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Clinical Oral Investigations Apr 2024This study was designed in two-legs. In the in vivo, we explored the potential of a rinse solution containing a combination (Comb) of 0.1 mg/mL CaneCPI-5...
Acquired enamel pellicle and biofilm engineering with a combination of acid-resistant proteins (CaneCPI-5, StN15, and Hemoglobin) for enhanced protection against dental caries - in vivo and in vitro investigations.
OBJECTIVE
This study was designed in two-legs. In the in vivo, we explored the potential of a rinse solution containing a combination (Comb) of 0.1 mg/mL CaneCPI-5 (sugarcane-derive cystatin), 1.88 × 10M StN15 (statherin-derived peptide) and 1.0 mg/mL hemoglobin (Hb) to change the protein profile of the acquired enamel pellicle(AEP) and the microbiome of the enamel biofilm. The in vitro, was designed to reveal the effects of Comb on the viability and bacterial composition of the microcosm biofilm, as well as on enamel demineralization.
MATERIALS AND METHODS
In vivo study, 10 participants rinsed (10mL,1 min) with either deionized water (HO-control) or Comb. AEP and biofilm were collected after 2 and 3 h, respectively, after rinsing. AEP samples underwent proteomics analysis, while biofilm microbiome was assessed via 16 S-rRNA Next Generation Sequencing(NGS). In vitro study, a microcosm biofilm protocol was employed. Ninety-six enamel specimens were treated with: 1)Phosphate-Buffered Solution-PBS(negative-control), 2)0.12%Chlorhexidine, 3)500ppmNaF and 4)Comb. Resazurin, colony-forming-units(CFU) and Transversal Microradiography(TMR) were performed.
RESULTS
The proteomic results revealed higher quantity of proteins in the Comb compared to control associated with immune system response and oral microbial adhesion. Microbiome showed a significant increase in bacteria linked to a healthy microbiota, in the Comb group. In the in vitro study, Comb group was only efficient in reducing mineral-loss and lesion-depth compared to the PBS.
CONCLUSIONS
The AEP modification altered the subsequent layers, affecting the initial process of bacterial adhesion of pathogenic and commensal bacteria, as well as enamel demineralization.
CLINICAL RELEVANCE
Comb group shows promise in shaping oral health by potentially introducing innovative approaches to prevent enamel demineralization and deter tooth decay.
Topics: Humans; Dental Pellicle; Dental Caries; Proteomics; Biofilms; Hemoglobins; Tooth Demineralization
PubMed: 38642171
DOI: 10.1007/s00784-024-05651-0 -
Regenerative Biomaterials 2024Eradicating biofouling from implant surfaces is essential in treating peri-implant infections, as it directly addresses the microbial source for infection and...
Eradicating biofouling from implant surfaces is essential in treating peri-implant infections, as it directly addresses the microbial source for infection and inflammation around dental implants. This controlled laboratory study examines the effectiveness of the four commercially available debridement solutions '(EDTA (Prefgel), NaOCl (Perisolv), HO (Sigma-Aldrich) and Chlorhexidine (GUM Paroex))' in removing the acquired pellicle, preventing pellicle re-formation and removing of a multi-species oral biofilm growing on a titanium implant surface, and compare the results with the effect of a novel formulation of a peroxide-activated 'Poloxamer gel (Nubone Clean)'. Evaluation of pellicle removal and re-formation was conducted using scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy and X-ray photoelectron spectroscopy to assess the surface morphology, elemental composition and chemical surface composition. Hydrophilicity was assessed through contact angle measurements. The multi-species biofilm model included , and , reflecting the natural oral microbiome's complexity. Biofilm biomass was quantified using safranin staining, biofilm viability was evaluated using confocal laser scanning microscopy, and SEM was used for morphological analyses of the biofilm. Results indicated that while no single agent completely eradicated the biofilm, the 'Poloxamer gel' activated with 'HO' exhibited promising results. It minimized re-contamination of the pellicle by significantly lowering the contact angle, indicating enhanced hydrophilicity. This combination also showed a notable reduction in carbon contaminants, suggesting the effective removal of organic residues from the titanium surface, in addition to effectively reducing viable bacterial counts. In conclusion, the 'Poloxamer gel + HO' combination emerged as a promising chemical decontamination strategy for peri-implant diseases. It underlines the importance of tailoring treatment methods to the unique microbial challenges in peri-implant diseases and the necessity of combining chemical decontaminating strategies with established mechanical cleaning procedures for optimal management of peri-implant diseases.
PubMed: 38435376
DOI: 10.1093/rb/rbae014 -
Caries Research Mar 2024The identification of acid-resistant proteins, including hemoglobin (Hb), within the acquired enamel pellicle (AEP) led to the proposition of the "acquired pellicle...
INTRODUCTION
The identification of acid-resistant proteins, including hemoglobin (Hb), within the acquired enamel pellicle (AEP) led to the proposition of the "acquired pellicle engineering" concept, which involves the modification of the AEP by incorporating specific proteins, presenting a novel strategy to prevent dental demineralization.
OBJECTIVE
Combining in vivo and in vitro proof-of-concept protocols we sought to reveal the impact of AEP engineering with Hb protein on the biofilm microbiome and enamel demineralization.
METHODS
In the in vivo studies, 10-volunteers, in 2 independent experiments, rinsed (10mL,1min) with: deionized water-negative control or 1.0mg/mL Hb. The AEP and biofilm formed along 2 or 3h, respectively, were collected. AEP was analyzed by quantitative shotgun-label-free proteomics and biofilm by 16S-rRNA Next-Generation-Sequencing (NGS). In vitro study, a microcosm biofilm protocol was employed. Seventy-two bovine enamel specimens were treated with: 1)Phosphate-Buffered Solution-PBS, 2)0.12% Chlorhexidine, 3)500ppm NaF; 4)1.0mg/mL Hb; 5)2.0mg/mL Hb, and 6)4.0mg/mL Hb. The biofilm was cultivated for 5-days. Resazurin, colony-forming-units(CFU) and Transversal Microradiography(TMR) were performed.
RESULTS
Proteomics and NGS analysis revealed that Hb increased proteins with antioxidant, antimicrobial, acid-resistance, hydroxyapatite-affinity, calcium-binding properties and showed a reduction in oral pathogenic bacteria. In vitro experiments demonstrated that the lowest Hb concentration was the most effective in reducing bacterial activity, CFU and enamel demineralization compared to PBS.
CONCLUSION
These findings suggest that Hb could be incorporated into anticaries dental products to modify the oral microbiome and control caries, highlighting its potential for AEP and biofilm microbiome engineering.
PubMed: 38432208
DOI: 10.1159/000537976 -
Journal of Dentistry Apr 2024This study evaluated the effect of administration of trans-resveratrol-containing orodispersible tablets on the protein composition of the AEP and on blood plasma... (Randomized Controlled Trial)
Randomized Controlled Trial
OBJECTIVE
This study evaluated the effect of administration of trans-resveratrol-containing orodispersible tablets on the protein composition of the AEP and on blood plasma trans-resveratrol concentrations.
METHODS
Ten volunteers participated in two crossover double-blind phases. In each phase, after dental prophylaxis, they received a trans-resveratrol (15 mg) orodispersible tablet, or a placebo tablet (without actives). The AEP formed after 120 min was collected with electrode filter papers soaked in 3 % citric acid. Blood samples were collected 30, 45, 60 and 120 min after the use of the tablet. After protein extraction, AEP samples were analyzed by shotgun labelfree quantitative proteomics and plasma samples were analyzed by high-performance liquid chromatography (HPLC).
RESULTS
Eight hundred and two proteins were identified in the AEP. Among them, 336 and 213 were unique to the trans-resveratrol and control groups, respectively, while 253 were common to both groups. Proteins with important functions in the AEP had increased expression in the trans-resveratroltreated group, such as neutrophil defensins, S100 protein isoforms, lysozyme C, cystatin-D, mucin-7, alphaamylase, albumin, haptoglobin and statherin. Trans-resveratrol was detected in the plasma at all the times evaluated, with the peak at 30 min.
CONCLUSIONS
The administration of trans-resveratrol in sublingual orodispersible tablets was effective both to increase the bioavailability of the polyphenol and the expression of antibacterial and acid-resistant proteins in the AEP, which might benefit oral and general health.
Topics: Humans; Dental Pellicle; Proteins; Resveratrol; Cross-Over Studies; Double-Blind Method
PubMed: 38367826
DOI: 10.1016/j.jdent.2024.104876 -
Technology and Health Care : Official... Jan 2024Commercially available oral rinses contain active ingredients with concentration that is claimed by manufacturers to be effective as antiplaque agent. To date there has...
The commercially available oral rinse vs. curcumin photosensitizers in an artificial mouth model mimicking their use after meals on early colonizers single species biofilm: An in vitro study.
BACKGROUND
Commercially available oral rinses contain active ingredients with concentration that is claimed by manufacturers to be effective as antiplaque agent. To date there has been no mention of the effect of oral rinse on the adherence of early plaque colonizers in plaque formation and the concentration to be used before/after meals.
OBJECTIVE
The chief aim of the study was to evaluate microbial retention on the salivary pellicle on treatment with oral rinses (CHX & EO)/PS (mimicking after meals use of mouth wash/PS).
METHODS
Noordini's Artifical Mouth model was used for developing the single species biofilm with early microbial colonizers of oral biofilm (A. viscosus, Strep. mitis and Strep. sanguinis respectively). The microbial retention on use of oral rinses comprising of CHX and EO as an active ingredients respectively was compared with Curcumin PS. For evaluating the microbial retention, the pellicle with microbial inoculation was developed on the glass beads in the mouth model. Subsequently the respective single specie biofilm was exposed to the mouth wash and PS after inoculation. It mimicked as use of mouth wash/PS after meals. The bacterial count in the dental biofilm was evaluated on serial dilution (CFU/ml). Sterile deionized water was used as a negative control. For qualitative analysis, Scanning electron microscope (SEM) was used to evaluate the microbial count.
RESULTS
From the data it was observed that for the treatment of single species experimental biofilm with commercially available mouth rinses (CHX & EO) and PS (curcumin), there was significant retention for S.mitis, S.sanguinis and A.viscosus. There was no significant difference observed between PS and CHX treated single species biofilm. Whereas a significant difference was observed between EO treated biofilms and CHX/PS treated biofilms (p⩽ 0.05).
CONCLUSION
It can be concluded from the results that curcumin PS and CHX should not be used after meals whereas EO containing mouth rinse can be used to maintain the oral mocroflora.
PubMed: 38339947
DOI: 10.3233/THC-231814 -
Archives of Oral Biology Apr 2024To study the effects of carbon dots (CDs), in combination with phytosphingosine (PHS), against acid-induced demineralization of hydroxyapatite in vitro.
OBJECTIVES
To study the effects of carbon dots (CDs), in combination with phytosphingosine (PHS), against acid-induced demineralization of hydroxyapatite in vitro.
METHODS
CDs were generated from citric acid and urea by microwave heating. Transmission electron microscope (TEM), FT-IR, and fluorescence intensity were used to characterize the CDs. A hydroxyapatite (HAp) model was used to investigate the protective effects of CDs, PHS, and their combinations with and without a salivary pellicle against acid-induced demineralization in vitro. Ca release as a parameter to evaluate the inhibition of demineralization was measured by capillary electrophoresis. The interactions between CDs, PHS, and HAp discs were investigated using a fluorescence detector.
RESULTS
Uniform-sized CDs were synthesized, showing typical optical characteristics. CDs exhibited no inhibition of acid-induced demineralization in vitro, in contrast to PHS. Notably, a pre-coating of CDs increased the protective effects of PHS against acid-induced demineralization, which was not disturbed by the presence of a salivary pellicle and Tween 20. Scanning electron microscope (SEM) confirmed the binding and layers formed of both CDs and PHS to the HAp surfaces. Based on fluorescence spectra CDs binding to HAp seemed to be dependent on Ca and PO interactions.
CONCLUSIONS
CDs combined with PHS showed protective effects against acid-induced demineralization of HAp discs in vitro.
Topics: Humans; Durapatite; Carbon; Spectroscopy, Fourier Transform Infrared; Tooth Demineralization; Sphingosine
PubMed: 38335699
DOI: 10.1016/j.archoralbio.2024.105911 -
Caries Research 2024Erosive tooth wear is a highly prevalent dental condition that is modified by the ever-present salivary pellicle. The aim of the present in situ study was to investigate...
INTRODUCTION
Erosive tooth wear is a highly prevalent dental condition that is modified by the ever-present salivary pellicle. The aim of the present in situ study was to investigate the effect of polyphenols on the ultrastructure of the pellicle formed on dentin in situ and a subsequent erosive challenge.
METHODS
The pellicle was formed on bovine dentin specimens for 3 min or 2 h in 3 subjects. After subjects rinsed with sterile water (negative control), 1% tannic acid, 1% hop extract, or tin/fluoride solution containing 800 ppm tin and 500 ppm fluoride (positive control), specimens were removed from the oral cavity. The erosive challenge was performed on half of the specimens with 1% citric acid, and all specimens were analyzed by transmission electron microscopy. Incorporation of tannic acid in the pellicle was investigated by fluorescence spectroscopy.
RESULTS
Compared to the negative control, ultrastructural analyses reveal a thicker and electron-denser pellicle after application of polyphenols, in which, according to spectroscopy, tannic acid is also incorporated. Application of citric acid resulted in demineralization of dentin, but to a lesser degree when the pellicle was pretreated with a tin/fluoride solution. The pellicle was more acid-resistant than the negative control when modified with polyphenols or tin/fluoride solution.
CONCLUSION
Polyphenols can have a substantial impact on the ultrastructure and acid resistance of the dentin pellicle, while the tin/fluoride solution showed explicit protection against erosive demineralization.
Topics: Humans; Animals; Cattle; Dental Enamel; Dental Pellicle; Fluorides; Tooth Erosion; Tin; Polyphenols; Tin Fluorides; Citric Acid; Dentin
PubMed: 38198764
DOI: 10.1159/000536199 -
Caries Research 2024This study investigated the changes in the acquired enamel pellicle (AEP) proteome when this integument is formed in vivo after treatment with sugarcane-derived cystatin...
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 × 10-5
M StN15 - group 4, or a blend of these - group 5. Following this, AEP formation occurred (2 h) and an enamel biopsy (10 µL, 0.01m 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, m
M ) 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.
Topics: Humans; Calcium; Dental Pellicle; Peptides; Proteome; Tooth Erosion; Hemoglobins
PubMed: 38198757
DOI: 10.1159/000536200 -
Dentistry Journal Dec 2023In this study, we examined the cytotoxic effects of six commercial children's mouthrinses (designated as #1, #2, #3, #4, #5, and #6) and four commercial children's...
In this study, we examined the cytotoxic effects of six commercial children's mouthrinses (designated as #1, #2, #3, #4, #5, and #6) and four commercial children's toothpastes (designated as #1, #2, #3, and #4) on primary human neonatal melanocytes that were used as a representative model for oral melanocytes. Mouthrinses diluted directly with culture medium (1:2, 1:5, 1:10, 1:100, and 1:1000) were added to monolayers of melanocytes for 2 min, followed by 24 h recovery, after which MTS cytotoxicity assay was conducted. The extracts of each toothpaste were prepared (50% /), diluted in culture medium (1:2, 1:5, 1:10, 1:50, 1:100, and 1:1000), and added to cell monolayers for 2 min (standard brushing time), followed by an analysis of cell viability after 24 h. Results showed that all mouthrinses except mouthrinse #4 showed significantly greater loss of cell viability, ascribed to cetylpyridinium chloride (CPC) that induced significant cytotoxicity to melanocytes (IC = 54.33 µM). In the case of toothpastes, the examination of cellular morphology showed that a 2 min exposure to all toothpaste extracts induced a concentration-dependent decline in cell viability, pronounced in toothpaste containing sodium lauryl sulfate (SLS) detergent. Further results suggested SLS to be the critical driver of cytotoxicity (IC = 317.73 µM). It is noteworthy that toothpaste #1 exhibited much lower levels of cytotoxicity compared to the other three toothpastes containing SLS. Taken together, these findings suggest that the melanocytotoxicity of children's mouthrinse (#4) and toothpaste (#1) is comparatively low. To the best of our knowledge, this is the first study to examine the impact of children's toothpastes and mouthrinses on neonatal primary human melanocytes. Future studies to investigate these findings in a realistic scenario replicating oral cavity conditions of the presence of microbiota, pellicle layer and saliva, and other cell types are warranted.
PubMed: 38132425
DOI: 10.3390/dj11120287 -
Pediatric Dentistry Nov 2023To evaluate, in vitro, whether the presence of the acquired pellicle (AP) interferes with the effect of 38 percent silver diamine fluoride (SDF) on the inhibition or...
To evaluate, in vitro, whether the presence of the acquired pellicle (AP) interferes with the effect of 38 percent silver diamine fluoride (SDF) on the inhibition or progression of carious dentin. Carious dentin specimens were divided into groups: C-no treatment control; FV-fluoride varnish; AP-acquired pellicle formation; SDF only, and APSDF-SDF plus acquired pellicle formation. After AP formation (AP and APSDF groups), all groups were subjected to pH cycling. Dentin mineral gain or loss was quantified using surface microhardness (SM). Concentrations of fluoride calcium (CaF₂) and fluorapatite (FAp) fluoride retained in dentin and fluoride concentrations in de- and remineralizing solutions were determined. Groups AP, FV, SDF, and APSDF showed reduced demineralization compared with the C group, but the AP group showed greater surface loss than the fluoridated groups. Groups FV, SDF, and APSDF showed greater mineral recovery than C and AP groups (P<0.05), but no differences were observed between the FV, SDF, and APSDF groups (P>0.05). The retained CaF₂ and FAp were greater in the groups treated with fluoride, but there was no difference between the SDF and APSDF groups. The presence of the acquired pellicle reduced the progression of carious dentin lesion but did not influence its remineralization when associated with silver diamine fluoride.
Topics: Humans; Fluorides, Topical; Fluorides; Dental Pellicle; Dentin; Dental Caries; Quaternary Ammonium Compounds; Minerals
PubMed: 38129757
DOI: No ID Found