Publications by authors named "Annalisa Palmieri"

86 Publications

Non-syndromic Cleft Palate: An Overview on Human Genetic and Environmental Risk Factors.

Front Cell Dev Biol 2020 20;8:592271. Epub 2020 Oct 20.

Department of Experimental, Diagnostic and Specialty Medicine, Alma Mater Studiorum - University of Bologna, Bologna, Italy.

The epithelial and mesenchymal cells involved in early embryonic facial development are guided by complex regulatory mechanisms. Any factor perturbing the growth, approach and fusion of the frontonasal and maxillary processes could result in orofacial clefts that represent the most common craniofacial malformations in humans. The rarest and, probably for this reason, the least studied form of cleft involves only the secondary palate, which is posterior to the incisive foramen. The etiology of cleft palate only is multifactorial and involves both genetic and environmental risk factors. The intention of this review is to give the reader an overview of the efforts made by researchers to shed light on the underlying causes of this birth defect. Most of the scientific papers suggesting potential environmental and genetic causes of non-syndromic cleft palate are summarized in this review, including genome-wide association and gene-environment interaction studies.
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http://dx.doi.org/10.3389/fcell.2020.592271DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7606870PMC
October 2020

Drug-Induced Gingival Overgrowth: A Pilot Study on the Effect of Diphenylhydantoin and Gabapentin on Human Gingival Fibroblasts.

Int J Environ Res Public Health 2020 11 7;17(21). Epub 2020 Nov 7.

Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, 44121 Ferrara, Italy.

Introduction: The administration of several classes of drugs can lead to the onset of gingival overgrowth: anticonvulsants, immunosuppressants, and calcium channel blockers. Among the anticonvulsants, the main drug associated with gingival overgrowth is diphenylhydantoin.

Materials And Methods: In this study, we compared the effects of diphenylhydantoin and gabapentin on 57 genes belonging to the "Extracellular Matrix and Adhesion Molecule" pathway, present in human fibroblasts of healthy volunteers.

Results: Both molecules induce the same gene expression profile in fibroblasts as well as a significant upregulation of genes involved in extracellular matrix deposition like COL4A1, ITGA7, and LAMB3. The two treatments also induced a significant downregulation of genes involved in the expression of extracellular matrix metalloproteases like MMP11, MMP15, MMP16, MMP24, and transmembrane receptor ITGB4.

Conclusions: Data recorded in our study confirmed the hypothesis of a direct action of these drugs at the periodontium level, inducing an increase in matrix production, a reduction in its degradation, and consequently resulting in gingival hyperplasia.
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http://dx.doi.org/10.3390/ijerph17218229DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7664415PMC
November 2020

Drug-Induced Gingival Overgrowth: The Effect of Cyclosporin A and Mycophenolate Mophetil on Human Gingival Fibroblasts.

Biomedicines 2020 Jul 17;8(7). Epub 2020 Jul 17.

Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, 44121 Ferrara, Italy.

Drug-induced gingival overgrowth may occur after a chronic administration of three classes of systemic drugs: Anticonvulsants, immunosuppressants, and calcium channel blockers. This study aimed to investigate how cyclosporin A and mycophenolate mophetil (immunosuppressive drugs) could interfere with human gingival fibroblasts functions, leading to gingival enlargement. Human gingival fibroblasts derived from the tissue of a 60-year-old female were cultured in a DMEME medium. A stock solution with 1 mg/mL of mycophenolate and 1 mg/mL of cyclosporine were prepared and dissolved in a DMEM medium to prepare a serial dilution at the concentrations of 5000, 2000, 1000, 500, and 100 ng/mL, for both treatments. Cell viability was measured using the PrestoBlue™ Reagent Protocol. Quantitative real-time RT-PCR was performed in order to analyze the expression of 57 genes coding for gingival fibroblasts "Extracellular Matrix and Adhesion Molecules". Mycophenolate and cyclosporine had no effect on fibroblast cell viability at 1000 ng/mL. Both the treatments showed similar effects on the expression profiling of treated cells: Downregulation of most extracellular matrix metalloproteases genes (, , , , ) was assessed, while , , , , , and were recorded to be upregulated in fibroblasts treated with immunosuppressive drugs. It has been demonstrated that gingival overgrowth can be caused by the chronic administration of cyclosporin A and mycophenolate mophetil. However, given the contrasting data of literature, further investigations are needed, making clear the possible effects of immunosuppressive drugs on fibroblasts.
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http://dx.doi.org/10.3390/biomedicines8070221DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7400382PMC
July 2020

Role of Cyclosporine in Gingival Hyperplasia: An In Vitro Study on Gingival Fibroblasts.

Int J Mol Sci 2020 Jan 16;21(2). Epub 2020 Jan 16.

Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, 44121 Ferrara, Italy.

Background: Gingival hyperplasia could occur after the administration of cyclosporine A. Up to 90% of the patients submitted to immunosuppressant drugs have been reported to suffer from this side effect. The role of fibroblasts in gingival hyperplasia has been widely discussed by literature, showing contrasting results. In order to demonstrate the effect of cyclosporine A on the extracellular matrix component of fibroblasts, we investigated the gene expression profile of human fibroblasts after cyclosporine A administration.

Materials And Methods: Primary gingival fibroblasts were stimulated with 1000 ng/mL cyclosporine A solution for 16 h. Gene expression levels of 57 genes belonging to the "Extracellular Matrix and Adhesion Molecules" pathway were analyzed using real-time PCR in treated cells, compared to untreated cells used as control.

Results: Expression levels of different genes were significantly de-regulated. The gene , which codes for the cell adhesion protein E-cadherin, showed up-regulation. Almost all the extracellular matrix metalloproteases showed down-regulation (, , , , , ). The administration of cyclosporine A was followed by down-regulation of other genes: , the transmembrane receptors and , and the basement membrane constituents and .

Conclusion: Data collected demonstrate that cyclosporine inhibits the secretion of matrix proteases, contributing to the accumulation of extracellular matrix components in the gingival connective tissue, causing gingival overgrowth. Patients affected by gingival overgrowth caused by cyclosporine A need to be further investigated in order to determine the role of this drug on fibroblasts.
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http://dx.doi.org/10.3390/ijms21020595DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7014429PMC
January 2020

ROCK1 is associated with non-syndromic cleft palate.

J Oral Pathol Med 2020 Feb 24;49(2):164-168. Epub 2019 Nov 24.

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy.

Background: Craniofacial morphogenesis is the result of an intricate multistep network of tightly controlled spatial and temporal signalling that involves several molecules and transcription factors organized into highly coordinated pathways. Any alteration in even one step of this delicate process can lead to congenital malformations such as cleft palate. One of the first steps in embryonal orofacial development is the migration of cells from the neural crests to the branchial arches. Next, the cells have to proliferate, differentiate, move and connect to each other in order to correctly form the palate. Cell contraction, promoted by the interaction of non-muscle myosin II and actin A, is a crucial step in morphogenesis and is regulated by ROCK1 protein.

Methods: A family-based association study was carried out in order to verify whether or not genetic variants of ROCK1 were associated with non-syndromic cleft palate (nsCP). Two cohorts from Italy and Iran, a total of 189 nsCP cases and their parents were enrolled.

Results: The rs35996865-G allele was under-transmitted in cases of nsCP [P = .006, odds ratio (OR) = 0.63 (95% CI 0.45-0.88)].

Conclusion: This investigation reveals for the first time data supporting a role for ROCK1 in nsCP aetiology.
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http://dx.doi.org/10.1111/jop.12973DOI Listing
February 2020

Drug-induced gingival hyperplasia: An in vitro study using amlodipine and human gingival fibroblasts.

Int J Immunopathol Pharmacol 2019 Jan-Dec;33:2058738419827746

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy.

Gingival overgrowth is a serious side effect that accompanies the use of amlodipine. Several conflicting theories have been proposed to explain the fibroblast's function in gingival overgrowth. To determine whether amlodipine alters the inflammatory responses, we investigated its effects on gingival fibroblast gene expression as compared with untreated cells. Fragments of gingival tissue of healthy volunteers (11 years old boy, 68 years old woman, and 20 years old men) were collected during operation. Gene expression of 29 genes was investigated in gingival fibroblast cell culture treated with amlodipine, compared with untreated cells. Among the studied genes, only 15 (, and ) were significantly deregulated. In particular, the most evident overexpressed genes in treated cells were and . These results seem to indicate a possible role of amlodipine in the inflammatory response of treated human gingival fibroblasts.
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http://dx.doi.org/10.1177/2058738419827746DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6822186PMC
March 2020

Non-syndromic cleft palate: Association analysis on three gene polymorphisms of the folate pathway in Asian and Italian populations.

Int J Immunopathol Pharmacol 2019 Jan-Dec;33:2058738419858572

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy.

Periconceptional folic acid supplementation can reduce the risk of inborn malformations, including orofacial clefts. Polymorphisms of MTHFR, TCN2, and CBS folate-related genes seem to modulate the risk of cleft lip with or without cleft palate (CL/P) in some populations. CL/P and cleft palate only (CPO) are different malformations that share several features and possibly etiological causes. In the present investigation, we conducted a family-based, candidate gene association study of non-syndromic CPO. Three single nucleotide polymorphisms, namely, rs1801133 of MTHFR, rs1801198 of TCN2, and rs4920037 of CBS, were investigated in a sample that included 129 Italian and 65 Asian families. No evidence of association between the three genotyped polymorphisms and CPO was found in the Italian and Asian cases, indeed the transmission disequilibrium test did not detect any asymmetry of transmission of alleles. This investigation, although with some limitation, further supports that CL/P and CPO diverge in their genetic background.
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http://dx.doi.org/10.1177/2058738419858572DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6822179PMC
March 2020

Copy number variation analysis of twin pairs discordant for cleft lip with or without cleft palate.

Int J Immunopathol Pharmacol 2019 Jan-Dec;33:2058738419855873

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy.

Non-syndromic cleft lip with or without cleft palate (nsCL/P) is a frequent orofacial malformation. The comparison of concordance rate observed in monozygotic and dizygotic twins supports high level of heritability and a strong genetic component. However, phenotype concordance for orofacial cleft in monozygotic twins is about 50%. The aim of the present investigation was to detect postzygotic events that may account for discordance in monozygotic twins. High-density SNP microarrays hybridization was used to genotype two pairs of monozygotic twins discordant for nsCL/P. Discordant SNP genotypes and copy number variants were analyzed to identify genetic differences responsible of phenotype discrepancy. A number of differences were observed, none involving known nsCL/P candidate genes or genomic regions. Considering the limitation of the study, related to the small sample size and to the large-scale investigation method, the results suggest that the detection of discordant events in other monozygotic twin pairs would be remarkable and warrant further investigations.
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http://dx.doi.org/10.1177/2058738419855873DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6822181PMC
March 2020

Berberine and exert a potential anticancer effect on colon cancer cells by acting on specific pathways.

Int J Immunopathol Pharmacol 2019 Jan-Dec;33:2058738419855567

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy.

Berberine (BBR) is a natural active principle with potential antitumor activity. The compound targets multiple cell signaling pathways, including proliferation, differentiation, and epithelial-mesenchymal transition. The aim of this study was to elucidate the mechanisms behind the anticancer activity of BBR by comparing the effects of purified BBR with those of the extract of , a medicinal plant that produces this metabolite. The expression levels of a panel of 44 selected genes in human colon adenocarcinoma (HCA-7) cell line were quantified by real-time polymerase chain reaction (PCR). BBR treatment resulted in a time- and dose-dependent down regulation of 33 genes differently involved in cell cycle, differentiation, and epithelial-mesenchymal transition. The trend was confirmed across the two types of treatment, the two time points, and the different absolute dosage of BBR. These findings suggest that the presence of BBR in extract significantly contributes to its antiproliferative activity.
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http://dx.doi.org/10.1177/2058738419855567DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6822188PMC
March 2020

The use of a new chemical device based on silver and cationic surfactants as a new approach for daily oral hygiene: A preliminary study on a group of periodontal patients.

Int J Immunopathol Pharmacol 2019 Jan-Dec;33:2058738419868101

Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy.

The aim of this study was to evaluate the abatement power of oral microbial loading of a new gel formulation based on the complex silver-2-mercaptobenzoate, chlorhexidine digluconate and didecyldimethylammonium chloride (named ADC) through polymerase chain reaction (PCR). The study sample consists of a group of 20 patients with chronic periodontal disease. Patients were over 25 years of age and did not undergo surgical or non-surgical periodontal treatment in the previous 6 months. The study sample was allotted into two groups of 10 patients each, homogeneous by age and sex. The test group received a bottle containing ADC gel, while the control group received an identical one containing placebo, similar to ADC in consistence, colour, taste and odour. Sub-gingival samples of four sites, one in each quadrant, of greatest probing depth in each patient were used. Microbiological analyses were performed at baseline and at day 15. Paired t test was performed to detect statistical significant reduction in total bacterial loading and oral pathogens in the study groups. The analysis showed a statistically significant reduction in the total bacterial loading evaluated pre- and post-treatment ( = 0.029) in the study groups. In the control group, the decrease in total bacterial loading was not significant ( = 0.279). Clinically, ADC gel does not have any side effects and discomfort such as pain, burning, tingling sensation or numbness and produces no adverse reactions in time. Our study aimed to evaluate the efficacy of a new chemical formulation with antibacterial properties to use for daily oral hygiene with a preliminary study. Our results showed a statistically significant reduction in total bacterial loading after treatment, but the limitations of our study do not allow us to demonstrate the clinical efficacy of the ADC gel.
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http://dx.doi.org/10.1177/2058738419868101DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6822178PMC
March 2020

Association between oral cleft and transcobalamin 2 polymorphism in a sample study from Nassiriya, Iraq.

Int J Immunopathol Pharmacol 2019 Jan-Dec;33:2058738419855571

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy.

Orofacial clefts are common congenital defects whose prevalence differs between geographical regions and ethnic groups. The inheritance is complex, involving the contribution of both genetic and environmental factors. The involvement of genes belonging to the folate pathway is still matter of debate, with strong evidences of association and conflicting results. After demonstrating the contribution, for a sample from the Italian population, of common mutations mapping on three genes of the folate pathway, our group tried to unravel their contribution in independent sample studies with different ethnicity. In the present investigation a set of 34 triads with oral cleft from Nassiriya, Iraq, has been genotyped for rs1801133 of MTHFR, rs1801198 of TCN2, and rs4920037 of CBS polymorphisms. Association analysis evidenced a decreased risk of cleft for children carrying the 667G allele at TCN2 gene ( = 0.02). This evidence further supported the relationship between polymorphisms of folate related genes and oral clefts, and outlined the relevance of studying populations having different ethnicity.
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http://dx.doi.org/10.1177/2058738419855571DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6822189PMC
March 2020

Molecular Aspects of Drug-Induced Gingival Overgrowth: An In Vitro Study on Amlodipine and Gingival Fibroblasts.

Int J Mol Sci 2019 Apr 25;20(8). Epub 2019 Apr 25.

Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, 44121 Ferrara, Italy.

Gingival overgrowth is a serious side effect that accompanies the use of amlodipine. Several conflicting theories have been proposed to explain the fibroblast's function in gingival overgrowth. To determine whether amlodipine alters the fibrotic response, we investigated its effects on treated gingival fibroblast gene expression as compared with untreated cells.

Materials And Methods: Fibroblasts from ATCC Cell Lines were incubated with amlodipine. The gene expression levels of 12 genes belonging to the "Extracellular Matrix and Adhesion Molecules" pathway was investigated in treated fibroblasts cell culture, as compared with untreated cells, by real time PCR.

Results: Most of the significant genes were up-regulated. (, , , , , , , ) except for , , , and , which were down-regulated.

Conclusion: These results seem to demonstrate that amlodipine has an effect on the extracellular matrix of gingival fibroblast. In the future, it would be interesting to understand the possible effect of the drug on fibroblasts of patients with amlodipine-induced gingival hyperplasia.
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http://dx.doi.org/10.3390/ijms20082047DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6514768PMC
April 2019

Reuterinos as adjuvant for peri-implant treatment: A pilot study.

Int J Immunopathol Pharmacol 2019 Jan-Dec;33:2058738419827745

5 Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy.

The objective of this study was to evaluate the effects of lozenges-containing Lactobacillus reuteri as an adjuvant treatment of peri-implant mucositis and to detect the level of L. reuteri colonization in the peri-implant tissues of treated patients. A total of 10 patients were selected. Subjects with at least one implant affected by peri-implant mucositis, with gingival index (GI) of ⩾2 in each quadrant, evaluated at the buccal aspect of all teeth. Patients included in the study were partially edentulous and had implants with mucositis or peri-implantitis. Implants with radiographic bone loss of ⩾5 mm and/or ⩾50% of the implant length were excluded, and only one implant per patient was included. Each patient received L. reuteri-containing lozenges. Microbiological sampling was performed at baseline and on day 28 and analysed by polymerase chain reaction (PCR). Our results indicate that the use of the probiotic did not influence the peri-implant microbiota in a statistically significant way, although there was a reduction in the number of periodontal and peri-implant species. The lack of statistically significant microbiological changes could be explained either by the small sample population or by the short evaluation period. Therefore, the poor colonization of L. reuteri in the peri-implant pockets can be explained by the different anatomical and histological characteristics of the interface of the dental-gingival unit with respect to the periodontal sulcus. The administration of a daily lozenge of L. reuteri for 4 weeks had a limited effect on the microbiological analysis. Probiotics provide an alternative therapeutic approach to consider in the prevention and treatment of peri-implant diseases, but further long-term prospective studies with standardized variables are needed.
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http://dx.doi.org/10.1177/2058738419827745DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6365990PMC
June 2019

Regenerative Dentistry and Stem Cells: A Multilineage Differentiation as a Safe and Useful Alternative Way of Harvesting and Selection Adipose Derived Mesenchymal Stem Cells.

Curr Drug Targets 2018 ;19(16):1991-1997

University of Ferrara, Italy.

Background: This review aims to address procedures and indications for the application of the adipose-derived stem cells (ADSCs) for regenerative dentistry. ADSCs have rarely been used in this particular field; conversely, experience from other clinical fields and basic research seems to recommend the suitability of this application.

Aims And Methods: We reviewed 32 out of 193 articles on Medline sorted by the relevance option. The main purpose of this paper is to perform a short review of the application of stem cells in regenerative dentistry, describing a multilineage differentiation as a safe and useful alternative way of harvesting and selection of ADSCs.

Results And Conclusion: The most common derivation of stem cells for regenerative dentistry is from the adipose tissue. There are conditions in which the levy adipose cannot be easily achieved, or where large amount of grafting is not needed. For this purpose, the possibility of selecting stromal stem cells directly from the lax subcutaneous connective tissue, preferably of the head region, would allow a technical simplification.
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http://dx.doi.org/10.2174/1389450119666180816122230DOI Listing
November 2019

A Noninvasive Test for MicroRNA Expression in Oral Squamous Cell Carcinoma.

Int J Mol Sci 2018 Jun 16;19(6). Epub 2018 Jun 16.

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, 40126 Bologna, Italy.

MicroRNAs have recently been proposed as non-invasive biomarkers in Oral Squamous Cell Carcinoma (OSCC). The aim of this study was to analyze the expression of a panel of miRNAs in epithelial cells collected by oral brushing from OSCCs from regenerative areas after OSCC surgical resection and from their respective normal distant mucosa. Oral brushing specimens were collected from 24 healthy donors, 14 OSCC patients with specimens from tumour and normal distant mucosa, and from 13 patients who had OSCC resection, with samples from regenerative areas after OSCC resection and normal distant mucosa. Expression levels of eight targets (miR-21, miR-375, miR-345, miR-181b, miR-146a, miR-649, miR-518b, and miR-191) were evaluated by real-time Polymerase Chain Reaction (PCR). A highly significant between-group difference was found for miR-21 (F = 6.58, < 0.001), miR-146a (F = 6.974, < 0.001), and miR-191 (F = 17.07, < 0.001). The major difference was observed between samples from healthy donors and from OSCC brushing, whereas no significant differences were observed between areas infiltrated by OSCC and their respective normal distant mucosa. Furthermore, altered expression of miR-146a and miR-191 was also observed in regenerative areas after OSCC resection.

Conclusions: Oral brushing could be proposed as a noninvasive method to study microRNA expression in oral mucosa in OSCC patients.
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http://dx.doi.org/10.3390/ijms19061789DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6032413PMC
June 2018

Fatigue resistance, electrochemical corrosion and biological response of Ti-15Mo with surface modified by amorphous TiO nanotubes layer.

J Biomed Mater Res B Appl Biomater 2019 01 4;107(1):86-96. Epub 2018 Mar 4.

University of São Paulo, São Carlos Institute of Chemistry, São Carlos, SP, Brazil.

The objective of this work was a systemic evaluation of the anodizing treatment in a β-type Ti-15Mo alloy to grow a TiO nanostructured layer for osseointegration improvement. The technical viability of the surface modification was assessed based on the resistance to mechanical fatigue, electrochemical corrosion, and biological response. By using an organic solution of NH F in ethylene glycol, a well-organized array of 90 nm diameter nanotubes was obtained with a potential of 40 V for 6 h, while undefined nanotubes of 25 nm diameter were formed with a potential of 20 V for 1 h. Nevertheless, the production of the 90 nm diameter nanotubes was followed by micrometer pits that significantly reduced the fatigue performance. The undefined nanotubes of 25 nm diameter, besides the greater cell viability and improved osteoblastic cell differentiation in comparison to the as-polished surface, were not deleterious to the fatigue and corrosion properties. This result strengthens the necessity of an overall evaluation of the anodizing treatment, particularly the fatigue resistance, before suggesting it for the design of implants. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 107B: 86-96, 2019.
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http://dx.doi.org/10.1002/jbm.b.34097DOI Listing
January 2019

Possible effect of SNAIL family transcriptional repressor 1 polymorphisms in non-syndromic cleft lip with or without cleft palate.

Clin Oral Investig 2018 Sep 27;22(7):2535-2541. Epub 2018 Jan 27.

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Via Belmeloro, 8, 40126, Bologna, Italy.

Objective: Orofacial development is a complex process subjected to failure impairing. Indeed, the cleft of the lip and/or of the palate is among the most frequent inborn malformations. The JARID2 gene has been suggested to be involved in non-syndromic cleft lip with or without cleft palate (nsCL/P) etiology. JARID2 interacts with the polycomb repressive complex 2 (PRC2) in regulating the expression patterns of developmental genes by modifying the chromatin state.

Materials And Methods: Genes coding for the PRC2 components, as well as other genes active in cell differentiation and embryonic development, were selected for a family-based association study to verify their involvement in nsCL/P. A total of 632 families from Italy and Asia participated to the study.

Results: Evidence of allelic association was found with polymorphisms of SNAI1; in particular, the rs16995010-G allele was undertransmitted to the nsCL/P cases [P = 0.004, odds ratio = 0.69 (95% C.I. 0.54-0.89)]. However, the adjusted significance value corrected for all the performed tests was P = 0.051.

Conclusions: The findings emerging by the present study suggest for the first time an involvement of SNAI1 in the nsCL/P onset.

Clinical Relevance: Interestingly, SNAI1 is known to promote epithelial to mesenchymal transition by repressing E-cadherin expression, but it needs an intact PRC2 to act this function. Alterations of this process could contribute to the complex etiology of nsCL/P.
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http://dx.doi.org/10.1007/s00784-018-2350-0DOI Listing
September 2018

Comparison of Bio-Revitalizing Injective Products: A Study on Skin Fibroblast Cultures.

Rejuvenation Res 2015 Jun 20;18(3):270-6. Epub 2015 May 20.

1 Department of Morphology, Surgery and Experimental Medicine, University of Ferrara , Ferrara, Italy .

Bio-revitalization is a commonly used technique in aesthetic medicine for improving skin quality and appearance by intra-dermal injection of hyaluronic acid (HA)-containing compounds. The present study compares different HA-containing injectables regarding their effects on cultured skin fibroblasts over time (24, 48, and 72 hr) by using RT-PCR and a panel of genes involved in dermal integrity. Human dermal fibroblasts were seeded on a layer of five different commercial medical devices containing 6.2 mg/mL 10 mg/mL 10%, 13 mg/mL and 20 mg/mL, respectively, of HA. The products differ not only in HA concentration but also in the content and quality of other ingredients; moreover, one of these products contained cross-linked HA. Differences among medical devices were found. In particular, HA concentration seems to be inversely correlated to elastin gene activation. Regarding the neutrophil elastase gene, the two medical devices with the higher concentration of HA displayed the greater effect. Genes encoding for hyaluronan synthase 1, hyaluronidase 1, and desmoplakin were enhanced, but the HA content of the different products did not seem to be directly related to gene activation. Therefore, the explanation for the differences must be studied further with respect to elements that are distinctive for each device. For the physician, it is important to choose which drugs or medical devices can be used and in what protocols. The present study performed a comparison that can be useful in better addressing the skin improvement therapies for aging and in its prevention.
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http://dx.doi.org/10.1089/rej.2014.1654DOI Listing
June 2015

p16(INK4) expression is not associated with human papillomavirus in oral lichen planus.

Oral Surg Oral Med Oral Pathol Oral Radiol 2014 Dec 16;118(6):694-702. Epub 2014 Sep 16.

Department of Biomedical and NeuroMotor Sciences (DiBiNeM), University of Bologna, and M. Malpighi Section of Pathology, Bellaria Hospital, Bologna, Italy.

Objective: Frequencies as high as 60% of overexpressed p16(INK4) were recently reported in lichen planus (LP). Because p16(INK4) overexpression may be a feature of human papilloma virus (HPV)-induced cancer, it has been postulated that LP may be somehow related to HPV. The present study is the first to evaluate both high p16(INK4) expression and HPV in patients with LP.

Study Design: Thirty-five consecutive biopsy specimens from patients with LP constituted the basis of the present study. Level of p16(INK4A) expression was evaluated in each sample by immunohistochemical analysis, and the presence of HPV DNA was tested by real-time polymerase chain reaction (PCR).

Results: p16(INK4) expression was detected in 26 specimens, whereas HPV was found in 4 lesions: 3 low-risk HPV and 1 high-risk HPV. All HPV-positive lesions also indicated p16(INK4A) overexpression, whereas 22 cases of overexpressed p16(INK4A) were HPV negative (Chi square 2.6; ns).

Conclusions: p16(INK4) overexpression is not correlated with HPV in patients with LP.
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http://dx.doi.org/10.1016/j.oooo.2014.09.004DOI Listing
December 2014

RFC1 and non-syndromic cleft lip with or without cleft palate: an association based study in Italy.

J Craniomaxillofac Surg 2014 Oct 2;42(7):1503-5. Epub 2014 May 2.

Department of Experimental, Diagnostic and Specialty Medicine, University di Bologna, Via Belmeloro 8, 40126 Bologna, Italy.

The molecular basis of orofacial development is largely unknown and needs to be unravelled. Non-syndromic cleft lip with or without cleft palate (NSCL/P) is the most common craniofacial malformation, with an incidence of about 1/700 live births, although variable according to ethnicity. Being a multifactorial disease, it arises as a result of an interplay between genetic and environmental factors. Several approaches have been developed to identify susceptibility genes. Genes belonging to the folate/homocysteine pathway are attracting increasing interest because folate supplementation before and during early pregnancy can reduce the risk of NSCL/P. We performed a family based association study in order to assess if a genetic variant of RFC1 could be involved in NSCL/P onset. We genotyped 404 unrelated probands and their relatives for three biallelic polymorphic variants (rs1051266, rs4818789 and rs3788205), that were selected because they produced conflicting results on previous investigations. Evidence of association was found between the investigated polymorphisms and NSCL/P in our sample of the Italian population, albeit with weak significance levels. Results from this investigation provided a support of previous studies suggesting a role of RFC1 in NSCL/P aetiology, reinforcing the concept that genetic predisposition to NSCL/P varies enormously within different ethnic groups.
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http://dx.doi.org/10.1016/j.jcms.2014.04.021DOI Listing
October 2014

Role of the MIR146A polymorphism in the origin and progression of oral squamous cell carcinoma.

Eur J Oral Sci 2014 Jun 10;122(3):198-201. Epub 2014 Mar 10.

Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy.

Gene expression and cell behavior are regulated by several factors, including small non-coding RNAs. MicroRNAs affecting cell growth, differentiation, and apoptosis are thought to play an important role in tumorigenesis. The levels of miR-146 appear to be associated with cancer development and progression, including that of oral squamous cell carcinoma. The aim of this investigation was to ascertain whether the single nucleotide polymorphism, rs2910164, mapping in the MIR146A gene, has a role in oral squamous cell carcinoma progression. A genetic association study was performed with a sample set of 346 oral squamous cell carcinomas collected in Italy. Our data indicate that the rs2910164 polymorphism is not associated with tumor development. However, a slight increase in the frequency of the variant allele was observed in Stage II tumors. Further investigations are needed to verify a possible role of the variant allele or rs2910164 in oral squamous cell carcinoma progression.
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http://dx.doi.org/10.1111/eos.12121DOI Listing
June 2014

Fibroblasts behavior after N-acetylcysteine and amino acids exposure: extracellular matrix gene expression.

Rejuvenation Res 2014 Jun 11;17(3):285-90. Epub 2014 Jun 11.

1 Department of Systems Medicine, University of Tor Vergata , Rome, Italy .

Reactive oxygen species (ROS) are chemically reactive molecules with impaired electrons that make them unstable and able to react easily with a great variety of molecules. The main targets of ROS are DNA, proteins, and membrane phospholipids. In the skin, ROS are able to affect the production of collagen and elastin, the main components of the extracellular matrix (ECM). This action contributes to the skin's aging. N-Acetylcysteine (NAC) is an acetylated cysteine residue with excellent anti-oxidant activity that boosts glutathione (GSH) levels. This study evaluates the effect of a solution of NAC and amino acids, which is used in aesthetic medicine as an intra-dermal injective treatment, on fibroblast behavior. To this aim, the expression levels of some ECM-related genes (HAS1, HYAL1 ELN, ELANE, MMP2, MMP3, MMP13, COL1A1, COL3A1) were analyzed on cultured dermal fibroblasts using real-time reverse transcription polymerase chain reaction (RT-PCR). All but two collagen genes were up-regulated after 24 hr of treatment.
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http://dx.doi.org/10.1089/rej.2013.1511DOI Listing
June 2014

Oral microflora and periodontal disease: new technology for diagnosis in dentistry.

Ann Stomatol (Roma) 2013 25;4(2):170-3. Epub 2013 Jun 25.

Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, Italy.

Periodontitis is a disease that affects and destroys the tissues that support teeth. Tissues damage results from a prolonged inflammatory response to an ecological shift in the composition of subgingival biofilms. Three bacterial species that constitute the red complex group, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola, are considered the main pathogens involved in periodontitis. In the present study a real-time PCR based assay was designed to detect and quantify red complex species, then used to investigate 146 periodontal pocket samples from 66 periodontitis patients and 80 controls. Results demonstrated a significant higher prevalence of red complex species and increased amount of P. gingivalis and T. denticola in periodontal pocket of periodontitis patients.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3755798PMC
August 2013

Microflora and periodontal disease.

Dent Res J (Isfahan) 2012 Dec;9(Suppl 2):S202-6

Department of Histology, Embryology and Applied Biology, Centre of Molecular Genetics, CARISBO Foundation, University of Bologna, Bologna, Italy.

Background: Periodontitis is a disease that affects and destroys the tissues that support teeth. Tissue damage results from a prolonged inflammatory response to an ecological shift in the composition of subgingival biofilms. Three bacterial species that constitute the red complex group, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola, are considered the main pathogens involved in periodontitis.

Materials And Methods: In the present study, a real-time polymerase chain reaction bases assay was designed to detect and quantify red complex species, then used to investigate 307 periodontal pocket samples from 127 periodontitis patients and 180 controls.

Results: Significant higher prevalence of red complex species and increased amount of P. gingivalis and T. denticola were detected in periodontal pocket of periodontitis patients.

Conclusions: Results demonstrated that the test is a valuable tool to improve diagnosis of periodontal disease.
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http://dx.doi.org/10.4103/1735-3327.109755DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3692174PMC
December 2012

IL6 and IL10 are genetic susceptibility factors of periodontal disease.

Dent Res J (Isfahan) 2012 Dec;9(Suppl 2):S197-201

Department of Histology, Embryology and Applied Biology, University of Bologna, Italy.

Background: Periodontitis is a disease mainly caused by a chronic infection of tissues that support the teeth. Several factors, such as diabetes, smoking and oral care, as well as genetic susceptibility can influence both the risk to develop periodontitis and its progression. The aim of the investigation was to test whether alleles of candidate genes were associated with periodontitis.

Materials And Methods: A case control study was performed with a cohort of 184 patients with chronic periodontitis and 231 healthy controls from the Italian population. A total of six single nucleotide polymorphisms from five candidate genes, i.e., IL1A, IL1B, IL6, IL10 and vitamin D receptor, were investigated.

Results: Evidence of association were obtained for rs1800795 mapping in IL6 (P value = 0.01) as well as for the rs1800872 mapping in IL10 (P = 0.04). The rarer variant allele lowered the risk to develop periodontitis at IL6 (Odds Ratio [OR] = 0.69 [95% confidence interval {CI} 0.51-0.93]) and increased the risk at IL10 (OR = 1.38 [95% CI 1.01-1.86]).

Conclusions: The present investigation indicated that polymorphisms of IL6 and IL10 constitute risk factors for chronic periodontitis, while there was no evidence implicating a specific IL1A or IL1B genotype.
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http://dx.doi.org/10.4103/1735-3327.109754DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3692173PMC
December 2012

Titanium nanotubes stimulate osteoblast differentiation of stem cells from pulp and adipose tissue.

Dent Res J (Isfahan) 2012 Dec;9(Suppl 2):S169-74

ENEA, IDROCOMB, C.R. Casaccia, Rome, Italy.

Background: Titanium is the gold standard among materials used for prosthetic devices because of its good mechanical and chemical properties. When exposed to oxygen, titanium becomes an oxide, anatase that is biocompatible and able to induce osseointegration.

Materials And Methods: IN THIS STUDY WE COMPARED THE EXPRESSION PROFILING OF STEM CELLS CULTIVATED ON TWO TYPES OF SURFACE: Pure titanium disk and nanotube titanium disk in order to detect if nanotube titanium instead (NTD) surface stimulates stem cells towards osteoblast differentiation.

Results: Stem cells cultivated on nanotube titanium disks showed the upregulation of bone-related genes RUNX2, FOSL1 and SPP1.

Conclusions: Results demonstrated that nanotube titanium disk surface is more osteo-induced surface compared to titanium disk, promoting the differentiation of mesenchymal stem cells in osteoblasts.
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http://dx.doi.org/10.4103/1735-3327.109745DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3692168PMC
December 2012

Titanium nanotubes activate genes related to bone formation in vitro.

Dent Res J (Isfahan) 2012 Dec;9(Suppl 2):S164-8

ENEA, IDROCOMB, C.R. Casaccia, Rome, Italy.

Background: Titanium is used worldwide to make osseointegrable devices, thanks to its favorable characteristics as mechanical proprieties and biocompatibility, demonstrated by in vivo studies with animal models and clinical trials over a forty-year period. However, the exact genetic effect of the titanium layer on cells is still not well characterized.

Materials And Methods: To investigate how titanium nanotubes stimulate osteoblasts differentiation and proliferation, some osteoblast genes (SP7, RUNX2, COL3A1, COL1A1, ALPL, SPP1 and FOSL1) were analyzed by quantitative Real Time RT- PCR.

Results: After 15 days, osteoblasts cultivated on titanium naotube showed the up-regulation of bone related genes SP7, ENG, FOSL1 and SPP1 and the down-regulation of RUNX2, COL3A1, COL1A1, and ALPL. After 30 days of treatment, the bone related genes SP7, ENG, FOSL1 and RUNX2 were up-regulated while COL3A1, COL1A1, ALPL and SPP1 were down-regulated.

Conclusions: Our results, demonstrates that titanium nanotubes can lead to osteoblast differentiation and extracellular matrix deposition and mineralization in dental pulp stem cells by the activation of osteoblast related genes SPP1, FOSL1 and RUNX2.
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http://dx.doi.org/10.4103/1735-3327.109736DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3692167PMC
December 2012

Lab-Test(®) 4: Dental caries and bacteriological analysis.

Dent Res J (Isfahan) 2012 Dec;9(Suppl 2):S139-41

Department of Histology, Embryology and Applied Biology, Centre of Molecular Genetics, CARISBO Foundation, University of Bologna, Bologna, Italy.

Dental caries is one of the most common infectious ultifactorial diseases worldwide, characterized by the progressive demineralization of the tooth, following the action of bacterial acid metabolism. The main factors predisposing the onset of the carious process are: 1) the presence of bacterial species able to lower the pH until critical values of 5.5, 2) the absence of adequate oral hygiene, 3) an inefficient immune response anti-caries, 4) the type of alimentary diet and 5) the structure of the teeth. Among the 200 bacterial species isolated from dental plaque the most pathogenic for dental caries are: Streptococcus mutans, Streptococcus sobrinus, Lactobacillus acidophilus, Actinomices viscusus and Bifidobacterium dentium. Our laboratory (LAB(®) s.r.l., Codigoro, Ferrara, Italy) has developed a test for absolute and relative quantification of the most common oral cariogenic bacteria. The test uses specific primers and probes for the amplification of bacteria genome sequences in Polymerase Chain Reaction Real Time. The results provide a profile of patient infection, helpful for improving the diagnosis and planning of preventive treatment to reduce the bacterial load.
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http://dx.doi.org/10.4103/1735-3327.109723DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3692161PMC
December 2012

Effect of titanium surface topographies on human bone marrow stem cells differentiation in vitro.

Odontology 2013 Jul 8;101(2):133-9. Epub 2012 Jun 8.

Dental School, University of Chieti-Pescara, Via F. Sciucchi 63, 66100, Chieti, Italy.

Coating characteristics of dental implants such as composition and topography regulate cell response during implant healing. The aim of this study was to assess how surface topography can affect osteogenic differentiation of mesenchymal stem cells (MSCs) by analyzing the expression levels of bone-related genes and MSCs marker. Thirty disk-shaped, commercially pure Grade 2 titanium samples (10 × 2 mm) with 3 different surface topographies (DENTSPLY-Friadent GmbH, Mannheim, Germany) were used in the present study: 10 Ti machined disks (control), 10 Ti sandblasted and acid-etched disks (DPS(®)) and 10 sandblasted and acid-etched disks at high temperature (Plus(®)). Samples were processed for real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis. By comparing machined and Plus(®) disks, quantitative real-time RT-PCR showed a significant reduction of the bone-related genes osteocalcin (BGLAP) and osteoblast transcriptional factor (RUNX2). The comparison between DPS(®) and Plus(®) disks showed a slight induction of all the genes examined (RUNX2, ALPL, COL1A1, COL3A1, ENG, FOSL1, SPP1, and SP7); only the expression of BGLAP remained stable. The present study, demonstrated that implant surface topography affects osteoblast gene expression. Indeed, Plus(®) surface produces an effect on MSCs in the late differentiation stages.
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http://dx.doi.org/10.1007/s10266-012-0067-0DOI Listing
July 2013

Anatase-based implants nanocoating on stem cells derived from adipose tissue.

Implant Dent 2012 Apr;21(2):118-23

Department of Maxillofacial Surgery, University of Ferrara, Ferrara, Italy.

Purpose: The aim of this study was to investigate the effect of a new anatase coating with antibacterial properties (Bactercline anatase coating [BAC]) on dental implants in the commitment of stem cells derived from adipose tissue to osteoblasts.

Materials And Methods: Using real-time reverse transcription polymerase chain reaction, the quantitative expression of specific genes, such as transcriptional factors (runx2 and sp7), bone-related genes (spp1, col1a1, col3a1, alpl, and fosl1), and mesenchymal stem cells marker (eng), was examined.

Results: BAC caused induction of bone-related genes such as sp7, fosl1, alpl, and spp1. In contrast, the expression of runx2, col3a1, and col1a1 was decreased in stem cells treated with BAC with respect to untreated cells.

Conclusion: The obtained results are relevant to better understand the molecular mechanism of bone regeneration and as a model for comparing other materials with similar clinical effects.
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http://dx.doi.org/10.1097/ID.0b013e31824bc948DOI Listing
April 2012