Biodegradable polymers are of interest for developing controlled protein drug delivery platforms. In this study, two poly (alpha-hydroxy) esters were formulated with Aerosol-OT, a surfactant stabilizer, to encapsulate the protein keratinocyte growth factor (KGF) for controlled release KGF is involved in a number of crucial biologic processes, most notably epithelial growth and repair. The concentration of KGF that caused a biological response in vitro was determined (optimally 10 ng/mL) and compared with the release of KGF from the two biodegradable polymer membrane formulations. Each polymer formulation released biologically relevant levels, 10 ng/mL of active KGF, although with different times release kinetics. The membrane composed of PLGA/AOT/KGF exhibited a faster release rate of KGF into solution after 120 h of degradation time than the release rate of the PLLA/AOT/KGF matrices. Cell seeding assays showed that both polymer matrices, when formulated with AOT, sustained cell growth. Time of Flight Secondary Ion Mass Spectrometry (ToF-SIMS) was used to characterize the distribution of AOT and KGF through the polymer membrane. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.
Aggregation patterns and fragmentation ion data from thin film preparations of the anionic surfactant sodium bis(2-ethylhexyl) sulfosuccinate (aka Aerosol-OT (AOT)) near the critical micelle concentration (CMC) in carbon tetrachloride were determined using time-of-flight secondary ion mass spectrometry (ToF-SIMS). Previous work using electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry to determine the chemical structure of AOT aggregates was compared to data from ToF-SIMS results from both positive and negative ion spectra. Quasi-molecular ions were detected for AOT in the positive and negative spectra at m/z 467 and 421, respectively, corresponding to [AOT+Na]+ and [AOT-Na]-. Repeating ion patterns assigned to AOT aggregates were detected in the positive spectra from n=3 to n=13, corresponding to the repeating series [AOTn+Na]+. A similar pattern [AOTn-Na]- was observed in the negative ion spectra from n=4 to n=14. ToF-SIMS analysis was also able to detect a previously unreported fragmentation pattern in the mass region below [AOT3+Na]+ when the film was cast from a solution with AOT concentration above the CMC. This pattern is observed starting at m/z 526 and continuing until the n=3 AOT is reached at m/z 1356 in the positive spectra. The pattern of ions is assigned to structures related to the sodium and sulfate ions from the headgroups of an aggregate of AOT molecules. The formation of the low mass pattern is shown to respond only to concentrations above the CMC, and allows for a more precise determination of CMC than previously reported methods. The CMC of AOT in carbon tetrachloride is shown to be between 2.0x10(-5) and 3.0x10(-5) molar.
Biodegradable polymers are of interest in developing strategies to control protein drug delivery. The protein that was used in this study is Keratinocyte Growth Factor (KGF) which is a protein involved in the re-epithelialization process. The protein is stabilized in the biodegradable polymer matrix during formulation and over the course of polymer degradation with the use of an ionic surfactant Aerosol-OT (AOT) which will encapsulate the protein in an aqueous environment. The release kinetics of the protein from the surface of these materials requires precise timing which is a crucial factor in the efficacy of this drug delivery system. Time of Flight Secondary Ion Mass Spectrometry (ToF-SIMS) was used in the same capacity to identify the molecular ion peak of the surfactant and polymer and use this to determine surface concentration. In the polymer matrix, the surfactant molecular ion peak was observed in the positive and negative mode at m/z 467 and 421, respectively. These peaks were determined to be [AOT + Na+] and [AOT−Na+]-. These methods are used to identify the surfactant and protein from the polymer matrix and are used to measure the rate of surface accumulation. The second step was to compare this accumulation rate with the release rate of the protein into an aqueous solution during the degradation of the biodegradable film. This rate is compared to that from fluorescence spectroscopy measurements using the protein autofluorescence from that released into aqueous solution.
This paper presents the first development of a mass-sensitive nanosensor for the isolation and quantitative analyses of engineered fullerene (C₆₀) nanoparticles, while excluding mixtures of structurally similar fullerenes. Amino-modified beta-cyclodextrin (β-CD-NH₂) was synthesized and confirmed by ¹HNMR as the host molecule to isolate the desired fullerene C₆₀. This was subsequently assembled onto the surfaces of gold-coated quartz crystal microbalance (QCM) electrodes using N-dicyclohexylcarbodiimide/N-hydroxysuccinimide (DCC/NHS) surface immobilization chemistry to create a selective molecular configuration described as (Au)-S-(CH₂)²-CONH-beta-CD sensor. The mass change on the sensor configuration on the QCM was monitored for selective quantitative analysis of fullerene C₆₀ from a C₆₀/C₇₀ mixture and soil samples. About ~10¹⁴-10¹⁶ C₆₀ particles/cm² were successfully quantified by QCM measurements. Continuous spike of 200 μL of 0.14 mg C₆₀ /mL produced changes in frequency (-Δf) that varied exponentially with concentration. FESEM and time-of-flight secondary-ion mass spectrometry confirmed the validity of sensor surface chemistry before and after exposure to fullerene C₆₀. The utility of this sensor for spiked real-world soil samples has been demonstrated. Comparable sensitivity was obtained using both the soil and purified toluene samples. This work demonstrates that the sensor has potential application in complex environmental matrices.
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