Microgel-Based Stretchable Reservoir Devices for Elongation Enhanced Small Molecule Release Rate.

Stretchable poly(N-isopropylacrylamide)-co-acrylic acid (pNIPAm-co-10% AAc) microgel-based reservoir devices were fabricated and used to control the release rate of the small molecule model drug tris(4-(dimethylamino)phenyl)methylium chloride (crystal violet, CV) to solution by varying the Au layer thickness coating the microgels and device elongation. Specifically, we showed that CV could be loaded into the microgel layer of the devices via electrostatic interactions at pH 6.5, and the release could be triggered upon exposure to a pH 3.0 solution, which breaks the microgel-CV electrostatic interactions. We demonstrated that the rate of release could be increased by decreasing the Au layer thickness coating microgels and by stretching, that is, thin Au and high elongation promoted the relatively fast release of CV from the device. We found that the Au overlayer thickness (and porosity) dominated the observed release rate profiles when the device was not stretched (or at low elongation), while elongation-induced cracks dominated the release rate at high elongation. We also showed that the CV release kinetics could transition from low ("off") to high ("on"), which enhanced when the devices are stretched. This behavior could be exploited in the future for autonomous release systems that release small molecules when stretched by natural processes, for example, movement of joints and muscles.

Synthesis of Co3O4/graphene nanocomposite using paraffin wax for adsorption of methyl violet in water.

This study discusses the use of Co3O4 impregnated graphene (CoOIG) as an efficient adsorbent for the removal of methyl violet (MV) dye from wastewater. CoOIG nanocomposites have been prepared by pyrolyzing paraffin wax with cobalt acetate. The synthesised nanocomposite was characterised by X-ray diffraction, field emission scanning electron microscope, transmission electron microscope, Fourier transform infrared spectroscope, Raman spectroscopy, and Brunauer-Emmett-Teller isotherm studies. The above studies indicate that the composites have cobalt oxide nanoparticles of size 51-58 nm embedded in the graphene nanoparticles. The adsorption studies were conducted with various parameters, pH, temperature and initial dye concentration, adsorbent dosage and contact time by the batch method. The adsorption of MV dye by the adsorbent CoOIG was about 90% initially at 15 min and 98% dye removal at pH 5. The data were fitted in Langmuir, Freundlich, Temkin, and Dubinin-Radushkevich and Sips isotherm models. Various thermodynamic parameters like Gibbs free energy, enthalpy, and entropy of the on-going adsorption process have also been calculated.

In vivo multispectral photoacoustic and photothermal flow cytometry with multicolor dyes: a potential for real-time assessment of circulation, dye-cell interaction, and blood volume.

Recently, photoacoustic (PA) flow cytometry (PAFC) has been developed for in vivo detection of circulating tumor cells and bacteria targeted by nanoparticles. Here, we propose multispectral PAFC with multiple dyes having distinctive absorption spectra as multicolor PA contrast agents. As a first step of our proof-of-concept, we characterized high-speed PAFC capability to monitor the clearance of three dyes (Indocyanine Green [ICG], Methylene Blue [MB], and Trypan Blue [TB]) in an animal model in vivo and in real time. We observed strong dynamic PA signal fluctuations, which can be associated with interactions of dyes with circulating blood cells and plasma proteins. PAFC demonstrated enumeration of circulating red and white blood cells labeled with ICG and MB, respectively, and detection of rare dead cells uptaking TB directly in bloodstream. The possibility for accurate measurements of various dye concentrations including Crystal Violet and Brilliant Green were verified in vitro using complementary to PAFC photothermal (PT) technique and spectrophotometry under batch and flow conditions. We further analyze the potential of integrated PAFC/PT spectroscopy with multiple dyes for rapid and accurate measurements of circulating blood volume without a priori information on hemoglobin content, which is impossible with existing optical techniques. This is important in many medical conditions including surgery and trauma with extensive blood loss, rapid fluid administration, and transfusion of red blood cells. The potential for developing a robust clinical PAFC prototype that is safe for human, and its applications for studying the liver function are further highlighted.

Photophysical, photochemical, and tumor-selectivity properties of bromine derivatives of rhodamine-123.

The conceptual basis for the development of mitochondrial targeting as a novel therapeutic strategy for both chemotherapy and photochemotherapy of neoplastic diseases rests on the observation that enhanced mitochondrial membrane potential is a common tumor cell phenotype. The potential of this strategy is highlighted by the fact that the toxic effects associated with a number of cationic dyes known to localize in energized cell mitochondria are much more pronounced in tumor cells than in normal cells. Here we evaluate the phototoxic properties of four bromine derivatives of rhodamine-123 toward human uterine sarcoma (MES-SA) and green monkey kidney (CV-1) cells and compare the degrees of tumor cell selectivity associated with these dyes with those associated with two model mitochondrial triarylmethanes (crystal violet and ethyl violet). Selective phototoxicity toward tumor cells was found to be highly dependent upon the lipophilic/hydrophilic character of the cationic photosensitizer. Our experimental data have indicated that the probability of success of mitochondrial targeting in (photo)chemotherapy of neoplastic diseases is higher when the octan-1-ol/water partition coefficient of the drug candidate falls within approximately two orders of magnitude from that of the prototypical mitochondria-specific dye rhodamine-123.

An in vitro study of biological safety of condoms and their additives.

The use of condoms to prevent sexually transmitted diseases, especially HIV, is widely encouraged. Condoms contain latex, nonspermicidal lubricants (such as dimethylsiliconium) and other nonspecified compounds, such as colorants and flavorings. Latex causes allergy reaction in susceptible individuals but little is known regarding the cytotoxic effects of other additives. The objective of this study was to develop a sensitive in vitro system to determine the toxic effects of condom material. The modified L929 FDA method and a more specific cell type, such as the cervical epithelial tumor cell line HeLa, was used. Lubricated (LC), lubricated and flavored (LFC), and lubricated, flavored and colored condoms (LFCC) were evaluated. Washings containing condom surface material were prepared by washing condom fragments in medium for different time intervals. Changes in cell number, viability and lysosome integrity in the L929 and HeLa cell lines was determined using the Crystal Violet, MTT and Neutral Red assays, respectively. The condom type affected cell viability and lysosome integrity, with LC inducing an increase in cell viability and LFC a decrease in lysosome integrity. The HeLa cell line in combination with the MTT and NR assay was themost sensitive in vitro system to determine the toxic effects of condom material.

Local pharmaceutical release from a new hydrogel implant.

BACKGROUND: Solid hydrogel polymers can act as reservoirs for controlled drug release. The object of this study was to quantify release kinetics for a single example of a class of uniquely structured hydrogels. The polymer of this study belongs to a class of permanent implants that release pharmaceuticals by diffusion from an entangled cross-linked matrix rather than by absorption of the implant by the body. The cross-linked matrix (CLM) of this study is biocompatible and polymerized in situ, forming a solid that is mechanically bonded to the implant site. It can seal tissue as well as deliver drugs at predetermined rates. We evaluated dye and antibiotic egress and assessed release kinetics and retention of antibiotic activity following elution from the CLM. MATERIALS AND METHODS: The prepolymerized test CLM was prepared in two ratios, 6 and 20% prepolymer, in an activating aqueous solution. Aqueous solutions were prepared from dyes of varying molecular weights. Aliquots of prepolymerized test CLM were allowed to solidify and dye release into the supernatant was quantified by spectrophotometry over a 168-h period. Antibiotic solutions were also employed to form solid CLMs. Tetracycline release over time was characterized by spectrophotometry. Antibiotic dosed solid CLMs were placed on agar plates streaked with Escherichia coli and incubated. Growth inhibition was assessed for each antibiotic. RESULTS: In the test CLM, dye and antibiotic release were found to be inversely related to molecular weight and consistent with a diffusion model. CLMs formed from aqueous solutions containing higher molecular weight dyes and antibiotics released those constituents more slowly than lower molecular weight constituents. This finding, as well as the effect on release rate under varying prepolymer concentration, was consistent with a diffusion release mechanism. Antibiotic released from the tissue sealant was shown to be potent by consistent inhibition of E. coli. CONCLUSIONS: Pharmaceutical release by a representative CLM was found to be controllable by varying the concentration of the pharmaceutical in the activating aqueous solution. The polymerization and release mechanisms did not degrade antibiotic biologic activity. CLMs may be a general class of biocompatible polymers that can locally deliver clinically useful biologics, the release kinetics of which are unaffected by the variability of implant absorption/inflammation mechanisms.

Effect of molecular structure on the performance of triarylmethane dyes as therapeutic agents for photochemical purging of autologous bone marrow grafts from residual tumor cells.

Extensively conjugated cationic molecules with appropriate structural features naturally accumulate into the mitochondria of living cells, a phenomenon typically more prominent in tumor than in normal cells. Because a variety of tumor cells also retain pertinent cationic structures for longer periods of time compared with normal cells, mitochondrial targeting has been proposed as a selective therapeutic strategy of relevance for both chemotherapy and photochemotherapy of neoplastic diseases. Here we report that the triarylmethane dye crystal violet stains cell mitochondria with efficiency and selectivity, and is a promising candidate for photochemotherapy applications. Crystal violet exhibits pronounced phototoxicity toward L1210 leukemia cells but comparatively small toxic effects toward normal hematopoietic cells (murine granulocyte-macrophage progenitors, CFU-GM). On the basis of a comparative examination of chemical, photochemical, and phototoxic properties of crystal violet and other triarylmethane dyes, we have identified interdependencies between molecular structure, and selective phototoxicity toward tumor cells. These structure-activity relationships represent useful guidelines for the development of novel purging protocols to promote selective elimination of residual tumor cells from autologous bone marrow grafts with minimum toxicity to normal hematopoietic stem cells.

Diisopropylglutathione ester protects A549 cells from the cytotoxic effects of sulphur mustard.

The A549 cell line was used to assess the ability of diisopropylglutathione (DIPE) to protect against a 100 microM challenge dose of sulphur mustard (HD) using gentian violet (GV), thiazolyl blue (MTT) and neutral red (NR) assays as indicators of cell culture viability. As part of a continuing study of the efficacy of protective nucleophiles as candidate treatments for HD poisoning, several different combinations of protectant and HD were used to determine the optimal means of protecting A549 cells from the effects of HD. It was found that DIPE (4 mM) could protect cells against the effects of HD though for optimal effect, DIPE had to be present at the time of HD challenge. Cultures protected with DIPE were up to 2.9-fold more viable than HD exposed cells 48 h after HD challenge when using the GV, MTT and NR assays to assess viability. Observations by phase contrast microscopy of GV stained cultures confirmed these findings. Pretreating A549 cultures with DIPE for 1 h followed by its removal prior to HD challenge did maintain cell viability, though at a relatively low level (only up to 1.4-fold more viable than HD only exposed cells). DIPE was also able to protect HD exposed A549 cultures when added to cell cultures at intervals of up to 12 to 15 min after the initial HD exposure, though viability tended to decrease over this period, so that at 1 h, addition of DIPE did not maintain the viability of the cultures. This is the first such report of the anti-HD protectant properties of DIPE in A549 cells. It is concluded that the protection observed against HD is probably largely due to extracellular inactivation of HD by DIPE.

Water protective effect of barrier creams and moisturizing creams: a new in vivo test method.

Frequent exposure to water is an important risk factor for the development of irritant hand eczema. Evaluation of cream efficacy in protecting against water is difficult. A new non-irritant, non-invasive method is introduced, based on evaluation of colour intensities when an aqueous solution of crystal violet is applied to the skin, after pretreatment with different creams. By skin reflectance measurement, differences in colour intensity were objectified. Measurements of transepidermal water loss were also performed, 2 barrier creams and a moisturizer were tested on the dorsal and volar aspects of the hands. One barrier cream contained silicone, the other solid particles. The moisturizer had a high content of lipid. In one experiment, the immediate effect of the creams in protecting against water was evaluated. In a second experiment, the water resistancy of the creams was tested, using a standardized water immersion procedure (4 x 20 min). The barrier cream with particles gave the best immediate protection (dorsal 76%, volar 69%). The moisturizer was intermediately protective (dorsal 57%, volar 34%), while very little protection was found for the silicone-containing cream (dorsal 16%, volar 10%). The water immersion procedure resulted in only minor changes in protection for each cream. When comparing transepidermal water loss in the treated areas with pretreatment values, a reduction was recorded only for the particle-containing barrier cream. The colour method described may be used as a quick and easy test of the protective effect of creams against water.

Evaluation of barrier creams: an in vitro technique on human skin.

A method was developed to measure in vitro on human skin the effectiveness of barrier creams against three dyes (eosin, methyl-violet and oil red O) with different n-octanol/water partition coefficients (0.19, 29.8 and 165, respectively). Some galenic properties (water washability, water content and viscosity) of the products were also evaluated to try to understand the mechanisms of such a protection. The barrier creams were assayed by measurements of the dyes in the epidermis of protected skin samples after an application time of 30 min. Whereas some products showed some degree of protection, as claimed on the packaging, we demonstrated in several cases disagreement with the manufacturer's information. Surprisingly, petrolatum was found to provide the best protection of all tested products in our in vitro model. There was no correlation between the galenic parameters of the assayed products and the level of protections, indicating that neither the water content nor the consistence of the formulations influenced the protection effectiveness. In conclusion, regarding the possible skin effects of some irritants, our results stress that barrier creams should be used with caution, knowing the protection limits of some of the formulations marketed.