Suitability of PLLA as piezoelectric substrates for tissue engineering evidenced by microscopy techniques

Since the discovery of the piezoelectric character of bone, the suitability of some piezoelectric materials have been studied for bone repair; they are thought to act like transducers converting the mechanical energy of skeletal deformation in electrical stimuli capable of controlling osteogenic growth. The mechanisms underlying this process are far from being understood and systematic studies at a local scale are required. Atomic force microscopy (AFM) is a unique way to observe phenomena at the nanoscale and liquid imaging provides a unique tool to assess biological phenomena at the nanoscale. So in this study, aiming at a better understanding of the role of piezoelectricity in the osteogenic growth, the interaction between a poled piezoelectric material, in this case poly (L-lactic) acid and an adhesion promoting protein, the fibronectin, and bone-like cells is evaluated by scanning probe microscopy and confocal laser scanning microscopy (CLSM). © Microscopy Society of America 2012.(undefined

Multicaloric effect in a multiferroic composite of Gd-5(Si,Ge)(4) microparticles embedded into a ferroelectric PVDF matrix

CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOThe coupling between electric, magnetic and elastic features in multiferroic materials is an emerging field in materials science, with important applications on alternative solid-state cooling technologies, energy harvesting and sensors/actuators. In this direction, we developed a thorough investigation of a multiferroic composite, comprising magnetocaloric/magnetostrictive Gd5Si2.4Ge1.6 microparticles blended into a piezo- and pyroelectric poly(vinylidene) fluoride (PVDF) matrix. Using a simple solvent casting technique, the formation and stabilization of PVDF electroactive phases are improved when the filler content increases from 2 to 12 weight fraction (wt.%). This effect greatly contributes to the magnetoelectric (ME) coupling, with the ME coefficient alpha(ME) increasing from 0.3 V/cm.Oe to 2.2 V/cm.Oe, by increasing the amount of magnetic material. In addition, magnetic measurements revealed that the ME-coupling has influenced the magnetocaloric effect via a contribution from the electroactive polymer and hence leading to a multicaloric effect. These results contribute to the development of multifunctional systems for novel technologies.9CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO203180/2014-3This work is funded by FEDER funds through the COMPETE 2020 Programme and National Funds throught FCT -Portuguese Foundation for Science and Technology under the projects POCI-01-0145-FEDER-029454, POCI-01-0145-FEDER-032527 and UID/FIS/04564/2016. This work was also supported by NECL with the project NORTE-01-0145-FEDER-022096 and by the European Union Horizon -2020 research and innovation program under the Marie Sklodowska-Curie Grant Agreement No. 734801. The authors acknowledge K. Pirota and F. Beron for the FTIR measurements performed at Unicamp in Brazil. Special thanks to Dr. A. Aliev for help in automatizing the ME experiments at Amirkhanov Institute of Physics and the helpful discussions. Federal Fluminense University, Brazil, permanent address for MSR; Aveiro University, Portugal, temporary address during this work for MSR. VMA thanks the CNPq for the Grant No. 203180/2014-3. A.A. acknowledges Russian Science Foundation for support magnetoelectric studies (grant No. 18-79-10176). J.H. Belo thanks FCT for Grant No. SFRH/BD/88440/2012, Project PTDC/FISMA/31302/2017, and his contract No. DL57/2016 reference SFRH-BPD-87430/2012. Finally, the discussion of results has been made possible through the mobility grant provided by the 5 top 100 Russian Academic Excellence Project at the Immanuel Kant Baltic Federal University

Microfabrication of a biomimetic arcade-like electrospun scaffold for cartilage tissue engineering applications

Designing and fabricating hierarchical geometries for tissue engineering (TE) applications is the major challenge and also the biggest opportunity of regenerative medicine in recent years, being the in vitro recreation of the arcade-like cartilaginous tissue one of the most critical examples due to the current inefficient standard medical procedures and the lack of fabrication techniques capable of building scaffolds with the required architecture in a cost and time effective way. Taking this into account, we suggest a feasible and accurate methodology that uses a sequential adaptation of an electrospinning-electrospraying set up to construct a system comprising both fibres and sacrificial microparticles. Polycaprolactone (PCL) and polyethylene glycol were respectively used as bulk and sacrificial biomaterials, leading to a bi-layered PCL scaffold which presented not only a depth-dependent fibre orientation similar to natural cartilage, but also mechanical features and porosity compatible with cartilage TE approaches. In fact, cell viability studies confirmed the biocompatibility of the scaffold and its ability to guarantee suitable cell adhesion, proliferation and migration throughout the 3D anisotropic fibrous network. Additionally, likewise the natural anisotropic cartilage, the PCL scaffold was capable of inducing oriented cell-material interactions since the morphology, alignment and density of the chondrocytes changed relatively to the specific topographic cues of each electrospun layer.publishe

Efficiency of Spermatogonial Dedifferentiation during Aging

Adult stem cells are critical for tissue homeostasis; therefore, the mechanisms utilized to maintain an adequate stem cell pool are important for the survival of an individual. In Drosophila, one mechanism utilized to replace lost germline stem cells (GSCs) is dedifferentiation of early progenitor cells. However, the average number of male GSCs decreases with age, suggesting that stem cell replacement may become compromised in older flies.Using a temperature sensitive allelic combination of Stat92E to control dedifferentiation, we found that germline dedifferentiation is remarkably efficient in older males; somatic cells are also effectively replaced. Surprisingly, although the number of somatic cyst cells also declines with age, the proliferation rate of early somatic cells, including cyst stem cells (CySCs) increases.These data indicate that defects in spermatogonial dedifferentiation are not likely to contribute significantly to an aging-related decline in GSCs. In addition, our findings highlight differences in the ways GSCs and CySCs age. Strategies to initiate or enhance the ability of endogenous, differentiating progenitor cells to replace lost stem cells could provide a powerful and novel strategy for maintaining tissue homeostasis and an alternative to tissue replacement therapy in older individuals

Mild dyserythropoiesis and beta-like globin gene expression imbalance due to the loss of histone chaperone ASF1B

The expression of the human β-like globin genes follows a well-orchestrated developmental pattern, undergoing two essential switches, the first one during the first weeks of gestation (ε to γ), and the second one during the perinatal period (γ to β). The γ- to β-globin gene switching mechanism includes suppression of fetal (γ-globin, HbF) and activation of adult (β-globin, HbA) globin gene transcription. In hereditary persistence of fetal hemoglobin (HPFH), the γ-globin suppression mechanism is impaired leaving these individuals with unusual elevated levels of fetal hemoglobin (HbF) in adulthood. Recently, the transcription factors KLF1 and BCL11A have been established as master regulators of the γ- to β-globin switch. Previously, a genomic variant in the KLF1 gene, identified by linkage analysis performed on twenty-seven members of a Maltese family, was found to be associated with HPFH. However, variation in the levels of HbF among family members, and those from other reported families carrying genetic variants in KLF1, suggests additional contributors to globin switching. ASF1B was downregulated in the family members with HPFH. Here, we investigate the role of ASF1B in γ- to β-globin switching and erythropoiesis in vivo. Mouse-human interspecies ASF1B protein identity is 91.6%. By means of knockdown functional assays in human primary erythroid cultures and analysis of the erythroid lineage in Asf1b knockout mice, we provide evidence that ASF1B is a novel contributor to steady-state erythroid differentiation, and while its loss affects the balance of globin expression, it has no major role in hemoglobin switching

Pregnancy in the mature adult mouse does not alter the proportion of mammary epithelial stem/progenitor cells

Introduction In humans, an early full-term pregnancy reduces lifetime breast cancer risk by up to 50% whereas a later pregnancy (>35 years old) can increase lifetime risk. Several mechanisms have been suggested, including changes in levels of circulating hormones, changes in the way the breast responds to these hormones, changes in gene expression programmes which may alter susceptibility to transformation and changes to mammary stem cell numbers or behaviour. Previous studies have shown that the mammary tissue isolated from both virgin and parous mice has the ability to repopulate a cleared mammary fat pad in transplant experiments. Limited dilution transplant assays have demonstrated that early pregnancy (at 5 weeks of age) reduces stem/progenitor cell numbers in the mouse mammary epithelium by twofold. However, the effects on stem/progenitor cell numbers in the mammary epithelium of a pregnancy in older animals have not yet been tested. Methods Mice were put through a full-term pregnancy at 9 weeks of age, when the mammary epithelium is mature. The total mammary epithelium was purified from parous 7-week post-lactation and age-matched virgin mice and analysed by flow cytometry and limiting dilution cleared fat pad transplants. Results There were no significant differences in the proportions of different mammary epithelial cell populations or numbers of CD24+/Low Sca-1- CD49fHigh cells (stem cell enriched basal mammary epithelial compartment). There was no significant difference in stem/progenitor cell frequency based on limiting dilution transplants between the parous and age-matched virgin epithelium. Conclusions Although differences between parous and virgin mammary epithelium at later time points post lactation or following multiple pregnancies cannot be ruled out, there are no differences in stem/progenitor cell numbers between mammary epithelium isolated from parous animals which were mated at 9 weeks old and virgin animals. However, a recent report has suggested that animals that were mated at 5 weeks old have a twofold reduction in stem/progenitor cell numbers. This is of interest given the association between early, but not late, pregnancy and breast cancer risk reduction in humans. However, a mechanistic connection between stem cell numbers and breast cancer risk remains to be established

Impact of the hypoxic microenvironment on spermatogonial stem cells in culture

The stem cell niche plays a crucial role in the decision to either self-renew or differentiate. Recent observations lead to the hypothesis that O2 supply by blood and local O2 tension could be key components of the testicular niche of spermatogonial stem cells (SSCs). In this study, we investigated the impact of different hypoxic conditions (3.5%, 1%, and 0.1% O2 tension) on murine and human SSCs in culture. We observed a deleterious effect of severe hypoxia (1% O2 and 0.1% O2) on the capacity of murine SSCs to form germ cell clusters when plated at low density. Severe effects on SSCs proliferation occur at an O2 tension ≤1% and hypoxia was shown to induce a slight differentiation bias under 1% and 0.1% O2 conditions. Exposure to hypoxia did not appear to change the mitochondrial mass and the potential of membrane of mitochondria in SSCs, but induced the generation of mitochondrial ROS at 3.5% and 1% O2. In 3.5% O2 conditions, the capacity of SSCs to form colonies was maintained at the level of 21% O2 at low cell density, but it was impossible to amplify and maintain stem cell number in high cell density culture. In addition, we observed that 3.5% hypoxia did not improve the maintenance and propagation of human SSCs. Finally, our data tend to show that the transcription factors HIF-1α and HIF-2α are not involved in the SSCs cell autonomous response to hypoxia