The amniotic fluid-derived cells: the biomedical challenge for the third millennium

Human amniotic fluid cells (H-AFC) have been used as a diagnostic tool for the prenatal diagnosis of fetal genetic anomalies for more than 50 years

CLINICAL AND BACTERIOLOGICAL EVALUATION OF SURGICAL SKIN WOUNDS TREATED WITH OZONIZED OLIVE OIL: A PRELIMINARY STUDY

Since in the last years the overuse of antibiotics and the appearance of “multi-drug resistant” bacteria constantly increased, scientific research was driven towards the study of new antimicrobial, especially of natural origin. The aim of the study was to evaluate the bacteriological and healing activity of an ozonized olive oil (OOL) used to treat abdominal surgical skin wounds in dogs and cats. A randomized blind controlled clinical trial was performed at the School of Biosciences and Veterinary Medicine, University of Camerino. For assessing the clinical efficacy, the ASEPSIS Scale, the Wound Evaluation Scale (WES) [1] and the Total Bacterial Count (TBC) analysis were performed for each patient [2]. The degree of owner satisfaction and the possible highlight of complications or adverse effects, were also considered. Animals were randomly divided into two groups: treated group (GA), that received topical application of OOL twice a day for three consecutive days, beginning the day of surgery, and control group (GC) that did not receive any kind of treatment. Comparing the TBC at T0, T1 and T2 between GC and GA, no significant differences were revealed. Within GC, it was observed a not significant increase in the TBC comparing T0 vs T1, T1 vs T2 and T0 vs T2. In the GA, between T0-T1 and T0-T2 the TBC value increased, while the TBC between T1-T2 (t=0.59, P=0.56) decreased, but not significantly. Comparing the frequency of optimal healing of both groups at the same time, a significant difference was observed at T2 (P=0.034) in favor of GA. Regarding the WES, a significant increase of patients of GA with optimal healing process from T0 to T2 was observed, while in the GC the percentage of optimal healing decreased from T0 to T1 and from T0 to T2 and increased from T1 to T2, but not significantly. Considering ASEPSIS score, the difference between GA and GC was not significant at T0 (T=0.05; P=0.96), at T1 (T=0.29; P=0.77), and at T2 (T=1.17; P=0.24). At T2, GA showed a high number of patients with satisfactory healing, but without any significant differences between the times of observation. This study allows to conclude that the OOL could be a medical device applicable to skin wounds without any collateral effect, it facilitates the healing process with a relative short period of treatment (3 days) and the owners, also reported an excellent grade of applicability and compliance. The absence of significant results related to the decrease of TBC mean values could find its explanation on a correct surgery preparation with iodophor agents. The results obtained with this study shows slight different results from the conclusions of a similar trial conducted by Kim et al. [3]. The explanations could be related to the shorter period of treatment and to the wound location, always in the abdominal area, with the consequent biological and physiological factors. No adverse events were reported. Infection related to licking was the only complication affecting 10% of patients in GC and 12.5% in GA

Genotoxicity Assays Analysis for Carbon Nanotubes: Friends or Foes? Preliminary Results on Human Peripheral Peukocytes

The literature is full of information on the biological effects, both in vitro and in vivo, of Carbon Nanotubes (CNTs) and almost all the data published are in agreement on the toxicity related to physicochemical properties. The gap in the knowledge of the effects of CNTs on biological systems is instead evident when approaching the damage induced to DNA by CNTs. In fact, genotoxicity data is still missing. Is this due to a lack of interest in this field of research or it is triggered by the fact that standard procedures for genotoxicity assays are still to come for CNTs? Actually, the second question seems to be the more appropriate, but also for science, "in media stat virtus" remains valid. This brief review, including some preliminary results on human peripheral leukocytes, will try to highlight this problem as seen from the genotoxicological point of view.JRC.I.4-Nanobioscience

A Pilot Prospective Study Evaluating the Effect of Curcuma-Based Herbal Food Supplement on the Outcome of In Vitro Fertilization in Patients Testing Positive for Four Immunological Biomarkers

Background and Objectives: Inflammation and oxidative stress have been described to reduce the chance for pregnancy instauration and maintenance. NOFLAMOX, a recently developed herbal preparation with recognized antioxidant and anti-inflammatory properties, can represent an interesting treatment to increase the chance of pregnancy, both physiological or after in vitro fertilization (IVF). The aim of this study was to assess NOFLAMOX’s effect; a population with unexplained infertility was screened for the recently described IMMUNOX panel based on four immunological biomarkers with a prospective study approach. Materials and Methods: Patients with unexplained infertility and positive for at least one of the biomarkers of the IMMUNOX panel were included in this study and treated with NOFLAMOX for three months prior to an IVF cycle. Results: Eighty-six patients (n = 86) were screened with the IMMUNOX panel and the forty-seven (54.5%) found positive were included in this study. In more detail, 11 were positive for TNFα (23.4%), 18 (38.3%) for glycodelin (GLY), 29 (61.7%) for Total Oxidative Status (TOS), and 32 (68.1%) for Complement Activity Toxic Factor (CATF). After three months of treatment, a significant reduction in the number of IMMUNOX-positive patients was observable, with 26 patients who turned IMMUNOX-negative displaying a quantitative statistically significant variation of 100% (11/11), 38.9% (7/18), 65.5% (18/29), and 75% (24/32), for TNFα, glycodelin, TOS, and CATF, respectively. Followed in the subsequent IVF cycle, this NOFLAMOX-treated population showed a pregnancy rate of 42.3% compared to the 4.7% of the IMMUNOX-positive group of patients. Conclusions: Taken together, the results of this study suggest that NOFLAMOX could represent an interesting option for those patients with unexplained infertility of inflammatory/oxidative origin. Further studies are needed to confirm these results and explore possible strategies to restore fertility in women with immune-mediated sterility

Genotoxicity and Morphological Transformation Induced by Cobalt Nanoparticles and Cobalt Chloride: an In Vitro Study on Balb/3T3 Mouse Fibroblasts

Nanotechnology is a new emerging field that involves the development, manufacturing and measurement of materials and systems in the sub micron down to few nanometers (nm) range. Its development is expected to have a large socio-economical impact in practically all industrial activity fields. However, there is still a lack of information about potential risks of manufactured nanoparticles (mNP) for the environment and with respect to human health. In this work, we studied cytotoxicity, genotoxicity and morphological transformation of cobalt nanoparticles (Co (0)-nano) and cobalt ions (Co+2) in Balb/3T3 cell model. We also evaluated Co-nano dissolution in culture medium and cellular uptake of Co-nano and Co+2. Our results indicate that Co-nano are exerting a cytotoxicity potential (IC50: 20 µM after 2h of exposure and 10 µM at 24 and 72h). For the Co-nano genotoxicity the results show that indeed the Co-nano is genotoxic (>1 µM) but no evident dose-dependent relationship was observed. For the morphological transformation, Co-nano show a statistically significance increase in the formation of Type III foci. Moreover analyzing the internalization of Co-nano we observed a higher uptake if compared with Co+2. Our results indicated cytotoxicity, morphological transformation and genotoxicity induced by Co-nano, while for Co+2 we found cytotoxicity, morphological transformation and genotoxicity, assessed by Comet assay but no by micronucleus test.JRC.I.4-Nanobioscience

Carbon nanotubes induce oxidative DNA damage in RAW 264.7 cells.

The induction of DNA and chromosome damage following in vitro exposure to carbon nanotubes (CNT) was assessed on the murine macrophage cell line RAW 264.7 by means of the micronucleus (MN) and the comet assays. Exposures to two CNT preparations (single-walled CNT (SWCNT > 90%) and multiwalled CNT (MWCNT > 90%) were performed in increasing mass concentrations (0.01-100 microg/ml). The frequency of micronuclei was significantly increased in cells treated with SWCNT (at doses above 0.1 microg/ml), whereas MWCNT had the same effect at higher concentrations (1 microg/ml) (P < 0.05). The results of the comet assay revealed that the effects of treatment with SWCNT were detectable at all concentrations tested (1-100 microg/ml); oxidized purines increased significantly, whereas pyrimidines showed a significant increase (P < 0.001) only at the highest concentration (100 microg/ml). In cells treated with MWCNT, an increase in DNA migration due to the oxidative damage to purines was observed at a concentration of 1 and 10 microg/ml, whereas pyrimidines showed a significant increase only at the highest mass concentration tested. However, both SWCNT and MWCNT induced a statistically significant cytotoxic effect at the highest concentrations tested (P < 0.001). These findings suggest that both the MN and comet assays can reliably detect small amount of damaged DNA at both chromosome and nuclear levels in RAW 264.7 cells. Moreover, the modified version of the comet assay allows the specific detection of the induction of oxidative damage to DNA, which may be the underlying mechanism involved in the CNT-associated genotoxicity