Liver Cancer: Current and Future Trends Using Biomaterials

Hepatocellular carcinoma (HCC) is the fifth most common type of cancer diagnosed and the second leading cause of death worldwide. Despite advancement in current treatments for HCC, the prognosis for this cancer is still unfavorable. This comprehensive review article focuses on all the current technology that applies biomaterials to treat and study liver cancer, thus showing the versatility of biomaterials to be used as smart tools in this complex pathologic scenario. Specifically, after introducing the liver anatomy and pathology by focusing on the available treatments for HCC, this review summarizes the current biomaterial-based approaches for systemic delivery and implantable tools for locally administrating bioactive factors and provides a comprehensive discussion of the specific therapies and targeting agents to effciently deliver those factors. This review also highlights the novel application of biomaterials to study HCC, which includes hydrogels and scaffolds to tissue engineer 3D in vitro models representative of the tumor environment. Such models will serve to better understand the tumor biology and investigate new therapies for HCC. Special focus is given to innovative approaches, e.g., combined delivery therapies, and to alternative approaches—e.g., cell capture—as promising future trends in the application of biomaterials to treat HCC

Physical exercise for bone health in men with prostate cancer receiving androgen deprivation therapy: a systematic review

PURPOSE: Androgen deprivation therapy (ADT) is a treatment used in men with prostate cancer (PCa); however it is responsible for many adverse effects, with negative impact on quality of life. ADT causes loss of bone mineral density (BMD) and skeletal muscle mass, alteration of body composition, and cognitive function, which altogether lead to increased risk of accidental falls and fractures. This systematic review analyses the effectiveness of physical exercise (PE) in preventing accidental falls and fractures and reducing the loss of BMD in men with PCa receiving ADT.METHODS: We searched MEDLINE, EMBASE, CINAHL, and the Cochrane Library for articles between database inception and September 2, 2020. Eligible studies included randomized controlled trials (RCTs) investigating the effects of exercise on bone health in men with PCa receiving ADT.RESULTS: Nine RCTs were included. Experimental PE consisted in multicomponent programmes that involved aerobic, resistance, impact-loading exercise, and football training. None of the RCTs investigated the risk of accidental falls and fractures, while two trials reported beneficial effects of PE on lumbar spine, hip, and femoral shaft BMD. No further significant difference was detected in the outcomes investigated.CONCLUSION: Evidence of the effectiveness of PE to prevent the risk of accidental falls and fractures and BMD loss is lacking. Nevertheless, clinical guidelines recommend PE as a part of the clinical management of men with PCa receiving ADT due to its known numerous health benefits. Research should focus on PE strategies to prevent accidental falls, a clinically relevant outcome in this vulnerable population.TRIAL REGISTRATION: The study protocol was registered with International Prospective Register of Systematic Reviews (PROSPERO, number CRD 42020158444 ) on 04/28/2020

Nutlin-loaded magnetic solid lipid nanoparticles for targeted glioblastoma treatment

Aim: Glioblastoma multiforme is one of the deadliest forms of cancer, and current treatments are limited to palliative cares. The present study proposes a nanotechnology-based solution able to improve both drug efficacy and its delivery efficiency. Materials & methods: Nutlin-3a and superparamagnetic nanoparticles were encapsulated in solid lipid nanoparticles, and the obtained nanovectors (nutlin-loaded magnetic solid lipid nanoparticle [Nut-Mag-SLNs]) were characterized by analyzing both their physicochemical properties and their effects on U-87 MG glioblastoma cells. Results: Nut-Mag-SLNs showed good colloidal stability, the ability to cross an in vitro blood–brain barrier model, and a superior pro-apoptotic activity toward glioblastoma cells with respect to the free drug. Conclusion: Nut-Mag-SLNs represent a promising multifunctional nanoplatform for the treatment of glioblastoma multiforme

Tympanic Membrane Collagen Expression by Dynamically Cultured Human Mesenchymal Stromal Cell/Star-Branched Poly(ε-Caprolactone) Nonwoven Constructs

The tympanic membrane (TM) primes the sound transmission mechanism due to special fibrous layers mainly of collagens II, III, and IV as a product of TM fibroblasts, while type I is less represented. In this study, human mesenchymal stromal cells (hMSCs) were cultured on star-branched poly("-caprolactone) (*PCL)-based nonwovens using a TM bioreactor and proper dierentiating factors to induce the expression of the TM collagen types. The cell cultures were carried out for one week under static and dynamic conditions. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) were used to assess collagen expression. A Finite Element Model was applied to calculate the stress distribution on the scaolds under dynamic culture. Nanohydroxyapatite (HA) was used as a filler to change density and tensile strength of *PCL scaolds. In dynamically cultured *PCL constructs, fibroblast surface marker was overexpressed, and collagen type II was revealed via IHC. Collagen types I, III and IV were also detected. Von Mises stress maps showed that during the bioreactor motion, the maximum stress in *PCL was double that in HA/*PCL scaolds. By using a *PCL nonwoven scaold, with suitable physico-mechanical properties, an oscillatory culture, and proper dierentiative factors, hMSCs were committed into fibroblast lineage-producing TM-like collagens

Barium titanate nanoparticles and hypergravity stimulation improve differentiation of mesenchymal stem cells into osteoblasts.

BACKGROUND: Enhancement of the osteogenic potential of mesenchymal stem cells (MSCs) is highly desirable in the field of bone regeneration. This paper proposes a new approach for the improvement of osteogenesis combining hypergravity with osteoinductive nanoparticles (NPs). MATERIALS AND METHODS: In this study, we aimed to investigate the combined effects of hypergravity and barium titanate NPs (BTNPs) on the osteogenic differentiation of rat MSCs, and the hypergravity effects on NP internalization. To obtain the hypergravity condition, we used a large-diameter centrifuge in the presence of a BTNP-doped culture medium. We analyzed cell morphology and NP internalization with immunofluorescent staining and coherent anti-Stokes Raman scattering, respectively. Moreover, cell differentiation was evaluated both at the gene level with quantitative real-time reverse-transcription polymerase chain reaction and at the protein level with Western blotting. RESULTS: Following a 20 g treatment, we found alterations in cytoskeleton conformation, cellular shape and morphology, as well as a significant increment of expression of osteoblastic markers both at the gene and protein levels, jointly pointing to a substantial increment of NP uptake. Taken together, our findings suggest a synergistic effect of hypergravity and BTNPs in the enhancement of the osteogenic differentiation of MSCs. CONCLUSION: The obtained results could become useful in the design of new approaches in bone-tissue engineering, as well as for in vitro drug-delivery strategies where an increment of nanocarrier internalization could result in a higher drug uptake by cell and/or tissue constructs

Growing bone tissue-engineered niches with graded osteogenicity: an in vitro method for biomimetic construct assembly

The traditional bone tissue-engineering approach exploits mesenchymal stem cells ( MSCs) to be seeded once only on three-dimensional (3D) scaffolds, hence, differentiated for a certain period of time and resulting in a homogeneous osteoblast population at the endpoint. However, after achieving terminal osteodifferentiation, cell viability is usually markedly compromised. On the other hand, naturally occurring osteogenesis results from the coexistence of MSC progenies at distinct differentiative stages in the same microenvironment. This diversification also enables long-term viability of the mature tissue. We report an easy and tunable in vitro method to engineer simple osteogenic cell niches in a biomimetic fashion. The niches were grown via periodic reseeding of undifferentiated MSCs on MSC/scaffold constructs, the latter undergoing osteogenic commitment. Timefractioning of the seeded cell number during differentiation time of the constructs allowed graded osteogenic cell populations to be grown together on the same scaffolds (i.e., not only terminally differentiated osteoblasts). In such cell-dynamic systems, the overall differentiative stage of the constructs could also be tuned by varying the cell density seeded at each inoculation. In this way, we generated two different biomimetic niche models able to host good reservoirs of preosteoblasts and other osteoprogenitors after 21 culture days. At that time, the niche type resulting in 40.8% of immature osteogenic progenies and only 59.2% of mature osteoblasts showed a calcium content comparable to the constructs obtained with the traditional culture method (i.e., 100.03 – 29.30 vs. 78.51 – 28.50 pg/cell, respectively; p = not significant), the latter colonized only by fully differentiated osteoblasts showing exhausted viability. This assembly method for tissue-engineered constructs enabled a set of important parameters, such as viability, colonization, and osteogenic yield of the MSCs to be balanced on 3D scaffolds, thus achieving biomimetic in vitro models with graded osteogenicity, which are more complex and reliable than those currently used by tissue engineers

Effect of dipyridamole on gap junctions regulation in diseased myocardium

Gap junctions (GJ) mediate electrical coupling between cardiac myocytes, allowing the spreading of the electrical wave responsible for synchronized contraction [1]. GJ function can be regulated by modulation of connexon densities on membranes, connexin (Cx) phosphorylation, trafficking and degradation. Recent studies showed that adenosine involves Cx43 turn-over in A1 receptor-dependent manner [2], and dipyridamole increases GJ coupling and amount of Cx43 in endothelial cells [3]. As the abnormalities in GJ organization and regulation have been implicated in diseased myocardium [1], the aim of the present study was to assess the regional expression of molecules involved in GJ regulation in a model of left ventricular disfunction (LVD). For this purpose the distribution and quantitative expression of Cx43, its phosphorylated form pS368-Cx43, PKC phosphorylated substrates, RhoA and A receptors, were investigated in experimental models of right ventricular-pacing induced LVD, undergoing concomitant dipyridamole therapy or placebo, and compared with healty myocardium obtained from sham operated minipigs. Results demonstrates that an altered pattern of factors involved in Cx43-made GJ regulation is present in ventricular myocardium with left ventricular dysfunction. Moreover, the dipyridamole treatment, that results in an improvement of heart function, seems to act also modulating expression and activation of these factors

Dipyridamole increases Cx43 expression in heart muscle cells through Adenosine 2A receptor/PKC pathway

Cx43, a predominant connexin in the heart, forms gap junctions (GJs) that facilitate electrical cell-cell coupling and hemichannels that represent a pathway for the exchange of ions and metabolites between cytoplasm and the extracellular milieu. Our recent results (1) demonstrated that an altered distribution and quantitative expression of factors involved in Cx43-made GJ regulation as Cx43, its phosphorylated form pS368-Cx43, PKC phosphorylated substrates, and adenosine 2A receptor (A2AR) are present in ventricular myocardium with left ventricular dysfunction. Moreover, dipyridamole treatment, which shows a mild protective role on left ventricular function, seems to act through re-modulating the expression and activation of these factors. The role of these factors on signal transduction cascade triggered by dipyridamole was evaluated in this study by pharmacological and immunoistochemical experiments using the rat cardiomyoblast cell line H9c2. The treatment of H9c2 cells with dipyridamole enhanced the expression of Cx43, A2AR and PKC activity while induced a decrease of pS368-Cx43. Interestingly, we found that the A2AR activation was a prerequisite for the effects of dipyridamole, in fact, the pre-treatment with CSC, a selective A2AR receptor antagonist, abolished its effects on the expression of these factors

Geostatistics – a tool applied to the distribution of Legionella pneumophila in a hospital water system

[b]Introduction.[/b] Legionnaires' disease is normally acquired by inhalation of legionellae from a contaminated environmental source. Water systems of large buildings, such as hospitals, are often contaminated with legionellae and therefore represent a potential risk for the hospital population. The aim of this study was to evaluate the potential contamination of [i]Legionella pneumophila[/i] (LP) in a large hospital in Italy through georeferential statistical analysis to assess the possible sources of dispersion and, consequently, the risk of exposure for both health care staff and patients. [b]Materials and Method. [/b]LP serogroups 1 and 2–14 distribution was considered in the wards housed on two consecutive floors of the hospital building. On the basis of information provided by 53 bacteriological analysis, a 'random' grid of points was chosen and spatial geostatistics or [i]FAIk Kriging[/i] was applied and compared with the results of classical statistical analysis. [b]Results[/b]. Over 50% of the examined samples were positive for [i]Legionella pneumophila[/i]. LP 1 was isolated in 69% of samples from the ground floor and in 60% of sample from the first floor; LP 2–14 in 36% of sample from the ground floor and 24% from the first. The iso-estimation maps show clearly the most contaminated pipe and the difference in the diffusion of the different [i]L. pneumophila[/i] serogroups. [b]Conclusion.[/b] Experimental work has demonstrated that geostatistical methods applied to the microbiological analysis of water matrices allows a better modeling of the phenomenon under study, a greater potential for risk management and a greater choice of methods of prevention and environmental recovery to be put in place with respect to the classical statistical analysis

Connexin 26 expression in mammalian cardiomyocytes

Connexins (Cxs) are a family of membrane-spanning proteins named according to their molecular weight. They have been known to form membrane channels mediating cell-cell communication, which play an essential role in the propagation of electrical activity throughout the heart. So far, expression of seven isoforms, namely Cx30.2, Cx37, Cx40, Cx43, Cx45, Cx46 and Cx57, have been found in cardiac myocytes (1,2). Cx26 has been described in a number of tissues but not yet in the heart, and its mutations are frequently associated with deafness and skin diseases (3,4). To our knowledge, the expression of Cx26 also in human, pig, rat and mouse cardiomyocytes has been demonstrated for the first time in the present study. Interestingly, this Cx was found as scattered throughout cell cytoplasm but not at level of the intercalated disks where the other cardiac Cxs are mainly located. Furthermore, in cardiomyocytes of a pig model of left ventricular dysfunction (LVD), Cx26 expression was modulated and dipyridamole treatment, which was previously demonstrated to have a protective action on left ventricular function (5), was associated to an increased Cx26 expression. Dipyridamole induced the same effect in cardiac rat cell line H9c2. For our study, paraffin embedded sections of human auricle, pig ventricle, mouse whole heart and H9c2 cells were used. Several methods were employed to test the expression of Cx26. In particular, different immunohistochemical and molecular biology techniques were performed by using two types of primary anti-Cx26 antibodies to ascertain the specificity of cardiomyocyte immunopositivity for Cx26 avoiding analysis-dependent artifacts