Protective Ag :TiO2 thin films for pressure sensors in orthopedic prosthesis: the importance of composition, structural and morphological features on the biological response of the coatings

DC reactive magnetron sputtered Ag:TiO2 nanocomposite thin films were developed to be used as protective coatings in pressure sensor devices. The coatings, with Ag content varying from 0 to about 30 at.%, were prepared and characterized in order to study their biological response. The as-deposited samples were annealed in vacuum at 500 °C in order to evaluate the influence of their morphological and structural differences over the response elicited upon contact with simulated bodily fluids and cultured human cells, as well as selected microorganisms. The results showed that the annealing treatment produced less porous films with an enhanced structure, with a significant reduction in structural defects and improved crystallinity. Additionally, samples with higher Ag contents (≥12.8 at.%) exhibited Ag agglomerates/clusters at the surface, a result anticipated from the XRD data. The crystallization of the TiO2 matrix was also observed by XRD analysis, albeit delayed by the dispersion of Ag into the matrix. Biological characterization showed that the antimicrobial activity and cytotoxicity of the coatings were directly related with their composition, closely followed by the particular structural and morphological features, namely those resulting from annealing process.This research is partially sponsored by FEDER funds through the program COMPETE—Programa Operacional Factores de Competitividade and by national funds through FCT—Fundação para a Ciência e a Tecnologia, under the projects PEst-C/EME/UI0285/2011, PTDC/SAU-ENB/116850/2010, PTDC/CTM-NAN/112574/2009P. T Matamá acknowledges FCT for Grant SFRH/BPD/47555/2008

Targeted Imaging of Urothelium Carcinoma in Human Bladders by an ICG pHLIP Peptide Ex vivo

Bladder cancer is the fifth most common in incidence and one of the most expensive cancers to treat. Early detection greatly improves the chances of survival and bladder preservation. The pH Low Insertion Peptide (pHLIP® peptide) conjugated with a near infrared fluorescent dye (ICG) targets low extracellular pH allowing visualization of malignant lesions in human bladder carcinoma ex vivo. Cystectomy specimens obtained after radical surgery were immediately irrigated with non-buffered saline and instilled with a solution of the ICG pHLIP® construct, incubated, and rinsed. Bladders were subsequently opened and imaged, the fluorescent spots were marked, and a standard pathological analysis was carried out to establish the correlation between ICG pHLIP® imaging and white light pathological assessment. Accurate targeting of bladder lesions was achieved with a sensitivity of 97%. Specificity is 100%, but reduced to 80%, if targeting of necrotic tissue from previous transurethral resections or chemotherapy are considered as false positives. ICG pHLIP® imaging agent marked high grade urothelial carcinomas, both muscle invasive and non-muscle invasive. Carcinoma in situ (CIS) was accurately diagnosed in 11 cases, whereas only 4 cases were seen using white light, so imaging with the ICG pHLIP® peptide offers improved early diagnosis of bladder cancers, and may also enable new treatment alternatives

Comparative Study of Tumor Targeting and Biodistribution of pH (Low) Insertion Peptides (pHLIP® Peptides) Conjugated with Different Fluorescent Dyes

Purpose Acidification of extracellular space promotes tumor development, progression, and invasiveness. pH (low) insertion peptides (pHLIP® peptides) belong to the class of pH-sensitive membrane peptides, which target acidic tumors and deliver imaging and/or therapeutic agents to cancer cells within tumors. Procedures Ex vivo fluorescent imaging of tissue and organs collected at various time points after administration of different pHLIP® variants conjugated with fluorescent dyes of various polarity was performed. Methods of multivariate statistical analyses were employed to establish classification between fluorescently labeled pHLIP® variants in multidimensional space of spectral parameters. Results The fluorescently labeled pHLIP® variants were classified based on their biodistribution profile and ability of targeting of primary tumors. Also, submillimeter-sized metastatic lesions in lungs were identified by ex vivo imaging after intravenous administration of fluorescent pHLIP® peptide. Conclusions Different cargo molecules conjugated with pHLIP® peptides can alter biodistribution and tumor targeting. The obtained knowledge is essential for the design of novel pHLIP®-based diagnostic and therapeutic agents targeting primary tumors and metastatic lesions

Aquatic Toxicity of Photocatalyst Nanoparticles to Green Microalgae Chlorella vulgaris

In the last years, nanoparticles such as TiO2, ZnO, NiO, CuO and Fe2O3 were mainly used in wastewater applications. In addition to the positive aspects concerning using nanoparticles in the advanced oxidation process of wastewater containing pollutants, the impact of these nanoparticles on the environment must also be investigated. The toxicity of nanoparticles is generally investigated by the nanomaterials’ effect on green algae, especially on Chlorella vulgaris. In this review, several aspects are reviewed: the Chlorella vulgaris culture monitoring and growth parameters, the effect of different nanoparticles on Chlorella vulgaris, the toxicity of photocatalyst nanoparticles, and the mechanism of photocatalyst during oxidative stress on the photosynthetic mechanism of Chlorella vulgaris. The Bold basal medium (BBM) is generally recognized as an excellent standard cultivation medium for Chlorella vulgaris in the known environmental conditions such as temperature in the range 20–30 °C and light intensity of around 150 μE·m2·s−1 under a 16/8 h light/dark cycle. The nanoparticles synthesis methods influence the particle size, morphology, density, surface area to generate growth inhibition and further algal deaths at the nanoparticle-dependent concentration. Moreover, the results revealed that nanoparticles caused a more potent inhibitory effect on microalgal growth and severely disrupted algal cells’ membranes

Toxicity of a Binary Mixture of TiO2 and Imidacloprid Applied to Chlorella vulgaris

Nanoparticles have applications in various fields such as manufacturing and materials synthesis, the environment, electronics, energy harvesting, and medicine. Besides many applications of nanoparticles, further research is required for toxic environmental effect investigation. The toxic effect of titanium dioxide nanoparticles on the physiology of the green alga Chlorella vulgaris was studied with a widely used pesticide, imidacloprid (IMD). Chlorella vulgaris was exposed for 120 h in Bold’s basal medium to different toxic compounds, such as (i) a high concentration of TiO2 nanoparticles, 150–2000 mg/L, usually optimised in the photocatalytic degradation of wastewater, (ii) an extremely toxic pesticide for the aquatic environment, imidacloprid, in concentrations ranging from 5 to 40 mg/L, (iii) TiO2 nanoparticles combined with imidacloprid, usually used in a photocatalytic system. The results show that the TiO2 nanoparticles and IMD inhibited Chlorella vulgaris cell growth and decreased the biovolume by approximately 80% when 2 g/L TiO2 was used, meaning that the cells devised a mechanism to cope with a potentially stressful situation; 120 h of Chlorella vulgaris exposure to 40 mg/L of IMD resulted in a 16% decreased cell diameter and a 41% decrease in cell volume relative to the control sample, associated with the toxic effect of pesticides on the cells. Our study confirms the toxicity of nanoparticles through algal growth inhibition with an effective concentration (EC50) value measured after 72 h of 388.14 mg/L for TiO2 and 13 mg/L for IMD in a single-toxic system. The EC50 of TiO2 slowly decreased from 258.42 to 311.11 mg/L when IMD from 5 to 20 mg/L was added to the binary-toxic system. The concentration of TiO2 in the binary-toxic system did not change the EC50 for IMD, and its value was 0.019 g/L. The photodegradation process of imidacloprid (range of 5–40 mg/L) was also investigated in the algal medium incubated with 150–600 mg/L of titanium dioxide

pH (Low) Insertion Peptide targets 4T1 mammary tumors

Malignant tumors exhibit elevated uptake and use of glucose, as well as metabolite buildup and hypoxia due to inadequate blood supply, leading to tumor acidosis. The extracellular acidity is linked to tumor progression. The most effective pH-sensitive tumor targeting agents should sense pH at the surface of cancer cells, where it is expected to be the lowest. pHLIPs® (pH (Low) Insertion Peptides) belong to the class of pH-sensitive agents capable of delivering imaging and/or therapeutic agents to cancer cells within tumors. Here, we investigated targeting of highly metastatic 4T1 mammary tumors in BALB/c mice with three Alexa546 labeled pHLIP variants including well-characterized WT-pHLIP and, recently introduced, Var3-and Var7-pHLIPs

PHLIP-FIRE, a cell insertion-triggered fluorescent probe for imaging tumors demonstrates targeted cargo delivery in vivo

We have developed an improved tool for imaging acidic tumors by reporting the insertion of a transmembrane helix: the pHLIP-Fluorescence Insertion REporter (pHLIP-FIRE). In acidic tissues, such as tumors, peptides in the pHLIP family insert as α-helices across cell membranes. The cell-inserting end of the pHLIP-FIRE peptide has a fluorophore-fluorophore or fluorophore-quencher pair. A pair member is released by disulfide cleavage after insertion into the reducing environment inside a cell, resulting in dequenching of the probe. Thus, the fluorescence of the pHLIP-FIRE probe is enhanced upon cell-insertion in the targeted tissues but is suppressed elsewhere due to quenching. Targeting studies in mice bearing breast tumors show strong signaling by pHLIP-FIRE, with a contrast index of ∼17, demonstrating (i) direct imaging of pHLIP insertion and (ii) cargo translocation in vivo. Imaging and targeted cargo delivery should each have clinical applications

Surface plasmon resonance effect on the optical properties of TiO2 doped by noble metals nanoparticles

Nanocomposite thin films composed of a TiO2 matrix doped with noble metals nanoparticles (MNPS), Au and Ag, were deposited on Si (100) and glass substrates by dc magnetron sputtering. The samples were annealed in a protective atmosphere at temperatures ranging from 200 to 700 ºC. The main goal of this work is to characterize and compare the Surface Plasmon Resonance (SPR) behaviour in both systems. The studies have been focused on the growth of the nanoclusters and on their role on the optical properties of the films. Size, shape and distribution of the nanoclusters embedded on the titanium oxide dielectric matrix are reported as key factors on the SPR behaviour in both systems (Au:TiO2 and Ag:TiO2). The MNPs grew due to diffusion mechanisms, which were led by the annealing treatments, even at the highest annealing temperatures. Evidences of the correlation between the nanocomposite film structural changes and the evolution of the optical properties due to the SPR activity are reported. The SPR phenomenon manifests itself as a wide band on the visible range on the absorption spectra, and it is confirmed by an important change on the surface colour tones of the samples.This research is sponsored by FEDER funds through the program COMPETE - Programa Operacional Factores de Competitividade and by national funds through FCT - Fundação para a Ciência e a Tecnologia, under the project PTDC/CTM/ 70037/2006 70037/20B and also UB-SCT (Barcelona) for the TEM characterization

Targeting Breast Tumors with pH (Low) Insertion Peptides

Extracellular acidity is associated with tumor progression. Elevated glycolysis and acidosis promote the appearance of aggressive malignant cells with enhanced multidrug resistance. Thus, targeting of tumor acidity can open new avenues in diagnosis and treatment of aggressive tumors and targeting metastatic cancers cells within a tumor. pH (low) insertion peptides (pHLIPs) belong to the class of pH-sensitive agents capable of delivering imaging and/or therapeutic agents to cancer cells within tumors. Here, we investigated targeting of highly metastatic 4T1 mammary tumors and spontaneous breast tumors in FVB/N-Tg (MMTV-PyMT)­634Mul transgenic mice with three fluorescently labeled pHLIP variants including well-characterized WT-pHLIP and, recently introduced, Var3- and Var7-pHLIPs. The Var3- and Var7-pHLIPs constructs have faster blood clearance than the parent WT-pHLIP. All pHLIPs demonstrated excellent targeting of the above breast tumor models with tumor accumulation increasing over 4 h postinjection. Staining of nonmalignant stromal tissues in transgenic mice was minimal. The pHLIPs distribution in tumors showed colocalization with 2-deoxyglucose and the hypoxia marker, Pimonidazole. The highest degree of colocalization of fluorescent pHLIPs was shown to be with lactate dehydrogenase A, which is related to lactate production and acidification of tumors. In sum, the pHLIP-based targeting of breast cancer presents an opportunity to monitor metabolic changes, and to selectively deliver imaging and therapeutic agents to tumors