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1.
Arch Oral Biol ; 165: 106016, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38838515

RESUMEN

OBJECTIVES: To evaluate in vivo 1) the bioavailability of trans-resveratrol when administered through sublingual capsules; 2) the effect of resveratrol on the protein composition of the acquired enamel pellicle (AEP). DESIGN: Ten volunteers received a sublingual capsule containing 50 mg of trans-resveratrol. Unstimulated saliva was then collected after 0, 30, 60, and 120 min and AEP was collected after 120 min following administration of the capsule. In the next week, the volunteers received a placebo sublingual capsule, and saliva and AEP were collected again. Saliva samples were analyzed for free trans-resveratrol using high-performance liquid chromatopgraphy (HPLC), and AEP samples were subjected to proteomic analysis (nLC-ESI-MS/MS). RESULTS: Trans-resveratrol was detected in saliva at all the time points evaluated, with the peak at 30 min. A total of 242 proteins were identified in both groups. Ninety-six proteins were increased and 23 proteins were decreased in the Resveratrol group. Among the up-regulated proteins, isoforms of cystatins, PRPs, Mucin-7, Histatin-1, Lactotrasnferrin and Lysozyme-C were increased and the isoforms of Protein S100, Neutrophil defensins, Albumin, PRPs, and, Statherin were decreased in Resveratrol group. CONCLUSION: The sublingual capsule is effective at increasing the bioavailability of trans-resveratrol in saliva. Several proteins involved in important processes to maintain systemic and oral health homeostasis were identified. These proteins differently expressed due to the presence of trans-resveratrol deserve attention for future studies, since they have important functions, mainly related to antimicrobial action.


Asunto(s)
Cápsulas , Película Dental , Resveratrol , Saliva , Humanos , Resveratrol/farmacología , Resveratrol/farmacocinética , Resveratrol/administración & dosificación , Saliva/metabolismo , Saliva/química , Masculino , Adulto , Película Dental/metabolismo , Película Dental/química , Cromatografía Líquida de Alta Presión , Femenino , Disponibilidad Biológica , Estilbenos/farmacocinética , Estilbenos/farmacología , Estilbenos/administración & dosificación , Proteómica , Espectrometría de Masas en Tándem , Proteínas y Péptidos Salivales/metabolismo
2.
Cell Biochem Funct ; 42(1): e3936, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38269522

RESUMEN

Resistance training (RT) with blood flow restriction (BFR) or high intensity (HI) are effective to increase muscle mass. To understand this effect, techniques known as "omics" are used to identify possible biomarkers. This study analyzed the salivary proteomic profile of healthy individuals trained before and after two RT protocols both designed with eight exercises for upper- and lower-limbs, one performed at low percentage of one-maximum repetition (%1RM) with BFR technique, and other at high %1RM (HI) without BRF technique. Four healthy males between 18 and 28 years participated in the study. Stimulated saliva was collected before (BBFR/BHI) and immediately after (ABFR/AHI) the two RT protocols. All protein-related processing was performed using label-free proteomic. The difference in expression between groups was expressed as p < .05 for downregulated proteins and 1-p > .95 for upregulated proteins. There was difference in salivary flow between ABFR and BBFR (p = .005). For HI, 87 proteins were found after the practice and 119 before. Three hemoglobin isoforms were increased in AHI compared with BHI. In the BFR comparison, 105 proteins were identified after (ABFR) and 70 before (BBFR). Among those increased ABFR, we highlight five hemoglobin isoforms and Deleted in malignant brain tumors 1 protein. Between ABFR and AHI, 17 isoforms of histones, Transaldolase, Transketolase, Glyceraldehyde-3-phosphate dehydrogenase, and Antileukoproteinase were decreased ABFR. For HI, there was an increase in proteins related to oxidative stress and metabolism of the musculoskeletal system, compared with BFR. HI seems to induce higher anabolic signaling to muscle mass increase and antiatherosclerotic effects.


Asunto(s)
Entrenamiento de Fuerza , Masculino , Humanos , Proteómica , Histonas , Hemoglobinas , Isoformas de Proteínas
3.
Caries Res ; 58(2): 90-107, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38198757

RESUMEN

INTRODUCTION: This study investigated the changes in the acquired enamel pellicle (AEP) proteome when this integument is formed in vivo after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), and a statherin-derived peptide (StN15), or their combination and then exposed to an intrinsic acid challenge. The effectiveness of these treatments in preventing intrinsic erosion was also evaluated. METHODS: Ten volunteers, after prophylaxis, in 5 crossover phases, rinsed with the following solutions (10 mL, 1 min): control (deionized water-H2O) - group 1, 0.1 mg/mL CaneCPI-5 - group 2, 1.0 mg/mL HB - group 3, 1.88 × 10-5M StN15 - group 4, or a blend of these - group 5. Following this, AEP formation occurred (2 h) and an enamel biopsy (10 µL, 0.01 m HCl, pH 2.0, 10 s) was conducted on one incisor. The biopsy acid was then analyzed for calcium (Arsenazo method). The vestibular surfaces of the other teeth were treated with the same acid. Acid-resistant proteins in the residual AEP were then collected and analyzed quantitatively via proteomics. RESULTS: Compared to control, treatment with the proteins/peptide, mixed or isolated, markedly enhanced acid-resistant proteins in the AEP. Notable increases occurred in pyruvate kinase PKM (11-fold, CaneCPI-5), immunoglobulins and submaxillary gland androgen-regulated protein 3B (4-fold, StN15), Hb, and lysozyme C (2-fold, StN15). Additionally, a range of proteins not commonly identified in the AEP but known to bind calcium or other proteins were identified in groups treated with the tested proteins/peptide either in isolation or as a mixture. The mean (SD, mM) calcium concentrations released from enamel were 3.67 ± 1.48a, 3.11 ± 0.72a, 1.94 ± 0.57b, 2.37 ± 0.90a, and 2.38 ± 0.45a for groups 1-5, respectively (RM-ANOVA/Tukey, p < 0.05). CONCLUSIONS: Our findings demonstrate that all treatments, whether using a combination of proteins/peptides or in isolation, enhanced acid-resistant proteins in the AEP. However, only HB showed effectiveness in protecting against intrinsic erosive demineralization. These results pave the way for innovative preventive methods against intrinsic erosion, using "acquired pellicle engineering" techniques.


Asunto(s)
Calcio , Erosión de los Dientes , Humanos , Calcio/metabolismo , Película Dental , Péptidos , Proteoma , Erosión de los Dientes/prevención & control , Hemoglobinas/metabolismo
4.
J Dent ; 141: 104810, 2024 02.
Artículo en Inglés | MEDLINE | ID: mdl-38110112

RESUMEN

OBJECTIVE: The aim of this study was to determine the effect of different concentrations of resveratrol in protecting enamel against initial dental erosion in vitro. METHODS: Ninety bovine enamel samples (4 × 4 mm) were divided into six groups: Phosphate buffered saline (negative control; PBS), Commercial solution (Elmex Erosion Protection™; positive control) and resveratrol at 4 different concentrations (1, 10, 100 or 400 µg/mL). Initially, the samples were incubated in saliva for the formation of the acquired pellicle (250 µL, 1 h, 37 °C, 250 rpm). Afterward, the samples were incubated in the respective treatments (250 µL, 1 min, 37 °C, 250 rpm) and then reincubated in saliva (250 µL, 1 h, 37 °C, 250 rpm). Finally, the samples were subjected to an erosive challenge by incubating in 1 % citric acid (1 mL, pH 3.5, 1 min, 25 °C, 250 rpm). The percentage surface microhardness change (% SMC) was assessed using a microhardness tester. Data were analyzed by Kruskal-Wallis and Dunn's tests (p < 0.05). RESULTS: The treatments with Elmex™ and resveratrol (1, 10 and 100 µg/mL) significantly protected enamel compared to the negative control, without significant differences among them. However, the group treated with the highest resveratrol concentration (400 µg/mL) did not show a significant difference from the negative control. CONCLUSIONS: Resveratrol at concentrations ranging from 1 to 100 µg/ml was effective in preventing loss of enamel surface microhardness. CLINICAL SIGNIFICANCE: This result suggests a potential new direction for the development of dental products based on resveratrol for the prevention of dental erosion.


Asunto(s)
Erosión de los Dientes , Animales , Bovinos , Resveratrol/farmacología , Erosión de los Dientes/prevención & control , Esmalte Dental , Película Dental , Saliva
5.
J Dent ; 139: 104724, 2023 12.
Artículo en Inglés | MEDLINE | ID: mdl-37797812

RESUMEN

OBJECTIVE: To evaluate the difference in the proteomic profile of stimulated saliva in patients with gastroesophageal reflux disease (GERD) with (GE) and without (GNE) erosive tooth wear (ETW), regarding both human and bacterial proteins. METHODS: Stimulated saliva (SS) was collected from 16 patients (8/group). Samples were centrifuged at 4.500 g for 15 min under refrigeration to remove all debris. The supernatant from each saliva sample was taken and frozen at -80 °C. After extracting the proteins, they were submitted to reverse phase liquid chromatography and mass spectrometry (nLC-ESI-MS/MS). Label-free proteomic quantification was performed using Protein Lynx Global Service (PLGS) software (p < 0.05) for human and bacterial proteins. RESULTS: In total, 67 human proteins were common for GNE and GE groups. GNE group presented, compared to GE group, increase in proteins that confer antimicrobial and acid resistant properties, such as cystatins, histatin and immunoglobulins. However, GNE group had a marked decrease in subunits of hemoglobin (α, ß and delta). Regarding bacterial proteins, for SS, 7 and 10 unique proteins were identified in the GE and GNE groups, respectively. They are related to protein synthesis and energy metabolism and interact with human proteins typically found in saliva and supramolecular complexes of the acquired pellicle. CONCLUSIONS: Our data indicate that the stimulation of the salivary flow increases acid resistant and antimicrobial proteins in saliva, which might protect against ETW. CLINICAL SIGNIFICANCE: This pioneer study showed important differences in the human and bacterial proteome of SS in patients with GERD with or without ETW.


Asunto(s)
Antiinfecciosos , Reflujo Gastroesofágico , Atrición Dental , Erosión de los Dientes , Desgaste de los Dientes , Humanos , Saliva/química , Espectrometría de Masas en Tándem , Proteómica , Proteoma , Proteínas Bacterianas
6.
Eur J Oral Sci ; 131(4): e12944, 2023 08.
Artículo en Inglés | MEDLINE | ID: mdl-37414726

RESUMEN

Acquired enamel pellicle plays an important role in the pathogenesis of early childhood caries (ECC), working as a protective interface between the tooth and the oral cavity. The aim of this cross-sectional in vivo proteomic study was to compare the acquired enamel pellicle protein profile of 3-5-year-old children with ECC (n = 10) and caries-free children (n = 10). Acquired enamel pellicle samples were collected and processed for proteomic analysis (nLC-ESI-MS/MS). In total, 241 proteins were identified. Basic salivary proline-rich protein 1 and 2, Cystatin-B, and SA were found only in the caries free group. When comparing caries free and ECC groups, lower protein levels were found in the caries free group for hemoglobin subunit beta, delta, epsilon, gamma-2, globin domain-containing protein and gamma-1, neutrophil defensin 3, serum albumin, protein S100-A8, and S100-A9. The proteins histatin-1, statherin, salivary acidic proline-rich phosphoprotein ½, proline-rich protein 4, submaxillary gland androgen-regulated protein 3B, alpha-amylase 1 and 2B were found at higher levels in the caries free group. The exclusive and the proteins found at higher levels in the caries free group might have protective functions that play a role in the prevention of caries, besides providing important insights to be evaluated in future studies for the possible development of new therapeutic strategies for ECC.


Asunto(s)
Caries Dental , Espectrometría de Masas en Tándem , Preescolar , Humanos , Película Dental/metabolismo , Proteómica , Estudios Transversales , Fosfoproteínas/metabolismo , Prolina/metabolismo , Proteínas y Péptidos Salivales/metabolismo , Saliva
7.
Biomedicines ; 11(5)2023 May 04.
Artículo en Inglés | MEDLINE | ID: mdl-37239031

RESUMEN

Phytocystatins are proteinaceous competitive inhibitors of cysteine peptidases involved in physiological and defensive roles in plants. Their application as potential therapeutics for human disorders has been suggested, and the hunt for novel cystatin variants in different plants, such as maqui (Aristotelia chilensis), is pertinent. Being an understudied species, the biotechnological potential of maqui proteins is little understood. In the present study, we constructed a transcriptome of maqui plantlets using next-generation sequencing, in which we found six cystatin sequences. Five of them were cloned and recombinantly expressed. Inhibition assays were performed against papain and human cathepsins B and L. Maquicystatins can inhibit the proteases in nanomolar order, except MaquiCPIs 4 and 5, which inhibit cathepsin B in micromolar order. This suggests maquicystatins' potential use for treating human diseases. In addition, since we previously demonstrated the efficacy of a sugarcane-derived cystatin to protect dental enamel, we tested the ability of MaquiCPI-3 to protect both dentin and enamel. Both were protected by this protein (by One-way ANOVA and Tukey's Multiple Comparisons Test, p < 0.05), suggesting its potential usage in dental products.

8.
J Mech Behav Biomed Mater ; 141: 105782, 2023 05.
Artículo en Inglés | MEDLINE | ID: mdl-36934688

RESUMEN

The sugarcane cystatin 5 (CaneCPI-5) showed protection against erosion and erosive tooth wear (ETW) under several protocols. However, evaluating these conditions in vivo is hard due to the lack of a suitable device. The aim of this study was to use clinically the relative surface reflection intensity (%SRI) by the Reflectometer Optipen to assess the acquired pellicle engineering with CaneCPI-5 rinse for the prevention of initial erosion in vivo. Nine volunteers were distributed in three cross-over phases, according to the rinse used, as follows: 1) Deionized water (negative control); 2) Elmex® (800 ppm Sn2+, 500 ppm F-; positive control); 3) 0.1 mg/mL CaneCPI-5. The following experimental steps were performed: Initially, the volunteers received prophylaxis and the initial %SRI was performed. Subsequently, they rinsed with the solutions (10 mL; 1min), followed by the formation of the acquired enamel pellicle (AEP; 120min). After, the erosive challenge with citric acid 1%, pH 2.5 (10 µL; 10s) was performed (in isolation) on the buccal surface of the maxillary central incisors (right and left). The calcium present in the acid was analyzed by Arsenazo III method. Finally, the final %SRI was performed. Data were analyzed by Kruskal-Wallis/Dunn's tests and Spearman's correlation were used (p < 0.05). For both variables, the negative control led to significantly less protection (lower reflectivity and higher calcium release) in comparison with the other groups. The best protection (higher reflectivity and lower calcium release) was observed for the Elmex® and the CaneCPI-5 groups, with no significant differences between them (p < 0.05). There was a significant correlation between both analyzes. The Reflectometer Optipen demonstrated to be a good device to assess clinically. Moreover, CaneCPI-5 rinse proved effective through acquired pellicle engineering against initial erosion in vivo.


Asunto(s)
Cistatinas , Saccharum , Erosión de los Dientes , Humanos , Erosión de los Dientes/prevención & control , Calcio
9.
Arch Oral Biol ; 148: 105643, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36773559

RESUMEN

OBJECTIVES: This study evaluated the protective impact of acquired enamel pellicle (AEP) engineering with statherin-derived peptide (StatpSpS), considering different AEP formation times. DESIGN: A total of 120 native human enamel specimens were divided into 2 main groups: 1) No AEP engineering and 2) AEP engineering with StatpSpS (pretreatment for 1 min; 37 °C, under agitation). Each group was further divided into 4 subgroups: No pellicle, or 1, 60-and-120 min AEP formation times (human saliva; 37 °C). The specimens were then subjected to an erosive challenge (1% citric acid; pH 3.6; 1 min; 25 °C). This procedure was repeated for 5 cycles. Relative surface reflection intensity (%SRI) was measured and scanning electron microscopy (SEM) of the enamel surface was done. RESULTS: All AEP engineering groups protected against initial dental erosion in comparison with No pellicle (p < 0.001), likewise all groups with AEP, independent of engineering or formation times (p 0.001). Furthermore, engineering with StatpSpS even without the presence of AEP protected the enamel when compared to the No engineering/No pellicle group (p < 0.0001). No difference was observed regarding the protection from the different AEP formation times (p > 0.05). Regarding the SEM analysis, in the "No AEP engineering & No AEP" group, a more severe effect of citric acid was observed, with more enamel prism heads and scratches on the surface when compared with the other groups. CONCLUSIONS: AEP provides almost instant protection at formation times even as short as 1 min, protecting the native enamel against erosion. Treatment with StatpSpS by itself provides similar protection as the AEP.


Asunto(s)
Erosión de los Dientes , Humanos , Película Dental , Erosión de los Dientes/prevención & control , Esmalte Dental , Péptidos/farmacología , Ácido Cítrico/farmacología
10.
Braz. oral res. (Online) ; 37: e085, 2023. tab, graf
Artículo en Inglés | LILACS-Express | LILACS, BBO - Odontología | ID: biblio-1513886

RESUMEN

Abstract The objective of this study was to compare the protein profile of the acquired enamel pellicle (AEP) formed in vivo in patients with or without gastroesophageal reflux disease (GERD), and with or without erosive tooth wear (ETW). Twenty-four volunteers were divided into 3 groups: 1) GERD and ETW; 2) GERD without ETW; and 3) control (without GERD). The AEP formed 120 min after prophylaxis was collected from the lingual/palatal surfaces. The samples were subjected to mass spectrometry (nLC-ESI-MS/MS) and label-free quantification by Protein Lynx Global Service software. A total of 213 proteins were identified, or 119, 92 and 106 from each group, respectively. Group 2 showed a high number of phosphorylated and calcium-binding proteins. Twenty-three proteins were found in all the groups, including 14-3-3 protein zeta/delta and 1-phosphatidylinositol. Several intracellular proteins that join saliva after the exfoliation of oral mucosa cells might have the potential to bind hydroxyapatite, or participate in forming supramolecular aggregates that bind to precursor proteins in the AEP. Proteins might play a central role in protecting the dental surface against acid dissolution.

11.
J. appl. oral sci ; 31: e20230036, 2023. graf
Artículo en Inglés | LILACS-Express | LILACS | ID: biblio-1440408

RESUMEN

Abstract Fluoride (F) has been widely used to control dental caries, and studies suggest beneficial effects against diabetes when a low dose of F is added to the drinking water (10 mgF/L). Objectives This study evaluated metabolic changes in pancreatic islets of NOD mice exposed to low doses of F and the main pathways altered by the treatment. Methodology In total, 42 female NOD mice were randomly divided into two groups, considering the concentration of F administered in the drinking water for 14 weeks: 0 or 10 mgF/L. After the experimental period, the pancreas was collected for morphological and immunohistochemical analysis, and the islets for proteomic analysis. Results In the morphological and immunohistochemical analysis, no significant differences were found in the percentage of cells labelled for insulin, glucagon, and acetylated histone H3, although the treated group had higher percentages than the control group. Moreover, no significant differences were found for the mean percentages of pancreatic areas occupied by islets and for the pancreatic inflammatory infiltrate between the control and treated groups. Proteomic analysis showed large increases in histones H3 and, to a lesser extent, in histone acetyltransferases, concomitant with a decrease in enzymes involved in the formation of acetyl-CoA, besides many changes in proteins involved in several metabolic pathways, especially energy metabolism. The conjunction analysis of these data showed an attempt by the organism to maintain protein synthesis in the islets, even with the dramatic changes in energy metabolism. Conclusion Our data suggests epigenetic alterations in the islets of NOD mice exposed to F levels comparable to those found in public supply water consumed by humans.

12.
Braz. dent. sci ; 26(1): 1-10, 2023. tab, ilus
Artículo en Inglés | LILACS, BBO - Odontología | ID: biblio-1411442

RESUMEN

Objective: this study aimed to evaluate knowledge and attitudes of professional wine tasters about erosive tooth wear. Material and Methods: two questionnaires were applied to 56 wine tasters during the 25th National Evaluation of Wines - Safra 2017 in Bento Gonçalves, Rio Grande do Sul. Descriptive data analysis and the Fisher's exact test were performed (p < 0.05). Results: for the analysis, 56 volunteers aged 25-76 years were included. Most of the volunteers reported attending dentist once a year (55.35%) and their teeth become sorer after ingesting liquids or acid foods (32.14%) over time. The majority (82.14%) believe wine tasting may have teeth effects and have already heard about erosive tooth wear (51.78%). However, when asked about symptomatology, more than half (53.57%) reported not to know. Brushing teeth immediately after ingesting something (37.5%) and increasing the brushing frequency (33.92%) were the most cited treatment options. The habit of using mouthwash solutions was significantly associated with the professionals age (p = 0.039). Conclusion: the volunteers reported a high intake of acid substances and, although most of them reported to consult dentist regularly and have already heard about erosive tooth wear, knowledge about this condition is still limited among these professionals. (AU)


Objetivo: o objetivo desse estudo foi avaliar o conhecimento e as atitudes de provadores de vinho profissionais relacionadas ao desgaste dentário erosivo. Material e Métodos: dois questionários foram aplicados a 56 provadores de vinho durante a 25ª Avaliação Nacional de Vinhos - Safra 2017 no município de Bento Gonçalves, Rio Grande do Sul. Foi realizada a análise descritiva dos dados e o teste Exato de Fisher (p < 0.05). Resultados: na análise foram incluídos 56 voluntários com idade entre 25 e 76 anos. A maioria dos voluntários relatou frequentar o dentista uma vez por ano (55,35%) e que seus dentes, com o passar do tempo, tornaram-se mais doloridos ao ingerir líquidos ou alimentos ácidos (32,14%). Mais da metade (82,14%) acreditam que as provas de vinho podem causar algum efeito sobre os dentes e já ouviram falar em desgaste dentário erosivo (51,78%). Porém, quando questionados sobre a sintomatologia, a maioria (53,57%) afirmou não ter conhecimento. Escovar os dentes imediatamente após ingerir algo (37,5%) e aumentar a frequência de escovação (33,92%) foram as opções mais compreendidas como tratamento. Utilizar soluções para bochecho esteve significativamente associado à idade dos profissionais (p=0,039). Conclusão: provadores de vinho profissionais relataram alta ingestão de substâncias ácidas e, embora a maioria consulte pelo menos uma vez por ano o dentista e já tenha ouvido falar a respeito do desgaste dentário erosivo, o conhecimento sobre essa condição ainda é limitado entre esses profissionais. (AU)


Asunto(s)
Humanos , Vino , Conocimientos, Actitudes y Práctica en Salud , Salud Bucal , Conocimiento , Desgaste de los Dientes
13.
Metabolites ; 12(11)2022 Nov 10.
Artículo en Inglés | MEDLINE | ID: mdl-36355174

RESUMEN

Obesity and pregnancy may have synergistic effects on periodontal condition, and proteomics could be an ideal approach to highlight the pathophysiological mechanisms associated with these outcomes. This study analyzed the salivary proteomics related to obesity and periodontitis in women during pregnancy (T1) and after delivery (T2). Initially, 126 women were recruited and forty were allocated into groups: with obesity and periodontitis (OP); with obesity, but without periodontitis (OWP); with normal BMI, but with periodontitis (NP); with normal BMI and without periodontitis (NWP). Whole-mouth saliva was collected in T1 and T2, and proteins were extracted and individually processed by label-free proteomics (nLC-ESI-MS/MS). The up-regulations of Heat shock 70 kDa protein 1A, 1B, and 1-like were related to both obesity and periodontitis, separately. Albumin and Thioredoxin were up-regulated in periodontitis cases, while Cystatins (mainly S, SA, SN) and Lactotransferrin were down-regulated. The high abundances of Submaxillary gland androgen-regulated protein 3B, Protein S100-A8, Matrix metalloproteinase-9, Heat shock 70 kDa protein 2 and 6, Putative Heat shock 70 kDa protein 7, Heat shock 71 kDa protein, Haptoglobin and Plastin-1 were significant in the combination of obesity and periodontitis. Obesity and periodontitis remarkably altered the proteome of the saliva during pregnancy with substantial alterations after delivery.

14.
Clin Oral Investig ; 26(1): 225-258, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34052889

RESUMEN

OBJECTIVES: Salivary glands are affected during radiotherapy in the head and neck region, leading to a reduction in salivary flow and changes its composition. Besides negatively affecting the oral soft tissues, this can also lead to dental impairment. Thus, we evaluated the effect of radiotherapy in the proteomic profile of the saliva in patients with head and neck cancer (HNC). MATERIALS AND METHODS: HNC patients had their saliva collected before (BRT), during (2-5 weeks; DRT), and after (3-4 months; ART) radiotherapy. Saliva was also collected from healthy volunteers (control; C). Samples were processed for proteomic analysis. RESULTS: In total, 1055 proteins were identified, among which 46 were common to all groups, while 86, 86, 286, and 395 were exclusively found in C, BRT, DRT, and ART, respectively. Remarkably, alpha-enolase was increased 35-fold DRT compared with BRT, while proline-rich proteins were decreased. ART there was a 16-fold increase in scaffold attachment factor-B1 and a 3-fold decrease in alpha-enolase and several cystatins. When compared with C, salivary proteins of BRT patients showed increases cystatin-C, lysozyme C, histatin-1, and proline-rich proteins CONCLUSION/CLINICAL REVELANCE: Both HNC and radiotherapy remarkably change the salivary protein composition. Altogether, our results, for the first time, suggest investigating alpha-enolase levels in saliva DRT in future studies as a possible biomarker and strategy to predict the efficiency of the treatment. Moreover, our data provide important insights for designing dental products that are more effective for these patients and contribute to a better understanding of the progressive changes in salivary proteins induced by radiotherapy. Graphical abstract.


Asunto(s)
Neoplasias de Cabeza y Cuello , Proteoma , Neoplasias de Cabeza y Cuello/radioterapia , Humanos , Proteómica , Saliva , Proteínas y Péptidos Salivales
15.
J Dent ; 116: 103874, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34798481

RESUMEN

OBJECTIVE: To study the proteomic alterations in the initial AEP after rinsing with CaneCPI-5, StN15 or Hb or their combination. MATERIALS AND METHODS: In five crossover phases, after prophylaxis, 10 volunteers in 5 consecutive days, rinsed (10 mL, 1 min) with the following solutions: deionized water (H2O- negative control- 1), 0.1 mg/mL CaneCPI-5 (2), 1.88×10-5 M StN15 (3), 1.0 mg/mL Hb (4) or their combination (5). The AEP formed after 3 min was collected with electrode filter papers soaked in 3% citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. RESULTS: Rinsing with the proteins/peptide increased the amounts of proteins in the AEP. The total numbers of proteins identified after rinsing with CaneCPI-5, StN15, Hb or their combination versus water, were 131, 167, 148 and 142, respectively. The treatment with the proteins/peptide or their combination increased proteins that bind calcium, phosphate and interact with distinct proteins, as well as proteins with antimicrobial and acid-resistant properties, such as, Cornifin-B (7.7, 12.6, and 4.3-fold for CaneCPI-5, StN15 and Hb, respectively), isoforms of Cystatin (2.2-2.4-fold for CaneCPI-5 and StN15), Proline-rich-protein 4 (4.3-fold; StN15), Histatin-1 (2.8-fold; StN15) and Hemoglobin (7.7-25-fold for Hb and Combination). Immunoglobulin, Keratin and Histone were exclusively identified upon treatment with the proteins/peptide, alone or combined. CONCLUSION: Rinsing with proteins/peptide, alone or combined, increased protective proteins in the initial AEP. CLINICAL RELEVANCE: Our results suggest that rinsing with the proteins/peptide or their combination increases the proteins capable of enhancing the protective function of the basal layer of AEP.


Asunto(s)
Proteínas , Proteómica , Película Dental/química , Humanos , Péptidos , Agua
16.
Caries Res ; 55(4): 333-340, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34344000

RESUMEN

Changes in the proteomic profile of the acquired enamel pellicle (AEP) formed for 3 min or 2 h after rinsing with a peptide containing the 15 N-terminal residues of statherin, with serines 2 and 3 phosphorylated (StatpSpS), were evaluated. Nine volunteers participated in 2 consecutive days. Each day, after professional tooth cleaning, they rinsed for 1 min with 10 mL of phosphate buffer containing 1.88 × 10-5 M StatpSpS or phosphate buffer only (control). The acquired pellicle formed on enamel after 3 min or 2 h was collected with electrode filter papers soaked in 3% citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics. In the 3-min AEP, 19 and 131 proteins were uniquely identified in the StatpSpS and control groups, respectively. Proteins typically found in the AEP were only found in the latter. Only 2 proteins (neutrophil defensins) were increased upon treatment with StatpSpS, while 65 proteins (among which are several typical AEP proteins) were decreased. In the 2-h AEP, 50 and 108 proteins were uniquely found in StatpSpS and control groups, respectively. Hemoglobin subunits and isoforms of keratin were only found in the StatpSpS group, while cystatin-C, cathepsin D, and cathepsin G, isoforms of heat shock 70 and protocadherin were exclusively found in the control group. In addition, 23 proteins were increased upon treatment with StatpSpS, among which are histatin-1, serum albumin, and isoforms of neutrophil defensin and keratin, while 77 were decreased, most of them were typical AEP proteins. In both evaluated periods, rinsing with StatpSpS profoundly changed the proteomic profile of the AEP, which might impact the protective role of this integument against carious or erosive demineralization. This study provides important insights on the dynamics of the protein composition of the AEP along time, after rinsing with a solution containing StatpSpS.


Asunto(s)
Proteoma , Proteómica , Esmalte Dental , Película Dental , Humanos , Péptidos
17.
Oral Oncol ; 118: 105315, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33985911

RESUMEN

Stimulation of saliva production is an alternative to improve the quality of life of patients treated by radiotherapy. However, there is no information about changes in the salivary proteome of stimulated and unstimulated saliva in these patients. OBJECTIVES: Thus, we evaluated the difference in the proteomic profile of stimulated and unstimulated saliva in patients with head and neck cancer (HNC) treated by radiotherapy. METHODS: Stimulated and unstimulated saliva were collected from 9 patients with HNC before (BRT), during (DRT; 2-5 weeks) and after (ART; 3-4 months) treatment. Healthy patients paired by age and gender also had their saliva collected (C; control group). The stimulated and unstimulated salivary flow were evaluated (p < 0.05). Salivary proteins were extracted and processed for shotgun proteomic analysis. RESULTS: Significant differences were observed between stimulated and unstimulated salivary flows for C and BRT (p greater than 0.001), but not for DRT and ART. Proteins involved with apoptosis, antibacterial and acid-resistance were decreased in stimulated saliva in comparison to unstimulated saliva DRT and ART. Isoforms of keratins were not identified in control and BRT. CONCLUSION: there is a marked difference in the protein profile of stimulated and unstimulated salivary flows in HNC patients treated by radiotherapy. In addition, saliva stimulation in patients with HNC decreases important proteins involved with dental protection. The unstimulated salivary flow seems to be the best alternative to search for biomarkers. Our results contribute in an unprecedented way to understand the changes in the salivary proteome of different flows in HNC patients undergoing radiotherapy treatment.


Asunto(s)
Neoplasias de Cabeza y Cuello , Proteoma , Saliva , Neoplasias de Cabeza y Cuello/radioterapia , Humanos , Proteómica , Calidad de Vida , Xerostomía
18.
J Dent ; 108: 103642, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33757866

RESUMEN

OBJECTIVES: To evaluate in vivo the proteomic profile of the acquired enamel pellicle (AEP) in patients with head and neck cancer (HNC) before, during and after radiotherapy. METHODS: Nine patients, after prophylaxis, had their AEPs collected before (BRT), during (DRT; 2-5 weeks) and after (ART; 3-4 months) radiotherapy. AEP was also collected from nine healthy patients (Control). The proteins were extracted in biological triplicate and processed by label-free proteomics. RESULTS: Statherin was increased more than 9-fold and several hemoglobin subunits were increased more than 5-fold DRT compared to BRT, while lactotransferrin, proline-rich proteins, cystatins, neutrophil defensins 1 and 3 and histatin-1 were decreased. ART, there was an increase in lactotransferrin and several isoforms of histones, while statherin and alpha-amylase proteins were decreased. MOAP-1 was exclusively found ART in comparison to BRT. When compared to Control, AEP of patients BRT showed an increase in proteins related to the perception of bitter taste, mucin-7 and alpha-amylases, while cystatin-S was decreased. CONCLUSIONS: HNC and radiotherapy remarkably altered the proteome of the AEP. Antibacterial and acid-resistant proteins were decreased during radiotherapy. CLINICAL SIGNIFICANCE: Our results provide important information for designing more effective dental products for these patients, in addition to contributing to a better understanding of the differential protective roles of the AEP proteins during radiotherapy. Moreover, some proteins identified in the AEP after radiotherapy may serve as prognostic markers for survival of HNC patients.


Asunto(s)
Proteínas del Esmalte Dental , Neoplasias de Cabeza y Cuello , Película Dental , Neoplasias de Cabeza y Cuello/radioterapia , Humanos , Proteoma , Proteómica , Saliva , Proteínas y Péptidos Salivales
19.
Arch Oral Biol ; 121: 104952, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33186792

RESUMEN

OBJECTIVE: Salivary proteomic analysis may help to understand physiopathological changes in crack cocaine dependents. This study aimed to compare the salivary protein profile between crack cocaine dependents and non-drug users. DESIGN: Nine heavy smokers and alcohol consumers men admitted to rehab due to crack cocaine abuse and nine non-drug users age-matched men were evaluated. Unstimulated whole saliva was collected. Proteomic analysis was performed by mass spectrometer. Data were processed using ProteinLynx GlobalServer software. Results were obtained by searching the Homo sapiens database from the UniProt catalog. The search tool IBI-IMIM was used to identify proteins candidates for biomarkers. RESULTS: The mean age of crack cocaine and control groups was 36.89 ±â€¯7.78 and 35.78 ±â€¯6.68 years, respectively. 458 salivary proteins were identified in both groups; 305 proteins in the crack cocaine group. Among the 68 proteins presented in both groups, 29 were down-regulated (i.e. "Statherin" and "Transforming growth factor-beta-induced protein ig-h3" were down-regulated at least 10-fold) and 27 up-regulated (i.e. "Negative elongation factor" was up-regulated 19-fold) in the crack cocaine group compared to controls. 90 out of the 458 proteins found in the proteomic analysis were identified as candidates for biomarkers of diseases. Among these, 65 (72.22 %) were detected in the crack cocaine group. CONCLUSION: Crack cocaine dependents with chronic alcohol and tobacco use have a higher number of proteins in saliva compared to non-drug users. 22.3 % of salivary proteins present in crack cocaine dependents were present in controls; 3.9 % of them were expressed in similar quantity.


Asunto(s)
Trastornos Relacionados con Cocaína/metabolismo , Cocaína Crack , Proteoma/análisis , Saliva/química , Proteínas y Péptidos Salivales/análisis , Adulto , Alcoholismo , Estudios de Casos y Controles , Humanos , Masculino , Proteómica , Fumar
20.
J Dent ; 102: 103478, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-32950632

RESUMEN

OBJECTIVES: To evaluate, in vivo: 1) proteomic alterations in the acquired enamel pellicle (AEP) after treatment with sugarcane-derived cystatin (CaneCPI-5), hemoglobin (HB), statherin-derived peptide (StN15) or their combination before the formation of the AEP and subsequent erosive challenge; 2) the protection of these treatments against erosive demnineralization. MATERIALS AND METHODS: In 5 crossover phases, after prophylaxis, 10 volunteers rinsed (10 mL, 1 min) with: deionized water-1, 0.1 mg/mL CaneCPI-5-2, 1.0 mg/mL HB-3, 1.88 × 10-5 M StN15-4 or their combination-5. AEP was formed (2 h) and enamel biopsy (10 µL, 1%citric acid, pH 2.5, 10 s) was performed on one incisor for calcium analysis. The same acid was applied on the vestibular surfaces of the remaining teeth. The acid-resistant proteins within the remaining AEP were collected. Samples were quantitatively analyzed by label-free proteomics. RESULTS: Treatment with the proteins/peptide, isolated or combined, increased several acid-resistant proteins in the AEP, compared with control. The highest increases were seen for PRPs (32-fold, StN15), profilin (15-fold, combination), alpha-amylase (9-fold; StN15), keratins (8-fold, CaneCPI-5 and HB), Histatin-1 (7-fold, StN15), immunoglobulins (6.5-fold, StN15), lactotransferrin (4-fold, CaneCPI-5), cystatins, lysozyme, protein S-100-A9 and actins (3.5-fold, StN15), serum albumin (3.5-fold, CaneCPI-5 and HB) and hemoglobin (3-fold, StN15). Annexin, calmodulin, keratin, tubulin and cystatins were identified exclusively upon treatment with the proteins/peptide, alone or combined. Groups 2, 3 and 4 had significantly lower Ca released from enamel compared to group 1 (Kruskal-Wallis/Dunn's, p < 0.05). CONCLUSIONS: Treatment with CaneCPI-5, HB or StN15 remarkably increases acid-resistant proteins in the AEP, protecting against erosion. CLINICAL SIGNIFICANCE: Our results show, for the first time, that treatment with proteins/peptide remarkably increases acid-resistant proteins in the AEP, protecting against erosive demineralization. These findings open an avenue for a new preventive approach for erosive demineralization, employing acquired pellicle engineering procedures that may in the future be incorporated into dental products.


Asunto(s)
Desmineralización Dental , Erosión de los Dientes , Esmalte Dental , Película Dental , Humanos , Péptidos , Proteómica , Desmineralización Dental/prevención & control
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