45 results on '"Mussano, F."'
Search Results
2. Early Biological Response of a Ultra-Hydrophilic Implant Surface Activated by Salts and Dry Technology: In Vitro Study on MC3T3-E1 Osteogenic Cell Line
- Author
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Canullo L, Chinigò G, Raffone C, Mussano F, Genova T, Gianfreda F, Bollero P, and Antonacci D
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Implant surface ,Chemistry ,Osteogenic cell ,In vitro study ,Line (text file) ,Mc3t3 e1 ,Biomedical engineering - Abstract
New implant surfaces created by sandblasting and acid etching enhance osseointegration processes. Surface energy seems to be an aspect of paramount importance in the first phase of healing, supporting protein adsorption and cell adhesion, proliferation, differentiation and bone mineralization. New methods were introduced to preserve over time and improve surface energy such as wet storage or bioactivation through salts coating created with dry technology. Purpose of this study was to evaluate the osteogenic response of pre-osteoblast lineage cells to dry bioactivated surface. MC3T3-E1 osteogenic cell line were cultured on ABT (SLA surface) and NANO (SLA surface with dry bioactive technology)., cell adhesion assay, proliferation assay and cell morphology were performed. Despite both surface treatments were able to support regular morphology; cell adhesion and proliferation were statistically improved ( p.value
- Published
- 2021
3. Healing properties of implants inserted concomitantly with anorganic bovine bone. A histomorphometric human study
- Author
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Menicucci, G, Mussano, F, Schierano, G, Rizzati, A, Aimetti, M, Gassino, G, Traini, T, and Carossa, S
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- 2013
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4. Cytokine, Chemokine, and Growth Factor Profile Characterization of Undifferentiated and Osteoinduced Human Adipose-Derived Stem Cells
- Author
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Mussano, F., primary, Genova, T., additional, Corsalini, M., additional, Schierano, G., additional, Pettini, F., additional, Di Venere, D., additional, and Carossa, S., additional
- Published
- 2017
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5. Role of surface finishing on the in vitro biological properties of a silicon nitride–titanium nitride (Si3N4–TiN) composite
- Author
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Mussano, F., primary, Genova, T., additional, Rivolo, P., additional, Mandracci, P., additional, Munaron, L., additional, Faga, M. G., additional, and Carossa, S., additional
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- 2016
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6. Si-Based Amorphous Thin Films for the Performance Improvement of Dental Prostheses and Implants
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Mandracci, Pietro, Gazia, R., Mussano, F., Carossa, S., and Pirri, Candido
- Published
- 2012
7. Role of surface finishing on the in vitro biological properties of a silicon nitride-titanium nitride (SiN-TiN) composite.
- Author
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Mussano, F., Genova, T., Rivolo, P., Mandracci, P., Munaron, L., Faga, M., and Carossa, S.
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- *
SILICON nitride , *TITANIUM nitride , *SURFACE finishing , *METALLIC composites , *MECHANICAL properties of metals , *STRENGTH of materials - Abstract
Silicon nitride (SiN) has been introduced clinically as an orthopedic biomaterial for interbody fusion devices and in joint replacements. However, the production of complex shapes through conventional mechanical machining is difficult and expensive and limits interesting applications. Thus, several electrically conductive reinforcements to the SiN matrix, like TiN, have been proposed, generating composites suitable to be wrought by electrical discharge machining (EDM). In this study, SiN-TiN with high strength, low density, and good electric conductivity wrought by EDM was studied. The role of surface finishing was investigated comparing the interface generated during the EDM process to that resulting from further polishing. The different topographical features were assessed by electron microscopy, energy dispersive X-ray spectrometry, and profilometry. Surface wettability was also determined based on the measurement of the OCA of water and diiodomethane. The biological responses induced in MC3T3 cells, a widely diffused osteoblast model, were correlated with the surface pattern. The unpolished samples could promote better cell viability, with a more relevant effect on the cytoskeleton arrangement as highlighted by numerous cytoplasmic extensions and filopodia-like structures and the high number of focal adhesions, while MC3T3 cells grown on polished SiN-TiN specimens displayed a flat morphology. In addition, the unpolished SiN-TiN increased osteocalcin production and calcium deposition. Taken together, these data support the biocompatibility and in vitro osteogenic properties of the electroconductive SiN-TiN investigated. Further in vivo studies are required to explore the possible use of bone implants directly obtained by EDM. [ABSTRACT FROM AUTHOR]
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- 2017
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8. Biological components in a standardized derivative of bovine colostrum
- Author
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Sacerdote, P., primary, Mussano, F., additional, Franchi, S., additional, Panerai, A.E., additional, Bussolati, G., additional, Carossa, S., additional, Bartorelli, A., additional, and Bussolati, B., additional
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- 2013
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9. The role of alumina-zirconia loading on the mechanical and biological properties of UHMWPE for biomedical applications
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Valentina Strongone, Federico Mussano, C. De Rosa, Finizia Auriemma, Donatella Duraccio, Tullio Genova, Giulio Malucelli, M.G. Faga, Duraccio, D., Strongone, V., Malucelli, G., Auriemma, F., De Rosa, C., Mussano, F. D., Genova, T., and Faga, M. G.
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biomedical applications ,Materials science ,UHMWPE ,Composite number ,Compression molding ,Ceramics and Composite ,02 engineering and technology ,engineering.material ,010402 general chemistry ,01 natural sciences ,composites ,Industrial and Manufacturing Engineering ,Biological property ,Filler (materials) ,Homogenizer ,Cubic zirconia ,Mechanics of Material ,Composite material ,Elastic modulus ,Mechanical Engineering ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Mechanics of Materials ,Ceramics and Composites ,engineering ,Wetting ,0210 nano-technology - Abstract
Different amounts of Alumina Toughened Zirconia (80–20 wt%) (ATZ) were dispersed in UHMWPE matrix with the purpose of obtaining a material easy to be processed, inexpensive, with suitable mechanical properties and good cells interaction. The UHMWPE/ATZ composites were prepared by using a homogenizer, followed by compression molding. All the composites show an increase in elastic modulus and hardness respect to neat UHMWPE. The composite with the lowest amount of ATZ (i.e. 2.5 wt%) also displays an increase of the yield stress. The biological behavior is not directly correlated neither to wettability nor to microroughness, while it seems that the distribution of ATZ filler plays an important role.
- Published
- 2019
10. Dysregulation of FLVCR1a-dependent mitochondrial calcium handling in neural progenitors causes congenital hydrocephalus.
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Bertino F, Mukherjee D, Bonora M, Bagowski C, Nardelli J, Metani L, Zanin Venturini DI, Chianese D, Santander N, Salaroglio IC, Hentschel A, Quarta E, Genova T, McKinney AA, Allocco AL, Fiorito V, Petrillo S, Ammirata G, De Giorgio F, Dennis E, Allington G, Maier F, Shoukier M, Gloning KP, Munaron L, Mussano F, Salsano E, Pareyson D, di Rocco M, Altruda F, Panagiotakos G, Kahle KT, Gressens P, Riganti C, Pinton PP, Roos A, Arnold T, Tolosano E, and Chiabrando D
- Subjects
- Animals, Humans, Mice, Neurogenesis genetics, Inositol 1,4,5-Trisphosphate Receptors metabolism, Inositol 1,4,5-Trisphosphate Receptors genetics, Neural Stem Cells metabolism, Neural Stem Cells pathology, Mitochondria metabolism, Hydrocephalus metabolism, Hydrocephalus genetics, Hydrocephalus pathology, Calcium metabolism, Receptors, Virus metabolism, Receptors, Virus genetics, Membrane Transport Proteins metabolism, Membrane Transport Proteins genetics
- Abstract
Congenital hydrocephalus (CH), occurring in approximately 1/1,000 live births, represents an important clinical challenge due to the limited knowledge of underlying molecular mechanisms. The discovery of novel CH genes is thus essential to shed light on the intricate processes responsible for ventricular dilatation in CH. Here, we identify FLVCR1 (feline leukemia virus subgroup C receptor 1) as a gene responsible for a severe form of CH in humans and mice. Mechanistically, our data reveal that the full-length isoform encoded by the FLVCR1 gene, FLVCR1a, interacts with the IP3R3-VDAC complex located on mitochondria-associated membranes (MAMs) that controls mitochondrial calcium handling. Loss of Flvcr1a in mouse neural progenitor cells (NPCs) affects mitochondrial calcium levels and energy metabolism, leading to defective cortical neurogenesis and brain ventricle enlargement. These data point to defective NPCs calcium handling and metabolic activity as one of the pathogenetic mechanisms driving CH., Competing Interests: Declaration of interests E.T., V.F., D.Chiabrando, S.P., F.B., and A.L.A. are inventors in a patent filed by the University of Torino, not related to the research reported here., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
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- 2024
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11. Impact of Inflammatory Markers and Senescence-Associated Secretory Phenotype in the Gingival Crevicular Fluid on the Outcomes of Periodontal Regeneration.
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Baima G, Romano F, Franco F, Roato I, Mussano F, Berta GN, and Aimetti M
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- Humans, Male, Female, Middle Aged, Adult, Regeneration, Matrix Metalloproteinase 8 metabolism, Matrix Metalloproteinase 8 genetics, Phenotype, Matrix Metalloproteinase 9 metabolism, Matrix Metalloproteinase 9 genetics, Inflammation metabolism, Treatment Outcome, Interleukin-1beta metabolism, Aged, Gingival Crevicular Fluid metabolism, Biomarkers
- Abstract
The aim of this study was to test the molecular expression profile (senescence-associated secretory phenotype; SASP) in gingival crevicular fluid (GCF) prior to surgery in relation to the distribution of clinical success of periodontal regeneration. Forty consecutive patients presenting sites with residual probing pocket depth (PPD) ≥ 6 mm and intrabony defects ≥ 3 mm were treated through a minimally invasive surgical technique. Pre-operatively, GCF was sampled for inflammatory biomarker analysis related to SASP [interleukin (IL)-1β, IL-6, and IL-12; matrix-metalloproteinases (MMP)-8 and -9]. Better or worse responders were classified depending on the achievement of a composite outcome measure at 1-year [COM; PPD ≤ 4 mm and clinical attachment gain (CAL) gain ≥ 3 mm]. Correlation analyses and logistic regression models were performed. Periodontal regeneration led to significant improvements in mean clinical and radiographic parameters. Teeth achieving COM presented significantly lower amounts of SASP factors compared with non-successful teeth. Higher CAL gain, PPD reduction, and radiographic bone fill were negatively correlated with IL-1β and MMP-8 and -9 ( p < 0.001), while IL-12 showed a direct relationship with CAL gain ( p = 0.005) and PPD reduction ( p = 0.038). Sites expressing higher SASP expression in the GCF before periodontal regeneration achieved worse clinical and radiographic outcomes.
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- 2024
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12. Orally Derived Stem Cell-Based Therapy in Periodontal Regeneration: A Systematic Review and Meta-Analysis of Randomized Clinical Studies.
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Campagna A, Baima G, Romano F, Amoroso F, Mussano F, Oteri G, Aimetti M, and Peditto M
- Abstract
The present systematic review was performed to assess the application of orally derived stem cells in periodontal regenerative therapy, and because of this, the following PICO question was proposed: "In patients with periodontitis, can the adjunctive use of orally derived stem cells provide additional clinical and radiographic benefits for periodontal regeneration?". Randomized clinical studies were electronically and manually searched up until December 2023. Quantitative analyses were performed with the aim of evaluating the mean differences (MDs) between the treatment and control groups in terms of clinical attachment level (CAL) gain, probing pocket depth (PPD) reduction, gingival recession (GR), and radiographic bone gain (RBG) using random effect models. A total of seven studies were selected for the systematic review. Meta-analyses excluding studies with a high risk of bias highlighted a non-statistically significant result for the use of stem cells when compared to the control groups in terms of CAL gain [MD = 1.05; 95% CI (-0.88, 2.97) p = 0.29] and PPD reduction [MD = 1.32; 95% CI (-0.25, 2.88) p = 0.10]. The same also applied to GR [MD = -0.08; 95% CI (-0.79, 0.63) p = 0.83] and RBG [MD = 0.50; 95% CI (-0.88, 1.88) p = 0.48]. Based on the high heterogeneity, there is not enough evidence to consider the adjunctive application of orally derived mesenchymal stem cells as a preferential approach for periodontal regenerative treatment, as compared to standard procedures.
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- 2024
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13. Vacuum Plasma Treatment Device for Enhancing Fibroblast Activity on Machined and Rough Titanium Surfaces.
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Canullo L, Genova T, Chinigò G, Iacono R, Pesce P, Menini M, and Mussano F
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This study was conducted to compare the effects of an innovative plasma surface treatment device that does not need a gas supply for titanium disks with two different surface topographies: the prototypical machined surface (MAC) and one of the most diffused roughened ones (SL) obtained through grit blasting and acid etching. A total of 200-MAC and 200-SL titanium disks were used. Each group of disks was divided into four sub-groups of 40 samples each that were subjected to five different tests. Among these, 150-MAC and 150-SL were considered the test group, and they were treated with plasma for 15, 30, and 60 s after being removed from the sterile packaging. On the other hand, 50-MAC and 50-SL were considered the control group, and they were only removed from sterile plastic vials. The samples were analyzed to evaluate the capability of the plasma treatment in influencing protein adsorption, cell adhesion, proliferation, and microbial growth on the test group disks when compared to the untreated disks. Protein adsorption was significantly enhanced after 20 min of plasma treatment for 15 and 30 s on the MAC and SL disks. Plasma treatment for 15 and 30 s significantly increased the level of adhesion in both treated samples after 30 min. Furthermore, the MAC samples showed a significant increase in cell adhesion 4 h after plasma treatment for 15 s. The SEM analysis highlighted that, on the treated samples (especially on the MAC disks), the cells with a polygonal and flat shape prevailed, while the fusiform- and globular-shaped cells were rare. The encouraging results obtained further confirm the effectiveness of plasma treatments on cell adhesion and fibroblast activity.
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- 2024
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14. Efficacy of a Solution Containing 33% Trichloroacetic Acid and Hydrogen Peroxide in Decontaminating Machined vs. Sand-Blasted Acid-Etched Titanium Surfaces.
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Baima G, Romano F, Roato I, Mosca Balma A, Pedraza R, Faga MG, Amoroso F, Orrico C, Genova T, Aimetti M, and Mussano F
- Abstract
This in vitro study assessed the efficacy of a solution containing 33% trichloroacetic acid (CCl
3 COOH; TCA) and hydrogen peroxide (H2 O2 ) in decontaminating machined (MAC) and sand-blasted acid-etched (SBAE) titanium surfaces. A total of 80 titanium disks were prepared (40 MAC and 40 SBAE). Streptococcus sanguinis and Enterococcus faecalis strains were incubated on 36 samples, while the remaining 44 were kept as controls. Roughness analysis and scanning electron microscopy were used to evaluate the surface features before and after TCAH2 O2 treatment. The viability of human adipose-derived mesenchymal stem cells (ASCs) after TCAH2 O2 decontamination was assessed with a chemiluminescent assay along with cell morphology through fluorescent staining. TCAH2 O2 preserved the surface topography of MAC and SBAE specimens. It also effectively eradicated bacteria on both types of specimens without altering the surface roughness ( p > 0.05). Also, no significant differences in protein adsorption between the pristine and TCAH2 O2 -treated surfaces were found ( p = 0.71 and p = 0.94). While ASC proliferation remained unchanged on MAC surfaces, a decrease was observed on the decontaminated SBAE specimens at 24 and 48 h ( p < 0.05), with no difference at 72 h ( p > 0.05). Cell morphology showed no significant changes after 72 h on both surface types even after decontamination. This study suggests TCAH2 O2 as a promising decontamination agent for titanium surfaces, with potential implications for peri-implant health and treatment outcomes.- Published
- 2024
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15. Denosumab and Zoledronic Acid Differently Affect Circulating Immune Subsets: A Possible Role in the Onset of MRONJ.
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Roato I, Pavone L, Pedraza R, Bosso I, Baima G, Erovigni F, and Mussano F
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- Humans, Female, Zoledronic Acid therapeutic use, Denosumab pharmacology, Denosumab therapeutic use, Cyclins, Bone Neoplasms drug therapy, Breast Neoplasms drug therapy, Respiratory Distress Syndrome chemically induced
- Abstract
This work investigated whether the anti-resorptive drugs (ARDs) zoledronic acid (Zol) and denosumab (Dmab) affect differently the levels of circulating immune cell subsets, possibly predicting the risk of developing medication-related ONJ (MRONJ) during the first 18 months of treatment. Blood samples were collected from 10 bone metastatic breast cancer patients receiving cyclin inhibitors at 0, 6, 12, and 18 months from the beginning of Dmab or Zol treatment. Eight breast cancer patients already diagnosed with MRONJ and treated with cyclin inhibitors and ARDs were in the control group. PBMCs were isolated; the trend of circulating immune subsets during the ARD treatment was monitored, and 12 pro-inflammatory cytokines were analyzed in sera using flow cytometry. In Dmab-treated patients, activated T cells were stable or increased, as were the levels of IL-12, TNF-α, GM-CSF, IL-5, and IL-10, sustaining them. In Zol-treated patients, CD8+T cells decreased, and the level of IFN-γ was undetectable. γδT cells were not altered in Dmab-treated patients, while they dramatically decreased in Zol-treated patients. In the MRONJ control group, Zol-ONJ patients showed a reduction in activated T cells and γδT cells compared to Dmab-ONJ patients. Dmab was less immunosuppressive than Zol, not affecting γδT cells and increasing activated T cells.
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- 2023
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16. Senescent Markers Expressed by Periodontal Ligament-Derived Stem Cells (PDLSCs) Harvested from Patients with Periodontitis Can Be Rejuvenated by RG108.
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Roato I, Baima G, Orrico C, Mosca Balma A, Alotto D, Romano F, Ferracini R, Aimetti M, and Mussano F
- Abstract
Periodontal ligament (PDL) has become an elective source of mesenchymal stem cells (PDLSCs) in dentistry. This research aimed to compare healthy PDLSCs (hPDLSCs) and periodontitis PDLSCs (pPDLSCs) to ascertain any possible functional differences owing to their milieux of origin. Cells were tested in terms of colony-forming unit efficiency; multi differentiating capacity; immunophenotype, stemness, and senescent state were studied by flow cytometry, immunofluorescence, and β-galactosidase staining; gene expression using RT-PCR. Both hPDLSCs and pPDLSCs were comparable in terms of their immunophenotype and multilineage differentiation capabilities, but pPDLSCs showed a senescent phenotype more frequently. Thus, a selective small molecule inhibitor of DNA methyltransferase (DNMT), RG108, known for its effect on senescence, was used to possibly reverse this phenotype. RG108 did not affect the proliferation and apoptosis of PDLSCs, and it showed little effect on hPDLSCs, while a significant reduction of both p16 and p21 was detected along with an increase of SOX2 and OCT4 in pPDLSCs after treatment at 100 μM RG108. Moreover, the subset of PDLSCs co-expressing OCT4 and p21 decreased, and adipogenic potential increased in pPDLSCs after treatment. pPDLSCs displayed a senescent phenotype that could be reversed, opening new perspectives for the treatment of periodontitis.
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- 2023
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17. Mechanical and Biological Characterization of PMMA/Al 2 O 3 Composites for Dental Implant Abutments.
- Author
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Roato I, Genova T, Duraccio D, Ruffinatti FA, Zanin Venturini D, Di Maro M, Mosca Balma A, Pedraza R, Petrillo S, Chinigò G, Munaron L, Malucelli G, Faga MG, and Mussano F
- Abstract
The mechanical and biological behaviors of PMMA/Al
2 O3 composites incorporating 30 wt.%, 40 wt.%, and 50 wt.% of Al2 O3 were thoroughly characterized as regards to their possible application in implant-supported prostheses. The Al2 O3 particles accounted for an increase in the flexural modulus of PMMA. The highest value was recorded for the composite containing 40 wt.% Al2 O3 (4.50 GPa), which was about 18% higher than that of its unfilled counterpart (3.86 GPa). The Al2 O3 particles caused a decrease in the flexural strength of the composites, due to the presence of filler aggregates and voids, though it was still satisfactory for the intended application. The roughness (Ra) and water contact angle had the same trend, ranging from 1.94 µm and 77.2° for unfilled PMMA to 2.45 µm and 105.8° for the composite containing the highest alumina loading, respectively, hence influencing both the protein adsorption and cell adhesion. No cytotoxic effects were found, confirming that all the specimens are biocompatible and capable of sustaining cell growth and proliferation, without remarkable differences at 24 and 48 h. Finally, Al2 O3 was able to cause strong cell responses (cell orientation), thus guiding the tissue formation in contact with the composite itself and not enhancing its osteoconductive properties, supporting the PMMA composite's usage in the envisaged application.- Published
- 2023
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18. Bioactive glass for periodontal regeneration: a systematic review.
- Author
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Motta C, Cavagnetto D, Amoroso F, Baldi I, and Mussano F
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- Humans, Bone Regeneration, Dental Care, Durapatite, Biocompatible Materials therapeutic use, Furcation Defects
- Abstract
Background: One of the major clinical challenges of this age could be represented by the possibility to obtain a complete regeneration of infrabony defects. Over the past few years, numerous materials and different approaches have been developed to obtain bone and periodontal healing. Among all biomaterials, bioglasses (BG) are one of the most interesting due to their ability to form a highly reactive carbonate hydroxyapatite layer. Our aim was to systematically review the literature on the use and capability of BG for the treatment of periodontal defects and to perform a meta-analysis of their efficacy., Methods: A search of MEDLINE/PubMed, Cochrane Library, Embase and DOSS was conducted in March 2021 to identify randomized controlled trials (RCTs) using BG in the treatment of intrabony and furcation defects. Two reviewers selected the articles included in the study considering the inclusion criteria. The outcomes of interest were periodontal and bone regeneration in terms of decrease of probing depth (PD) and gain of clinical attachment level (CAL). A network meta-analysis (NMA) was fitted, according to the graph theory methodology, using a random effect model., Results: Through the digital search, 46 citations were identified. After duplicate removal and screening process, 20 articles were included. All RCTs were retrieved and rated following the Risk of bias 2 scale, revealing several potential sources of bias. The meta-analysis focused on the evaluation at 6 months, with 12 eligible articles for PD and 10 for CAL. As regards the PD at 6 months, AUTOGENOUS CORTICAL BONE, BIOGLASS and PLATELET RICH FIBRIN were more efficacious than open flap debridement alone, with a statistically significant standardized mean difference (SMD) equal to -1.57, -1.06 and - 2.89, respectively. As to CAL at 6 months, the effect of BIOGLASS is reduced and no longer significant (SMD = -0.19, p-value = 0.4) and curiously PLATELET RICH FIBRIN was more efficacious than OFD (SMD =-4.13, p-value < 0.001) in CAL gain, but in indirect evidence., Conclusions: The present review partially supports the clinical efficacy of BG in periodontal regeneration treatments for periodontal purposes. Indeed, the SMD of 0.5 to 1 in PD and CAL obtained with BG compared to OFD alone seem clinically insignificant even if it is statistically significant. Heterogeneity sources related to periodontal surgery are multiple, difficult to assess and likely hamper a quantitative assessment of BG efficacy., (© 2023. The Author(s).)
- Published
- 2023
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19. E15.5 Mouse Embryo Micro-CT Using a Bruker Skyscan 1172 Micro-CT.
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Astanina E, Petrillo S, Genova T, Mussano F, and Bussolino F
- Abstract
X-ray computed microtomography (µCT) is a powerful tool to reveal the 3D structure of tissues and organs. Compared with the traditional sectioning, staining, and microscopy image acquisition, it allows a better understanding of the morphology and a precise morphometric analysis. Here, we describe a method for 3D visualization and morphometric analysis by µCT scanning of the embryonic heart of iodine-stained E15.5 mouse embryos., Competing Interests: Competing interestsThe authors declare no conflict of interests., (Copyright © 2023 The Authors; exclusive licensee Bio-protocol LLC.)
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- 2023
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20. Immune Dysfunction in Medication-Related Osteonecrosis of the Jaw.
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Roato I, Mauceri R, Notaro V, Genova T, Fusco V, and Mussano F
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- Humans, Denosumab adverse effects, Diphosphonates adverse effects, Bisphosphonate-Associated Osteonecrosis of the Jaw drug therapy, Bone Density Conservation Agents adverse effects, Respiratory Distress Syndrome drug therapy
- Abstract
The pathogenesis of medication-related osteonecrosis of the jaw (MRONJ) is multifactorial and there is a substantial consensus on the role of antiresorptive drugs (ARDs), including bisphosphonates (BPs) and denosumab (Dmab), as one of the main determinants. The time exposure, cumulative dose and administration intensity of these drugs are critical parameters to be considered in the treatment of patients, as cancer patients show the highest incidence of MRONJ. BPs and Dmab have distinct mechanisms of action on bone, but they also exert different effects on immune subsets which interact with bone cells, thus contributing to the onset of MRONJ. Here, we summarized the main effects of ARDs on the different immune cell subsets, which consequently affect bone cells, particularly osteoclasts and osteoblasts. Data from animal models and MRONJ patients showed a deep interference of ARDs in modulating immune cells, even though a large part of the literature concerns the effects of BPs and there is a lack of data on Dmab, demonstrating the need to further studies.
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- 2023
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21. The Impact of Plasma Membrane Ion Channels on Bone Remodeling in Response to Mechanical Stress, Oxidative Imbalance, and Acidosis.
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Perin M, Chinigò G, Genova T, Mussano F, and Munaron L
- Abstract
The extracellular milieu is a rich source of different stimuli and stressors. Some of them depend on the chemical-physical features of the matrix, while others may come from the 'outer' environment, as in the case of mechanical loading applied on the bones. In addition to these forces, a plethora of chemical signals drives cell physiology and fate, possibly leading to dysfunctions when the homeostasis is disrupted. This variety of stimuli triggers different responses among the tissues: bones represent a particular milieu in which a fragile balance between mechanical and metabolic demands should be tuned and maintained by the concerted activity of cell biomolecules located at the interface between external and internal environments. Plasma membrane ion channels can be viewed as multifunctional protein machines that act as rapid and selective dual-nature hubs, sensors, and transducers. Here we focus on some multisensory ion channels (belonging to Piezo, TRP, ASIC/EnaC, P2XR, Connexin, and Pannexin families) actually or potentially playing a significant role in bone adaptation to three main stressors, mechanical forces, oxidative stress, and acidosis, through their effects on bone cells including mesenchymal stem cells, osteoblasts, osteoclasts, and osteocytes. Ion channel-mediated bone remodeling occurs in physiological processes, aging, and human diseases such as osteoporosis, cancer, and traumatic events.
- Published
- 2023
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22. Bacterial and Cellular Response to Yellow-Shaded Surface Modifications for Dental Implant Abutments.
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Genova T, Chinigò G, Munaron L, Rivolo P, Luganini A, Gribaudo G, Cavagnetto D, Mandracci P, and Mussano F
- Subjects
- Surface Properties, Titanium chemistry, Bacterial Adhesion, Cell Adhesion, Dental Implants
- Abstract
Dental implants have dramatically changed the rehabilitation procedures in dental prostheses but are hindered by the possible onset of peri-implantitis. This paper aims to assess whether an anodization process applied to clinically used surfaces could enhance the adhesion of fibroblasts and reduce bacterial adhesion using as a reference the untreated machined surface. To this purpose, four different surfaces were prepared: (i) machined (MAC), (ii) machined and anodized (Y-MAC), (iii) anodized after sand-blasting and acid etching treatment (Y-SL), and (iv) anodized after double acid etching (Y-DM). All specimens were characterized by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX). Moreover, the mean contact angle in both water and diiodomethane as well as surface free energy calculation was assessed. To evaluate changes in terms of biological responses, we investigated the adhesion of Streptococcus sanguinis ( S. sanguinis ) and Enterococcus faecalis ( E. faecalis ), fetal bovine serum (FBS) adsorption, and the early response of fibroblasts in terms of cell adhesion and viability. We found that the anodization reduced bacterial adhesion, while roughened surfaces outperformed the machined ones for protein adsorption, fibroblast adhesion, and viability independently of the treatment. It can be concluded that surface modification techniques such as anodization are valuable options to enhance the performance of dental implants.
- Published
- 2022
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23. Challenges of Periodontal Tissue Engineering: Increasing Biomimicry through 3D Printing and Controlled Dynamic Environment.
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Roato I, Masante B, Putame G, Massai D, and Mussano F
- Abstract
In recent years, tissue engineering studies have proposed several approaches to regenerate periodontium based on the use of three-dimensional (3D) tissue scaffolds alone or in association with periodontal ligament stem cells (PDLSCs). The rapid evolution of bioprinting has sped up classic regenerative medicine, making the fabrication of multilayered scaffolds-which are essential in targeting the periodontal ligament (PDL)-conceivable. Physiological mechanical loading is fundamental to generate this complex anatomical structure ex vivo. Indeed, loading induces the correct orientation of the fibers forming the PDL and maintains tissue homeostasis, whereas overloading or a failure to adapt to mechanical load can be at least in part responsible for a wrong tissue regeneration using PDLSCs. This review provides a brief overview of the most recent achievements in periodontal tissue engineering, with a particular focus on the use of PDLSCs, which are the best choice for regenerating PDL as well as alveolar bone and cementum. Different scaffolds associated with various manufacturing methods and data derived from the application of different mechanical loading protocols have been analyzed, demonstrating that periodontal tissue engineering represents a proof of concept with high potential for innovative therapies in the near future.
- Published
- 2022
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24. The TFEB-TGIF1 axis regulates EMT in mouse epicardial cells.
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Astanina E, Doronzo G, Corà D, Neri F, Oliviero S, Genova T, Mussano F, Middonti E, Vallariello E, Cencioni C, Valdembri D, Serini G, Limana F, Foglio E, Ballabio A, and Bussolino F
- Subjects
- Animals, Cells, Cultured, Mice, Organogenesis, Pericardium metabolism, Epithelial-Mesenchymal Transition physiology, Transforming Growth Factor beta metabolism
- Abstract
Epithelial-mesenchymal transition (EMT) is a complex and pivotal process involved in organogenesis and is related to several pathological processes, including cancer and fibrosis. During heart development, EMT mediates the conversion of epicardial cells into vascular smooth muscle cells and cardiac interstitial fibroblasts. Here, we show that the oncogenic transcription factor EB (TFEB) is a key regulator of EMT in epicardial cells and that its genetic overexpression in mouse epicardium is lethal due to heart defects linked to impaired EMT. TFEB specifically orchestrates the EMT-promoting function of transforming growth factor (TGF) β, and this effect results from activated transcription of thymine-guanine-interacting factor (TGIF)1, a TGFβ/Smad pathway repressor. The Tgif1 promoter is activated by TFEB, and in vitro and in vivo findings demonstrate its increased expression when Tfeb is overexpressed. Furthermore, Tfeb overexpression in vitro prevents TGFβ-induced EMT, and this effect is abolished by Tgif1 silencing. Tfeb loss of function, similar to that of Tgif1, sensitizes cells to TGFβ, inducing an EMT response to low doses of TGFβ. Together, our findings reveal an unexpected function of TFEB in regulating EMT, which might provide insights into injured heart repair and control of cancer progression., (© 2022. The Author(s).)
- Published
- 2022
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25. Plasma of Argon Treatment of the Implant Surface, Systematic Review of In Vitro Studies.
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Carossa M, Cavagnetto D, Mancini F, Mosca Balma A, and Mussano F
- Subjects
- Argon pharmacology, Cell Adhesion, Humans, Surface Properties, Osteoblasts, Titanium pharmacology
- Abstract
This paper aims to review the evidence of the cellular activity on titanium samples exposed to Plasma of Argon (PoA) treatment. A systematic review was carried out based on the PRISMA statement by searching the Cochrane Library, PubMed, Web of Science, EMBASE and Scopus, up to October 2020. Papers were selected according to PICOS format that is: Population (P): osteoblasts, fibroblasts, gingival cells; Intervention (I): PoA disinfection treatment; Comparison (C): untreated controls; Outcome (O): cell culture; Setting (S): in vitro assays. The quality assessment was performed according to the CRIS Guidelines (Checklist for Reporting In vitro Studies). A total of 661 articles were found, of which 16 were included. The quality assessment revealed an overall poor quality of the studies analyzed. In vitro studies on the potential of PoA showed a potential effect in promoting higher cell adhesion and protein adsorption in the earliest times (hours). This outcome was not so evident when later stages of cell growth on the surfaces were tested and compared to the control groups. Only one study was conducted in vivo on a human sample regarding abutment cleaning. No meta-analysis was conducted because of the variety of experimental settings, mixed methods and different cell lines studied. PoA seems to be effective in promoting cell adhesion and protein adsorption. The duration of this effect remains unclear. Further evidence is required to demonstrate the long-term efficacy of the treatment and to support the use of PoA treatment in clinical practice.
- Published
- 2022
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26. Disinfection and Biocompatibility of Titanium Surfaces Treated with Glycine Powder Airflow and Triple Antibiotic Mixture: An In Vitro Study.
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Alovisi M, Carossa M, Mandras N, Roana J, Costalonga M, Cavallo L, Pira E, Putzu MG, Bosio D, Roato I, Mussano F, and Scotti N
- Abstract
The aim of this in vitro study was to compare three disinfection protocols of biofilm-coated machined (MAC) and acid etched (SLA) commercial pure Grade 4 Titanium disks. Samples were infected with a vial of polymicrobial biofilm to simulate peri-implantitis in vitro. Seventeen MAC and twenty SLA titanium disks were randomly assigned to: (1) glycine powder air-flow (GYPAP) for 1 min; (2) a local delivered triple paste antibiotic composed by a gel mixture with ciprofloxacin, metronidazole, and clarithromycin (3MIX) for 1 h; and (3) a combination of both (GYPAP + 3MIX). Biocompatibility of the titanium disks after each treatment protocol was assessed by measurement of adhesion and growth of adipose-derived mesenchymal stem cells (ASCs) after 24 and 72 h. A confocal laser scanning microscope (CLSM) assessed the antibacterial effect of each treatment. Data of the antibacterial efficacy and cell viability were presented as mean with standard deviation and calculated by one-way ANOVA with multiple comparisons via Bonferroni tests. Results were considered significant with p < 0.05. The higher cell viability was achieved by the 3MIX and GYPAP combination on the SLA surfaces after 72 h. CLSM analysis showed a mean ratio of dead bacteria statistically higher in the 3MIX + GYPAP group compared with the GYPAP and 3MIX subgroups (p < 0.05). In conclusion, data showed that the combination of GYPAP and 3MIX could be preferred to the other protocols, especially in presence of SLA titanium surface.
- Published
- 2022
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27. CD73/Adenosine Pathway Involvement in the Interaction of Non-Small Cell Lung Cancer Stem Cells and Bone Cells in the Pre-Metastatic Niche.
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Bertolini G, Compagno M, Belisario DC, Bracci C, Genova T, Mussano F, Vitale M, Horenstein A, Malavasi F, Ferracini R, and Roato I
- Subjects
- Adenosine metabolism, Humans, Neoplastic Stem Cells metabolism, Receptor, Adenosine A2A metabolism, Carcinoma, Non-Small-Cell Lung, Lung Neoplasms
- Abstract
Adenosinergic signaling is an important regulator of tissue homeostasis and extracellular accumulation of adenosine (Ado) and is associated with different pathologies, such as cancer. In non-small-cell lung cancer (NSCLC), a subset of CD133/CXCR4+ cancer stem cell (CSCs) has been demonstrated to initiate bone metastases. Here we investigated how NSCLC CSCs interact with osteoclasts (OCs) and osteoblasts (OBs) by modulating Ado production and OC activity. We proved that CSC-spheres, generated in vitro from NSCLC cell lines, express CD38, PC-1, and CD73, enzymes of the non-canonical adenosinergic pathway, produce high level of Ado, and down-regulate A1R and A3R inhibitory receptors, while expressing A2AR and A2BR. To address the Ado role and modulation of the in-bone pre-metastatic niche, we performed co-cultures of CSC-spheres with OCs and OBs cells. Firstly, we verified that active OCs do not activate non-canonical the adenosinergic pathway, conversely to OBs. OCs co-cultured with CSC-spheres increase Ado production that is related to the OC resorption activity and contributes to T-cell suppression. Finally, we proved the efficacy of anti-CD73 agents in blocking NSCLC cell migration. Overall, we assessed the importance of adenosinergic signaling in the interaction between CSCs and OCs at the pre-metastatic niche, with therapeutic implications related to Ado production.
- Published
- 2022
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28. Endothelial Cells Promote Osteogenesis by Establishing a Functional and Metabolic Coupling With Human Mesenchymal Stem Cells.
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Petrillo S, Genova T, Chinigò G, Roato I, Scarpellino G, Kopecka J, Altruda F, Tolosano E, Riganti C, Mussano F, and Munaron L
- Abstract
Bone formation involves a complex crosstalk between endothelial cells (EC) and osteodifferentiating stem cells. This functional interplay is greatly mediated by the paracrine and autocrine action of soluble factors released at the vasculature-bone interface. This study elucidates the molecular and functional responses triggered by this intimate interaction. In this study, we showed that human dermal microvascular endothelial cells (HMEC) induced the expression of pro-angiogenic factors in stem cells from human exfoliated deciduous teeth (SHED) and sustain their osteo-differentiation at the same time. In contrast, osteodifferentiating SHED increased EC recruitment and promoted the formation of complex vascular networks. Moreover, HMEC enhanced anaerobic glycolysis in proliferating SHED without compromising their ability to undergo the oxidative metabolic shift required for adequate osteo-differentiation. Taken together, these findings provide novel insights into the molecular mechanism underlying the synergistic cooperation between EC and stem cells during bone tissue renewal., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Petrillo, Genova, Chinigò, Roato, Scarpellino, Kopecka, Altruda, Tolosano, Riganti, Mussano and Munaron.)
- Published
- 2022
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29. Endothelial Heme Dynamics Drive Cancer Cell Metabolism by Shaping the Tumor Microenvironment.
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Petrillo S, De Giorgio F, Kopecka J, Genova T, Fiorito V, Allocco AL, Bertino F, Chiabrando D, Mussano F, Altruda F, Munaron L, Riganti C, and Tolosano E
- Abstract
The crosstalk among cancer cells (CCs) and stromal cells within the tumor microenvironment (TME) has a prominent role in cancer progression. The significance of endothelial cells (ECs) in this scenario relies on multiple vascular functions. By forming new blood vessels, ECs support tumor growth. In addition to their angiogenic properties, tumor-associated ECs (TECs) establish a unique vascular niche that actively modulates cancer development by shuttling a selected pattern of factors and metabolites to the CC. The profile of secreted metabolites is strictly dependent on the metabolic status of the cell, which is markedly perturbed in TECs. Recent evidence highlights the involvement of heme metabolism in the regulation of energy metabolism in TECs. The present study shows that interfering with endothelial heme metabolism by targeting the cell membrane heme exporter Feline Leukemia Virus subgroup C Receptor 1a (FLVCR1a) in TECs, resulted in enhanced fatty acid oxidation (FAO). Moreover, FAO-derived acetyl-CoA was partly consumed through ketogenesis, resulting in ketone bodies (KBs) accumulation in FLVCR1a-deficient TECs. Finally, the results from this study also demonstrate that TECs-derived KBs can be secreted in the extracellular environment, inducing a metabolic rewiring in the CC. Taken together, these data may contribute to finding new metabolic vulnerabilities for cancer therapy.
- Published
- 2021
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30. Oral Cavity as a Source of Mesenchymal Stem Cells Useful for Regenerative Medicine in Dentistry.
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Roato I, Chinigò G, Genova T, Munaron L, and Mussano F
- Abstract
The use of mesenchymal stem cells (MSCs) for regenerative purposes has become common in a large variety of diseases. In the dental and maxillofacial field, there are emerging clinical needs that could benefit from MSC-based therapeutic approaches. Even though MSCs can be isolated from different tissues, such as bone marrow, adipose tissue, etc., and are known for their multilineage differentiation, their different anatomical origin can affect the capability to differentiate into a specific tissue. For instance, MSCs isolated from the oral cavity might be more effective than adipose-derived stem cells (ASCs) for the treatment of dental defects. Indeed, in the oral cavity, there are different sources of MSCs that have been individually proposed as promising candidates for tissue engineering protocols. The therapeutic strategy based on MSCs can be direct, by using cells as components of the tissue to be regenerated, or indirect, aimed at delivering local growth factors, cytokines, and chemokines produced by the MSCs. Here, the authors outline the major sources of mesenchymal stem cells attainable from the oral cavity and discuss their possible usage in some of the most compelling therapeutic frontiers, such as periodontal disease and dental pulp regeneration.
- Published
- 2021
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31. Biohybrid Bovine Bone Matrix for Controlled Release of Mesenchymal Stem/Stromal Cell Lyosecretome: A Device for Bone Regeneration.
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Bari E, Roato I, Perale G, Rossi F, Genova T, Mussano F, Ferracini R, Sorlini M, Torre ML, and Perteghella S
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- Animals, Bone Substitutes chemistry, Cattle, Cell Differentiation drug effects, Delayed-Action Preparations chemistry, Delayed-Action Preparations pharmacology, Extracellular Vesicles chemistry, Extracellular Vesicles genetics, Humans, Mesenchymal Stem Cells chemistry, Mesenchymal Stem Cells drug effects, Osteoblasts drug effects, Osteogenesis drug effects, Plastic Surgery Procedures methods, Bone Matrix chemistry, Bone Regeneration drug effects, Bone Substitutes pharmacology, Mesenchymal Stem Cell Transplantation
- Abstract
SmartBone
® (SB) is a biohybrid bone substitute advantageously proposed as a class III medical device for bone regeneration in reconstructive surgeries (oral, maxillofacial, orthopedic, and oncology). In the present study, a new strategy to improve SB osteoinductivity was developed. SB scaffolds were loaded with lyosecretome, a freeze-dried formulation of mesenchymal stem cell (MSC)-secretome, containing proteins and extracellular vesicles (EVs). Lyosecretome-loaded SB scaffolds (SBlyo) were prepared using an absorption method. A burst release of proteins and EVs (38% and 50% after 30 min, respectively) was observed, and then proteins were released more slowly with respect to EVs, most likely because they more strongly adsorbed onto the SB surface. In vitro tests were conducted using adipose tissue-derived stromal vascular fraction (SVF) plated on SB or SBlyo. After 14 days, significant cell proliferation improvement was observed on SBlyo with respect to SB, where cells filled the cavities between the native trabeculae. On SB, on the other hand, the process was still present, but tissue formation was less organized at 60 days. On both scaffolds, cells differentiated into osteoblasts and were able to mineralize after 60 days. Nonetheless, SBlyo showed a higher expression of osteoblast markers and a higher quantity of newly formed trabeculae than SB alone. The quantification analysis of the newly formed mineralized tissue and the immunohistochemical studies demonstrated that SBlyo induces bone formation more effectively. This osteoinductive effect is likely due to the osteogenic factors present in the lyosecretome, such as fibronectin, alpha-2-macroglobulin, apolipoprotein A, and TGF-β.- Published
- 2021
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32. Isolation and Characterization of Buccal Fat Pad and Dental Pulp MSCs from the Same Donor.
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Genova T, Cavagnetto D, Tasinato F, Petrillo S, Ruffinatti FA, Mela L, Carossa M, Munaron L, Roato I, and Mussano F
- Abstract
Mesenchymal stem cells (MSCs) can be harvested from different sites in the oral cavity, representing a reservoir of cells useful for regenerative purposes. As direct comparisons between at least two types of MSCs deriving from the same patient are surprisingly rare in scientific literature, we isolated and investigated the osteoinductive potential of dental pulp stem cells (DPSCs) and buccal fat pad stem cells (BFPSCs). MSCs were isolated from the third molar dental pulp and buccal fat pads of 12 patients. The number of viable cells was quantified through manual count. Proliferation and osteodifferentiation assays, flow cytometry analysis of cell phenotypes, and osteocalcin release in vitro were performed. The isolation of BFPSCs and DPSCs was successful in 7 out of 12 (58%) and 3 out of 12 (25%) of retrieved samples, respectively. The yield of cells expressing typical stem cell markers and the level of proliferation were higher in BFPSCs than in DPSCs. Both BFP-SCs and DPSCs differentiated into osteoblast-like cells and were able to release a mineralized matrix. The release of osteocalcin, albeit greater for BFPSCs, did not show any significant difference between BFPSCs and DPSCs. The yield of MSCs depends on their site of origin as well as on the protocol adopted for their isolation. Our data show that BFP is a valuable source for the derivation of MSCs that can be used for regenerative treatments.
- Published
- 2021
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33. Individual mandibular movement registration and reproduction using an optoeletronic jaw movement analyzer and a dedicated robot: a dental technique.
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Carossa M, Cavagnetto D, Ceruti P, Mussano F, and Carossa S
- Subjects
- Dental Articulators, Jaw Relation Record, Mandible, Movement, Reproduction, Robotics
- Abstract
Background: Fully adjustable articulators and pantographs record and reproduce individual mandibular movements. Although these instruments are accurate, they are operator-dependant and time-consuming. Pantographic recording is affected by inter and intra operator variability in the individuation of clinical reference points and afterwards in reading pantographic recording themselves. Finally only border movements can be reproduced., Methods: Bionic Jaw Motion system is based on two components: a jaw movement analyzer and a robotic device that accurately reproduces recorded movements. The jaw movement analyzer uses an optoelectronic motion system technology made of a high frequency filming camera that acquires 140frames per second and a custom designed software that recognizes and determines the relative distance at each point in time of markers with known geometries connected to each jaw. Circumferential modified retainers connect markers and do not cover any occlusal surfaces neither obstruct occlusion. The recording process takes 5 to 10 s. Mandibular movement performance requires six degrees of freedom of movement, 3 rotations and 3 translations. Other robots are based on the so-called delta mechanics that use several parallel effectors to perform desired movements in order to decompose a complex trajectory into multiple more simple linear movements. However, each parallel effector introduces mechanical inter-component tolerances and mathematical transformations that are required to transform a recorded movement into the combination of movements to be performed by each effector. Bionic Jaw Motion Robot works differently, owing to three motors that perform translational movements and three other motors that perform rotations as a gyroscope. This configuration requires less mechanical components thus reducing mechanical tolerances and production costs. Both the jaw movement analyzer and the robot quantify the movement of the mandible as a rigid body with six degrees of freedom. This represents an additional advantage as no mathematical transformation is needed for the robot to reproduce recorded movements., Results: Based on the described procedure, Bionic Jaw Motion provide accurate recording and reproduction of maxillomandibular relation in static and dynamic conditions., Conclusion: This robotic system represents an important advancement compared to available analogical and digital alternatives both in clinical and research contexts for cost reduction, precision and time saving opportunities.
- Published
- 2020
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34. Advances on Bone Substitutes through 3D Bioprinting.
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Genova T, Roato I, Carossa M, Motta C, Cavagnetto D, and Mussano F
- Subjects
- Humans, Bioprinting instrumentation, Bioprinting methods, Bone Regeneration, Bone Substitutes chemistry, Printing, Three-Dimensional instrumentation, Tissue Engineering instrumentation, Tissue Engineering methods, Tissue Scaffolds chemistry
- Abstract
Reconstruction of bony defects is challenging when conventional grafting methods are used because of their intrinsic limitations (biological cost and/or biological properties). Bone regeneration techniques are rapidly evolving since the introduction of three-dimensional (3D) bioprinting. Bone tissue engineering is a branch of regenerative medicine that aims to find new solutions to treat bone defects, which can be repaired by 3D printed living tissues. Its aim is to overcome the limitations of conventional treatment options by improving osteoinduction and osteoconduction. Several techniques of bone bioprinting have been developed: inkjet, extrusion, and light-based 3D printers are nowadays available. Bioinks, i.e., the printing materials, also presented an evolution over the years. It seems that these new technologies might be extremely promising for bone regeneration. The purpose of the present review is to give a comprehensive summary of the past, the present, and future developments of bone bioprinting and bioinks, focusing the attention on crucial aspects of bone bioprinting such as selecting cell sources and attaining a viable vascularization within the newly printed bone. The main bioprinters currently available on the market and their characteristics have been taken into consideration, as well.
- Published
- 2020
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35. Fibroblast Interaction with Different Abutment Surfaces: In Vitro Study.
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Canullo L, Genova T, Gross Trujillo E, Pradies G, Petrillo S, Muzzi M, Carossa S, and Mussano F
- Subjects
- Cell Adhesion, Cell Proliferation physiology, Cells, Cultured, Fibroblasts drug effects, Humans, In Vitro Techniques, Surface Properties, Argon pharmacology, Fibroblasts cytology, Titanium chemistry
- Abstract
Background: Attaining an effective mucosal attachment to the transmucosal part of the implant could protect the peri-implant bone., Aim: To evaluate if chair side surface treatments (plasma of Argon and ultraviolet light) may affect fibroblast adhesion on different titanium surfaces designed for soft tissue healing., Methods: Grade 5 titanium discs with four different surface topographies were subdivided into 3 groups: argon-plasma; ultraviolet light, and no treatment. Cell morphology and adhesion tests were performed at 20 min, 24 h, and 72 h., Results: Qualitative observation of the surfaces performed at the SEM was in accordance with the anticipated features. Roughness values ranged from smooth (MAC Sa = 0.2) to very rough (XA Sa = 21). At 20 min, all the untreated surfaces presented hemispherical cells with reduced filopodia, while the cells on treated samples were more spread with broad lamellipodia. However, these differences in spreading behavior disappeared at 24 h and 72 h. Argon-plasma, but not UV, significantly increased the number of fibroblasts independently of the surface type but only at 20 min. Statistically, there was no surface in combination with a treatment that favored a greater cellular adhesion., Conclusions: Data showed potential biological benefits of treating implant abutment surfaces with the plasma of argon in relation to early-stage cell adhesion.
- Published
- 2020
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36. The Crosstalk Between Osteodifferentiating Stem Cells and Endothelial Cells Promotes Angiogenesis and Bone Formation.
- Author
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Genova T, Petrillo S, Zicola E, Roato I, Ferracini R, Tolosano E, Altruda F, Carossa S, Mussano F, and Munaron L
- Abstract
The synergistic crosstalk between osteodifferentiating stem cells and endothelial cells (ECs) gained the deserved consideration, shedding light on the role of angiogenesis for bone formation and healing. A deep understanding of the molecular basis underlying the mutual influence of mesenchymal stem cells (MSCs) and ECs in the osteogenic process may help improve greatly bone regeneration. Here, the authors demonstrated that osteodifferentiating MSCs co-cultured with ECs promote angiogenesis and ECs recruitment. Moreover, through the use of 3D co-culture systems, we showed that ECs are in turn able to further stimulate the osteodifferentiation of MSCs, thus enhancing bone production. These findings highlighted the existence of a virtuous loop between MSCs and ECs that is central to the osteogenic process. Unraveling the molecular mechanisms governing the functional interaction MSCs and ECs holds great potential in the field of regenerative medicine., (Copyright © 2019 Genova, Petrillo, Zicola, Roato, Ferracini, Tolosano, Altruda, Carossa, Mussano and Munaron.)
- Published
- 2019
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37. Proanthocyanidin may improve the shear bond strength at the composites/dentine interface.
- Author
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Autieri G, Mussano F, Petruzzi M, Carossa M, Genova T, Corsalini M, and Carossa S
- Subjects
- Humans, Molar, Shear Strength, Composite Resins chemistry, Dental Bonding methods, Dental Cements chemistry, Dentin, Proanthocyanidins
- Abstract
TThis study aimed at assessing whether proanthocyanidin, a collagen cross-linker, affects the adhesion strength of resin composites on the dentine surface. Freshly extracted, caries-free, human molars (N=55) were embedded in transparent resin and bisected. The halves were then assigned to either a treated or a non-treated group, where the treatment consisted of a 10 min incubation in a 6.5% proanthocyanidin solution in PBS. A resin composite cylinder was polymerized perpendicularly to the dentinal surfaces and shear tests were made, using an Instron-like machine. The fracture surfaces were characterized by optical (Picro-Sirius Red stain) and electron microscopy (FESEM EDX analysis). Mean bond strength values were 10.73 MPa (SD 3.70) for the treated group and 8.69 MPa (SD 3.20) for the non-treated group (p less than 0.05 Students t-test). No constant fracture patterns could be found within the two groups. Proanthocyanidin treatment may improve the adhesion properties of the dentine-bonding interface.
- Published
- 2018
38. Osteogenic Differentiation Modulates the Cytokine, Chemokine, and Growth Factor Profile of ASCs and SHED.
- Author
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Mussano F, Genova T, Petrillo S, Roato I, Ferracini R, and Munaron L
- Subjects
- Adipocytes metabolism, Biomarkers, Cell Survival, Cells, Cultured, Humans, Immunophenotyping, Mesenchymal Stem Cells cytology, Mesenchymal Stem Cells metabolism, Phenotype, Tooth, Deciduous cytology, Adipose Tissue cytology, Cell Differentiation, Cytokines metabolism, Osteogenesis, Stem Cells cytology, Stem Cells metabolism, Tooth, Deciduous metabolism
- Abstract
Great efforts have been made to improve bone regeneration techniques owing to a growing variety of sources of stem cells suitable for autologous transplants. Specifically, adipose-derived stem cells (ASCs) and stems cells from human exfoliated deciduous teeth (SHED) hold great potential for bone tissue engineering and cell therapy. After a preliminary characterization of the main biomolecules ASCs and SHED released in their conditioned media, cells were kept both in normal and osteo-inducing conditions. Conventional assays were performed to prove their osteogenic potential such as quantitative real-time polymerase chain reaction (qRT-PCR) (for RUNX-2, collagen type I, osteopontin and osteonectin), alkaline phosphatase activity, osteocalcin production, and von Kossa staining. Conditioned media were tested again after the osteogenic induction and compared to maintaining condition both at base line and after 14 days of culture. The osteogenic condition inhibited the release of all the biomolecules, with the exception, concerning SHED, of growth-regulated alpha protein precursor (GROα), and, to a lesser extent, interleukin (IL)-8. In conclusion, our data support that undifferentiated ASCs and SHED may be preferable to committed ones for general cell therapy approaches, due to their higher paracrine activity. Osteoinduction significantly affects the cytokine, chemokine, and growth factor profile in a differential way, as SHED kept a more pronounced pro-angiogenic signature than ASCs., Competing Interests: The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.
- Published
- 2018
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39. Adipose-Derived Stromal Vascular Fraction/Xenohybrid Bone Scaffold: An Alternative Source for Bone Regeneration.
- Author
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Roato I, Belisario DC, Compagno M, Verderio L, Sighinolfi A, Mussano F, Genova T, Veneziano F, Pertici G, Perale G, and Ferracini R
- Abstract
Adipose tissue-derived stem cells (ASCs) are a promising tool for the treatment of bone diseases or skeletal lesions, thanks to their ability to potentially repair damaged tissue. One of the major limitations of ASCs is represented by the necessity to be isolated and expanded through in vitro culture; thus, a strong interest was generated by the adipose stromal vascular fraction (SVF), the noncultured fraction of ASCs. SVF is a heterogeneous cell population, directly obtained after collagenase treatment of adipose tissue. In order to investigate and compare the bone-regenerative potential of SVF and ASCs, they were plated on SmartBone®, a xenohybrid bone scaffold, already used in clinical practice with successful results. We showed that SVF plated on SmartBone, in the presence of osteogenic factors, had better osteoinductive capabilities than ASCs, in terms of differentiation into bone cells, mineralization, and secretion of soluble factors stimulating osteoblasts. Indeed, we observed an increasing area of new tissue over time, with and without OM. These data strongly support an innovative idea for the use of adipose SVF and bone scaffolds to promote tissue regeneration and repair, also thanks to an easier cell management preparation that allows a potentially larger use in clinical applications.
- Published
- 2018
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40. Apical periodontitis: preliminary assessment of microbiota by 16S rRNA high throughput amplicon target sequencing.
- Author
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Mussano F, Ferrocino I, Gavrilova N, Genova T, Dell'Acqua A, Cocolin L, and Carossa S
- Subjects
- Adult, Female, High-Throughput Nucleotide Sequencing methods, Humans, Male, Periapical Granuloma microbiology, Pilot Projects, Radicular Cyst microbiology, Young Adult, Microbiota genetics, Periapical Periodontitis microbiology, RNA, Ribosomal, 16S genetics
- Abstract
Background: Apical periodontitis includes periapical granulomas and radicular cysts, which are histologically distinguished by the absence and the presence of an epithelial lining, respectively. The main cause of apical periodontitis is the bacterial colonization of the root canal space. This research aimed at assessing whether and how periapical granulomas and radicular cysts differ in terms of microbiota using high throughput amplicon target sequencing (HTS) techniques., Methods: This study included 5 cases of Periapical Granulomas (PGs) and 5 cases of Radicular Cysts (RCs) selected on the base of histology out of 37 patients from January 2015 to February 2016. Complete medical history, panoramic radiograms (OPTs) and histologic records of each patient were assessed. Only lesions greater than 1 cm in diameter and developed in proximity to teeth with bad prognosis were included. The microbiota present in periapical granulomas and radicular cysts thus retrieved was finely characterized by pyrosequencing of the 16S rRNA genes., Results: The core of OTUs shared between periapical granulomas and radicular cysts was dominated by the presence of facultative anaerobes taxa such as: Lactococcus lactis, Propionibacterium acnes, Staphylococcus warneri, Acinetobacter johnsonii and Gemellales. L. lactis, the main OTUs of the entire datasets, was associated with periapical granuloma samples. Consistently with literature, the anaerobic taxa detected were most abundant in radicular cyst samples. Indeed, a higher abundance of presumptive predicted metabolic pathways related to Lipopolysaccharide biosynthesis was found in radicular cyst samples., Conclusions: The present pilot study confirmed the different microbial characterization of the two main apical periodontitis types and shade light on the possible role of L. lactis in periapical granulomas.
- Published
- 2018
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41. Heme accumulation in endothelial cells impairs angiogenesis by triggering paraptosis.
- Author
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Petrillo S, Chiabrando D, Genova T, Fiorito V, Ingoglia G, Vinchi F, Mussano F, Carossa S, Silengo L, Altruda F, Merlo GR, Munaron L, and Tolosano E
- Subjects
- Animals, Cells, Cultured, Endoplasmic Reticulum Stress genetics, Female, Heme genetics, Humans, Male, Membrane Transport Proteins deficiency, Membrane Transport Proteins genetics, Mice, Mice, Knockout, Neovascularization, Pathologic genetics, Neovascularization, Pathologic pathology, Receptors, Virus deficiency, Receptors, Virus genetics, Apoptosis genetics, Endothelial Cells metabolism, Heme metabolism, Membrane Transport Proteins metabolism, Neovascularization, Pathologic metabolism, Receptors, Virus metabolism
- Abstract
Heme is required for cell respiration and survival. Nevertheless, its intracellular levels need to be finely regulated to avoid heme excess, which may catalyze the production of reactive oxygen species (ROS) and promote cell death. Here, we show that alteration of heme homeostasis in endothelial cells due to the loss of the heme exporter FLVCR1a, results in impaired angiogenesis. In vitro, FLVCR1a silencing in endothelial cells causes defective tubulogenesis and poor viability due to intracellular heme accumulation. Consistently, endothelial-specific Flvcr1a knockout mice show aberrant angiogenesis responsible for hemorrhages and embryonic lethality. Importantly, we demonstrate that impaired heme export leads to endothelial cell death by paraptosis and provide evidence that endoplasmic reticulum (ER) stress precedes heme-induced paraptosis. These findings highlight a crucial role for the cytosolic heme pool in the control of endothelial cell survival and in the regulation of the angiogenic process. Interfering with endothelial heme export represents a valuable model for a deeper understanding of the molecular mechanisms underlying heme-triggered paraptosis and, in the future, might provide a novel tool for the modulation of angiogenesis in pathophysiologic conditions.
- Published
- 2018
- Full Text
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42. Nano-Pore Size of Alumina Affects Osteoblastic Response.
- Author
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Mussano F, Genova T, Serra FG, Carossa M, Munaron L, and Carossa S
- Subjects
- Animals, Cell Adhesion, Cell Culture Techniques methods, Cell Differentiation, Cell Line, Mice, Osteoblasts cytology, Osteoblasts physiology, Tissue Scaffolds adverse effects, Aluminum Oxide chemistry, Nanopores, Osteoblasts drug effects, Tissue Scaffolds chemistry
- Abstract
The rapid development and application of nanotechnology to biological interfaces has impacted the bone implant field, allowing researchers to finely modulate the interface between biomaterials and recipient tissues. In the present study, oxidative anodization was exploited to generate two alumina surfaces with different pore diameters. The former displayed surface pores in the mean range of 16-30 nm, while in the latter pores varied from to 65 to 89 nm. The samples were characterized by Field Emission Scanning Electron Microscopy (FESEM) and Energy Dispersive X-ray spectroscopy (EDX) analysis prior to being tested with pre-osteoblastic MC3T3-E1 cells. In vitro cell response was studied in terms of early cell adhesion, viability, and morphology, including focal adhesion quantification. Both the alumina samples promoted higher cell adhesion and viability than the control condition represented by the standard culture dish plastic. Osteogenic differentiation was assessed through alkaline phosphatase activity and extracellular calcium deposition, and it was found that of the two nano-surfaces, one was more efficient than the other. By comparing for the first time two nano-porous alumina surfaces with different pore diameters, our data supported the role of nano-topography in inducing cell response. Modulating a simple aspect of surface texture may become an attractive route for guiding bone healing and regeneration around implantable metals., Competing Interests: The authors declare no conflict of interest.
- Published
- 2018
- Full Text
- View/download PDF
43. An alumina toughened zirconia composite for dental implant application: in vivo animal results.
- Author
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Schierano G, Mussano F, Faga MG, Menicucci G, Manzella C, Sabione C, Genova T, von Degerfeld MM, Peirone B, Cassenti A, Cassoni P, and Carossa S
- Subjects
- Animals, Biomarkers metabolism, Bone Density drug effects, Bone and Bones cytology, Bone and Bones drug effects, Bone and Bones metabolism, Gene Expression Regulation drug effects, Osteogenesis drug effects, Osteogenesis genetics, Surface Properties, Swine, Swine, Miniature, Aluminum Oxide pharmacology, Dental Implants, Materials Testing methods, Zirconium pharmacology
- Abstract
Ceramic materials are widely used for biomedical applications because of their remarkable biological and mechanical properties. Composites made of alumina and zirconia are particularly interesting owing to their higher toughness with respect to the monolithic materials. On this basis, the present study is focused on the in vivo behavior of alumina toughened zirconia (ATZ) dental implants treated with a hydrothermal process. A minipig model was implemented to assess the bone healing through histology and mRNA expression at different time points (8, 14, 28, and 56 days). The novel ATZ implant was compared to a titanium clinical standard. The implants were analyzed in terms of microstructure and surface roughness before in vivo tests. The most interesting result deals with a statistically significant higher digital histology index for ATZ implants with respect to titanium standard at 56 days, which is an unprecedented finding, to the authors' knowledge. Even if further investigations are needed before proposing the clinical use in humans, the tested material proved to be a promising candidate among the possible ceramic dental implants.
- Published
- 2015
- Full Text
- View/download PDF
44. Guiding the osteogenic fate of mouse and human mesenchymal stem cells through feedback system control.
- Author
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Honda Y, Ding X, Mussano F, Wiberg A, Ho CM, and Nishimura I
- Subjects
- Animals, Bone Morphogenetic Proteins metabolism, Culture Media chemistry, Culture Media pharmacology, Humans, Mesenchymal Stem Cells drug effects, Mice, Signal Transduction drug effects, Smad Proteins metabolism, Cell Differentiation drug effects, Mesenchymal Stem Cells cytology, Mesenchymal Stem Cells metabolism, Models, Biological, Osteogenesis drug effects
- Abstract
Stem cell-based disease modeling presents unique opportunities for mechanistic elucidation and therapeutic targeting. The stable induction of fate-specific differentiation is an essential prerequisite for stem cell-based strategy. Bone morphogenetic protein 2 (BMP-2) initiates receptor-regulated Smad phosphorylation, leading to the osteogenic differentiation of mesenchymal stromal/stem cells (MSC) in vitro; however, it requires supra-physiological concentrations, presenting a bottleneck problem for large-scale drug screening. Here, we report the use of a double-objective feedback system control (FSC) with a differential evolution (DE) algorithm to identify osteogenic cocktails of extrinsic factors. Cocktails containing significantly reduced doses of BMP-2 in combination with physiologically relevant doses of dexamethasone, ascorbic acid, beta-glycerophosphate, heparin, retinoic acid and vitamin D achieved accelerated in vitro mineralization of mouse and human MSC. These results provide insight into constructive approaches of FSC to determine the applicable functional and physiological environment for MSC in disease modeling, drug screening and tissue engineering.
- Published
- 2013
- Full Text
- View/download PDF
45. Circadian rhythm and cartilage extracellular matrix genes in osseointegration: a genome-wide screening of implant failure by vitamin D deficiency.
- Author
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Mengatto CM, Mussano F, Honda Y, Colwell CS, and Nishimura I
- Subjects
- Animals, Circadian Rhythm physiology, Femur, Gene Expression Profiling, Gene Regulatory Networks, Genome, Genome-Wide Association Study, Implants, Experimental, Male, Oligonucleotide Array Sequence Analysis, Rats, Rats, Sprague-Dawley, Titanium, Cartilage ultrastructure, Circadian Rhythm genetics, Extracellular Matrix genetics, Osseointegration genetics, Prosthesis Failure etiology, Vitamin D Deficiency complications
- Abstract
Background: Successful dental and orthopedic implants require the establishment of an intimate association with bone tissue; however, the mechanistic explanation of how biological systems accomplish osseointegration is still incomplete. We sought to identify critical gene networks involved in osseointegration by exploring the implant failure model under vitamin D deficiency., Methodology: Adult male Sprague-Dawley rats were exposed to control or vitamin D-deficient diet prior to the osteotomy surgery in the femur bone and the placement of T-shaped Ti4Al6V implant. Two weeks after the osteotomy and implant placement, tissue formed at the osteotomy site or in the hollow chamber of T-shaped implant was harvested and total RNA was evaluated by whole genome microarray analyses., Principal Findings: Two-way ANOVA of microarray data identified 103 genes that were significantly (>2 fold) modulated by the implant placement and vitamin D deficiency. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses assigned the highest z-score to the circadian rhythm pathway including neuronal PAS domain 2 (NPAS2), and period homolog 2 (Per2). NPAS2 and Aryl hydrocarbon receptor nuclear translocator-like (ARNTL/Bmal 1) were upregulated around implant and diminished by vitamin D deficiency, whereas the expression pattern of Per2 was complementary. Hierarchical cluster analysis further revealed that NPAS2 was in a group predominantly composed of cartilage extracellular matrix (ECM) genes. Whereas the expression of bone ECM genes around implant was not significantly affected by vitamin D deficiency, cartilage ECM genes were modulated by the presence of the implant and vitamin D status. In a proof-of-concept in vitro study, the expression of cartilage type II and X collagens was found upregulated when mouse mesenchymal stem cells were cultured on implant disk with 1,25D supplementation., Conclusions: This study suggests that the circadian rhythm system and cartilage extracellular matrix may be involved in the establishment of osseointegration under vitamin D regulation.
- Published
- 2011
- Full Text
- View/download PDF
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