The Effect of Thermoplastic Elastomer and Fly Ash on the Properties of Polypropylene Composites with Long Glass Fibers.

A cost-effective solution to the problems that the automotive industry is facing nowadays regarding regulations on emissions and fuel efficiency is to achieve weight reduction of automobile parts. Glass fiber-reinforced polymers are regularly used to manufacture various components, and some parts may also contain thermoplastic elastomers for toughness improvement. This work aimed to investigate the effect of styrene-(ethylene-co-butylene)-styrene triblock copolymer (E) and treated fly ash (C) on the morphological, thermal, and mechanical properties of long glass fiber (G)-reinforced polypropylene (PP). Results showed that the composites obtained through melt processing methods presented similar thermal stability and improved (nano)mechanical properties compared to 25-30 wt.% G-reinforced PP composites (PP-25G/PP-30G). Specifically, the impact strength and surface hardness were greatly improved. The addition of 20 wt.% E led to a 25-39% increase in impact strength and surface elasticity, while the addition of 6.5 wt.% C led to a 16% increase in surface hardness. The composite based on 25 wt.% G, 6.5 wt.% C, and 20 wt.% E presented the best-balanced properties (8-17% increase in impact strength, 38-41% increase in axial strain, and 35% increase in surface hardness) compared with PP-30G/PP-25G. Structural and morphological analysis confirmed the presence of a strong interaction between the components that make the composites. Based on these results, the PP-G-E-C composites could be presented as a viable material for automotive applications.

Opinion of the Scientific Committee on health, environmental and emerging risks on the safety of titanium dioxide in toys.

The Opinion of the Scientific Committee on Health, Environmental and Emerging Risks advises the European Commission on whether the uses of titanium dioxide in toys and toy materials can be considered to be safe in light of the identified exposure, and the classification of titanium dioxide as carcinogenic category 2 after inhalation. Four toy products including casting kits, chalk, powder paints and white colour pencils containing various amounts of TiO2 as colouring agent were evaluated for inhalation risks. For the oral route, childrens' lip gloss/lipstick, finger paint and white colour pencils were evaluated. When it can be demonstrated with high certainty that no ultrafine fraction is present in pigmentary TiO2 preparations used in toys and toy materials, safe use with no or negligible risk for all products considered is indicated based on the exposure estimations of this Opinion. However, if an ultrafine fraction is assumed to be present, safe use is not indicated, except for white colour pencils.

Valorization of Polypropylene Waste in the Production of New Materials with Adequate Mechanical and Thermal Properties for Environmental Protection.

Innovative composites based on polypropylene waste impurified cu HDPE (PPW) combined with two thermoplastic block-copolymers, namely styrene-butadiene-styrene (SBSBC) and styrene-isoprene-styrene (SISBC) block-copolymers, and up to 10 wt% nano-clay, were obtained by melt blending. SBSBC and SISBC with almost the same content of polystyrene (30 wt%) were synthesized by anionic sequential polymerization and used as compatibilizers for PPW. Optical microscopy evaluation of the PPW composites showed that the n-clay was encapsulated into the elastomer. Addition of n-clay, together with SBSBC or SISBC, increased the interphase surface of the components in the PPW composites and enhanced the superficial area/volume ratio, which led to a recycled material with improved performance. The data resulting from differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), mechanical evaluation, and dynamic mechanical analysis (DMA) revealed that PPW reinforcement with n-clay and styrene-diene block-copolymers allows the obtaining of composites with favorable mechanical and thermal properties, and excellent impact strength for potential engineering applications.

Wood Surface Modification with Hybrid Materials Based on Multi-Walled Carbon Nanotubes.

In this work, new treatments based on multi-walled carbon nanotubes (MWCNTs), MWCNTs decorated with zinc oxide (ZnO), MWCNTs decorated with hydroxyapatite (HAp) and MWCNTs decorated with silver (Ag) nanoparticles dispersed in PHBHV solution are proposed for improving sound oak wood properties. We hypothesize that the solutions containing decorated MWCNTs will be more efficient as wood consolidants, not only because of the improved mechanical properties of the treated wood but also because of the hydrophobic layer created on the wood surface. In order to test these hypotheses, the treatments' potential was investigated by a number of complex methods, such as colorimetric parameter measurements, water absorption tests, mechanical tests, artificial aging and antifungal tests. The data confirm that the treated wood materials have moderate stability, and the color differences are not perceived with the naked eye. A significant improvement of the treated samples was observed by water absorption, humidity and mechanical tests compared to untreated wood. The best results were obtained for samples treated by brushing with solutions based on decorated CNTs, which confirms that a uniform and thicker layer is needed on the surface to ensure better protection. The wood behavior with accelerated aging revealed that the control sample degraded faster compared to the other treated samples. Antifungal tests showed that higher growth inhibition was obtained for samples treated with 0.2% MWCNTs_ZnO + PHBHV. Considering all of the obtained results, it can be concluded that the most effective treatment was MWCNTs_ZnO + PHBHV at a nanocomposite concentration of 0.2%, applied by brushing. Thus, wood protection against mold and fungi will be achieved, simultaneously ensuring improved mechanical strength and water barrier properties and therefore maintaining the structural integrity of sound oak wood over time.

The comparative effects of Resveratrol and Curcumin in combination with photodynamic therapy.

Background and aim: Photodynamic therapy, PDT, is a promising option among the local treatments with oncolytic potential. Although the basic principle is simple, its intricate mechanisms allow for a broad range of optimization methods. The purpose of this study was to assess the effects of Resveratrol and Curcumin as adjuvants of PDT on experimental tumors. Methods: Sixty-six Wistar male rats were divided into 11 groups: control, Curcumin (CUR), Resveratrol (RES) alone or followed by irradiation (IR) (CUR+IR and RES+IR, respectively), 5,10,15,20-tetra-sulphonato-phenyl-porphyrin (TSPP), TSPP+IR (PDT), and CUR or RES administered prior to or after PDT (CUR+TSPP+IR, RES+TSPP+IR, TSPP+IR+CUR, TSPP+IR+RES). Results: Both CUR and RES significantly decreased lipid peroxidation, while RES also showed an increase in glutathione (GSH) levels, especially when it was administered before PDT (p<0.01). Both antioxidants decreased cyclooxygenase (COX)2 expression, to a minimum when they administered prior to PDT (p<0.001 and p<0.01) while nitric oxide synthase (NOS)2 expression diminished in the combined regimen, particularly in RES associated with PDT. CUR and RES induced similar changes in terms of cell death, but CUR seemed to be more efficient on tumor necrosis and showed a higher apoptotic index when was administered after PDT (p<0.001). Conclusion: Both RES and CUR in association with PDT decreased oxidative stress, diminished the COX2 and NOS2 expressions and increased cell death by positively influencing the necrotic rate and apoptotic index, particularly when CUR was administered after PDT. The results show that CUR is a promising class to study in PDT optimization and further invites to exploit its promises.

Biocompatible and Antimicrobial Cellulose Acetate-Collagen Films Containing MWCNTs Decorated with TiO2 Nanoparticles for Potential Biomedical Applications.

This research focuses on the synthesis of multi-walled carbon nanotubes (MWCNTs) decorated with TiO2 nanoparticles (NPs) and incorporated in cellulose acetate-collagen film in order to obtain a new biomaterial with potential biomedical applications and improved antimicrobial activity. The successful decoration of the MWCNTs with TiO2 NPs was confirmed by several structural and morphological analysis, such as Fourier transformed infrared spectroscopy, Raman spectroscopy, X-ray diffraction and transmission electron microscopy. The obtained nanocomposites were further incorporated into cellulose acetate-collagen films, at different concentrations and absorption kinetics, antimicrobial activity and in vitro biocompatibility of the obtained films was investigated. The antimicrobial tests sustained that the presence of the nanocomposites into the polymeric matrix is an important aspect in increasing and maintaining the antimicrobial activity of the polymeric wound dressings over time. The biocompatibility and cytotoxicity of the obtained films was evaluated using cellular viability/proliferation assay and fluorescent microscopy which revealed the ability of the obtained materials as potential wound dressing biomaterial.

Preparation and Biocompatibility of Poly Methyl Methacrylate (PMMA)-Mesoporous Bioactive Glass (MBG) Composite Scaffolds.

In recent years, the rising number of bone diseases which affect millions of people worldwide has led to an increased demand for materials with restoring and augmentation properties that can be used in therapies for bone pathologies. In this work, PMMA- MBG composite scaffolds containing ceria (0, 1, 3 mol%) were obtained by the phase separation method. The obtained composite scaffolds were characterized by X-ray diffraction, infrared spectroscopy, and scanning electron microscopy. UV-Vis measurement and EDX analysis confirmed the presence of cerium ions in the composite scaffolds. Evaluation of the in-vitro biocompatibility using MTT assay showed that composite scaffold containing 1 mol% of ceria presented higher viability than control cells (100%) for concentrations ranging between 5 and 50% after 96 h of incubation.

Multifunctional Composite Coatings Based on Photoactive Metal-Oxide Nanopowders (MgO/TiO2) in Hydrophobic Polymer Matrix for Stone Heritage Conservation.

Multifunctional composite coatings composed of metal oxide nanoparticles dispersed in polymer matrices are an advanced solution to solve the problem of stone heritage deterioration. Their innovative design is meant to be stable, durable, transparent, easy to apply and remove, non-toxic, hydrophobic, and permeable. Coating formulations for the protection of buildings and monuments have been intensively researched lately. Such formulations are based on multifunctional composite coatings incorporating metal oxides. The present work aims to combine the hydrophobic properties of sodium polyacrylate (NaPAC16) with the antimicrobial effectiveness, with promising antimicrobial results even in the absence of light, and good compatibility of MgO (a safe to use, low cost and environmentally friendly material) and TiO2 (with antibacterial and antifungal properties), in order to develop coatings for stone materials protection. MgO (pure phase periclase) and TiO2 (pure phase anatase) nanopowders were prepared through sol-gel method, specifically routes. Aqueous dispersions of hydrophobically modified polymer (NaPAC16, polyacrylic acid sodium salt) and MgO/TiO2 nanopowders were deposited through layer-by-layer dip coating technique on glass slides and through immersion on stone fragments closely resembling the mosaic stone from the fourth century AD Roman Mosaic Edifice, from Constanta, Romania. The oxide nanopowders were characterized by: Thermal analysis (TG/DTA), scanning electron microscopy (SEM), X-ray diffraction (XRD), BET specific surface area and porosity, and UV-Vis spectroscopy for band gap determination. An aqueous dispersion of modified polyacrylate polymer and oxide nanopowders was deposited on different substrates (glass slides, red bricks, gypsum mortars). Film hydrophobicity was verified by contact angle measurements. The colour parameters were evaluated. Photocatalytic and antimicrobial activity of the powders and composite coatings were tested.

Hybrid Materials Based on Multi-Walled Carbon Nanotubes and Nanoparticles with Antimicrobial Properties.

In this study, multi-walled carbon nanotubes (MWCNTs) were decorated with different types of nanoparticles (NPs) in order to obtain hybrid materials with improved antimicrobial activity. Structural and morphological analysis, such as Fourier transformed infrared spectroscopy, Raman spectroscopy, X-ray diffraction, transmission electron microscopy, environmental scanning electron microscopy/energy-dispersive X-ray spectroscopy and the Brunauer-Emmett-Teller technique were used in order to investigate the decoration of the nanotubes with NPs. Analysis of the decorated nanotubes showed a narrow size distribution of NPs, 7-13 nm for the nanotubes decorated with zinc oxide (ZnO) NPs, 15-33 nm for the nanotubes decorated with silver (Ag) NPs and 20-35 nm for the nanotubes decorated with hydroxyapatite (HAp) NPs, respectively. The dispersion in water of the obtained nanomaterials was improved for all the decorated MWCNTs, as revealed by the relative absorbance variation in time of the water-dispersed nanomaterials. The obtained nanomaterials showed a good antimicrobial activity; however, the presence of the NPs on the surface of MWCNTs improved the nanocomposites' activity. The presence of ZnO and Ag nanoparticles enhanced the antimicrobial properties of the material, in clinically relevant microbial strains. Our data proves that such composite nanomaterials are efficient antimicrobial agents, suitable for the therapy of severe infection and biofilms.

In Vitro Cytotoxic Protective Effect of Alginate-Encapsulated Capsaicin Might Improve Skin Side Effects Associated with the Topical Application of Capsaicin.

Chronic neuropathic pain, particularly peripheral pain, is a cause of great concern for diabetic patients. Current treatments include numerous agents such as capsaicinoids, a known deterrent of neuropathic pain despite the inconvenience associated with local side effects. In this context, the current work aims to elucidate the potential mechanisms involved in cytotoxicity by capsaicin and proposes an efficient formulation of capsaicin in alginate microcapsules, which significantly reduces side effects from capsaicin topical administration. For this, human dermal fibroblast cells were treated with alginate-microencapsulated capsaicin extracts and screened for potential cytotoxic effects produced by the treatment. Cell viability and morphology were examined, as well as oxidative stress status and anti-inflammatory potential. Our results show that the alginate encapsulated formulation of capsaicin exerted lower cytotoxic effects on human dermal fibroblasts as measured by cell viability and reactive oxygen species (ROS) production. Furthermore, the expression profiles of inflammatory cytokines were significantly altered by the treatment as compared with the control culture.

Development of thermoplastic composites based on recycled polypropylene and waste printed circuit boards.

In the last several years, the electronic waste, especially printed circuit boards have significantly increased over the world, generating one of the highest rates of solid waste. The recycling process of the printed circuit boards implies mainly the recovery of metals and glass fibers, while the reuse of the polymeric support has remained largely in the phase of research. In this paper, the non-metallic part of printed circuit boards was used as filler (up to 30%), but also to improve the fire resistance of thermoplastic composites based on recycled polypropylene and diene block-copolymers. The synergy between the elastic effect of elastomers and the reinforcing effect of the waste powder into the thermoplastic matrix was studied by mechanical and dynamo-mechanical analysis, X-ray diffraction, optical microscopy, micro-calorimetry and thermo-gravimetrical analysis. Improved mechanical properties, especially impact strength was observed. The compatibization of components considering the interactions between the ethylene-butylene blocks from the hydrogenated and maleinized styrene-butadiene block-copolymer and recycled polypropylene, respectively between the MA groups and the functionalities of the waste powder, evidenced by FTIR, was highlighted by changes in the X-ray pattern and an increased fire resistance and thermal stability.

Nanomaterials Used in Conservation and Restoration of Cultural Heritage: An Up-to-Date Overview.

In the last few years, the preservation of cultural heritage has become an important issue globally, due to the fact that artifacts and monuments are continually threatened by degradation. It is thus very important to find adequate consolidators that are capable of saving and maintaining the natural aspect of these objects. This study aims to provide an updated survey of the main nanomaterials used for the conservation and restoration of cultural heritage. In the last few years, besides the classic nanomaterials used in this field, such as metal nanoparticles (copper and silver) and metal oxides (zinc and aluminum), hydroxyapatite and carbonated derivatives, tubular nanomaterials (such as carbon nanotubes) have been used as a potential consolidate material of cultural heritage. Tubular nanomaterials have attracted attention for use in different fields due to their structures, as well as their ability to present multiple walls. These nanotubes have the necessary properties in preserving cultural heritage, such as superior mechanical and elastic strength (even higher than steel), high hydrophobicity (with a contact angle up to 140°), optical properties (high photodegradation protection), large specific surface area (from 50 to 1315 m2/g, depending on the number of walls) for absorption of other nanomaterials and relatively good biocompatibility.

The in vivo modulatory effects of Cornus mas extract on photodynamic therapy in experimental tumors.

BACKGROUND: Photodynamic therapy (PDT) is a treatment of cancer due to its ability to induce cell death, oxidative stress and acute inflammatory reaction in targeted sites. To optimize the effect of PDT the addition of some compounds with supplementary cytotoxic effect on tumor cells was tried. METHODS: The study was performed on 35 Wistar male albino rats with Walker 256 carcinosarcoma. The animals were randomly assigned in seven groups (n = 5) and treated as follows: group 1 - control; group 2 - Cornus mas (CM) extract 15 mg/kg b.w., administered for 7 days; group 3 - CM extract administered for 7 days followed by irradiation (CM + IR); group 4 - one dose of tetra-p-sulfonato-phenyl-porphyrin (TSPP) 10 mg/kg b.w.; group 5 - TSPP + IR; group 6 - CM extract administered daily for 7 days before TSPP and IR (CM + TSPP + IR); group 7 - TSPP + IR followed by CM administered for 7 days (TSPP + IR + CM). RESULTS: The results showed that MDA and GSSG levels increased after PDT in parallel with the increasing of COX-2 expression and DNA damage. Apoptotic and necrotic index enhanced in TSPP + IR, effect improved by CM association before PDT. CM + TSPP + IR regimen also induced more intense inflammatory reactions, increased COX-2 expression, determined DNA damage, apoptosis and necrosis, compared to the TSPP + IR + CM group. Both combined therapeutic regimens reduced MDA levels in tumor tissue, especially CM + TSPP + IR and increased the antioxidant defense and iNOS expression. CONCLUSIONS: Our results demonstrated that CM associated before PDT had beneficial effects in PDT and may represent a promising option in PDT strategies.

Capsaicin: Physicochemical properties, cutaneous reactions and potential applications in painful and inflammatory conditions.

Capsaicin is a natural protoalkaloid recognized as the main pungent component in hot peppers (Capsicum annuum L.). The capsaicin receptor is highly expressed in the unmyelinated type C nerve fibers originating from small diameter sensory neurons in dorsal root ganglia and cranial nerve ganglia correspondents. Capsaicin and related vanilloids have a variety of effects on primary sensory neurons function, from sensory neuron excitation characterized by local burning sensation and neurogenic inflammation, followed by conduction blockage accompanied by reversible ultrastructural changes of peripheral nociceptive endings (desensitization), going as far as irreversible degenerative changes (neurotoxicity). The main role in capsaicin-induced neurogenic inflammation relies on the capsaicin sensitive, small diameter primary sensory neurons, therefore its evaluation could be used as a diagnostic instrument in functional alterations of cutaneous sensory nerve fibers. Moreover, capsaicin-induced desensitization and neurotoxicity explain the analgesic/anti-nociceptive and anti-inflammatory effects of topical capsaicin and its potential use in the management of painful and inflammatory conditions. In this study, we describe the effects of capsaicin on neurogenic inflammation and nociception, as well as its potential diagnostic value and therapeutic impact in various conditions involving impairment of sensory nerve fibers.

Methods of synthesis, properties and biomedical applications of polyhydroxyalkanoates: a review.

This study aims to provide an overview of the main polyhydroxyalkanoates (PHAs) used in medical applications. In this review, it has been demonstrated that the properties of PHAs can be controlled both by varying the concentration of units in the copolymer and the substrate for PHA production. Another way of controlling the mechanical properties of PHAs is varying the 3HV content, such as the case of P(3HB-co-3HV). A higher 3HV content in the structure of this polyester will lead to a lower cristallinity and, therefore, to greater flexibility, strength and elongation at break. PHAs are biocompatible, completely biodegradable, and non-toxic. Considering the state of the art technologies and their ideal properties, PHAs (especially the short-chain-length PHA polymers) can be tailored for specific medical applications, such as surgical suture, scaffolds, grafts and heart valves or drug delivery systems. Nowaday, random copolyesters of 3HB and 4HB (P(3HB-co-4HB)) are used for the development of biodegradable implants loaded with antibiotics for therapeutic treatment of chronic osteomyelitis. It is also believed that these biodegradable materials can be efficient alternatives for reducing the pollution produced by the medical waste consisting of replacing plastic handles, packs, syringes or tubes that are trashed into the homes, clinics and hospitals around the world.

Unveiling Ga(III) phthalocyanine-a different photosensitizer in neuroblastoma cellular model.

Phthalocyanines (Pc) and their metallated derivatives are strongly considered for photodynamic therapy (PDT) possessing unique properties as possible new photosensitizers (PS). We have used toxicological assessments, real-time monitoring of cellular impedance, and imagistic measurements for assessing the in vitro dark toxicity and PDT efficacy of Ga(III)-Pc in SHSy5Y neuroblastoma cells. We have established the non-toxic concentration range of Ga(III)-Pc, a compound which shows a high intracellular accumulation, with perinuclear distribution in confocal microscopy. By choosing Ga(III)Pc non-toxic dose, we performed in vitro experimental PDT hampering cellular proliferation. Our proposed Ga(III)-Pc could complete a future PS panel for neuroblastoma alternate therapy.

Combined regimen of photodynamic therapy mediated by Gallium phthalocyanine chloride and Metformin enhances anti-melanoma efficacy.

BACKGROUND: Melanoma therapy is challenging, especially in advanced cases, due to multiple developed tumor defense mechanisms. Photodynamic therapy (PDT) might represent an adjuvant treatment, because of its bimodal action: tumor destruction and immune system awakening. In this study, a combination of PDT mediated by a metal substituted phthalocyanine-Gallium phthalocyanine chloride (GaPc) and Metformin was used against melanoma. The study aimed to: (1) find the anti-melanoma efficacy of GaPc-PDT, (2) assess possible beneficial effects of Metformin addition to PDT, (3) uncover some of the mechanisms underlining cell killing and anti-angiogenic effects. METHODS: Two human lightly pigmented melanoma cell lines: WM35 and M1/15 subjected to previous Metformin exposure were treated by GaPc-PDT. Cell viability, death mechanism, cytoskeleton alterations, oxidative damage, were assessed by means of colorimetry, flowcytometry, confocal microscopy, spectrophotometry, ELISA, Western Blotting. RESULTS: GaPc proved an efficient photosensitizer. Metformin addition enhanced cell killing by mechanisms dependent on the cell line, namely apoptosis in the metastatic M1/15 and necrosis in the radial growth phase, WM35. Cell death mechanism relied on the inhibition of nuclear transcription factor (NF)-κB activation and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) sensitization, leading to TRAIL and TNF-α induced apoptosis. Metformin diminished the anti-angiogenic effect of PDT. CONCLUSIONS: Metformin addition to GaPc-PDT increased tumor cell killing through enhanced oxidative damage and induction of proapoptotic mechanisms, but altered PDT anti-angiogenic effects. GENERAL SIGNIFICANCE: Combination of Metformin and PDT might represent a solution to enhance the efficacy, leading to a potential adjuvant role of PDT in melanoma therapy.

Toxicological and efficacy assessment of post-transition metal (Indium) phthalocyanine for photodynamic therapy in neuroblastoma.

Metallo-phthalocyanines due to their photophysical characteristics as high yield of triplet state and long lifetimes, appear to be good candidates for photodynamic therapy (PDT). Complexes with diamagnetic metals such as Zn2+, Al3+ Ga3+ and In3+meet such requirements and are recognized as potential PDT agents. Clinically, Photofrin® PDT in neuroblastoma therapy proved in pediatric subjects diagnosed with progressive/recurrent malignant brain tumors increased progression free survival and overall survival outcome. Our study focuses on the dark toxicity testing of a Chloro-Indium-phthalocyanine photosensitizer (In-Pc) upon SH-SY5Y neuroblastoma cell line and its experimental in vitro PDT. Upon testing, In-Pc has shown a relatively high singlet oxygen quantum yield within the cells subjected to PDT (0.553), and 50 µg/mL IC50. Classical toxicological and efficacy assessment were completed with dynamic cellular impedance measurement methodology. Using this technology we have shown that long time incubation of neuroblastoma cell lines in In-Pc (over 5 days) does not significantly hinder cell proliferation when concentration are ≤ 10 µg/mL. When irradiating neuroblastoma cells loaded with non-toxic concentration of In-Pc, 50% of cells entered apoptosis. Transmission electron microscopy has confirmed apoptotic characteristics of cells. Investigating the proliferative capacity of the in vitro treated cells we have shown that cells that "escape" the irradiation protocol, present a reduced proliferative capacity. In conclusion, In-Pc represents another photosensitizer that can display sound PDT properties enhancing neuroblastoma therapy armentarium.

Melanogenesis and DNA damage following photodynamic therapy in melanoma with two meso-substituted porphyrins.

Melanoma, a cancer derived from melanocytes is very difficult to treat, especially in advanced cases. There are several encouraging studies of the efficacy of photodynamic therapy (PDT) in melanoma. However, PDT has to overcome the main defense mechanisms like: defects in the apoptotic pathways, pigmentation, sequestration of the photosensitisers (PS) inside melanosomes and increased oxidative stress defense. Two meso-substituted porphyrins, meso-5,10,15,20-tetrakis (4-hydroxyphenyl) porphyrin (THOPP) and meso-5-(4-hydroxyphenyl)-10, 15, 20-tris (4-methoxyphenyl) porphyrin (THOMPP) were used as PS to investigate several mechanisms underlining the PDT anti-melanoma effects, on a lightly pigmented melanoma cell line (WM35), in vitro. γH2AX foci formation (a measure of DNA double strand brakes) was used for the assessment of DNA damage by means of immune-fluorescence and western blot. Cytoskeleton alterations were detected by phalloidin staining. Tyrosinase activity and melanin pigment were quantified by spectrophotometry, tyrosinase protein by western blot, total peroxidase activity by resorurfin reaction (Amplex Red). PDT induced high levels of DNA damage, cytoskeleton alterations and enhanced pigmentogenesis. THOPP mediated PDT was the most efficient. The melanogenesis stimulated by PDT was directly correlated to the PDT induced cellular damage and provided no protection against therapy. Thus, PDT induced melanogenesis combined with severe DNA damage was able to overcome the mechanisms of resistance and increased the efficiency of PDT in WM35 melanoma cells. These results are encouraging for a possible use of PDT, as an adjuvant therapy in lightly pigmented melanomas.

Impact of multicellular tumor spheroids as an in vivo­like tumor model on anticancer drug response.

The incidence of colorectal cancer is higher in men than in women, amounting to 15% of cancer-related diseases as a whole. As such, undesirable effects, arising from the administration of current chemotherapeutic agents (the FOLFIRI/FOLFOX combinations), which are exerted on the remaining non-cancerous tissues and/or cells, have contributed to the occurrence of resistance to multiple drugs, thus markedly reducing their efficacy. However, the delivery of chemotherapeutic agents may be improved and their action may be more selectively targeted to diseased tissues/cells by means of developing biotechnologies and nano­techniques. Thus, the current focus is on creating biological tissue and related tumor models, by means of three­dimensional (3D) spheres, in an attempt to bridge the gap between results obtained in the pre­clinical phase and promising outcomes obtained in clinical trials. For this purpose, the characterization and use of so­called 'multicellular tumor spheroids', may prove to be invaluable. In this study, we focus on describing the efficacy of a model 3D system as compared to the traditional 2D tumor spheres in determining drug response, highlighting a potentially greater effect of the drugs following the encapsulation of respective liposomes. The results obtained demonstrate the successful preparation of a suspension of liposomes loaded with folinic acid, oxaliplatin and 5­fluorouracil (5­FU), and loaded with meso­tetra (4­sulfonatophenyl) porphyrin. Following its use on HT­29 colorectal cancer cells, an important comparative reduction was noted in the viability of the HT­29 cells, demonstrating the efficacy of multicellular tumor spheroids carrying liposomes loaded with therapeutic drugs. These findings indicate that the method of drug encapsulation in liposomes may improve the treatment efficacy of chemotherapeutic agents.