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298 results on '"Becchetti, Ennio"'

15. Effects of Titanium Surface Roughness on Mesenchymal Stem Cell Commitment and Differentiation Signaling.

Author

Balloni, Stefania, Calvi, Edoardo Maria, Damiani, Francesco, Bistoni, Giovanni, Calvitti, Mario, Locci, Paola, Becchetti, Ennio, and Marinucci, Lorella

Subjects
TITANIUM, EXTRACELLULAR matrix proteins, ELECTRON microscopy, MESSENGER RNA, GENE expression, GENETIC regulation, CELL growth, STEM cells, CELL adhesion, CELL communication
Abstract

Purpose: Human mesenchymal stem cells (hMSCs) are primary cells capable of differentiating to osteocytic lineage when stimulated under appropriate conditions. This study examined changes in hMSC morphology, proliferation, and gene expression after growth on machined or dual acid-etched (AE) titanium surfaces. Materials and Methods: hMSCs, isolated from adult human bone marrow, were cultured on titanium surfaces. The two specimens of titanium surfaces in this study included machined and AE titanium disks. Cell morphology was evaluated by scanning electron microscopy, and cell proliferation and collagen synthesis were estimated by measuring the amount of 3H-thymidine incorporation into DNA and ³H-proline incorporation into collagen fibers. Alkaline phosphatase (ALP) activity was determined by measuring the release of p-nitrophenol from disodium p-nitrophenyl phosphate. Changes in gene expression for bone morphogenetic protein-2 (BMP-2), Runx2 type II, Osterix (Osx), osteopontin, type I collagen, ALP, osteocalcin, and bone sialoprotein were determined by reverse-transcriptase polymerase chain reaction after 22 days of in vitro culture in osteogenic medium. Results: The two substrates had no significant effects on cell adhesion and proliferation. Morphologic characteristics were observed by scanning electron microscopy. hMSCs on the machined surface spread more and were flatter than cells cultured on the AE surface. Osteopontin mRNA expression was similar on all surfaces, and the other mRNA transcripts were increased in hMSC cultured on AE surface. In particular, BMP-2, Runx2, and Osx, three osteogenic factors that induce the progressive differentiation of multipotent mesenchymal cells into osteoblasts, were expressed more on AE titanium than on machined titanium. Collagen and ALP assays confirmed the highest level of mRNA transcripts correlated with increases in these proteins. Conclusion: These results showed that an AE titanium surface stimulated the expression of markers of osteoblastic phenotype more than a machined titanium surface. [ABSTRACT FROM AUTHOR]

Published
2009

16. Effect of Titanium Surface Roughness on Human Osteoblast Proliferation and Gene Expression In Vitro.

Author

Marinucci, Lorella, Balloni, Stefania, Becchetti, Ennio, Belcastro, Salvatore, Guerra, Mario, Calvitti, Mario, Lull, Cinzia, Calvi, Edoardo Maria, and Locci, Paola

Subjects
BONE cells, CELL proliferation, GENE expression, TITANIUM, SURFACE roughness, EXTRACELLULAR matrix, REVERSE transcriptase, POLYMERASE chain reaction
Abstract

Purpose: Cell proliferation and extracellular matrix formation are primary events in bone formation. At the dental implant-tissue interface, implant surface roughness modulates osteoblast functions. The aim of the present in vitro study was to investigate the effect of varying surface roughness of titanium implant material on cell proliferation and mRNA expression of specific markers of osteoblast phenotype. Materials and Methods: Primary cultures of osteoblasts derived from human mandibular bone were cultured on titanium surfaces. Three titanium surfaces were studied: machined titanium, micro-sandblasted titanium, and macro-sandblasted titanium (average surface roughnesses of 0.5 and 3 μm, respectively). Cell morphology was estimated by scanning electron microscope analysis and cell proliferation by measuring the amount of 3H-thymidine incorporation into DNA. mRNA expression of osteonectin, osteopontin, bone sialoprotein (BSP), and Runx2, which are markers of osteoblastic phenotype, were determined by reverse transcriptase polymerase chain reaction (RT-PCR) analysis. Results: Human osteoblasts cultured on machined titanium spread more and were flatter than cells cultured on rough titanium. All blasted surfaces showed significantly higher DNA synthesis than the machined surfaces. Osteonectin mRNA expression was similar on all surfaces. Other mRNA transcripts were increased in osteoblasts cultured on rough titanium surfaces, particularly the macro-sandblasted surface. Conclusions: An average surface roughness of 3 μm (macro-sandblasted titanium) is more suitable than an average surface roughness of 0.5 μm (micro-sandblasted titanium) in favoring osteoblast differentiation in vitro. [ABSTRACT FROM AUTHOR]

Published
2006

17. Effects of cadmium on viability and function of porcine pre-pubertal Sertoli cells

Author

Luca, Giovanni, Lilli, Cinzia, Bellucci, Catia, Mancuso, Francesca, Calvitti, Mario, Arato, Iva, Falabella, G., Aglietti, Maria Chiara, Becchetti, Ennio, Lumare, A., Calafiore, Riccardo, and Bodo, Maria

Subjects
Cadmium, Sertoli, function
Abstract

Cadmium, an ubiquitous environmental pollutant mainly used for industrial purposes, is highly associated with reproductive toxicity. Sertoli cells (SC), by providing an appropriate microenvironment for the development of germ cells, play a pivotal role on spermatogenesis regulation (Geoffroy-Siraudin et al. 2012). Aim of our investigation was to assess the effects of cadmium on high mammalian SC viability and function. Porcine pre-pubertal SC were isolated, according to previously established methods (Fallarino et al. 2009) and treated with 3 different concentrations (5-10-15 μM) of cadmium chloride. The evaluation of SC function in terms of inhibin B and anti-Müllerian hormone (AMH) secretion showed a significant decrease in all SC treated conditions respect as compared to SC control. On the contrary, evaluation of the FSH-R integrity on SC surface, in terms of 17-b-estradiol production under FSH stimulation, showed no difference between SC control and 5 μM cadmium treated SC monolayers in comparison to 10 and 15μM cadmium treated SC monolayers, where FSH-R was impaired. In addition, the apoptotic test showed a significant increase of early (ANNEXIN V-/Propidium Iodide+) (AV-/PI+) and late apoptotic cells (AV+/ PI+) in all cadmium treated SC conditions in comparison with SC control. In conclusion, our data demonstrate that cadmium, even at low dose, exerts toxic effects on Sertoli cells function possibly adversely affecting the spermatogenesis., Italian Journal of Anatomy and Embryology, Vol 117, No 2 (Supplement) 2012

Published
2013
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23. I tessuti connettivi

Author

Becchetti, Ennio and Calvitti, Mario

Subjects
tessuti connettivi, Istologia
Published
2012

24. Sertoli Cells as potential drug carriers: uptake and stability

Author

Giovagnoli, Stefano, Luca, Giovanni, Mancuso, Francesca, Calvitti, Mario, Piroddi, M., Schoubben, Aurelie Marie Madeleine, Blasi, Paolo, Galli, Francesco, Moretti, Massimo, Becchetti, Ennio, Ricci, Maurizio, and Calafiore, Riccardo

Published
2012

28. Xenograft of free or microencapsulated Sertoli cells as a potential therapy for experimental Type 2 Diabetes Mellitus

Author

Luca, Giovanni, Calvitti, Mario, Mancuso, Francesca, Murdolo, G., Arato, Iva, Falabella, G., Tortoioli, C., Lilli, Cinzia, Bellucci, Catia, Baroni, Tiziano, Bodo, Maria, Becchetti, Ennio, and Calafiore, Riccardo

Subjects
xenograft, Sertoli cell, Type 2 diabetes
Abstract

Introduction and Aim. Type 2 Diabetes Mellitus (T2DM), one of the world’s most important, common and costly diseases associated with devastating complications, is caused by insulin resistance mainly due to a chronic inflammation of the visceral adipose tissue with local and systemic increases in proinflammatory cytokines and adipokines such as tumor necrosis factor-α (TNF-α) and IL-6. Sertoli cells (SC), considered mere mechanical cell aids, have been recently revisited with respect to their functional competence showing that these cells may provide immunomodulatory and trophic factors that are able to ameliorate survival and development of different cell types and constitute an immuno-protective shield for transplantation in many diseases such as Type 1 Diabetes Mellitus. Aim of this work was to verify if the injection of free (subcutaneously) or microencapsulated (intraperitoneally) SC would reverse hyperglycaemia in db/db mice spontaneous T2DM Materials and Methods. Porcine pre-pubertal SC were isolated, according to previously established methods, after finely chopping the retrieval testicles, with double enzymatic digestion with Collagenase P and a mixed solution of trypsin and DNase I. SC enveloped in Barium alginate-based microcapsules (Ba-SCMCs) were prepared according to our method, by a mono air-jet device system. Free SC and Ba-SCMCs were examined as far as: (a) SC morphology by light microscopy; (b) SC viability, by fluorescence microscopy after staining with ethidium bromide and fluorescein diacetate; (c) SC in vitro function (α-aromatase activity and IGF-I secretion); (d) reversal of T2DM in spontaneous diabetes db/db mice, were concerned. Results. Ba-SCMCs showed excellent features in terms of size, morphology, sphericity and coalescence. SC viability, both in free and microencapsulated SC, was very high (over 90%). Very good α-aromatase activity and IGF-I secretion were associated with the examined SC preparations. Both subcutaneous free SC injection and intraperitoneal transplantation of Ba-SCMCs demonstrated a significant reduction, in 60% of the treated mice, of HbA1c (6.6 % vs 8.8 %) with a normalization of intraperitoneal glucose tolerance test. Conclusions. SC may be enveloped in Ba-SCMCs with no loss of their functional properties and morphology. Xenograft of SC, both free and enveloped in barium alginate microcapsules, induced an important reduction of HbA1c blood levels with a normalization of glucose tolerance test (IPGTT). This result might open new perspectives for the future therapy of human T2DM., Italian Journal of Anatomy and Embryology, Vol 116, No 1 (Supplement) 2011

Published
2011
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31. Xenograft of microencapsulated Sertoli cells as a therapy for experimental Laron syndrome

Author

Luca, Giovanni, Calvitti, Mario, Falabella, Giulia, Bellucci, Catia, Mancuso, Francesca, Berryman, Darlene, Arato, Iva, List, Edward, Becchetti, Ennio, Kopchick, John, Baroni, Tiziano, and Calafiore, Riccardo

Subjects
Laron, Sertoli, transplantation
Abstract

Italian Journal of Anatomy and Embryology, Vol 115, No 1/2 (Supplement) 2010

Published
2010

47. Tessuti connettivi

Author

Becchetti, Ennio and Calvitti, Mario

Subjects
tessuti connettivi, matrice extracellulare
Published
2007

50. New insights on Pancreatic Islet Encapsulation

Author

Blasi, Paolo, Giovagnoli, Stefano, Ricci, Maurizio, Luca, Giovanni, Mancuso, Francesca, Calvitti, Mario, Becchetti, Ennio, Calafiore, Riccardo, and Rossi, Carlo

Subjects
cell incapsulation, alginate, coherent microcapsules
Published
2007

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