Loredana Petecchia - Academia.edu (original) (raw)

Papers by Loredana Petecchia

Research paper thumbnail of Smoke exposure, wheezing, and asthma development: A systematic review and meta-analysis in unselected birth cohorts

Pediatric Pulmonology, Mar 20, 2014

The understanding of the role of smoking exposure in the induction of wheezing and asthma in chil... more The understanding of the role of smoking exposure in the induction of wheezing and asthma in children is important for prevention. A systematic review of literature and a meta-analysis were conducted to identify studies on unselected prospective birth cohorts. The effect of exposure to maternal/parental smoking on the induction of current wheezing or asthma was evaluated in children aged 6 months, <6 years, and ≥6 years. Pooled odds ratios (OR) with 95% confidence intervals (CI) were estimated. We identified 43 papers. Exposure to maternal prenatal smoking was associated with an increased risk of wheezing in <6-year-olds (OR 1.36; 95% CI: 1.19-1.55) and of wheezing or asthma in ≥6-year-olds (OR: 1.22, 95% CI: 1.03-1.44). A positive association (OR: 1.24, 95% CI: 1.11-1.38) was also found in the only three studies that evaluated exposure to maternal prenatal smoking alone. Postnatal exposures to maternal/parental smoking were associated with wheezing in <6-year-olds (OR: 1.21; 95% CI: 1.13-1.31 and OR: 1.30; 95% CI: 1.13-1.51), although it was often impossible to separate the role of postnatal from that of prenatal exposure; data in schoolchildren are limited and this precluded a meta-analysis. No clear association was found between exclusive postnatal exposure and wheezing or asthma. We confirmed an important role of prenatal exposure to maternal smoking on the induction of wheezing and asthma in offspring, particularly in the first years of life. More studies with a consistent number of subjects only exposed to smoke postnatally are needed to better investigate the harmful effects on the induction of wheezing or asthma, particularly in schoolchildren. Pediatr Pulmonol. 2015; 50:353-362. © 2014 Wiley Periodicals, Inc.

Research paper thumbnail of Bradykinin phosphorylates mitogen-activated protein kinases in human lung fibroblasts through Epidermal Growth Factor Receptor transactivation

Research paper thumbnail of Bradykinin-induced fibroblast proliferation and myofibroblast differentiation via B2 receptor pathway

Research paper thumbnail of A biophysical approach to quantify skeletal stem cells trans-differentiation as a model for the study of osteoporosis

Biophysical Chemistry, 2017

The stroma of human bone marrow contains a population of skeletal stem cells (hBM-MSC) which are ... more The stroma of human bone marrow contains a population of skeletal stem cells (hBM-MSC) which are common ancestors, among the others, of osteoblasts and adipocytes. It has been proposed that the imbalance between hBM-MSC osteogenesis and adipogenesis, which naturally accompanies bone marrow senescence, may contribute to the development of bone-associated diseases, like osteoporosis. The possibility to reproduce this mechanism in vitro has been demonstrated, providing a good model to disclose the details of the complex bone-fat generation homeostasis. Nevertheless, the lack of a simple approach to quantitatively assess the actual stage of a cellular population hindered the adoption of this in vitro model. In this work, the direct differentiation of hBM-MSCs towards a single (osteo or adipo) lineage was characterized using quantitative biophysical and biological approaches, together with the parallel process of trans-differentiation from one lineage to the other. The results confirm that the original plasticity of hBM-MSCs is maintained along the initial stages of the differentiation, showing that in vitro conversion of pre-osteoblasts into adipocytes and, vice versa, of pre-adipocytes into osteoblasts is extremely efficient, comparable with the direct differentiation. Moreover, a method based on digital holography is proposed, providing a quantitative indication of the phenotype stage along differentiation.

Research paper thumbnail of Effects of neoglycosylated collagen matrices on neuroblastoma and human stem cells: a new perspective for neuro-regeneration?

Research paper thumbnail of Biophysical characterization of nanostructured TiO2 as a good substrate for hBM-MSC adhesion, growth and differentiation

Experimental Cell Research, 2017

ABSTRACT Mesenchymal stem cells from human bone marrow (hBM‐MSC) are widely utilized for clinical... more ABSTRACT Mesenchymal stem cells from human bone marrow (hBM‐MSC) are widely utilized for clinical applications involving bone healing. In this context, their use has been often optimized in association to variously designed titanium substrates, being this material of great use in orthopaedic implants. According to recent findings, the ability of hBM‐MSC to differentiate towards a specific lineage is not only driven by biochemical signals, but physical stimuli, such as rigidity or roughness of the substrate, can also support a commitment towards osteogenic differentiation. Moreover, the presence of features with defined dimensional scales, in particular nanometer‐size, also proved to elicit specific biological effects. Here we evaluated the effectiveness of a nano‐patterned titanium surface in sustaining hBM‐MSC adhesion, growth and differentiation by means of a panel of biophysical tools: morphometry, electrophysiology, intracellular calcium measurements and immunocytochemistry. The results substantiate the idea that this micro‐textured titanium dioxide is a good surface for growth and differentiation of hBM‐MSC and it exhibits a stimulating action mainly in the initial period of differentiation. Moreover, the basal concentration of free cytosolic Calcium [Ca2+]i is confirmed to be a good hallmark of the hBM‐MSC maturation stage. The study could provide relevant hints to help improving the biocompatibility and osteointegration potential of clinical titanium implants. HIGHLIGHTSThe presence of nanoscale TiO2 surface features enhances hBM‐MSC osteogenesis.Ca2+ homeostasis changes are involved in surface‐driven hBM‐MSC lineage commitment.2D and 3D morphological parameters provide insights into hBM‐MSC adhesion phase.

Research paper thumbnail of Orally exhaled nitric oxide as a marker of inflammation in childhood asthma

Research paper thumbnail of Bradykinin transactivates epidermal growth factor receptor in human lung fibroblasts

Research paper thumbnail of Bradykinin (BK) induces human airway fibroblast (Fb) proliferation and Myofibroblast (MyoFb) differentiation via ERK1/2 MAPK pathway

Research paper thumbnail of Pulmonary , gastrointestinal and urogenital pharmacology Bradykinin-induced asthmatic fi broblast / myo fi broblast activities via bradykinin B 2 receptor and different MAPK pathways

Bradykinin drives normal lung fibroblasts into myofibroblasts, induces fibroblast proliferation a... more Bradykinin drives normal lung fibroblasts into myofibroblasts, induces fibroblast proliferation and activates mitogen activated protein kinase pathways (MAPK) but its effects on bronchial fibroblasts from asthmatics (HBAFb) have not been yet studied. We studied bradykinin-induced fibroblast proliferation and differentiation and the related intracellular mechanisms in HBAFb compared to normal bronchial fibroblasts (HNBFb). Bradykinin-stimulated HBAFb and HNBFb were used to assess: bradykinin B2 receptor expression by Western blot analysis; cell proliferation by [ H] thymidine incorporation; α-smooth muscle actin (SMA) expression/polymerization by Western blot and immunofluorescence; epidermal growth factor (EGF) receptor, extracellular-regulated kinase (ERK) 1/2 and p38 MAPK activation by immunoprecipitation and Western blot, respectively. Constitutive bradykinin B2 receptor and α-SMA expression was higher in HBAFb as compared to HNBFb. Bradykinin increased bradykinin B2 receptor exp...

Research paper thumbnail of The effect of pulsed electromagnetic field exposure on osteoinduction of human mesenchymal stem cells cultured on nano-TiO2 surfaces

PloS one, 2018

Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) are considered a great promise in the... more Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) are considered a great promise in the repair and regeneration of bone. Considerable efforts have been oriented towards uncovering the best strategy to promote stem cells osteogenic differentiation. In previous studies, hBM-MSCs exposed to physical stimuli such as pulsed electromagnetic fields (PEMFs) or directly seeded on nanostructured titanium surfaces (TiO2) were shown to improve their differentiation to osteoblasts in osteogenic condition. In the present study, the effect of a daily PEMF-exposure on osteogenic differentiation of hBM-MSCs seeded onto nanostructured TiO2 (with clusters under 100 nm of dimension) was investigated. TiO2-seeded cells were exposed to PEMF (magnetic field intensity: 2 mT; intensity of induced electric field: 5 mV; frequency: 75 Hz) and examined in terms of cell physiology modifications and osteogenic differentiation. Results showed that PEMF exposure affected TiO2-seeded cells osteogenesis by ...

Research paper thumbnail of The effect of transforming growth factor (TGFf)-β1 and (TGF)-β2 on nasal polyp fibroblast activities involved upper airway remodeling : Modulation by fluticasone propionate

Research paper thumbnail of Bradykinin b2 receptor expression in the bronchial mucosa of allergic asthmatics: the role of NF-kB

Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology, Jan 20, 2015

Bradykinin (BK) mediates acute allergic asthma and airway remodeling. Nuclear factor-kappa B (NF-... more Bradykinin (BK) mediates acute allergic asthma and airway remodeling. Nuclear factor-kappa B (NF-kB) is potentially involved in BK B2 receptor (B2R) regulation. In this observational cross-sectional study B2R and NF-kB expression was evaluated in bronchial biopsies from mild asthmatics (after diluent/allergen challenge) and healthy controls examining the role of NF-kB in B2R expression in primary human fibroblasts from normal and asthmatic subjects (HNBFb and HABFb). B2R and NF-kB (total and nuclear) expression was analyzed by immunohistochemistry in biopsies from 10 mild intermittent asthmatics (48 hours after diluent/allergen challenge) and 10 controls undergoing bronchoscopy. B2R co-localization in 5B5(+) and αSMA(+) mesenchymal cells was studied by immunofluorescence/confocal microscopy, and B2R expression in HABFb/HNBFb incubated with interleukin (IL)-4/IL-13 with/without BK, and after NF-kB inhibitor, by Western blotting. Bronchial mucosa B2R and nuclear NF-kB expression was h...

Research paper thumbnail of Expression of vascular remodelling markers in relation to bradykinin receptors in asthma and COPD

Thorax, 2013

Background Vascular remodelling plays a central role in asthma and chronic obstructive pulmonary ... more Background Vascular remodelling plays a central role in asthma and chronic obstructive pulmonary disease (COPD). Bradykinin (BK) is a vasoactive proinflammatory peptide mediating acute responses in asthma. We investigated the role of angiogenic factors in relation to BK receptors in asthma and COPD. Methods Bronchial biopsies from 33 patients with COPD, 24 old (≥50 years) patients with (≥50 years) asthma, 18 old control smokers, 11 old control nonsmokers, 15 young (≤40yrs) patients with (≤40 years) asthma and 10 young control non-smokers were immunostained for CD31, vascular endothelial growth factor-A (VEGF-A), angiogenin and BK receptors (B2R and B1R). Fibroblast and endothelial co-localisation of relevant molecules were performed by immunofluorescence. BK-induced VEGF-A and angiogenin release was studied (ELISA) in bronchial fibroblasts from subjects with asthma and COPD. Results In bronchial lamina propria of old patients with asthma, CD31 and VEGF-A + cell numbers were higher than old control non-smokers (p<0.05). Angiogenin + , B2R + and B1R + cell numbers in old patients with asthma were higher than in old control non-smokers, control smokers and patients with COPD (p<0.01). Angiogenin + cell numbers were higher in patients with COPD than both old control groups (p<0.05). In all patients with asthma the number of B2R + cells was positively related to the numbers of B1R + (r s =0.43), angiogenin + (r s =0.42) and CD31 cells (r s =0.46) (p<0.01). Angiogenin + cell numbers were negatively related to forced expiratory volume in 1 s (r s =−0.415, p=0.008). Double immunofluorescence revealed that CD31 cells of capillary vessels coexpressed B2R and that fibroblasts coexpressed B2R, VEGF-A and angiogenin. BK (10 −6 M) induced significant angiogenin release in fibroblasts from asthma and to a lesser extent in COPD. Conclusions Unlike COPD, this study suggests the involvement of BK receptors in bronchial vascular remodelling in asthma.

Research paper thumbnail of Pharmacological Modulation of the Bradykinin-Induced Differentiation of Human Lung Fibroblasts: Effects of Budesonide and Formoterol

Journal of Asthma, 2012

Objective. Bradykinin (BK) induces differentiation of lung fibroblasts into myofibroblasts, which... more Objective. Bradykinin (BK) induces differentiation of lung fibroblasts into myofibroblasts, which play an important role in extracellular matrix remodeling in the airways of asthmatic patients. It is unclear whether this process is affected by antiasthma therapies. Here, we evaluated whether a glucocorticoid, budesonide (BUD), and a long-acting /?2-agonist, formoterol (FM), either alone or in combination, modified BK-induced lung fibroblast differentiation, and affected the BK-activated intracellular signaling pathways. Methods. Human fetal lung fibroblasts were incubated with BUD (0.001-0.1 nM) and/or FM (0.0001-0.1 |iM) before exposure to BK (0.1 or 1 nM). Fibroblast differentiation into a-smooth-muscleactin-positive (a-SMA^) myofibroblasts, BK2 receptor (B2R) expression, extracellular signal-regulated kinase 1/2 (ERK 1/2) phosphorylation (p-ERKl/2), intracellular Ca^"*" concentration (fCa^^],), and p65 nuclear factor kappa B translocation were evaluated. Resuhs. BUD (0.1 \M) and FM (0.1 |j,M), either alone or in combination, completely inhibited BK-induced a-SMA protein expression and decreased the numbers of a-SMAf ibroblasts, with a clear reduction in a-SMA stress fibers organization. BUD also completely inhibited the increase of B2R, whereas FM with or without BUD had no effect. BK-induced increases of [Ca^"'"]; and p-ERKl/2 were significantly reduced to similar levels by BUD and FM, either alone or in combination, whereas p65 translocation was completely inhibited by all treatments. Conclusion. Both BUD and FM, either alone or in combination, effectively inhibited the BK-induced differentiation of fibroblasts into a-SMA^ myofibroblasts and the intracellular signaling pathways involved in fibroblast activation. These results suggest that BUD and FM combination therapy has potential to inhibit fibroblastdependent matrix remodeling in the airways of asthmatic patients. Keywords a-smooth muscle actin (a-SMA), bradykinin B2 receptor (B2R), human lung fibroblasts, tnitogen-activated protein kinases (MAPKs)

Research paper thumbnail of Bradykinin-induced asthmatic fibroblast/myofibroblast activities via bradykinin B2 receptor and different MAPK pathways

European Journal of Pharmacology, 2013

Bradykinin drives normal lung fibroblasts into myofibroblasts, induces fibroblast proliferation a... more Bradykinin drives normal lung fibroblasts into myofibroblasts, induces fibroblast proliferation and activates mitogen activated protein kinase pathways (MAPK) but its effects on bronchial fibroblasts from asthmatics (HBAFb) have not been yet studied. We studied bradykinin-induced fibroblast proliferation and differentiation and the related intracellular mechanisms in HBAFb compared to normal bronchial fibroblasts (HNBFb). Bradykinin-stimulated HBAFb and HNBFb were used to assess: bradykinin B2 receptor expression by Western blot analysis; cell proliferation by [(3)H] thymidine incorporation; α-smooth muscle actin (SMA) expression/polymerization by Western blot and immunofluorescence; epidermal growth factor (EGF) receptor, extracellular-regulated kinase (ERK) 1/2 and p38 MAPK activation by immunoprecipitation and Western blot, respectively. Constitutive bradykinin B2 receptor and α-SMA expression was higher in HBAFb as compared to HNBFb. Bradykinin increased bradykinin B2 receptor expression in HBAFb. Bradykinin, via bradykinin B2 receptor, significantly increased fibroblast proliferation at lower concentration (10(-11)M) and α-SMA expression/polymerization at higher concentration (10(-6)M) in both cells. Bradykinin increased ERK1/2 and p38 phosphorylation via bradykinin B2 receptor; EGF receptor inhibitor AG1478 and panmetalloproteinase inhibitor GM6001 blocked bradykinin-induced ERK1/2 activation but not p38 phosphorylation. Bradykinin, via bradykinin B2 receptor, induced EGF receptor phosphorylation that was suppressed by AG1478. In HBAFb AG1478, GM6001, the ERK1/2-inhibitor U0126 and the p38 inhibitor SB203580 suppressed bradykinin-induced cell proliferation, but only SB203580 reduced myofibroblast differentiation. These data indicate that bradykinin is actively involved in asthmatic bronchial fibroblast proliferation and differentiation, through MAPK pathways and EGF receptor transactivation, by which bradykinin may contribute to airway remodeling in asthma, opening new horizons for potential therapeutic implications in asthmatic patients.

Research paper thumbnail of Bronchial Airway Epithelial Cell Damage Following Exposure to Cigarette Smoke Includes Disassembly of Tight Junction Components Mediated by the Extracellular Signal-Regulated Kinase 1/2 Pathway

Chest, 2009

Background: Through a variety of biochemical mechanisms, cigarette smoke (CS) may damage airway e... more Background: Through a variety of biochemical mechanisms, cigarette smoke (CS) may damage airway epithelium, altering its normal structure and function. Injury to epithelium may include changes in tight junction (TJ) integrity with impairment of epithelial barrier function. Methods and results: To study the effect of the exposure to CS condensate (CSC) on TJ integrity, two human bronchial epithelial cell lines (HBECs), BEAS-2B and 16HBEI40-, were used. Exposure of the two HBECs to CSC resulted in a time-dependent and concentration-dependent disassembly of TJs, which were already detectable at 24 h at all the CSC concentrations tested (5%,10%, and 20%), associated with changes in cell shape, suggesting cell damage. However, a significant inhibition of cell growth and an increase in DNA fragmentation were detected only at the highest CSC concentration tested (20%) at 48 and 72 h, respectively. The involvement of epidermal growth factor receptor (EGFR)-extracellular signal-regulated kinase (ERK) 1/2 cascade in CSC-induced damage was shown by the observation that exposure to CSC (5%) induced a marked phosphorylation of ERKI/2, already detectable after 5-min incubation and confirmed by the demonstration that not only ERKl/2 phosphorylation but also CSC-induced TJ disassembly and DNA fragmentation were partially inhibited by a mitogen-activated protein kinase kinase inhibitor (UOI26) and completely blocked by a EGFR inhibitor (AGI478). Conclusion: CSC-induced damage to airway epithelium includes disassembly of TJs, modulated through the EGFR-ERKI/2 signaling pathway.

Research paper thumbnail of Controlled single-cell cyclic compression and transcription analysis: A pilot study

Biophysical Chemistry, 2017

• Controlled dynamic compression setup with single-cell sensitivity • qPCR protocol able to retri... more • Controlled dynamic compression setup with single-cell sensitivity • qPCR protocol able to retrieve targets expression with single-cell sensitivity • Workflow tested on bone cells, physiologically subjected to mechanical stimuli

Research paper thumbnail of Development of label-free biophysical markers in osteogenic maturation

Journal of the Mechanical Behavior of Biomedical Materials

Research paper thumbnail of Osteogenic Differentiation of MSC through Calcium Signaling Activation: Transcriptomics and Functional Analysis

PLOS ONE, 2016

The culture of progenitor mesenchymal stem cells (MSC) onto osteoconductive materials to induce a... more The culture of progenitor mesenchymal stem cells (MSC) onto osteoconductive materials to induce a proper osteogenic differentiation and mineralized matrix regeneration represents a promising and widely diffused experimental approach for tissue-engineering (TE) applications in orthopaedics. Among modern biomaterials, calcium phosphates represent the best bone substitutes, due to their chemical features emulating the mineral phase of bone tissue. Although many studies on stem cells differentiation mechanisms have been performed involving calcium-based scaffolds, results often focus on highlighting production of in vitro bone matrix markers and in vivo tissue ingrowth, while information related to the biomolecular mechanisms involved in the early cellular calcium-mediated differentiation is not well elucidated yet. Genetic programs for osteogenesis have been just partially deciphered, and the description of the different molecules and pathways operative in these differentiations is far from complete, as well as the activity of calcium in this process. The present work aims to shed light on the involvement of extracellular calcium in MSC differentiation: a better understanding of the early stage osteogenic differentiation program of MSC seeded on calcium-based biomaterials is required in order to develop optimal strategies to promote osteogenesis through the use of new generation osteoconductive scaffolds. A wide spectrum of analysis has been performed on time-dependent series: gene expression profiles are obtained from samples (MSC seeded on calcium-based scaffolds), together with related microRNAs expression and in vivo functional validation. On this basis, and relying on literature knowledge, hypotheses are made on the biomolecular players activated by the biomaterial calcium-phosphate component. Interestingly, a key role of miR-138 was highlighted, whose inhibition markedly increases osteogenic differentiation in vitro and enhance ectopic bone formation in vivo. Moreover, there is evidence that Ca-P substrate triggers osteogenic differentiation through genes (SMAD and RAS family) that are typically regulated during dexamethasone (DEX) induced differentiation.

Research paper thumbnail of Smoke exposure, wheezing, and asthma development: A systematic review and meta-analysis in unselected birth cohorts

Pediatric Pulmonology, Mar 20, 2014

The understanding of the role of smoking exposure in the induction of wheezing and asthma in chil... more The understanding of the role of smoking exposure in the induction of wheezing and asthma in children is important for prevention. A systematic review of literature and a meta-analysis were conducted to identify studies on unselected prospective birth cohorts. The effect of exposure to maternal/parental smoking on the induction of current wheezing or asthma was evaluated in children aged 6 months, &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;6 years, and ≥6 years. Pooled odds ratios (OR) with 95% confidence intervals (CI) were estimated. We identified 43 papers. Exposure to maternal prenatal smoking was associated with an increased risk of wheezing in &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;6-year-olds (OR 1.36; 95% CI: 1.19-1.55) and of wheezing or asthma in ≥6-year-olds (OR: 1.22, 95% CI: 1.03-1.44). A positive association (OR: 1.24, 95% CI: 1.11-1.38) was also found in the only three studies that evaluated exposure to maternal prenatal smoking alone. Postnatal exposures to maternal/parental smoking were associated with wheezing in &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;6-year-olds (OR: 1.21; 95% CI: 1.13-1.31 and OR: 1.30; 95% CI: 1.13-1.51), although it was often impossible to separate the role of postnatal from that of prenatal exposure; data in schoolchildren are limited and this precluded a meta-analysis. No clear association was found between exclusive postnatal exposure and wheezing or asthma. We confirmed an important role of prenatal exposure to maternal smoking on the induction of wheezing and asthma in offspring, particularly in the first years of life. More studies with a consistent number of subjects only exposed to smoke postnatally are needed to better investigate the harmful effects on the induction of wheezing or asthma, particularly in schoolchildren. Pediatr Pulmonol. 2015; 50:353-362. © 2014 Wiley Periodicals, Inc.

Research paper thumbnail of Bradykinin phosphorylates mitogen-activated protein kinases in human lung fibroblasts through Epidermal Growth Factor Receptor transactivation

Research paper thumbnail of Bradykinin-induced fibroblast proliferation and myofibroblast differentiation via B2 receptor pathway

Research paper thumbnail of A biophysical approach to quantify skeletal stem cells trans-differentiation as a model for the study of osteoporosis

Biophysical Chemistry, 2017

The stroma of human bone marrow contains a population of skeletal stem cells (hBM-MSC) which are ... more The stroma of human bone marrow contains a population of skeletal stem cells (hBM-MSC) which are common ancestors, among the others, of osteoblasts and adipocytes. It has been proposed that the imbalance between hBM-MSC osteogenesis and adipogenesis, which naturally accompanies bone marrow senescence, may contribute to the development of bone-associated diseases, like osteoporosis. The possibility to reproduce this mechanism in vitro has been demonstrated, providing a good model to disclose the details of the complex bone-fat generation homeostasis. Nevertheless, the lack of a simple approach to quantitatively assess the actual stage of a cellular population hindered the adoption of this in vitro model. In this work, the direct differentiation of hBM-MSCs towards a single (osteo or adipo) lineage was characterized using quantitative biophysical and biological approaches, together with the parallel process of trans-differentiation from one lineage to the other. The results confirm that the original plasticity of hBM-MSCs is maintained along the initial stages of the differentiation, showing that in vitro conversion of pre-osteoblasts into adipocytes and, vice versa, of pre-adipocytes into osteoblasts is extremely efficient, comparable with the direct differentiation. Moreover, a method based on digital holography is proposed, providing a quantitative indication of the phenotype stage along differentiation.

Research paper thumbnail of Effects of neoglycosylated collagen matrices on neuroblastoma and human stem cells: a new perspective for neuro-regeneration?

Research paper thumbnail of Biophysical characterization of nanostructured TiO2 as a good substrate for hBM-MSC adhesion, growth and differentiation

Experimental Cell Research, 2017

ABSTRACT Mesenchymal stem cells from human bone marrow (hBM‐MSC) are widely utilized for clinical... more ABSTRACT Mesenchymal stem cells from human bone marrow (hBM‐MSC) are widely utilized for clinical applications involving bone healing. In this context, their use has been often optimized in association to variously designed titanium substrates, being this material of great use in orthopaedic implants. According to recent findings, the ability of hBM‐MSC to differentiate towards a specific lineage is not only driven by biochemical signals, but physical stimuli, such as rigidity or roughness of the substrate, can also support a commitment towards osteogenic differentiation. Moreover, the presence of features with defined dimensional scales, in particular nanometer‐size, also proved to elicit specific biological effects. Here we evaluated the effectiveness of a nano‐patterned titanium surface in sustaining hBM‐MSC adhesion, growth and differentiation by means of a panel of biophysical tools: morphometry, electrophysiology, intracellular calcium measurements and immunocytochemistry. The results substantiate the idea that this micro‐textured titanium dioxide is a good surface for growth and differentiation of hBM‐MSC and it exhibits a stimulating action mainly in the initial period of differentiation. Moreover, the basal concentration of free cytosolic Calcium [Ca2+]i is confirmed to be a good hallmark of the hBM‐MSC maturation stage. The study could provide relevant hints to help improving the biocompatibility and osteointegration potential of clinical titanium implants. HIGHLIGHTSThe presence of nanoscale TiO2 surface features enhances hBM‐MSC osteogenesis.Ca2+ homeostasis changes are involved in surface‐driven hBM‐MSC lineage commitment.2D and 3D morphological parameters provide insights into hBM‐MSC adhesion phase.

Research paper thumbnail of Orally exhaled nitric oxide as a marker of inflammation in childhood asthma

Research paper thumbnail of Bradykinin transactivates epidermal growth factor receptor in human lung fibroblasts

Research paper thumbnail of Bradykinin (BK) induces human airway fibroblast (Fb) proliferation and Myofibroblast (MyoFb) differentiation via ERK1/2 MAPK pathway

Research paper thumbnail of Pulmonary , gastrointestinal and urogenital pharmacology Bradykinin-induced asthmatic fi broblast / myo fi broblast activities via bradykinin B 2 receptor and different MAPK pathways

Bradykinin drives normal lung fibroblasts into myofibroblasts, induces fibroblast proliferation a... more Bradykinin drives normal lung fibroblasts into myofibroblasts, induces fibroblast proliferation and activates mitogen activated protein kinase pathways (MAPK) but its effects on bronchial fibroblasts from asthmatics (HBAFb) have not been yet studied. We studied bradykinin-induced fibroblast proliferation and differentiation and the related intracellular mechanisms in HBAFb compared to normal bronchial fibroblasts (HNBFb). Bradykinin-stimulated HBAFb and HNBFb were used to assess: bradykinin B2 receptor expression by Western blot analysis; cell proliferation by [ H] thymidine incorporation; α-smooth muscle actin (SMA) expression/polymerization by Western blot and immunofluorescence; epidermal growth factor (EGF) receptor, extracellular-regulated kinase (ERK) 1/2 and p38 MAPK activation by immunoprecipitation and Western blot, respectively. Constitutive bradykinin B2 receptor and α-SMA expression was higher in HBAFb as compared to HNBFb. Bradykinin increased bradykinin B2 receptor exp...

Research paper thumbnail of The effect of pulsed electromagnetic field exposure on osteoinduction of human mesenchymal stem cells cultured on nano-TiO2 surfaces

PloS one, 2018

Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) are considered a great promise in the... more Human bone marrow-derived mesenchymal stem cells (hBM-MSCs) are considered a great promise in the repair and regeneration of bone. Considerable efforts have been oriented towards uncovering the best strategy to promote stem cells osteogenic differentiation. In previous studies, hBM-MSCs exposed to physical stimuli such as pulsed electromagnetic fields (PEMFs) or directly seeded on nanostructured titanium surfaces (TiO2) were shown to improve their differentiation to osteoblasts in osteogenic condition. In the present study, the effect of a daily PEMF-exposure on osteogenic differentiation of hBM-MSCs seeded onto nanostructured TiO2 (with clusters under 100 nm of dimension) was investigated. TiO2-seeded cells were exposed to PEMF (magnetic field intensity: 2 mT; intensity of induced electric field: 5 mV; frequency: 75 Hz) and examined in terms of cell physiology modifications and osteogenic differentiation. Results showed that PEMF exposure affected TiO2-seeded cells osteogenesis by ...

Research paper thumbnail of The effect of transforming growth factor (TGFf)-β1 and (TGF)-β2 on nasal polyp fibroblast activities involved upper airway remodeling : Modulation by fluticasone propionate

Research paper thumbnail of Bradykinin b2 receptor expression in the bronchial mucosa of allergic asthmatics: the role of NF-kB

Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology, Jan 20, 2015

Bradykinin (BK) mediates acute allergic asthma and airway remodeling. Nuclear factor-kappa B (NF-... more Bradykinin (BK) mediates acute allergic asthma and airway remodeling. Nuclear factor-kappa B (NF-kB) is potentially involved in BK B2 receptor (B2R) regulation. In this observational cross-sectional study B2R and NF-kB expression was evaluated in bronchial biopsies from mild asthmatics (after diluent/allergen challenge) and healthy controls examining the role of NF-kB in B2R expression in primary human fibroblasts from normal and asthmatic subjects (HNBFb and HABFb). B2R and NF-kB (total and nuclear) expression was analyzed by immunohistochemistry in biopsies from 10 mild intermittent asthmatics (48 hours after diluent/allergen challenge) and 10 controls undergoing bronchoscopy. B2R co-localization in 5B5(+) and αSMA(+) mesenchymal cells was studied by immunofluorescence/confocal microscopy, and B2R expression in HABFb/HNBFb incubated with interleukin (IL)-4/IL-13 with/without BK, and after NF-kB inhibitor, by Western blotting. Bronchial mucosa B2R and nuclear NF-kB expression was h...

Research paper thumbnail of Expression of vascular remodelling markers in relation to bradykinin receptors in asthma and COPD

Thorax, 2013

Background Vascular remodelling plays a central role in asthma and chronic obstructive pulmonary ... more Background Vascular remodelling plays a central role in asthma and chronic obstructive pulmonary disease (COPD). Bradykinin (BK) is a vasoactive proinflammatory peptide mediating acute responses in asthma. We investigated the role of angiogenic factors in relation to BK receptors in asthma and COPD. Methods Bronchial biopsies from 33 patients with COPD, 24 old (≥50 years) patients with (≥50 years) asthma, 18 old control smokers, 11 old control nonsmokers, 15 young (≤40yrs) patients with (≤40 years) asthma and 10 young control non-smokers were immunostained for CD31, vascular endothelial growth factor-A (VEGF-A), angiogenin and BK receptors (B2R and B1R). Fibroblast and endothelial co-localisation of relevant molecules were performed by immunofluorescence. BK-induced VEGF-A and angiogenin release was studied (ELISA) in bronchial fibroblasts from subjects with asthma and COPD. Results In bronchial lamina propria of old patients with asthma, CD31 and VEGF-A + cell numbers were higher than old control non-smokers (p<0.05). Angiogenin + , B2R + and B1R + cell numbers in old patients with asthma were higher than in old control non-smokers, control smokers and patients with COPD (p<0.01). Angiogenin + cell numbers were higher in patients with COPD than both old control groups (p<0.05). In all patients with asthma the number of B2R + cells was positively related to the numbers of B1R + (r s =0.43), angiogenin + (r s =0.42) and CD31 cells (r s =0.46) (p<0.01). Angiogenin + cell numbers were negatively related to forced expiratory volume in 1 s (r s =−0.415, p=0.008). Double immunofluorescence revealed that CD31 cells of capillary vessels coexpressed B2R and that fibroblasts coexpressed B2R, VEGF-A and angiogenin. BK (10 −6 M) induced significant angiogenin release in fibroblasts from asthma and to a lesser extent in COPD. Conclusions Unlike COPD, this study suggests the involvement of BK receptors in bronchial vascular remodelling in asthma.

Research paper thumbnail of Pharmacological Modulation of the Bradykinin-Induced Differentiation of Human Lung Fibroblasts: Effects of Budesonide and Formoterol

Journal of Asthma, 2012

Objective. Bradykinin (BK) induces differentiation of lung fibroblasts into myofibroblasts, which... more Objective. Bradykinin (BK) induces differentiation of lung fibroblasts into myofibroblasts, which play an important role in extracellular matrix remodeling in the airways of asthmatic patients. It is unclear whether this process is affected by antiasthma therapies. Here, we evaluated whether a glucocorticoid, budesonide (BUD), and a long-acting /?2-agonist, formoterol (FM), either alone or in combination, modified BK-induced lung fibroblast differentiation, and affected the BK-activated intracellular signaling pathways. Methods. Human fetal lung fibroblasts were incubated with BUD (0.001-0.1 nM) and/or FM (0.0001-0.1 |iM) before exposure to BK (0.1 or 1 nM). Fibroblast differentiation into a-smooth-muscleactin-positive (a-SMA^) myofibroblasts, BK2 receptor (B2R) expression, extracellular signal-regulated kinase 1/2 (ERK 1/2) phosphorylation (p-ERKl/2), intracellular Ca^"*" concentration (fCa^^],), and p65 nuclear factor kappa B translocation were evaluated. Resuhs. BUD (0.1 \M) and FM (0.1 |j,M), either alone or in combination, completely inhibited BK-induced a-SMA protein expression and decreased the numbers of a-SMAf ibroblasts, with a clear reduction in a-SMA stress fibers organization. BUD also completely inhibited the increase of B2R, whereas FM with or without BUD had no effect. BK-induced increases of [Ca^"'"]; and p-ERKl/2 were significantly reduced to similar levels by BUD and FM, either alone or in combination, whereas p65 translocation was completely inhibited by all treatments. Conclusion. Both BUD and FM, either alone or in combination, effectively inhibited the BK-induced differentiation of fibroblasts into a-SMA^ myofibroblasts and the intracellular signaling pathways involved in fibroblast activation. These results suggest that BUD and FM combination therapy has potential to inhibit fibroblastdependent matrix remodeling in the airways of asthmatic patients. Keywords a-smooth muscle actin (a-SMA), bradykinin B2 receptor (B2R), human lung fibroblasts, tnitogen-activated protein kinases (MAPKs)

Research paper thumbnail of Bradykinin-induced asthmatic fibroblast/myofibroblast activities via bradykinin B2 receptor and different MAPK pathways

European Journal of Pharmacology, 2013

Bradykinin drives normal lung fibroblasts into myofibroblasts, induces fibroblast proliferation a... more Bradykinin drives normal lung fibroblasts into myofibroblasts, induces fibroblast proliferation and activates mitogen activated protein kinase pathways (MAPK) but its effects on bronchial fibroblasts from asthmatics (HBAFb) have not been yet studied. We studied bradykinin-induced fibroblast proliferation and differentiation and the related intracellular mechanisms in HBAFb compared to normal bronchial fibroblasts (HNBFb). Bradykinin-stimulated HBAFb and HNBFb were used to assess: bradykinin B2 receptor expression by Western blot analysis; cell proliferation by [(3)H] thymidine incorporation; α-smooth muscle actin (SMA) expression/polymerization by Western blot and immunofluorescence; epidermal growth factor (EGF) receptor, extracellular-regulated kinase (ERK) 1/2 and p38 MAPK activation by immunoprecipitation and Western blot, respectively. Constitutive bradykinin B2 receptor and α-SMA expression was higher in HBAFb as compared to HNBFb. Bradykinin increased bradykinin B2 receptor expression in HBAFb. Bradykinin, via bradykinin B2 receptor, significantly increased fibroblast proliferation at lower concentration (10(-11)M) and α-SMA expression/polymerization at higher concentration (10(-6)M) in both cells. Bradykinin increased ERK1/2 and p38 phosphorylation via bradykinin B2 receptor; EGF receptor inhibitor AG1478 and panmetalloproteinase inhibitor GM6001 blocked bradykinin-induced ERK1/2 activation but not p38 phosphorylation. Bradykinin, via bradykinin B2 receptor, induced EGF receptor phosphorylation that was suppressed by AG1478. In HBAFb AG1478, GM6001, the ERK1/2-inhibitor U0126 and the p38 inhibitor SB203580 suppressed bradykinin-induced cell proliferation, but only SB203580 reduced myofibroblast differentiation. These data indicate that bradykinin is actively involved in asthmatic bronchial fibroblast proliferation and differentiation, through MAPK pathways and EGF receptor transactivation, by which bradykinin may contribute to airway remodeling in asthma, opening new horizons for potential therapeutic implications in asthmatic patients.

Research paper thumbnail of Bronchial Airway Epithelial Cell Damage Following Exposure to Cigarette Smoke Includes Disassembly of Tight Junction Components Mediated by the Extracellular Signal-Regulated Kinase 1/2 Pathway

Chest, 2009

Background: Through a variety of biochemical mechanisms, cigarette smoke (CS) may damage airway e... more Background: Through a variety of biochemical mechanisms, cigarette smoke (CS) may damage airway epithelium, altering its normal structure and function. Injury to epithelium may include changes in tight junction (TJ) integrity with impairment of epithelial barrier function. Methods and results: To study the effect of the exposure to CS condensate (CSC) on TJ integrity, two human bronchial epithelial cell lines (HBECs), BEAS-2B and 16HBEI40-, were used. Exposure of the two HBECs to CSC resulted in a time-dependent and concentration-dependent disassembly of TJs, which were already detectable at 24 h at all the CSC concentrations tested (5%,10%, and 20%), associated with changes in cell shape, suggesting cell damage. However, a significant inhibition of cell growth and an increase in DNA fragmentation were detected only at the highest CSC concentration tested (20%) at 48 and 72 h, respectively. The involvement of epidermal growth factor receptor (EGFR)-extracellular signal-regulated kinase (ERK) 1/2 cascade in CSC-induced damage was shown by the observation that exposure to CSC (5%) induced a marked phosphorylation of ERKI/2, already detectable after 5-min incubation and confirmed by the demonstration that not only ERKl/2 phosphorylation but also CSC-induced TJ disassembly and DNA fragmentation were partially inhibited by a mitogen-activated protein kinase kinase inhibitor (UOI26) and completely blocked by a EGFR inhibitor (AGI478). Conclusion: CSC-induced damage to airway epithelium includes disassembly of TJs, modulated through the EGFR-ERKI/2 signaling pathway.

Research paper thumbnail of Controlled single-cell cyclic compression and transcription analysis: A pilot study

Biophysical Chemistry, 2017

• Controlled dynamic compression setup with single-cell sensitivity • qPCR protocol able to retri... more • Controlled dynamic compression setup with single-cell sensitivity • qPCR protocol able to retrieve targets expression with single-cell sensitivity • Workflow tested on bone cells, physiologically subjected to mechanical stimuli

Research paper thumbnail of Development of label-free biophysical markers in osteogenic maturation

Journal of the Mechanical Behavior of Biomedical Materials

Research paper thumbnail of Osteogenic Differentiation of MSC through Calcium Signaling Activation: Transcriptomics and Functional Analysis

PLOS ONE, 2016

The culture of progenitor mesenchymal stem cells (MSC) onto osteoconductive materials to induce a... more The culture of progenitor mesenchymal stem cells (MSC) onto osteoconductive materials to induce a proper osteogenic differentiation and mineralized matrix regeneration represents a promising and widely diffused experimental approach for tissue-engineering (TE) applications in orthopaedics. Among modern biomaterials, calcium phosphates represent the best bone substitutes, due to their chemical features emulating the mineral phase of bone tissue. Although many studies on stem cells differentiation mechanisms have been performed involving calcium-based scaffolds, results often focus on highlighting production of in vitro bone matrix markers and in vivo tissue ingrowth, while information related to the biomolecular mechanisms involved in the early cellular calcium-mediated differentiation is not well elucidated yet. Genetic programs for osteogenesis have been just partially deciphered, and the description of the different molecules and pathways operative in these differentiations is far from complete, as well as the activity of calcium in this process. The present work aims to shed light on the involvement of extracellular calcium in MSC differentiation: a better understanding of the early stage osteogenic differentiation program of MSC seeded on calcium-based biomaterials is required in order to develop optimal strategies to promote osteogenesis through the use of new generation osteoconductive scaffolds. A wide spectrum of analysis has been performed on time-dependent series: gene expression profiles are obtained from samples (MSC seeded on calcium-based scaffolds), together with related microRNAs expression and in vivo functional validation. On this basis, and relying on literature knowledge, hypotheses are made on the biomolecular players activated by the biomaterial calcium-phosphate component. Interestingly, a key role of miR-138 was highlighted, whose inhibition markedly increases osteogenic differentiation in vitro and enhance ectopic bone formation in vivo. Moreover, there is evidence that Ca-P substrate triggers osteogenic differentiation through genes (SMAD and RAS family) that are typically regulated during dexamethasone (DEX) induced differentiation.