RESUMO
Herein, smart coatings based on photo-responsive polymer nanocapsules (NC) and deposited by laser evaporation are presented. These systems combine remotely controllable release and high encapsulation efficiency of nanoparticles with the easy handling and safety of macroscopic substrates. In particular, azobenzene-based NC loaded with active molecules (thyme oil and coumarin 6) were deposited through Matrix-Assisted Pulsed Laser Evaporation (MAPLE) on flat inorganic (KBr) and organic (polyethylene, PE) and 3D (acrylate-based micro-needle array) substrates. SEM analyses highlighted the versatility and performance of MAPLE in the fabrication of the designed smart coatings. DLS analyses, performed on both MAPLE- and drop casting-deposited NC, demonstrated the remarkable adhesion achieved with MAPLE. Finally, thyme oil and coumarin 6 release experiments further demonstrated that MAPLE is a promising technique for the realization of photo-responsive coatings on various substrates.
RESUMO
ß-glucosidase (BG) plays a key role in determining the efficiency of the enzymatic complex cellulase for the degradation of cellulose into sugars. It hydrolyses the cellobiose, an inhibitor of the enzymatic complex. Therefore, the immobilization of BG is a great challenge for the industrial application of cellulases. Cellulases usually contains a BG amount insufficient to avoid inhibition by cellobiose. Here the BG was immobilized by matrix assisted pulsed laser evaporation (MAPLE) technique. The frozen matrix was composed of water, water/m-DOPA and water/m-DOPA/quinone. The effect of the excipients on the final conformation of the enzyme after the MAPLE processing was determined. The enzyme secondary structure was studied by FTIR analysis. The catalytic performances of the deposited films were tested in the cellobiose hydrolysis reaction. The results demonstrate that the presence of the oxidized form of m-DOPA, the O-quinone form, can protect the protein native structure, with the laser inducing little or no damage. In fact, only the samples deposited from this target preserved the secondary structure of the polypeptide chain and allowed a complete hydrolysis of cellobiose for four consecutive runs, showing a high operational stability of the biocatalyst.
Assuntos
Benzoquinonas/metabolismo , Di-Hidroxifenilalanina/análogos & derivados , Di-Hidroxifenilalanina/metabolismo , beta-Glucosidase/metabolismo , Catálise , Celobiose/metabolismo , Celulase/metabolismo , Enzimas Imobilizadas/metabolismo , Concentração de Íons de Hidrogênio , Hidrólise , Complexos Multienzimáticos , Quinonas/metabolismo , TemperaturaRESUMO
The deposition of thin films of poly(D,L-lactide) (PDLLA) by using the matrix-assisted pulsed laser evaporation (MAPLE) technique is investigated. PDLLA is a highly biocompatible and biodegradable polymer, with wide applicability in the biomedical field. The laser wavelength used in the MAPLE process is optimized to obtain a good-quality deposition. The structure of the polymer film is analyzed by Fourier transform infrared spectroscopy (FTIR). It is found that the chemical structure of PDLLA undergoes little or no damage during deposition with near-infrared laser radiation (1064 nm). It is thus confirmed that at this wavelength, the MAPLE technique can be applied for fragile biopolymer molecules, which are easily damaged by other laser radiations (UV radiation). This method allows future development of tailored polymer coatings for biomedical applications.
Assuntos
Materiais Revestidos Biocompatíveis/química , Cristalização/métodos , Gases/química , Lasers , Poliésteres/química , Materiais Revestidos Biocompatíveis/efeitos da radiação , Gases/efeitos da radiação , Raios Infravermelhos , Teste de Materiais , Poliésteres/efeitos da radiaçãoRESUMO
Matrix Assisted Pulsed Laser Evaporation (MAPLE) is a thin film deposition technique derived from Pulsed Laser Deposition (PLD) for deposition of delicate (polymers, complex biological molecules, etc.) materials in undamaged form. The main difference of MAPLE technique with respect to PLD is the target: it is a frozen solution or suspension of the (guest) molecules to be deposited in a volatile substance (matrix). Since laser beam energy is mainly absorbed by the matrix, damages to the delicate guest molecules are avoided, or at least reduced. Lipase, an enzyme catalyzing reactions borne by triglycerides, has been used in biosensors for detection of ß-hydroxyacid esters and triglycerides in blood serum. Enzymes immobilization on a substrate is therefore required. In this paper we show that it is possible, using MAPLE technique, to deposit lipase on a substrate, as shown by AFM observation, preserving its conformational structure, as shown by FTIR analysis.
RESUMO
The use of bioabsorbable polymers in applications as temporary structural function, recovering damage in live tissues, is a promising research area. Membranes of poly(lactic acid) (PLA) may act as support to adhesion and cellular invasion or as devices for guided tissue regeneration (GTR). In this study, the same casting technique used to prepare membranes was used to prepare PLA tubes. These tubes can be used for tests in nerve guided regeneration (NGR). To improve flexibility of the device, a bioabsorbable plasticizer was added to the polymer. The initial results showed that the proposed technique allowed the preparation of flexible tubes that can be used for NGR.
Assuntos
Implantes Absorvíveis , Intubação/instrumentação , Regeneração Nervosa , Adesão Celular , Desenho de Equipamento , Humanos , Ácido Láctico/química , Membranas Artificiais , Neurônios/citologia , Plastificantes/química , Maleabilidade , Poliésteres , Polímeros/química , Propriedades de SuperfícieRESUMO
The use of biodegradable polyesters as temporary structural supports in the recuperation of damaged live tissue is a promising area of research. Poly(L-lactic acid) (PLLA) membranes can act as a support for cell fixation and growth or as a barrier against soft tissues invasion in recuperating bone tissues. In this work, five different types of PLLA membranes, which varied in their polymer-solvent ratio and their content of plasticizer were studied. For the study in vivo, 6 mm diameter disks were inserted subcutaneously in the dorsal region of 15 Wistar rats, and the reactions on rats were studied 15 days later. In another series of experiments the samples were immersed in phosphate buffer, pH 7.4 at 37 degrees C, for 30 days. Membranes without plasticizer were morphologically dense and did not allow cell invasion nor tissue adherence, in contrast to membranes with plasticizer. While porosity enhanced cell fixation and growth, it made the membrane more fragile mechanically when compared to membranes without pores.
RESUMO
The development of biodegradable materials has lead to renewed interest in the study of their interactions with the host organism in order to make the resulting products appropriate for use as temporary materials in protheses. Poly L-(lactic acid)(PLLA)-based biodegradable devices have been used for several purposes. The physical properties of these materials can be modified by the addition of a plasticizer, such as the triethylcitrate, to provide flexibility and porosity to the implants and enhance control of the polymer degradation time. In this work we examined the biological properties of a PLLA porous membrane containing 7% triethylcitrate, by assessing the process of degradation and the interaction with dermal tissue. Samples of skin obtained from female Wistar rats 2-180 days after implantation with PLLA-based membrane were processed for light microscopy and scanning electron microscopy. The membranes became surrounded by a delicate network of connective tissue which gradually invaded the membrane structure. Polymer degradation began with the appearance of radial fractures in the globular units of the biodegradable membrane, especially by 90 and 180 days after implantation.
RESUMO
Bioabsorbable materials have been widely used in the repair of damaged tissue as well as in the controlled release of drugs and as a supports for cultured cells. The degradation time of poly-L-(lactic acid) (PLLA) may be controlled by altering the polymer porosity through the addition of the plasticizer triethylcitrate. This in turn influences the extent cellular infiltration. In this study, we examined the degradation of PLLA membranes containing different concentrations of plasticizer. PLLA discs were implanted subcutaneouly in rats and withdrawn 2, 14 and 60 days after implantation. The samples were processed for light microscopy and scanning electron microscopy (SEM). Polymer degradation was proportional to the concentration of plasticizer, indicating that triethylcitrate could affect the degradation time of the implants, without damaging the polymer biocompatibility.