Publications by authors named "Daniele Bani"

149 Publications

Human Recombinant Relaxin (Serelaxin) as Anti-fibrotic Agent: Pharmacology, Limitations and Actual Perspectives.

Curr Mol Med 2021 Mar 8. Epub 2021 Mar 8.

Department of Experimental & Clinical Medicine, Section of Anatomy & Histology, Research Unit of Histology & Embryology, University of Florence, Florence. Italy.

Relaxin (recombinant human relaxin-2 hormone; RLX-2; serelaxin) had raised expectations as a new medication for fibrotic diseases. A plethora of in vitro and in vivo studies have offered convincing demonstrations that relaxin promotes remodelling of connective tissue extracellular matrix mediated by inhibition of multiple fibrogenic pathways, especially the downstream signalling of transforming growth factor (TGF)-β1, a major pro-fibrotic cytokine, and the recruitment and activation of myofibroblast, the main fibrosis-generating cells. However, all clinical trials with relaxin in patients with fibrotic diseases gave inconclusive results. In this review, we have summarized the molecular mechanisms of fibrosis, highlighting those which can be effectively targeted by relaxin. Then, we have performed a critical reappraisal of the clinical trials performed to-date with relaxin as anti-fibrotic drug, in order to highlight their key points of strength and weakness and to identify some future opportunities for the therapeutic use of relaxin, or its analogues, in fibrotic diseases and pathologic scarring which, in our opinion, deserve to be investigated.
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http://dx.doi.org/10.2174/1566524021666210309113650DOI Listing
March 2021

Co-Delivery of Berberine Chloride and Tariquidar in Nanoliposomes Enhanced Intracellular Berberine Chloride in a Doxorubicin-Resistant K562 Cell Line Due to P-gp Overexpression.

Pharmaceutics 2021 Feb 26;13(3). Epub 2021 Feb 26.

Department of Chemistry "Ugo Schiff", University of Florence, via Ugo Schiff 6, 50019 Sesto Fiorentino, Italy.

The MDR phenomenon has become a major obstacle in the treatment of cancers, and among the strategies to reverse it, the inhibition of P-gp function and expression is essential to increase for effective anticancer drugs. In the present paper, the co-delivery of berberine chloride and tariquidar loaded nanoliposomes was investigated with the aim of enhancing solubility and improving desired effects for the antineoplastic drug and the P-gp inhibitor. Developed nanoliposomes were loaded with the electron-dense enzyme horseradish peroxidase, and analyzed by TEM to investigate their ability to enter in both K562 and K562/DOXO cell lines. Receptor-mediated endocytosis was evidenced for both cell lines. Nanoliposomes were loaded with tariquidar, berberine chloride, or both, maintaining chemical and physical characteristics-i.e., size, homogeneity, and encapsulation efficiency-and high suitability for parenteral administration. Tariquidar was able to reverse the MDR in the K562/DOXO cell line. Tariquidar- and berberine chloride-loaded nanoliposomes showed a significant increase of berberine chloride accumulation in tumor cells, which could be correlated with resensitization of the resistant cells to the antitumor agent. These results suggest that the co-delivery of the P-gp inhibitor, tariquidar, and the cytotoxicity inducer, berberine chloride, looks like a promising approach to overcome the MDR.
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http://dx.doi.org/10.3390/pharmaceutics13030306DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8025904PMC
February 2021

Enhanced Antitumoral Activity and Photoacoustic Imaging Properties of AuNP-Enriched Endothelial Colony Forming Cells on Melanoma.

Adv Sci (Weinh) 2021 Feb 21;8(4):2001175. Epub 2020 Dec 21.

Department of Experimental and Clinical Biomedical Sciences University of Florence Florence 50134 Italy.

Near infrared (NIR)-resonant gold nanoparticles (AuNPs) hold great promise in cancer diagnostics and treatment. However, translating the theranostic potential of AuNPs into clinical applications still remains a challenge due to the difficulty to improve the efficiency and specificity of tumor delivery in vivo as well as the clearance from liver and spleen to avoid off target toxicity. In this study, endothelial colony forming cells (ECFCs) are exploited as vehicles to deliver AuNPs to tumors. It is first demonstrated that ECFCs display a great capability to intake AuNPs without losing viability, and exert antitumor activity per se. Using a human melanoma xenograft mouse model, it is next demonstrated that AuNP-loaded ECFCs retain their capacity to migrate to tumor sites in vivo 1 day after injection and stay in the tumor mass for more than 1 week. In addition, it is demonstrated that ECFC-loaded AuNPs are efficiently cleared by the liver over time and do not elicit any sign of damage to healthy tissue.
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http://dx.doi.org/10.1002/advs.202001175DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7887578PMC
February 2021

Alexander Disease Modeling in Zebrafish: An In Vivo System Suitable to Perform Drug Screening.

Genes (Basel) 2020 Dec 11;11(12). Epub 2020 Dec 11.

Department of Earth, Environment and Life Sciences (DISTAV), University of Genoa, 16132 Genoa, Italy.

Alexander disease (AxD) is a rare astrogliopathy caused by heterozygous mutations, either inherited or arising de novo, on the glial fibrillary acid protein (GFAP) gene (17q21). Mutations in the GFAP gene make the protein prone to forming aggregates which, together with heat-shock protein 27 (HSP27), αB-crystallin, ubiquitin, and proteasome, contribute to form Rosenthal fibers causing a toxic effect on the cell. Unfortunately, no pharmacological treatment is available yet, except for symptom reduction therapies, and patients undergo a progressive worsening of the disease. The aim of this study was the production of a zebrafish model for AxD, to have a system suitable for drug screening more complex than cell cultures. To this aim, embryos expressing the human gene carrying the most severe p.R239C under the control of the zebrafish gene promoter underwent functional validation to assess several features already observed in in vitro and other in vivo models of AxD, such as the localization of mutant GFAP inclusions, the ultrastructural analysis of cells expressing mutant GFAP, the effects of treatments with ceftriaxone, and the heat shock response. Our results confirm that zebrafish is a suitable model both to study the molecular pathogenesis of mutations and to perform pharmacological screenings, likely useful for the search of therapies for AxD.
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http://dx.doi.org/10.3390/genes11121490DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7764705PMC
December 2020

Extra virgin olive oil and related by-products (Olea europaea L.) as natural sources of phenolic compounds for abdominal pain relief in gastrointestinal disorders in rats.

Food Funct 2020 Dec 25;11(12):10423-10435. Epub 2020 Nov 25.

Department of Neuroscience, Psychology, Drug Research and Child Health - NEUROFARBA - Pharmacology and Toxicology Section, University of Florence, Viale Pieraccini 6, 50139, Florence, Italy.

Management of abdominal pain, a common symptom of IBDs and IBS, is still a clinical problem. Extra virgin olive oil (EVOO), a main component of the Mediterranean diet, shows positive effects on chronic inflammation in IBDs. In this study, the effect of the oral administration of EVOO (3 mL) and two olive milling by-products, DPA (300 mg kg) and DRF (300 mg kg), on preventing the development of abdominal pain in a DNBS-induced colitis model in rats was evaluated. The doses were chosen with the aim of simulating a plausible daily intake in humans. DPA and EVOO treatments significantly reduced the abdominal viscero-motor response to colon-rectal distension at 2 and 3 mL of balloon distension volume, both 7 and 14 days after the DNBS-injection. DRF showed efficacy in the reduction of visceral hypersensitivity only with 3 mL balloon inflation. In awake animals, DPA and DRF reduced pain perception (evaluated as abdominal withdrawal reflex) with all balloon distension volumes, while EVOO was effective only with higher distension volumes. Fourteen days after the DNBS-injection, all samples reduced the macroscopic intestinal damage (quantified as the macroscopic damage score) also showing, at the microscopic level, a reduction of the inflammatory infiltrate (quantified by hematoxylin and eosin analysis), fibrosis (highlighted by picrosirius red staining), the increase in mast cells and their degranulation (analyzed by triptase immunohistochemistry). This is the first report on the promotion of abdominal pain relief in a rat model obtained administering EVOO and two derived by-products. Our results suggest a protective role of phenol-rich EVOO and milling by-products, which may be proposed as food ingredients for novel functional foods.
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http://dx.doi.org/10.1039/d0fo02293dDOI Listing
December 2020

and Can Predict the Efficacy of Adjuvant Fluoropyrimidine-Based Chemotherapy in Colorectal Cancer Patients.

Oncol Res 2021 Mar 18;28(6):631-644. Epub 2020 Nov 18.

Department of Health Sciences, University of FlorenceFlorenceItaly.

The benefit of adjuvant chemotherapy in the early stages of colorectal cancer (CRC) is still disappointing and the prediction of treatment outcome quite difficult. Recently, through a transcriptomic approach, we evidenced a role of and gene expression in predicting response to fluoropyrimidine-based adjuvant chemotherapy in stage III CRC patients. Thus, the aim of this study was to validate in an independent cohort of stages IIIII CRC patients our previous findings. and mRNA expression levels were evaluated in 74 formalin-fixed paraffin-embedded tumor and matched normal mucosa samples obtained by stages IIIII CRC patients treated with fluoropyrimidine-based adjuvant chemotherapy. PININ, the protein encoded by , was immunohistochemically evaluated in 15 tumor and corresponding normal mucosa samples, selected on the basis of a low, medium, or high mRNA expression tumor/mucosa ratio. and mRNA mean expression levels were significantly higher in tumor compared with normal tissues. Patients with high or tumor mRNA levels according to ROC-based cutoffs showed a shorter disease-free survival (DFS) compared with patients with low tumor mRNA gene expression. Also, patients with tumor mRNA expression values of both genes below the identified cutoffs had a significantly longer DFS compared with patients with the expression of one or both genes above the cutoffs. In a representative large cohort of stages IIIII CRC untreated patients retrieved from GEO datasets, no difference in DFS was observed between patients with high and low or gene expression levels. These data confirm our previous findings and underscore the relevance of and expression in predicting DFS in early stages of CRC treated with fluoropyrimidine-based adjuvant chemotherapy. If further validated in a prospective case series, both biomarkers could be used to identify patients who benefit from this treatment and to offer alternative chemotherapy regimens to potential unresponsive patients. In relation to the suggested biological role of and in CRC, they might also be exploited as potential therapeutic targets.
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http://dx.doi.org/10.3727/096504020X16056983169118DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7962934PMC
March 2021

Evolution and new frontiers of histology in bio-medical research.

Microsc Res Tech 2021 Feb 11;84(2):217-237. Epub 2020 Sep 11.

Biomedical and Neuromotor Sciences, Alma Mater University Bologna, Bologna, Italy.

Histology refers to the study of the morphology of cells within their natural tissue environment. As a bio-medical discipline, it dates back to the development of first microscopes which allowed to override the physical visual limitation of the human eye. Since the first observations, it was understood that cell shape predicts function and, therefore, shape alterations can identify and explain dysfunction and diseases. The advancements in morphological investigation techniques have allowed to extend our understanding of the shape-function relationships close to the molecular level of organization of tissues, as well as to derive reliable data not only from fixed, and hence static, biological samples but also living cells and tissues and even for extended time periods. These modern approaches, which encompass quantitative microscopy, precision microscopy, and dynamic microscopy, represent the new frontier of morphology. This article summarizes how the microscopy techniques have evolved to properly face the challenges of biomedical sciences, thus transforming histology from a merely qualitative discipline, which played an ancillary role to traditional "major" sciences such as anatomy, to a modern experimental science capable of driving knowledge progress in biology and medicine.
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http://dx.doi.org/10.1002/jemt.23579DOI Listing
February 2021

Human Relaxin-2 (Serelaxin) Attenuates Oxidative Stress in Cardiac Muscle Cells Exposed In Vitro to Hypoxia-Reoxygenation. Evidence for the Involvement of Reduced Glutathione Up-Regulation.

Antioxidants (Basel) 2020 Aug 21;9(9). Epub 2020 Aug 21.

Department of Experimental & Clinical Medicine, Research Unit of Histology & Embryology, University of Florence, viale G. Pieraccini 6, 50139 Florence, Italy.

Serelaxin (RLX) designates the pharmaceutical form of the human natural hormone relaxin-2 that has been shown to markedly reduce tissue and cell damage induced by hypoxia and reoxygenation (HR). The evidence that RLX exerts similar protective effects on different organs and cells at relatively low, nanomolar concentrations suggests that it specifically targets a common pathogenic mechanism of HR-induced damage, namely oxidative stress. In this study we offer experimental evidence that RLX (17 nmol L-1), added to the medium of HR-exposed H9c2 rat cardiac muscle cells, significantly reduces cell oxidative damage, mitochondrial dysfunction and apoptosis. These effects appear to rely on the up-regulation of the cellular availability of reduced glutathione (GSH), a ubiquitous endogenous antioxidant metabolite. Conversely, superoxide dismutase activity was not influenced by RLX, which, however, was not endowed with chemical antioxidant properties. Taken together, these findings verify the major pharmacological role of RLX in the protection against HR-induced oxidative stress, and shed first light on its mechanisms of action.
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http://dx.doi.org/10.3390/antiox9090774DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555919PMC
August 2020

Th17 lymphocyte-dependent degradation of joint cartilage by synovial fibroblasts in a humanized mouse model of arthritis and reversal by secukinumab.

Eur J Immunol 2021 01 9;51(1):220-230. Epub 2020 Aug 9.

Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, Florence, Italy.

How T-helper (Th) lymphocyte subpopulations identified in synovial fluid from patients with juvenile idiopathic arthritis (JIA) (Th17, classic Th1, or nonclassic Th1) drive joint damage is of great interest for the possible use of biological drugs that inhibit the specific cytokines. Our objective was to clarify the role of such Th subpopulations in the pathogenesis of articular cartilage destruction by synovial fibroblasts (SFbs), and the effect of Th17 blockage in an animal model. SFbs were isolated from healthy subjects and patients with JIA, and peripheral blood Th lymphocytes subsets were obtained from healthy subjects. Fragments of human cartilage from healthy subjects in a collagen matrix containing JIA or normal SFbs grafted underskin in SCID mice were used to measure cartilage degradation under the effects of Th supernatants. JIA SFbs overexpress MMP9 and MMP2 and Th17 induce both MMPs in normal SFbs, while nonclassic Th1 upregulate urokinase plasminogen activator (uPA) activity. In vitro invasive phenotype of normal SFbs is stimulated with conditioned medium of Th17 and nonclassic-Th1. In the in vivo "inverse wrap" model, normal SFbs stimulated with supernatants of Th17-lymphocytes and nonclassic Th1 produced a cartilage invasion and degradation similar to JIA SFbs. Secukinumab inhibits the cartilage damage triggered by factors produced by Th17.
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http://dx.doi.org/10.1002/eji.202048773DOI Listing
January 2021

Germline Mutation in Gene Associated with Loss of Heterozygosity: Usefulness of Next-Generation Sequencing in the Genetic Screening of Patients with Pheochromocytoma.

Int J Endocrinol 2020 30;2020:3671396. Epub 2020 May 30.

Dept. of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy.

The genetic approach of pheochromocytomas and paragangliomas has changed in the last two decades. Nowadays, we know that more than 40% of patients have a germline mutation in one of the susceptibility genes identified to date. Our aim is to underline how genetic diagnosis by next-generation sequencing (NGS) can improve the management of patients affected by pheochromocytomas and paragangliomas in our routine diagnostic screening. We reported a case presentation and next-generation sequencing analysis supported by in silico studies and evaluation of mitochondrial status in tissue. A 46-year-old male affected by a left secreting pheochromocytoma underwent surgery in 2017. After surgery, the normetanephrine levels decreased very slowly and a suspected abdominal lymph node was detected. We found a novel germline gene mutation, c.4052C > T, p. Pro1351Leu associated with tumor loss of heterozygosity, and resulted likely-pathogenetic by in silico studies. This mutation was also associated with an increased number of mitochondria through the electron microscopy compared with wild-type tissues as suggestive for mitochondria neoformation compensatory to the mitochondrial autophagic figures observed. Our results underline the usefulness of next-generation sequencing in the presence of multiple tumor predisposition genes and how, at the same time, its use may result challenging for the clinicians. To date, performing the genetic analysis according to the latest Consensus Statement is mandatory in patients affected by PHEO/PGL.
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http://dx.doi.org/10.1155/2020/3671396DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7277055PMC
May 2020

Pomegranate Mesocarp against Colitis-Induced Visceral Pain in Rats: Effects of a Decoction and Its Fractions.

Int J Mol Sci 2020 Jun 17;21(12). Epub 2020 Jun 17.

Department of Neuroscience, Psychology, Drug Research and Child Health-NEUROFARBA-Pharmacology and Toxicology Section, University of Florence, Viale Pieraccini 6, 50139 Florence, Italy.

The management of chronic visceral pain related to Inflammatory Bowel Diseases or Irritable Bowel Syndrome is still a clinical problem and new therapeutic strategies continue to be investigated. In the present study, the efficacy of a pomegranate decoction and of its polysaccharide and ellagitannin components in preventing the development of colitis-induced abdominal pain in rats was evaluated. After colitis induction by 2,4-dinitrobenzenesulfonic acid (DNBS), the pomegranate decoction (300 mg kg), polysaccharides (300 mg kg), and ellagitannins (45 mg kg) were orally administered for 14 days. Repeated treatment with decoction reduced visceral hypersensitivity in the colitic animals both at 7 and 14 days. Similar efficacy was shown by polysaccharides, but with lower potency. Ellagitannins administered at dose equivalent to decoction content showed higher efficacy in reducing the development of visceral pain. Macroscopic and microscopic evaluations performed on the colon 14 days after the damage showed that all three preparations reduced the overall amount of mast cells, the number of degranulated mast cells, and the density of collagen fibers in the mucosal stroma. Although ellagitannins seem to be responsible for most of the beneficial effects of pomegranate on DNBS-induced colitis, the polysaccharides support and enhance its effect. Therefore, pomegranate mesocarp preparations could represent a complementary approach to conventional therapies for promoting abdominal pain relief.
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http://dx.doi.org/10.3390/ijms21124304DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7353021PMC
June 2020

Recombinant human H2 relaxin (serelaxin) as a cardiovascular drug: aiming at the right target.

Authors:
Daniele Bani

Drug Discov Today 2020 07 29;25(7):1239-1244. Epub 2020 Apr 29.

Research Unit of Histology & Embryology, Department of Experimental & Clinical Medicine University of Florence, Florence, Italy. Electronic address:

Serelaxin (recombinant human relaxin-2 hormone; RLX-2) had raised expectations as a new medication for cardiovascular diseases. Evidence from preclinical studies indicated that serelaxin has chronotropic, inotropic, and anti-arrhythmic actions on the myocardium and cardioprotective effects mediated by vasodilation, angiogenesis, and inhibition of inflammation and fibrosis. However, clinical trials with serelaxin in patients with acute heart failure (AHF) gave inconclusive results. A critical reappraisal of the comprehensive cardiovascular actions of serelaxin clearly delineates acute myocardial infarction (AMI) as a feasible therapeutic target. Serelaxin acts at multiple levels on the pathogenic mechanisms of AMI and previous in vivo studies suggest that its administration at reperfusion affords myocardial salvage. Thus, serelaxin could be an effective adjunctive medical therapy to coronary angioplasty.
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http://dx.doi.org/10.1016/j.drudis.2020.04.014DOI Listing
July 2020

Testosterone improves muscle fiber asset and exercise performance in a metabolic syndrome model.

J Endocrinol 2020 05;245(2):259-279

Sexual Medicine and Andrology Unit, Department of Biomedical, Experimental and Clinical Sciences 'Mario Serio', University of Florence, Florence, Italy.

Lifestyle modifications, including physical exercise (PhyEx), are well-known treatments for metabolic syndrome (MetS), a cluster of metabolic and cardiovascular risk factors often associated to hypogonadism. Given the trophic role of testosterone on skeletal muscle (SkM), this study was aimed at evaluating the effects of testosterone treatment on SkM metabolism and exercise performance in male rabbits with high-fat diet (HFD)-induced MetS. HFD rabbits, treated or not with testosterone (30 mg/kg/week) for 12 weeks, were compared to regular diet animals (RD). A subset of each group was exercise-trained for 12 weeks. HFD increased type-II (fast, glycolytic) and decreased type-I (slow, oxidative) muscle fibers compared to RD as evaluated by RT-PCR and histochemistry. Testosterone reverted these effects, also inducing the expression of mitochondrial respiration enzymes and normalizing HFD-induced mitochondrial cristae reduction. Moreover, testosterone significantly increased the expression of myogenic/differentiation markers and genes related to glucidic/lipid metabolism. At the end of the PhyEx protocol, when compared to RD, HFD rabbits showed a significant reduction of running distance and running time, while testosterone counteracted this effect, also decreasing lactate production. In the trained groups, muscle histology showed a significant reduction of oxidative fibers in HFD compared to RD and the positive effect of testosterone in maintaining oxidative metabolism, as also demonstrated by analyzing mitochondrial ultrastructure, succinate dehydrogenase activity and ATP production. Our results indicate that testosterone could be useful to promote oxidative muscle metabolism altered by MetS, thus improving exercise performance. Conversely, testosterone administration to otherwise eugonadal rabbits (RD) only increased muscle fiber diameter but not endurance performance.
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http://dx.doi.org/10.1530/JOE-19-0532DOI Listing
May 2020

uPAR Knockout Results in a Deep Glycolytic and OXPHOS Reprogramming in Melanoma and Colon Carcinoma Cell Lines.

Cells 2020 01 28;9(2). Epub 2020 Jan 28.

Department of Experimental and Clinical Biomedical Sciences, University of Florence, Viale G.B. Morgagni 50, 50134 Firenze, Italy.

Urokinase Plasminogen Activator (uPA) Receptor (uPAR) is a well-known GPI-anchored three-domain membrane protein with pro-tumor roles largely shown in all the malignant tumors where it is over-expressed. Here we have exploited the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 gene knock out approach to investigate its role in the oxidative metabolism in human melanoma and colon cancer as the consequences of its irreversible loss. Knocking out , a uPAR-encoding gene, in A375p, A375M6 and HCT116, which are two human melanoma and a colon carcinoma, respectively, we have observed an increased number of mitochondria in the two melanoma cell lines, while we evidenced an immature biogenesis of mitochondria in the colon carcinoma culture. Such biological diversity is, however, reflected in a significant enhancement of the mitochondrial spare respiratory capacity, fueled by an increased expression of GLS2, and in a decreased glycolysis paired with an increased secretion of lactate by all uPAR KO cells. We speculated that this discrepancy might be explained by an impaired ratio between LDHA and LDHB.
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http://dx.doi.org/10.3390/cells9020308DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7072355PMC
January 2020

Effect of Unloading Condition on the Healing Process and Effectiveness of Platelet Rich Plasma as a Countermeasure: Study on In Vivo and In Vitro Wound Healing Models.

Int J Mol Sci 2020 Jan 9;21(2). Epub 2020 Jan 9.

ASA campus Joint Laboratory, ASA Res. Div., Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, 50139 Florence, Italy.

Wound healing is a very complex process that allows organisms to survive injuries. It is strictly regulated by a number of biochemical and physical factors, mechanical forces included. Studying wound healing in space is interesting for two main reasons: (i) defining tools, procedures, and protocols to manage serious wounds and burns eventually occurring in future long-lasting space exploration missions, without the possibility of timely medical evacuation to Earth; (ii) understanding the role of gravity and mechanical factors in the healing process and scarring, thus contributing to unravelling the mechanisms underlying the switching between perfect regeneration and imperfect repair with scarring. In the study presented here, a new in vivo sutured wound healing model in the leech () has been used to evaluate the effect of unloading conditions on the healing process and the effectiveness of platelet rich plasma (PRP) as a countermeasure. The results reveal that microgravity caused a healing delay and structural alterations in the repair tissue, which were prevented by PRP treatment. Moreover, investigating the effects of microgravity and PRP on an in vitro wound healing model, it was found that PRP is able to counteract the microgravity-induced impairment in fibroblast migration to the wound site. This could be one of the mechanisms underlying the effectiveness of PRP in preventing healing impairment in unloading conditions.
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http://dx.doi.org/10.3390/ijms21020407DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013931PMC
January 2020

Development and Percutaneous Permeation Study of Escinosomes, Escin-Based Nanovesicles Loaded with Berberine Chloride.

Pharmaceutics 2019 Dec 15;11(12). Epub 2019 Dec 15.

Department of Chemistry, University of Florence, Via Ugo Schiff 6, 50019 Sesto Fiorentino (FI), Italy.

Escin is a natural saponin, clinically used for the anti-edematous and anti-inflammatory effects. The aim of the study was to explore the possibility of converting escin into vesicle bilayer-forming component. The hyaluronidase inhibition activity of escin was evaluated after its formulation in escinosomes. Berberine chloride, a natural quaternary isoquinoline alkaloid isolated from several medicinal plants that is traditionally used for various skin conditions was loaded in the vesicles. The developed nanovesicles were characterized in terms of diameter, polydispersity, ζ-potential, deformability, recovery, encapsulation efficiency, stability, and release kinetics. Nanovesicle permeation properties through artificial membranes and rabbit ear skin were investigated using skin-PAMPA and Franz cells were also evaluated. Escinosomes, made of phosphatidylcholine and escin, were loaded with berberine chloride. These nanovesicles displayed the best characteristics for skin application, particularly optimal polydispersity (0.17) and deformability, high negative ζ-potential value, great encapsulation efficiency (about 67%), high stability, and the best release properties of berberine chloride (about 75% after 24 h). In conclusion, escinosomes seem to be new vesicular carriers, capable to maintain escin properties such as hyaluronidase inhibition activity, and able to load other active molecules such as berberine chloride, in order to enhance or expand the activity of the loaded drug.
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http://dx.doi.org/10.3390/pharmaceutics11120682DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6955842PMC
December 2019

Liver haploinsufficiency of RuvBL1 causes hepatic insulin resistance and enhances hepatocellular carcinoma progression.

Int J Cancer 2020 06 28;146(12):3410-3422. Epub 2019 Nov 28.

Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, Florence, Italy.

RuvBL1 is an AAA+ ATPase whose expression in hepatocellular carcinoma (HCC) correlates with a poor prognosis. In vitro models suggest that targeting RuvBL1 could be an effective strategy against HCC. However, the role of RuvBL1 in the onset and progression of HCC remains unknown. To address this question, we developed a RuvBL1 mouse model and evaluated the outcome of DEN-induced liver carcinogenesis up to 12 months of progression. We found that RuvBL1 haploinsufficiency initially delayed the onset of liver cancer, due to a reduced hepatocyte turnover in RuvBL1 mice. However, RuvBL1 mice eventually developed HCC nodules that, with aging, grew larger than in the control mice. Moreover, RuvBL1 mice developed hepatic insulin resistance and impaired glucose homeostasis. We could determine that RuvBL1 regulates insulin signaling through the Akt/mTOR pathway in liver physiology in vivo as well as in normal hepatocytic and HCC cells in vitro. Whole transcriptome analysis of mice livers confirmed the major role of RuvBL1 in the regulation of hepatic glucose metabolism. Finally, RuvBL1 expression was found significantly correlated to glucose metabolism and mTOR signaling by bioinformatic analysis of human HCC sample from the publicly available TGCA database. These data uncover a role of RuvBL1 at the intersection of liver metabolism, hepatocyte proliferation and HCC development, providing a molecular rationale for its overexpression in liver cancer.
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http://dx.doi.org/10.1002/ijc.32787DOI Listing
June 2020

Photonic Therapy in Periodontal Diseases an Overview with Appraisal of the Literature and Reasoned Treatment Recommendations.

Int J Mol Sci 2019 Sep 24;20(19). Epub 2019 Sep 24.

Department of Experimental and Clinical Medicine, Research Unit of Histology & Embryology, University of Florence, viale G. Pieraccini 6, 50139 Florence, Italy.

Recent reviews and meta-analyses of the literature over the past quarter-century have failed to provide enough evidence to prove or disprove the actual utility of photonic therapy in periodontitis, alone or adjunctive to conventional approaches. This apparent paradox has been explained by the many physical, molecular, biological, anatomical, and technical variables of photonic treatments, which can differ in light-emitting devices (laser or LED), wavelengths, irradiation power and modes, clinical objectives, follow-up times, disease grading, and assessment methods. This multi-faceted, controversial scenario has led practitioners to underestimate the actual potential of photonic therapy in periodontal diseases. In this critical appraisal of the literature, we have briefly summarized the main photonic therapies and instruments used in Periodontology, highlighting their main characteristics and limitations. Then, we have tried to identify and discuss the key methodological issues which can have an impact on the outcome of photonic therapies. Our main goal was to identify the best parameters, settings, and methodologies to perform effective periodontal photonic treatments and to extrapolate some recommendations for clinical use. Should these recommendations find a consensus among periodontologists and be adopted in future clinical studies, they will hopefully contribute to dissipate the present confusion and uncertainty on this complex matter.
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http://dx.doi.org/10.3390/ijms20194741DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6801906PMC
September 2019

Different Antioxidant Efficacy of Two Mn-Containing Superoxide Anion Scavengers on Hypoxia/Reoxygenation-Exposed Cardiac Muscle Cells.

Sci Rep 2019 07 16;9(1):10320. Epub 2019 Jul 16.

Department of Experimental & Clinical Medicine, Research Unit of Histology & Embryology, University of Florence, viale G. Pieraccini 6, 50139, Florence, Italy.

Oxidative stress due to excess superoxide anion ([Formula: see text]) produced by dysfunctional mitochondria is a key pathogenic event of aging and ischemia-reperfusion diseases. Here, a new [Formula: see text]-scavenging Mn complex with a new polyamino-polycarboxylate macrocycle (4,10-dimethyl-1,4,7,10-tetraazacyclododecane-1,7-diacetate) containing 2 quinoline units (MnQ2), designed to improve complex stability and cell permeability, was compared to parental Mn complex with methyls replacing quinolines (MnM2). MnQ2 was more stable than MnM2 (log K = 19.56(8) vs. 14.73(2) for the equilibrium Mn + L, where L = Q2 and M2) due to the involvement of quinoline in metal binding and to the hydrophobic features of the ligand which improve metal desolvation upon complexation. As oxidative stress model, H9c2 rat cardiomyoblasts were subjected to hypoxia-reoxygenation. MnQ2 and MnM2 (10 μmol L) were added at reoxygenation for 1 or 2 h. The more lipophilic MnQ2 showed more rapid cell and mitochondrial penetration than MnM2. Both MnQ2 and MnM2 abated endogenous ROS and mitochondrial [Formula: see text], decreased cell lipid peroxidation, reduced mitochondrial dysfunction, in terms of efficiency of the respiratory chain and preservation of membrane potential (Δψ) and permeability, decreased the activation of pro-apoptotic caspases 9 and 3, and increased cell viability. Of note, MnQ2 was more effective than MnM2 to exert cytoprotective anti-oxidant effects in the short term. Compounds with redox-inert Zn replacing the functional Mn were ineffective. This study provides clues which further our understanding of the structure-activity relationships of Mn-chelates and suggests that Mn-polyamino-polycarboxylate macrocycles could be developed as new anti-oxidant drugs.
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http://dx.doi.org/10.1038/s41598-019-46476-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6635543PMC
July 2019

Serelaxin (recombinant human relaxin-2) treatment affects the endogenous synthesis of long chain poly-unsaturated fatty acids and induces substantial alterations of lipidome and metabolome profiles in rat cardiac tissue.

Pharmacol Res 2019 06 5;144:51-65. Epub 2019 Apr 5.

Cellular and Molecular Cardiology Unit and Department of Cardiology, Institute of Biomedical Research (IDIS-SERGAS), Travesía Choupana s/n, 15706 Santiago de Compostela, Spain; CIBERCV, Institute of Health Carlos III, C/ Monforte de Lemos 3-5, Pabellón 11. Planta 0, 28029 Madrid, Spain.

Background And Purpose: Recombinant human relaxin-2, serelaxin, is being proved as a novel drug with therapeutic efficacy in some cardiovascular diseases, especially heart failure, a disease whose physiopathology and course are firmly correlated with important alterations in cardiac metabolism. The aim of our present work was to investigate changes in the cardiac metabolome following relaxin-2 treatment.

Experimental Approach: Sprague-Dawley rats were treated with human recombinant relaxin-2 using osmotic minipumps at a dose of 0.4 mg/kg/day for 2 weeks. Body composition was measured with a nuclear magnetic resonance imaging system seven days after surgery and on the final day of the experiment. The last two days of treatment, respiratory quotient, locomotor activity and energy expenditure were measured with a calorimetric system. The plasma levels of relaxin-2, total cholesterol, high- and low- density lipoproteins (HDL, LDL), triglycerides and the hepatic enzymes glutamic-pyruvic transaminase (GTP) and gamma-glutamyltransferase (GGT) levels were analyzed. The metabolic profiling of both atria from relaxin-2-treated and control rats was carried out using two separate ultra-high performance liquid chromatography (UHPLC)-Time of Flight-MS based platforms analyzing methanol and chloroform/methanol extracts combined with a UHPLC-single quadrupole-MS based platform used to analyze aminoacids and with a methanol/water extract platform that covered polar metabolites. Identified ion features in the methanol extract platform included fatty acids, acyl carnitines, bile acids, monoacylglycerophospholipids, monoetherglycerophospholipids, free sphingoid bases, and oxidized fatty acids. The chloroform / methanol extract platform provided coverage over glycerolipids, cholesterol esters, sphingolipids, diacylglycerophospholipids, and acyl-ether-glycerophospholipids. Gene expression levels of the adipokines adiponectin, leptin and nesfatin-1 in visceral adipose tissue and cardiac gene expression levels of key enzymes of desaturation and elongation of n-6 and n-3 PUFAs were assessed by Real Time-PCR.

Key Results: Twenty-eight metabolites out of three hundred sixty-two were significantly altered by human relaxin-2. These included fifteen glycerophospholipids: three phosphatidylethanolamines (PE) and twelve phosphatidylcholines (PC); eight sphingolipids: three ceramides (Cer) and five sphingomyelins (SM); and also five aminoacids and one carboxylic acid. Interestingly, the majority of changes correspond to lipid classes, twelve of them polyunsaturated diacylglycerophosphatidylcholines with long acyl chains, containing mainly docosahexaenoic acid (22:6) and arachidonic acid (20:4). Atrial levels of Elovl5 (Elongation of very long chain fatty acids protein 5), Fads1 (Δ5-fatty acid desaturase) and Fads2 (Δ6-fatty acid desaturase), key enzymes of elongation and desaturation of n-6 and n-3 PUFAs like arachidonic acid and DHA, respectively, were significantly increased by relaxin-2 treatment. Atrial tissues from rats treated with relaxin-2 showed a significant increase in the mRNA levels of Srebf1, a transcription factor that activates the gene expression of Elovl5, Fads1 and Fads2. The treatment with relaxin-2 significantly decreased the visceral fat mRNA expression levels of adiponectin, leptin and nesfatin-1, adipokines known to exert an important influence on the regulation of cardiovascular function.

Conclusion And Implications: Serelaxin (human recombinant relaxin-2) treatment induces significant changes in cardiac major components of the membrane lipid bilayer such as glycerophospholipids and sphingolipids, known to have structural roles but also very relevant regulatory effects in cardiac function. Serelaxin induced also modifications in several aminoacids of high influence in cardiac energy metabolism regulation. Our results highlight the need to further understand the role of relaxin-2 in the regulation of cardiac energy metabolism, in the context of the therapeutic strategies for the treatment of cardiometabolic pathologies as heart failure.
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http://dx.doi.org/10.1016/j.phrs.2019.04.009DOI Listing
June 2019

Glutamate Receptor-Mediated Neurotoxicity in a Model of Ethanol Dependence and Withdrawal in Rat Organotypic Hippocampal Slice Cultures.

Front Neurosci 2018 24;12:1053. Epub 2019 Jan 24.

Section of Clinical Pharmacology and Oncology, Department of Health Sciences, University of Florence, Florence, Italy.

Long-term alcohol use can lead to alterations in brain structure and functions and, in some cases, to neurodegeneration. Several mechanisms have been proposed to explain ethanol (EtOH)-related brain injury. One of the most relevant mechanisms of alcohol-induced neurodegeneration involves glutamatergic transmission, but their exact role is not yet fully understood. We investigated the neurochemical mechanisms underlying the toxicity induced by EtOH dependence and/or withdrawal by exposing rat organotypic hippocampal slices to EtOH (100-300 mM) for 7 days and then incubating the slices in EtOH-free medium for the subsequent 24 h. EtOH withdrawal led to a dose-dependent CA1 pyramidal cell injury, as detected with propidium iodide fluorescence. Electron microscopy of hippocampal slices revealed that not only EtOH withdrawal but also 7 days chronic EtOH exposure elicited signs of apoptotic cell death in CA1 pyramidal cells. These data were supported by electrophysiological recordings of spontaneus Excitatory Post Synaptic Currents (sEPSCs) from CA1 pyramidal cells. The average amplitude of sEPSCs in slices treated with EtOH for 7 days was significantly increased, and even more so during the first 30 min of EtOH withdrawal, suggesting that the initial phase of the neurodegenerative process could be due to an excitotoxic mechanism. We then analyzed the expression levels of presynaptic (vGlut1, vGlut2, CB1 receptor, synaptophysin) and postsynaptic (PSD95, GluN1, GluN2A, GluN2B, GluA1, GluA2, mGluR1 and mGluR5) proteins after 7 days EtOH incubation or after EtOH withdrawal. We found that only GluA1 and mGluR5 expression levels were significantly increased after EtOH withdrawal and, in neuroprotection experiments, we observed that AMPA and mGluR5 antagonists attenuated EtOH withdrawal-induced toxicity. These data suggest that chronic EtOH treatment promotes abnormal synaptic transmission that may lead to CA1 pyramidal cell death after EtOH withdrawal through glutamate receptors and increased excitotoxicity.
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http://dx.doi.org/10.3389/fnins.2018.01053DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6353783PMC
January 2019

Neurotoxicity of Unconjugated Bilirubin in Mature and Immature Rat Organotypic Hippocampal Slice Cultures.

Neonatology 2019 15;115(3):217-225. Epub 2019 Jan 15.

Department of Neurosciences, Psychology, Drug Research and Child Health, University of Florence, Florence, Italy.

Background: The physiopathology of bilirubin-induced neurological disorders is not completely understood.

Objectives: The aim of our study was to assess the effect on bilirubin neurotoxicity of the maturity or immaturity of exposed cells, the influence of different unconjugated bilirubin (UCB) and human serum albumin (HSA) concentrations, and time of UCB exposure.

Methods: Organotypic hippocampal slices were exposed for 48 h to different UCB and HSA concentrations after 14 (mature) or 7 (immature) days of in vitro culture. Immature slices were also exposed to UCB and HSA for 72 h. The different effects of exposure time to UCB on neurons and astrocytes were evaluated.

Results: We found that 48 h of UCB exposure was neurotoxic for mature rat organotypic hippocampal slices while 72 h of exposure was neurotoxic for immature slices. Forty-eight-hour UCB exposure was toxic for astrocytes but not for neurons, while 72-h exposure was toxic for both astrocytes and neurons. HSA prevented UCB toxicity when the UCB:HSA molar ratio was ≤1 in both mature and immature slices.

Conclusions: We confirmed UCB neurotoxicity in mature and immature rat hippocampal slices, although immature ones were more resistant. HSA was effective in preventing UCB neurotoxicity in both mature and immature rat hippocampal slices.
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http://dx.doi.org/10.1159/000494101DOI Listing
December 2019

Appropriate laser wavelengths for photodynamic therapy with methylene blue.

Lasers Med Sci 2018 Nov 30;33(8):1837-1838. Epub 2018 Jun 30.

Department of Experimental and Clinical Medicine, Research Unit of Histology and Embryology, University of Florence, Viale G.Pieraccini 6, 50139, Florence, Italy.

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http://dx.doi.org/10.1007/s10103-018-2566-xDOI Listing
November 2018

INT-767 prevents NASH and promotes visceral fat brown adipogenesis and mitochondrial function.

J Endocrinol 2018 08;238(2):107-127

Sexual Medicine and Andrology UnitDepartment of Experimental and Clinical Biomedical Sciences 'Mario Serio', University of Florence, Florence, Italy

The bile acid receptors, farnesoid X receptor (FXR) and Takeda G-protein-coupled receptor 5 (TGR5), regulate multiple pathways, including glucose and lipid metabolism. In a rabbit model of high-fat diet (HFD)-induced metabolic syndrome, long-term treatment with the dual FXR/TGR5 agonist INT-767 reduces visceral adipose tissue accumulation, hypercholesterolemia and nonalcoholic steatohepatitis. INT-767 significantly improves the hallmarks of insulin resistance in visceral adipose tissue (VAT) and induces mitochondrial and brown fat-specific markers. VAT preadipocytes isolated from INT-767-treated rabbits, compared to preadipocytes from HFD, show increased mRNA expression of brown adipogenesis markers. In addition, INT-767 induces improved mitochondrial ultrastructure and dynamic, reduced superoxide production and improved insulin signaling and lipid handling in preadipocytes. Both and treatments with INT-767 counteract, in preadipocytes, the HFD-induced alterations by upregulating genes related to mitochondrial biogenesis and function. In preadipocytes, INT-767 behaves mainly as a TGR5 agonist, directly activating dose dependently the cAMP/PKA pathway. However, experiments also suggest that FXR activation by INT-767 contributes to the insulin signaling improvement. INT-767 treatment counteracts HFD-induced liver histological alterations and normalizes the increased pro-inflammatory genes. INT-767 also induces a significant reduction of fatty acid synthesis and fibrosis markers, while increasing lipid handling, insulin signaling and mitochondrial markers. In conclusion, INT-767 significantly counteracts HFD-induced liver and fat alterations, restoring insulin sensitivity and prompting preadipocytes differentiation toward a metabolically healthy phenotype.
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http://dx.doi.org/10.1530/JOE-17-0557DOI Listing
August 2018

Gold Nanoparticles Functionalized with RGD-Semipeptides: A Simple yet Highly Effective Targeting System for α β Integrins.

Chemistry 2018 Aug 16;24(46):12093-12100. Epub 2018 Jul 16.

Center for Biomolecular Nanotechnologies@UniLe, Istituto Italiano di Tecnologia (IIT), Via Barsanti, 73010, Arnesano, Lecce, Italy.

Effective and selective targeting of the α β integrin subtype is of high relevance in cancer research for the development of therapeutic systems with improved efficacy and of diagnostic imaging probes. We report here a new class of highly selective, α β -targeted gold nanoparticles (AuNPs), which carry cyclic 4-aminoproline-RGD semipeptides (cAmpRGD) as the targeting moiety immobilized at low surface density on the poly(ethylene glycol) (PEG)-based nanoparticle coating. We show that these nanoparticles are potent inhibitors of the integrin-mediated melanoma tumor cell adhesion to vitronectin and are selectively internalized via receptor-mediated endocytosis. Furthermore, we have developed bifunctional cAmpRGD-functionalized AuNPs by conjugation of a fluorophore (FAM or TAMRA) to a separate set of reactive groups on the PEG-based coating. These bifunctional AuNPs not only recapitulate the binding properties of cAmpRGD-AuNPs but also can be visualized via confocal laser microscopy, allowing direct observation of nanoparticle internalization. The peculiar molecular design of these nanoparticles and their precisely defined architecture at the molecular level accounts for their selective integrin binding with very low nonspecific background.
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http://dx.doi.org/10.1002/chem.201801823DOI Listing
August 2018

Model lipid bilayers mimic non-specific interactions of gold nanoparticles with macrophage plasma membranes.

J Colloid Interface Sci 2018 Apr 6;516:284-294. Epub 2018 Feb 6.

Department of Chemistry and CSGI, University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino, Florence, Italy.

Understanding the interaction between nanomaterials and biological interfaces is a key unmet goal that still hampers clinical translation of nanomedicine. Here we investigate and compare non-specific interaction of gold nanoparticles (AuNPs) with synthetic lipid and wild type macrophage membranes. A comprehensive data set was generated by systematically varying the structural and physicochemical properties of the AuNPs (size, shape, charge, surface functionalization) and of the synthetic membranes (composition, fluidity, bending properties and surface charge), which allowed to unveil the matching conditions for the interaction of the AuNPs with macrophage plasma membranes in vitro. This effort directly proved for the first time that synthetic bilayers can be set to mimic and predict with high fidelity key aspects of nanoparticle interaction with macrophage eukaryotic plasma membranes. It then allowed to model the experimental observations according to classical interface thermodynamics and in turn determine the paramount role played by non-specific contributions, primarily electrostatic, Van der Waals and bending energy, in driving nanoparticle-plasma membrane interactions.
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http://dx.doi.org/10.1016/j.jcis.2018.01.064DOI Listing
April 2018

Treatment of severe periodontitis with a laser and light-emitting diode (LED) procedure adjunctive to scaling and root planing: a double-blind, randomized, single-center, split-mouth clinical trial investigating its efficacy and patient-reported outcomes at 1 year.

Lasers Med Sci 2018 Jul 18;33(5):991-1002. Epub 2018 Jan 18.

Department of Experimental and Clinical Medicine, Research Unit of Histology & Embryology, University of Florence, Viale G. Pieraccini 6, 50139, Florence, Italy.

Broad methodological heterogeneity makes the literature on the clinical effects of laser treatment in periodontitis, both as monotherapy and adjunct to non-surgical therapy, which is difficult to interpret. The present split-mouth study was performed: (i) to determine the efficacy and safety of a photoablative-photodynamic diode laser therapy, including antiseptic LED irradiation, in adjunct to scaling and root planing (iPAPD+SRP) vs. sham-treatment+SRP for the treatment of diffuse severe periodontitis and (ii) to estimate the patient-reported outcomes. Twenty-four patients with severe periodontitis were treated with iPAPD+SRP or sham-treatment+SRP. iPAPD+SRP consisted of the following: (1) intra-/extra-pocket de-epithelization with photoablative λ 810 nm laser, (2) disinfection with λ 405 nm LED, (3) SRP, and (4) 10 weekly antiseptic/anti-inflammatory photodynamic treatments with λ 635 nm laser and 0.1% toluidine blue as photosensitizer. Clinical and cytofluorescent periodontal markers and patient-reported results were analyzed. At 1-year follow-up, both groups showed a significant reduction of several severity markers of periodontitis, namely probing depth (PD) and bleeding on probing (BoP), as well as of bacteria, polymorphonuclear cells, erythrocytes and damaged epithelial cells in exfoliative samples, as compared with day 0. The quadrants subjected to iPAPD+SRP showed significantly better values of these parameters as well as of clinical attachment level (CAL) as compared with those undergoing sham-treatment+SRP. The patients' perceived pain/discomfort, and overall liking was also in favor of the iPAPD+SRP treatment. This study confirms the efficacy of combined phototherapy in adjunct to SRP which had emerged from previous clinical trials, extending its field of application to severe periodontitis.
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http://dx.doi.org/10.1007/s10103-018-2441-9DOI Listing
July 2018

Relaxin induces up-regulation of ADAM10 metalloprotease in RXFP1-expressing cells by PI3K/AKT signaling.

Mol Cell Endocrinol 2018 09 24;472:80-86. Epub 2017 Nov 24.

Research Unit of Histology & Embryology, Dept. Experimental & Clinical Medicine, University of Florence, Viale G.Pieraccini 6, 50139 Florence, Italy. Electronic address:

ADAM10 metalloprotease is required for activation of Notch-1, a transmembrane receptor regulating cell differentiation, proliferation and apoptosis, whose intracellular proteolytic fragment NICD mediates some key cardiovascular effects of the hormone relaxin (RLX). This study demonstrates the involvement of ADAM10 and PI3K/Akt signaling in mediating RLX-induced Notch-1 activation. H9c2 cardiomyocytes and NIH3T3 fibroblasts were incubated with human RLX-2 (17 nmol/l, 24 h) in presence or absence of the PI3K or Akt inhibitors wortmannin (WT, 100 nmol/l) and triciribine (TCN, 1 μmol/l). Cyclohexanedione-inactivated RLX (iRLX) served as negative control. RLX significantly increased Akt phosphorylation, ADAM10 and NICD expression, which were abolished by WT or TCN and did not occur with iRLX. These findings highlight a new receptor-specific signal transduction pathway of RLX.
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http://dx.doi.org/10.1016/j.mce.2017.11.021DOI Listing
September 2018

Effects of an Erbium:Yttrium-Aluminum-Garnet Laser and Ultrasonic Scaler on Titanium Dioxide-Coated Titanium Surfaces Contaminated With Subgingival Plaque: An In Vitro Study to Assess Post-Treatment Biocompatibility With Osteogenic Cells.

J Periodontol 2017 11 10;88(11):1211-1220. Epub 2017 Jul 10.

Department of Experimental and Clinical Medicine, Section of Anatomy and Histology, University of Florence, Florence, Italy.

Background: Effects of conventional ultrasonic scaler versus an erbium:yttrium-aluminum-garnet (Er:YAG) laser on titanium surfaces contaminated with subgingival plaque from patients with peri-implantitis are evaluated in terms of: 1) plaque and biocorroded titanium oxide coating removal; 2) surface change induction; and 3) residual biocompatibility toward osteoblasts.

Methods: Subgingival plaque-coated titanium disks with a moderately rough surface were fixed with ethanol and treated with an ultrasonic scaler (metal tip) or Er:YAG laser (20.3 or 38.2 J/cm) in non-contact mode. Fluorescent detection of residual plaque was performed. Disk surface morphology was evaluated by scanning electron microscopy. Viability, attachment, proliferation, and differentiation of Saos-2 osteoblasts on new and treated disks were assayed by propidium iodide/DNA stain assay and confocal microscopic analysis of cytoskeleton, Ki67, expression of osteopontin and alkaline phosphatase, and formation of mineralized nodules.

Results: Both methods resulted in effective debridement of treated surfaces, the plaque area being reduced to 11.7% with the ultrasonic scaler and ≤0.03% with the Er:YAG laser (38.2 J/cm). Ultrasound-treated disks showed marked surface changes, incomplete removal of the titanium dioxide (TiO) layer, and scanty plaque aggregates, whereas the Er:YAG laser (38.2 J/cm) completely stripped away the plaque and TiO layer, leaving a micropitted surface. Both treatments maintained a good biocompatibility of surfaces to Saos-2 osteoblasts. Air-water cooling kept disk temperature below the critical threshold of 47°C.

Conclusion: This study shows that an ultrasonic scaler with metal tip is less efficient than high-energy Er:YAG irradiation to remove the plaque and TiO layer on anodized disks, although both procedures appear capable of restoring an adequate osseoconductivity of treated surfaces.
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http://dx.doi.org/10.1902/jop.2017.170195DOI Listing
November 2017

Notch Signaling in Ischemic Damage and Fibrosis: Evidence and Clues from the Heart.

Front Pharmacol 2017 5;8:187. Epub 2017 Apr 5.

Research Unit of Histology and Embryology, Department of Experimental and Clinical Medicine, University of FlorenceFlorence, Italy.

Notch signaling is a major intercellular coordination mechanism highly conserved throughout evolution. In vertebrates, Notch signaling is physiologically involved in embryo development, including mesenchymal cell commitment, formation of heart tissues and angiogenesis. In post-natal life, Notch signaling is maintained as a key mechanism of cell-cell communication and its dysregulations have been found in pathological conditions such as ischemic and fibrotic diseases. In the heart, Notch takes part in the protective response to ischemia, being involved in pre- and post-conditioning, reduction of reperfusion-induced oxidative stress and myocardial damage, and cardiomyogenesis. Conceivably, the cardioprotective effects of Notch may depend on neo-angiogenesis, thus blunting lethal myocardial ischemia, as well as on direct stimulation of cardiac cells to increase their resistance to injury. Another post-developmental adaptation of Notch signaling is fibrosis: being involved in the orientation of mesenchymal cell fate, Notch can modulate the differentiation of pro-fibrotic myofibroblasts, e.g., by reducing the effects of the profibrotic cytokine TGF-β. In conclusion, Notch can regulate the interactions between heart muscle and stromal cells and switch cardiac repair from a pro-fibrotic default pathway to a pro-cardiogenic one. These features make Notch signaling a suitable target for new cardiotropic therapies.
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http://dx.doi.org/10.3389/fphar.2017.00187DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5381357PMC
April 2017