Composite Coatings of Chitosan and Silver Nanoparticles Obtained by Galvanic Deposition for Orthopedic Implants.

In this work, composite coatings of chitosan and silver nanoparticles were presented as an antibacterial coating for orthopedic implants. Coatings were deposited on AISI 304L using the galvanic deposition method. In galvanic deposition, the difference of the electrochemical redox potential between two metals (the substrate and a sacrificial anode) has the pivotal role in the process. In the coupling of these two metals a spontaneous redox reaction occurs and thus no external power supply is necessary. Using this process, a uniform deposition on the exposed area and a good adherence of the composite coating on the metallic substrate were achieved. Physical-chemical characterizations were carried out to evaluate morphology, chemical composition, and the presence of silver nanoparticles. These characterizations have shown the deposition of coatings with homogenous and porous surface structures with silver nanoparticles incorporated and distributed into the polymeric matrix. Corrosion tests were also carried out in a simulated body fluid at 37 °C in order to simulate the same physiological conditions. Corrosion potential and corrosion current density were obtained from the polarization curves by Tafel extrapolation. The results show an improvement in protection against corrosion phenomena compared to bare AISI 304L. Furthermore, the ability of the coating to release the Ag+ was evaluated in the simulated body fluid at 37 °C and it was found that the release mechanism switches from anomalous to diffusion controlled after 3 h.

c-FLIPL enhances anti-apoptotic Akt functions by modulation of Gsk3ß activity.

This corrects the article DOI: 10.1038/cdd.2010.65.

miR-221/222 overexpession in human glioblastoma increases invasiveness by targeting the protein phosphate PTPµ.

Glioblastoma is the most frequent brain tumor in adults and is the most lethal form of human cancer. Despite the improvements in treatments, survival of patients remains poor. In order to identify microRNAs (miRs) involved in glioma tumorigenesis, we evaluated, by a miRarray, differential expression of miRs in the tumorigenic glioma LN-18, LN-229 and U87MG cells compared with the non-tumorigenic T98G cells. Among different miRs we focused our attention on miR-221 and -222. We demonstrated the presence of a binding site for these two miRs in the 3' untranslated region of the protein tyrosine phosphatase µ (PTPµ). Previous studies indicated that PTPµ suppresses cell migration and is downregulated in glioblastoma. Significantly, we found that miR-221 and -222 overexpression induced a downregulation of PTPµ as analyzed by both western blot and real-time PCR. Furthermore, miR-222 and -221 induced an increase in cell migration and growth in soft agar in glioma cells. Interestingly, the re-expression of PTPµ gene was able to revert the miR-222 and -221 effects on cell migration. Furthermore, we found an inverse correlation between miR-221 and -222 and PTPµ in human glioma cancer samples. In conclusion, our results suggest that miR-221 and -222 regulate glioma tumorigenesis at least in part through the control of PTPµ protein expression.

c-FLIPL enhances anti-apoptotic Akt functions by modulation of Gsk3ß activity.

Akt is a serine-threonine kinase that has an important role in transducing survival signals. Akt also regulates a number of proteins involved in the apoptotic process. To find new Akt interactors, we performed a two-hybrid screening in yeast using full-length Akt cDNA as bait and a human cDNA heart library as prey. Among 200 clones obtained, two of them were identified as coding for the c-FLIP(L) protein. c-FLIP(L) is an endogenous inhibitor of death receptor-induced apoptosis through the caspase-8 pathway. Using co-immunoprecipitation experiments of either transfected or endogenous proteins, we confirmed the interaction between Akt and c-FLIP(L). Furthermore, we observed that c-FLIP(L) overexpression interferes with Gsk3-ß phosphorylation levels. Moreover, through its effects on Gsk3ß, c-FLIP(L) overexpression in cancer cells induced resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). This effect was mediated by the regulation of p27(Kip1) and caspase-3 expression. These results indicate the existence of a new mechanism of resistance to TRAIL in cancer cells, and unexpected functions of c-FLIP(L).

MicroRNA signatures of TRAIL resistance in human non-small cell lung cancer.

To define novel pathways that regulate susceptibility to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) in non-small cell lung cancer (NSCLC), we have performed genome-wide expression profiling of microRNAs (miRs). We show that in TRAIL-resistant NSCLC cells, levels of different miRs are increased, and in particular, miR-221 and -222. We demonstrate that these miRs impair TRAIL-dependent apoptosis by inhibiting the expression of key functional proteins. Indeed, transfection with anti-miR-221 and -222 rendered CALU-1-resistant cells sensitive to TRAIL. Conversely, H460-sensitive cells treated with -221 and -222 pre-miRs become resistant to TRAIL. miR-221 and -222 target the 3'-UTR of Kit and p27(kip1) mRNAs, but interfere with TRAIL signaling mainly through p27(kip1). In conclusion, we show that high expression levels of miR-221 and -222 are needed to maintain the TRAIL-resistant phenotype, thus making these miRs as promising therapeutic targets or diagnostic tool for TRAIL resistance in NSCLC.