The Osteoprint: A bioinspired two-photon polymerized 3-D structure for the enhancement of bone-like cell differentiation

The need for a better understanding of cell behavior and for exploiting cell functions in various healthcare applications has driven biomedical research to develop increasingly complex fabrication strategies to reproduce the natural biological microenvironment in vitro. Different approaches have led to the development of refined examples of 2- and 3-D structures able to sustain cellular proliferation, differentiation and functionality very similar to those normally occurring in living organisms. One such approach is two-photon polymerization. In this paper, we present a trabecula-like structure (which we have named “Osteoprint”) that resembles to the typical microenvironment of trabecular bone cells. Starting from microtomography images of the trabecular bone, we prepared several Osteoprints through two-photon polymerization and tested the behavior of SaOS-2 bone-like cells cultured on our structures. Interestingly, we found that Osteoprints deeply affect cellular behavior, determining an exit from the cell cycle and an enhancement of osteogenic differentiation. Indeed, we found an up-regulation of the genes involved in SaOS-2 cell maturation and an increase in hydroxyapatite production and accumulation upon SaOS-2 culture on the Osteoprints. The findings we obtained are extremely interesting, and open up new perspectives in “bioinspired” approaches for tissue engineering and regenerative medicine.     

Pelviperineoscopy. Preliminary study of feasibility on cadavers

OBJECTIVE: The traditional access of perineum for the treatment of the pelvic organ prolapse by vaginal route is probably responsible for the risk of mesh exposure and a longer convalescence. So, endoscopic access to perinemeum needs to be evaluated. PATIENTS AND METHODS: Feasibility study on cadavers. Details of the procedure: opening of the pararectal space by digital dissection first after incision on the level of the posterior commissure. Introduction of an optical trocart to the level of the perineum incision; dissection of pararectal space with optics and CO(2); individualization of the various elements; installation of a transobturator trocart and a transgluteal trocart; dissection of the rectovaginal septum and visualization of the sacrospinous ligament and pudendal nerve. The measured variables were: operational incidents, possibility of creation of working space, dissection of the rectovaginal septum; finally, visualization of the sacrospinous ligament and pudendal pedicle. RESULTS: On the 4 studied cadavers, we could carry out a dissection of pelvirectal space in all the cases. On the 8 pararectal fosses, in all the cases we could carry out a cavity of dissection and to open the recto vaginal septum, visualization of the sacrosciatic ligament and pudendal pedicle was possible in 6 cases out of 8. There were one rectal injury, two vaginal injuries and one lesion of the pudendal pedicle. DISCUSSION AND CONCLUSION: This endoscopic access allows in the majority of cases to see the structures necessary to the realization of a vaginal sacrospinofixation or the installation of posterior mesh without a colpotomy and a traumatic exposure. The incidents are probably due to our inexperience and should disappear in time. The pelvi-perineoscopy is an endoscopic access of perineum which should be evaluated.     

A New Anorganic Equine Bone Substitute for Oral Surgery: Structural Characterization and Regenerative Potential

Different xenogeneic inorganic bone substitutes are currently used as bone grafting materials in oral and maxillo-facial surgery. The aim of the present study was to determine the physicochemical properties and the in vivo performance of an anorganic equine bone (AEB) substitute. AEB is manufactured by applying a process involving heating at >300 °C with the aim of removing all the antigens and the organic components. AEB was structurally characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray fluorescence (XRF), and Fourier-transformed infrared (FT-IR) spectroscopy and compared to the anorganic bovine bone (ABB). In order to provide a preliminary evaluation of the in vivo performance of AEB, 18 bone defects were prepared and grafted with AEB (nine sites), or ABB (nine sites) used as a control, in nine Yucatan Minipigs. De novo bone formation, residual bone substitute, as well as local inflammatory and tissue effects were histologically evaluated at 30 and 90 days after implantation. The structural characterization showed that the surface morphology, particle size, chemical composition, and crystalline structure of AEB were similar to cancellous human bone. The histological examination of AEB showed a comparable pattern of newly formed bone and residual biomaterial to that of ABB. Overall, the structural data and pre-clinical evidence reported in the present study suggests that AEB can be effectively used as bone grafting material in oral surgery procedures.     

Can Fasting Curb the Metabolic Syndrome Epidemic?

Metabolic syndrome (MetS) represents a cluster of metabolic abnormalities that includes hypertension, central obesity, insulin resistance, and atherogenic dyslipidemia. Due to the high prevalence (around 1/3 of the world population) economic burden of MetS, there is a need for new dietary, lifestyle, and therapeutic options. Recently, fasting emerged as a dietary method proposed for controlling metabolic risk factors. Intermittent fasting (IF), or time-restricted feeding (TRF), describes an array of feeding patterns in which calorie intake is restricted to a specific time period. Hence, this review aimed to elucidate the latest data on MetS and explore the viability of simple management options, such as IF and TRF. Preclinical studies have shown how IF/TRF exerts beneficial effects on the gut microbiota, glucose and insulin metabolism, weight and visceral fat, and lipid metabolism. However, the results obtained from human studies are somewhat conflicting, as weight loss was achieved in all studies, whereas in some studies, there was no significant effect on insulin resistance, cholesterol/lipid metabolism, or blood pressure. Nevertheless, as only very few human studies were performed, there is a need for more randomized control trials on larger cohorts of patients with MetS to gather higher-yield evidence to clarify whether IF/TRF are suitable dietary patterns for this population.     

Malignant melanoma in HIV: Epidemiology,pathogenesis, and management

People affected by immunodeficiency, and especially those infected by HIV, are burdened by a higher risk of developing malignancies. It has been estimated that the incidence of melanoma in HIV‐infected people is 2.6‐fold higher than in uninfected ones. In this group of patients, melanoma shows a more aggressive phenotype and poorer survival rates compared to HIV‐negative people. Standard guidelines of diagnosis and care do not exist yet. Studies suggest high index of suspicion and a low threshold for biopsy in HIV‐positive patients regardless of their CD4⁺ count and the use of standard surgical margins for re‐excision procedures. In case of diagnosis of melanoma in HIV‐positive patients, a thorough search for metastatic disease is recommended because of the more aggressive course of this cancer in HIV‐positive patients. Moreover, to rapidly find out any recurrence or metastatic disease after treatment, these patients need a close follow‐up, every 3 months, for the first 2 years and at least twice yearly thereafter. Although surgery remains the main therapeutic option, application of immune checkpoint‐based immunotherapy is being studied and seems to be promising. The aim of this review is to present the current knowledge and future options for melanoma diagnosis and treatment in people living with HIV.     

Importation of mycobacteriosis with ornamental fish: Medico-legal implications

Mycobacterium fortuitum, as well as Mycobacterium marinum and Mycobacterium chelonae, are the etiological agents of fish Mycobacterioses. Mycobacteriosis has been reported to affect a wide range of freshwater and marine fish species, suggesting an ubiquitous distribution, and can cause zoonotic infections (known as "fish tank granuloma" or "swimming pool granuloma") in humans exposed to fish and contaminated water. Infection in human consists of nodular cutaneous lesions that can progress to tenosynovitis, arthritis, and osteomyelitis, depending on the immunological status. Authors describe some cases observed during routinary diagnostic activity in aquarium fish. Fish were sampled and histopathological, microbiological, and biomolecular exams were carried out. Histopathology showed systemic granulomatosis. Microbiological and biomolecular exams allowed us to identify the M. fortuitum as a main species. Finally, some considerations on the legal aspects of such disease are discussed.     

A role for all-trans-retinoic acid in the early steps of lymphatic vasculature development

The molecular mechanisms that regulate the earliest steps of lymphatic vascular system development are unknown. To identify regulators of lymphatic competence and commitment, we used an in vitro vascular assay with mouse embryonic stem cell-derived embryoid bodies (EBs). We found that incubation with retinoic acid (RA) and, more potently, with RA in combination with cAMP, induced the expression of the lymphatic competence marker LYVE-1 in the vascular structures of the EBs. This effect was dependent on RA receptor (RAR)-α and protein kinase A signaling. RA-cAMP incubation also promoted the development of CD31+/LYVE-1+/Prox1+ cell clusters. In situ studies revealed that RAR-α is expressed by endothelial cells of the cardinal vein in ED 9.5-11.5 mouse embryos. Timed exposure of mouse and Xenopus embryos to excess of RA upregulated LYVE-1 and VEGFR-3 on embryonic veins and increased formation of Prox1-positive lymphatic progenitors. These findings indicate that RA signaling mediates the earliest steps of lymphatic vasculature development.     

24-Hour QT variability in heart failure

Background

Previous studies have shown that increased temporal variability of repolarization, as reflected by QT interval variability measured for 10 minutes, predicted spontaneous ventricular arrhythmias in implantable cardioverter defribrillator patients, but it is unclear how these measures perform in 24-hour recordings.

Methods

Twenty-four-hour digital Holter recordings from 372 subjects with chronic heart failure enrolled in Gruppo Italiano per lo Studio della Sopravvivenza nell'Insufficienza Cardiaca, (GISSI) Heart Failure study were analyzed using a template-matching, semiautomatic algorithm to measure QT and heart rate time series in sequential 5-minute epochs for 24 hours. QT variability was expressed as normalized QT variance (QTVN) or as the log ratio of the QTVN over normalized heart rate variance (QT variability index, or QTVI).

Results

A pronounced diurnal variation was seen in both QTVI and QTVN. Both were lowest in the midnight to 6 am time frame and increased throughout the day, peaking at noon to 6 pm, then decreasing 6 pm to midnight. For QTVI, all 4 time points were significantly different (P < .0001). QT variability index correlated with heart rate (r = 0.38, P < .0001) and was significantly higher for those in higher New York Heart Association (NYHA) classes (r = 0.22, P = .0003). Normalized QT variance did not correlate with heart rate or NYHA but correlated negatively with serum potassium (r = −0.22, P = .0002) and manifested the greatest increase during midmorning hours.

Conclusions

Repolarization lability as reflected in QT variability has a pronounced diurnal variation and increases significantly after 6 am, the time of greatest arrhythmic risk. QT variability for 24 hours might improve risk prediction in chronic heart failure patients and should be tested in appropriate trials.