Proteomics of acquired pellicle in gastroesophageal reflux disease patients with or without erosive tooth wear

Martini, Tatiana; Ríos, Daniela; Cassiano, Luiza Paula Silva; Silva, Cíntia Maria de Souza; Taira, Éven Akemi; Ventura, Talita Mendes Oliveira; Pereira, Hugo Juarez Vieira; Magalhães, Ana Carolina; Carvalho, Thiago Saads; Baumann, Tommy; Lussi, Adrian; Oliveira, Ricardo Brandt de; Palma‐Dibb, Regina Guenka; Buzalaf, Marília Afonso Rabelo

doi:10.1016/j.jdent.2018.12.007

Cited by 37 publications

(52 citation statements)

References 38 publications

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“…Previous studies have demonstrated the importance of the protein composition of the AEP. Distinct protein profiles have been observed between GERD patients with and without erosive tooth wear [Martini et al, 2019]. Furthermore, statherin is reduced in the AEP in eroded teeth compared to healthy teeth in the same individual .…”

Section: Discussionmentioning

confidence: 99%

Differences in the Natural Enamel Surface and Acquired Enamel Pellicle following Exposure to Citric or Hydrochloric Acid

et al. 2020

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Objectives: The aim of this study was to investigate variations in the interaction between enamel, that is, the acquired enamel pellicle (AEP) and citric or hydrochloric acid. Materials and Methods: A 24-h AEP was formed on natural enamel specimens (n = 40) from pooled whole mouth human saliva. Samples were randomly allocated to citric (0.3%, pH 3.2) or hydrochloric (HCl) acid (0.01 M, pH 2.38) exposure for 30 or 300 s. The total protein concentration (TPC), and phosphorous and calcium concentrations of the pellicle were determined before and after acid exposure, and again after re-immersion in saliva. Surface roughness and tandem scanning confocal microscopy imaging were used to assess enamel changes. Results: After 300 s of citric acid exposure, the mean ± SD TPC reduced from 5.1 ± 1.1 to 3.5 ± 1.1 mg/mL (p < 0.05). In contrast, after 300 s of HCl exposure, the mean TPC did not reduce significantly from baseline (6.6 ± 1.1 to 5.7 ± 0.7 mg/mL) but was significantly reduced in the reformed pellicle to 4.9 ± 1.2 mg/mL (p < 0.001). This reduction occurred after significant release of calcium and phosphorous from the enamel surface (p < 0.001). Thirty seconds of exposure to either acid had no obvious effect on the AEP. The surface roughness of the enamel decreased after acid exposure but no differences between groups was observed. Conclusions: These findings indicate that citric acid interacted with proteins in the AEP upon contact, offering enamel protection. In contrast, HCl appeared to bypass the pellicle, and reduced protein was observed only after changes in the enamel chemical composition.

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Section: Discussionmentioning

confidence: 99%

Differences in the Natural Enamel Surface and Acquired Enamel Pellicle following Exposure to Citric or Hydrochloric Acid

et al. 2020

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“…The protocol of the study was approved by the Ethics Committees of Bauru and Ribeirão Preto School of Dentistry, University of São Paulo (CAAE 44007415.1.0000.5417 and 44007415.1.3001.5419, respectively). The same patients who had taken part in our previous AEP study [Martini et al, 2019] also took part in this study (n = 8 per group), after signing an informed consent. They were of both genders and between 20 and 60 years of age.…”

Section: Part 1: Clinical Study Ethical Aspects and Subjectsmentioning

confidence: 99%

“…Recently, we have found important alterations in the protein profile of the AEP of GERD patients with ETW, compared to those without ETW [Martini et al, 2019]. Interestingly, the proteome of AEP of GERD patients without ETW showed an increased expression of various subunits of hemoglobin (Hb), different isoforms of cystatin, as well as in albumin, when compared to GERD patients with ETW [Martini et al, 2019]. Among these proteins, the greatest increase was found for Hb (more than 3-fold).…”

Section: Introductionmentioning

confidence: 99%

Salivary Hemoglobin Protects against Erosive Tooth Wear in Gastric Reflux Patients

et al. 2020

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Objective: Saliva is the most important biological factor to protect against erosive tooth wear (ETW). Gastroesophageal reflux disease (GERD) patients have an increased risk of ETW due to the frequent presence of intrinsic acids in the oral cavity. Remarkably, not all GERD patients suffer from ETW, which might be due to differences in the composition of the saliva. Methods: This study compared the proteomic profile of saliva in patients (1) with GERD and ETW (basic erosive wear examination, BEWE, score ≥9; GE group) and (2) with GERD without ETW (BEWE = 0; GNE group) using shotgun label-free quantitative proteomic analysis nLC-ESI-MS/MS. The ability of hemoglobin (Hb) to protect against initial enamel erosion caused by a daily 10-s immersion of enamel specimens in 0.01 M HCl (pH 2.3) for 3 days was evaluated in vitro for proof of concept. Surface hardness change was used as response variable. Results: The differential expression of Hb subunits was significantly increased in the GNE group versus the GE group, in particular the Hb α-subunit that showed a >22-fold increase. Expressions of serum albumin (4.5-fold) and isoforms of cytoskeletal keratin type II (>3-fold) were also increased in the GNE group. Proteinase inhibitors, such as α1-antitrypsin and α2-macroglobulin, were only identified in the GNE group. In vitro, Hb (1.0 and 4.0 mg/mL) significantly reduced initial enamel erosion compared to a negative control after 3 days. Conclusions: Our results indicate that many proteins, with special emphasis on Hb, may be involved in the resistance of GERD patients to the occurrence of ETW. These proteins may be candidates for inclusion in dental products to protect against ETW.

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“…These remain in the AEP after exposure to acids 8 and are currently objects of great interest, since they might protect against dental caries and erosion. In the last few years, proteomic approaches have been used to identify these proteins 9 - 12 so that they can be added to dental products which, when applied, could modify the composition of the AEP, increasing its protective potential against acids. 13 …”

Section: Introductionmentioning

confidence: 99%

“…The amount of proteins that can be recovered is even smaller under in vitro condition, due to the absence of continuous salivary flow. Moreover, in the in vivo studies available so far, AEP samples collected from 8-10 volunteers are pooled in order to obtain enough proteins to be analyzed by mass spectrometry, 10 - 12 , 14 - 18 which does not allow proper assessment of the biological variation of the samples. In these in vivo studies, the collection of the AEP samples is done with filter paper soaked in 3% citric acid.…”

Section: Introductionmentioning

confidence: 99%

Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis

2020

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Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis Saliva is the major contributor for the protein composition of the acquired enamel pellicle (AEP), a bacteria-free organic layer formed by the selective adsorption of salivary proteins on the surface of the enamel. However, the amount of proteins that can be recovered is even smaller under in vitro condition, due to the absence of continuous salivary flow. Objective: This study developed an in vitro AEP protocol for proteomics analysis using a new formation technique with different collection solutions. Methodology: 432 bovine enamel specimens were prepared (4x4 mm) and divided into four groups (n=108). Unstimulated saliva was provided by nine subjects. The new AEP formation technique was based on saliva resupply by a new one every 30 min within 120 minutes at 37ºC under agitation. AEP was collected using an electrode filter paper soaked in the collection solutions according with the group: 1) 3% citric acid (CA); 2) 0.5% sodium dodecyl sulfate (SDS); 3) CA followed by SDS (CA+SDS); 4) SDS followed by CA (SDS+CA). The pellicles collected were processed for analysis through LC-ESI-MS/MS technique. Results: A total of 55 proteins were identified. The total numbers of proteins identified in each group were 40, 21, 28 and 41 for the groups CA, SDS, CA+SDS and SDS+CA, respectively. Twenty-three typical AEP proteins were identified in all groups, but Mucin was only found in CA and CA+SDS, while three types of PRP were not found in the SDS group. Moreover, a typical enamel protein, Enamelin, was identified in the CA+SDS group only. Conclusion: The new technique of the in vitro AEP formation through saliva replacement was essential for a higher number of the proteins identified. In addition, considering practicality, quantity and quality of identified proteins, citric acid seems to be the best solution to be used for collection of AEP proteins.

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Proteomics of acquired pellicle in gastroesophageal reflux disease patients with or without erosive tooth wear

Cited by 37 publications

References 38 publications

Differences in the Natural Enamel Surface and Acquired Enamel Pellicle following Exposure to Citric or Hydrochloric Acid

Differences in the Natural Enamel Surface and Acquired Enamel Pellicle following Exposure to Citric or Hydrochloric Acid

Salivary Hemoglobin Protects against Erosive Tooth Wear in Gastric Reflux Patients

Optimizing the formation of the acquired enamel pellicle in vitro for proteomic analysis

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