Dobles de Lípidos/química , Compuestos de Amonio Cuaternario/química , Coloides , Estabilidad de Medicamentos , Nefelometría y Turbidimetría , Transición de Fase , Temperatura
12.
J Nanobiotechnology ; 6: 6, 2008 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-18462496

RESUMEN

BACKGROUND: Particulate systems are well known to be able to deliver drugs with high efficiency and fewer adverse side effects, possibly by endocytosis of the drug carriers. On the other hand, cationic compounds and assemblies exhibit a general antimicrobial action. In this work, cationic nanoparticles built from drug, cationic lipid and polyelectrolytes are shown to be excellent and active carriers of amphotericin B against C. albicans. RESULTS: Assemblies of amphotericin B and cationic lipid at extreme drug to lipid molar ratios were wrapped by polyelectrolytes forming cationic nanoparticles of high colloid stability and fungicidal activity against Candida albicans. Experimental strategy involved dynamic light scattering for particle sizing, zeta-potential analysis, colloid stability, determination of AmB aggregation state by optical spectra and determination of activity against Candida albicans in vitro from cfu countings. CONCLUSION: Novel and effective cationic particles delivered amphotericin B to C. albicans in vitro with optimal efficiency seldom achieved from drug, cationic lipid or cationic polyelectrolyte in separate. The multiple assembly of antibiotic, cationic lipid and cationic polyelctrolyte, consecutively nanostructured in each particle produced a strategical and effective attack against the fungus cells.

13.
Int J Biol Macromol ; 115: 792-800, 2018 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-29702165

RESUMEN

This work was motivated by the need of stimuli responsive drug carriers, which can be activated by low cost non-invasive stimuli such as external magnetic field (EMF). Thus, novel antimicrobial materials based on xanthan gum (XG), magnetic nanoparticles (MNP), bovine serum albumin (BSA) and amoxicillin (Amox) were designed in order to promote the release of Amox under magnetic stimuli. Firstly, surfaces with different functionalities were prepared by sequential deposition of thin layers on Si wafers and characterized by means of ellipsometry and atomic force microscopy. Amox adsorbed preferentially onto XG or BSA films. In solution, favorable interactions between Amox and BSA were evidenced by substantial changes in the BSA secondary structure, as revealed by circular dichroism. Patches of XG and XG/MNP/BSA were immersed in 2 g L-1 Amox, yielding 10 ±â€¯3 and 17 ±â€¯4 µg/cm3 Amox loading, respectively. The inclusion of 0.2 wt% Fe3O4 in the patches and their exposure to EMF enabled in vitro release of Amox, at pH 5.5 and 0.02 mol L-1 NaCl, following the quasi-Fickian behavior. Amox diffused from XG/MNP/BSA patches in agar medium containing Staphylococcus aureus and Escherichia coli, inhibiting their growth. The inhibition of E. coli growth was particularly efficient under EMF.


Asunto(s)
Amoxicilina/química , Portadores de Fármacos/química , Liberación de Fármacos , Óxido Ferrosoférrico/química , Campos Magnéticos , Polisacáridos Bacterianos/química , Albúmina Sérica Bovina/química , Adsorción , Amoxicilina/farmacología , Animales , Antibacterianos/química , Antibacterianos/farmacología , Bovinos , Escherichia coli/efectos de los fármacos , Concentración de Iones de Hidrógeno , Staphylococcus aureus/efectos de los fármacos
14.
J Phys Chem B ; 111(29): 8520-6, 2007 Jul 26.
Artículo en Inglés | MEDLINE | ID: mdl-17407341

RESUMEN

Chitosan (CH) decorated polystyrene (PS) particles were synthesized within complexes of CH, a polycation under acid conditions, and tiny amounts of sodium dodecylsulfate (SDS). Particle characterization was performed by means of dynamic light scattering, zeta potential measurements, and transmission electron microscopy. All dispersions were stable in the ionic strength of 2.0 mol L-1 NaCl during 2 months. The outstanding colloidal stability was attributed to the presence of a hydrated CH layer around the particles. CH decorated PS particles were attached to atomic force microscopy cantilevers and probed against Si wafers in water and in NaCl 0.01 mol/L. The mean thickness of CH layer amounted to 35 +/- 11 and 16 +/- 6 nm, when the medium was water and NaCl 0.01 mol/L, respectively. Adsorption isotherm of hexokinase (HK) onto PS/CH particles studied by means of spectrophotometry showed three regions: an initial step; adsorption plateau and multilayer formation. Enzymatic activity of free HK and immobilized HK was monitored by means of spectrophotometry as a function of storing time and reuse. After 3 days, storing HK free in solution dramatically lost its catalytic properties. On the contrary, HK-covered PS/CH particles retained enzymatic activity over 1 month. Moreover, HK-covered PS/CH particles could be reused in the determination of glucose two times consecutively, without losing activity. These interesting findings were discussed in light of the role of water in enzyme conformation.


Asunto(s)
Quitosano/química , Enzimas Inmovilizadas/química , Hexoquinasa/química , Microesferas , Poliestirenos/química , Coloides/química , Conformación Proteica , Dodecil Sulfato de Sodio/química , Análisis Espectral , Agua/química
15.
J Colloid Interface Sci ; 313(2): 519-26, 2007 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-17532328

RESUMEN

The interaction between composite dipalmitoylphosphatidylcholine (DPPC)/dioctadecyldimethylammonium bromide (DODAB) bilayer vesicles in the gel state and silica is investigated over the 0-20% DODAB range from determination of adsorption curves, silica sedimentation, particle sizing and zeta-potentials. At 1 mg/mL silica, 0% DODAB, pH 6.3, over the 0-150 mM NaCl range of ionic strengths, high affinity adsorption curves were barely affected by ionic strength and all of them exhibited limiting adsorption values above the level expected for single bilayer deposition. At 1 mg/mL silica, 2% DODAB, pH 6.3 and 1 mM NaCl, high affinity adsorption curves fortuitously presented limiting adsorption indicative of one bilayer deposition on each silica particle. At %DODAB<2% or %DODAB>2%, limiting adsorption was above and below the level expected for bilayer deposition, respectively. Increasing %DODAB in the vesicle composition negatively modulated the limiting adsorption on silica despite the increasing surface charge on vesicles and electrostatic attraction between vesicles and particles. The results point out the difficulty of closed vesicle disruption (required for bilayer deposition from vesicles) when the bilayer is tightly packed in the rigid gel state and might be of interest for analytical applications of immobilized vesicles on silica.


Asunto(s)
1,2-Dipalmitoilfosfatidilcolina/química , Membrana Dobles de Lípidos/química , Compuestos de Amonio Cuaternario/química , Dióxido de Silicio/química , Adsorción , Geles/química , Concentración de Iones de Hidrógeno , Tamaño de la Partícula , Cloruro de Sodio/química
16.
Int J Biol Macromol ; 41(4): 404-9, 2007 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-17640725

RESUMEN

The adsorption behavior of horseradish peroxidase (HRP) onto hybrid particles of poly(methylmethacrylate) (PMMA) and carboxymethylcellulose (CMC) was investigated by means of spectrophotometry. Dispersions of PMMA/CMC particles were characterized by light scattering, zeta potential measurements and scanning electron microscopy before and after HRP adsorption. HRP adsorbed irreversibly onto PMMA/CMC particles; the adsorption isotherm showed an initial step and an adsorption plateau. The enzymatic activity of free HRP and immobilized HRP (plateau region) was monitored by means of spectrophotometry as a function of storing time. Upon adsorbing HRP there is little (up to 20%) or no reduction of enzymatic activity in comparison to that observed for free HRP in solution. After storing free HRP and HRP-covered PMMA/CMC particles for 18 days the level of enzymatic activity is kept. HRP-covered PMMA/CMC particles dispersions, which were dried and re-dispersed, retained 50% of their catalytic properties. These interesting findings were discussed in the light of a beneficial effect of a hydrated microenvironment for maintenance of enzyme conformation and activity.


Asunto(s)
Carboximetilcelulosa de Sodio/química , Peroxidasa de Rábano Silvestre/metabolismo , Peroxidasa de Rábano Silvestre/farmacocinética , Polimetil Metacrilato/química , Polisacáridos/química , Adsorción , Catálisis , Enzimas Inmovilizadas/química , Luz , Microscopía Electrónica de Rastreo , Oxidación-Reducción , Dispersión de Radiación , Espectrofotometría Ultravioleta , Factores de Tiempo
17.
Nanomaterials (Basel) ; 7(12)2017 Dec 02.
Artículo en Inglés | MEDLINE | ID: mdl-29207496

RESUMEN

Nanostructured particles of polystyrene sulfate (PSS) covered by a cationic lipid bilayer of dioctadecyldimethylammonium bromide (DODAB) incorporated gramicidin D (Gr) yielding optimal and broadened bactericidal activity against both Escherichia coli and Staphylococcus aureus. The adsorption of DODAB/Gr bilayer onto PSS nanoparticles (NPs) increased the zeta-average diameter by 8-10 nm, changed the zeta-potential of the NPs from negative to positive, and yielded a narrow size distributions for the PSS/DODAB/Gr NPs, which displayed broad and maximal microbicidal activity at very small concentrations of the antimicrobials, namely, 0.057 and 0.0057 mM DODAB and Gr, respectively. The results emphasized the advantages of highly-organized, nanostructured, and cationic particles to achieve hybrid combinations of antimicrobials with broad spectrum activity at considerably reduced DODAB and Gr concentrations.

18.
Biomimetics (Basel) ; 2(4)2017 Oct 20.
Artículo en Inglés | MEDLINE | ID: mdl-31105181

RESUMEN

The optimization of bilayer coverage on particles is important for a variety of biomedical applications, such as drug, vaccine, and genetic material delivery. This work aims at optimizing the deposition of cationic bilayers on silica over a range of experimental conditions for the intervening medium and two different assemblies for the cationic lipid, namely, lipid films or pre-formed lipid bilayer fragments. The lipid adsorption on silica in situ over a range of added lipid concentrations was determined from elemental analysis of carbon, hydrogen, and nitrogen and related to the colloidal stability, sizing, zeta potential, and polydispersity of the silica/lipid nanoparticles. Superior bilayer deposition took place from lipid films, whereas adsorption from pre-formed bilayer fragments yielded limiting adsorption below the levels expected for bilayer adsorption.

19.
Carbohydr Polym ; 165: 285-293, 2017 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-28363551

RESUMEN

Hydroxypropyl methylcellulose (HPMC) and xyloglucan (XG) crosslinked with citric acid over a range of HPMC/XG weight ratios formed sustainable blend films characterized by Fourier transform infrared spectroscopy, thermogravimetric analysis, scanning electron microscopy, tensile tests, circular dichroism and determination of inhibitory activity against Staphylococcus aureus and Escherichia coli. Both in solution and in the crosslinked films, HPMC chains lost the original ordered conformation upon interacting with XG, giving rise to an entropic gain. The highest values of tensile strength (25MPa) and Young's modulus (689MPa) occurred for the 50:50 HPMC/XG blend films. In vitro loading of gentamicin sulfate (GS) in the films amounted to 0.18±0.05 -0.37±0.05g of GS per g polymer. At pH 7.4 and 37°C, the GS release kinetics from the films fitted with the Korsmeyer-Peppas model revealed a non-Fickian release mechanism with diffusional coefficient n∼0.7. The cross-linked films of HPMC, XG and their blends loaded with GS showed outstanding antibacterial activity against Staphylococcus aureus and Escherichia coli, disclosing their potential for novel biomedical applications.


Asunto(s)
Antiinfecciosos/farmacología , Gentamicinas/farmacología , Glucanos/farmacología , Derivados de la Hipromelosa/farmacología , Xilanos/farmacología , Escherichia coli/efectos de los fármacos , Metilcelulosa , Staphylococcus aureus/efectos de los fármacos
20.
Cell Biochem Biophys ; 44(3): 446-52, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16679532

RESUMEN

Biomimetic particles supporting lipid bilayers are becoming increasingly important to isolate and reconstitute protein function. Cholera toxin (CT) from Vibrio cholerae, an 87-kDa AB5 hexameric protein, and its receptor, the monosialoganglioside GM1, a cell membrane glycolipid, self-assembled on phosphatidylcholine (PC) bilayer-covered silica particles at 1 CT/5 GM1 molar ratio in perfect agreement with literature. This receptor-ligand recognition represented a proof-of-concept that receptors in general can be isolated and their function reconstituted using biomimetic particles, i.e., bilayer-covered silica. After incubation of colloidal silica with small unilamellar PC vesicles in saline solution, pH 7.4, PC adsorption isotherms on silica from inorganic phosphorus analysis showed a high PC affinity for silica with maximal PC adsorption at bilayer deposition. At 0.3 mM PC, fluorescence of pyrene-labeled GM(1) showed that GM(1) incorporation in biomimetic particles increased as a function of particles concentration. At 1 mg/mL silica, receptor incorporation increased to a maximum of 40% at 0.2-0.3 mM PC and then decreased as a function of PC concentration. At 5 microM GM(1), 0.3 mM PC, and 1 mg/mL silica, CT binding increased as a function of CT concentration with a plateau at 2 mg bound CT/m2 silica, which corresponded to the 5 GM(1)/1 CT molar proportion and showed successful reconstitution of receptor-ligand interaction.


Asunto(s)
Toxina del Cólera/química , Gangliósido G(M1)/química , Membrana Dobles de Lípidos/química , Fosfolípidos/química , Receptores de Superficie Celular/fisiología , Adsorción , Animales , Biomimética/métodos , Bovinos , Fenómenos Químicos , Química Física , Calor , Control de Infecciones , Liposomas/química , Lípidos de la Membrana , Tamaño de la Partícula , Fosfatidilcolinas/química , Pirenos/química , Receptores de Superficie Celular/aislamiento & purificación , Dióxido de Silicio/química , Vibrio cholerae/enzimología
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