---
title: "Short-term effects of toothpaste formulations on salivary calcium: randomized crossover study"
id: "plos-one-0-short-term-effects-of-different-toothpaste-formulations-on-total-salivary"
canonical_url: "https://medichelpline.com/clinical-feed/plos-one-0-short-term-effects-of-different-toothpaste-formulations-on-total-salivary"
content_type: "clinical_feed_article"
specialty: "Dentistry"
source_name: "PLOS ONE (Medicine)"
source_url: "https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715"
published_at: "2026-09-17T14:00:00.000Z"
evidence_level: "Journal Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# Short-term effects of toothpaste formulations on salivary calcium: randomized crossover study
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/plos-one-0-short-term-effects-of-different-toothpaste-formulations-on-total-salivary
- **Specialty:** [Dentistry](https://medichelpline.com/clinical-feed/dentistry.md)
- **Primary Source:** PLOS ONE (Medicine)
- **Source URL:** [Original Journal Publication](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715)
- **Published At:** 2026-09-17T14:00:00.000Z
- **Evidence Rating:** Journal Feed
## Executive GIST (TL;DR)
- This randomized, three-treatment, three-period crossover trial evaluated immediate changes in **salivary calcium** after single use of three commercially available sodium fluoride toothpastes in 48 healthy adults. - Toothpastes tested were: a PVM/MA copolymer formulation (Sensodyne), a pyrophosphate plus zinc formulation (Dentalux), and a sodium bicarbonate formulation without anticalculus agents (Parodontax). - Unstimulated saliva was sampled at baseline (T0), immediately after brushing (T1), 30 minutes (T2), and 60 minutes (T3); calcium was measured by a colorimetric assay. - Linear mixed-effects models assessed effects of toothpaste formulation, time, treatment sequence, and the toothpaste-by-time interaction, with participant as a random effect. - Significant main effects were observed for **toothpaste formulation** and **measurement time**, and a significant toothpaste-by-time interaction; treatment sequence was not significant. - Parodontax produced significantly lower salivary calcium concentrations immediately after brushing compared with Dentalux and Sensodyne (Bonferroni-adjusted P < 0.001); differences diminished at 30 minutes and were absent at 60 minutes. - No significant differences were found between Dentalux (pyrophosphate/zinc) and Sensodyne (PVM/MA) at any time point. - Authors emphasize that products differ in multiple ingredients, so findings reflect product-level effects rather than isolated ingredient effects; they call for multicentre studies to confirm findings and determine clinical relevance. - Study was approved by Witten/Herdecke University ethics committee, enrolled 48 participants based on an a priori power calculation, and was retrospectively registered (DRKS00039385).
## Clinical Analysis & Structured Key Points
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Short-term effects of different toothpaste formulations on total salivary calcium: A randomized crossover study * Lina Aminy , Contributed equally to this work with: Lina Aminy, Mozhgan Bizhang Roles Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Project administration, Software, Writing – original draft, Writing – review & editing * E-mail: lina.aminy@uni-wh.de Affiliation Chair of Operative and Preventive Dentistry, Witten/Herdecke University, Witten, Germany [ ![ORCID logo](https://journals.plos.org/resource/img/orcid_16x16.png) https://orcid.org/0009-0004-6546-9923 ](https://orcid.org/0009-0004-6546-9923 "ORCID Registry") ⨯ * Mozhgan Bizhang , Contributed equally to this work with: Lina Aminy, Mozhgan Bizhang Roles Conceptualization, Formal analysis, Methodology, Project administration, Resources, Supervision, Validation, Visualization, Writing – original draft, Writing – review & editing Affiliation Chair of Operative and Preventive Dentistry, Witten/Herdecke University, Witten, Germany ⨯ * Claudia Klinger, Roles Formal analysis, Project administration, Validation, Writing – review & editing Affiliation Division of Functional Genomics, Chair of Biochemistry and Molecular Medicine, ZBAF, Witten/Herdecke University, Witten, Germany ⨯ * Stefan Zimmer, Roles Writing – review & editing Affiliation Chair of Operative and Preventive Dentistry, Witten/Herdecke University, Witten, Germany [ ![ORCID logo](https://journals.plos.org/resource/img/orcid_16x16.png) https://orcid.org/0000-0001-6697-2548 ](https://orcid.org/0000-0001-6697-2548 "ORCID Registry") ⨯ * Andreas Savelsbergh Roles Conceptualization, Formal analysis, Methodology, Project administration, Resources, Supervision, Validation, Visualization, Writing – review & editing Affiliation Division of Functional Genomics, Chair of Biochemistry and Molecular Medicine, ZBAF, Witten/Herdecke University, Witten, Germany [ ![ORCID logo](https://journals.plos.org/resource/img/orcid_16x16.png) https://orcid.org/0000-0002-6136-3230 ](https://orcid.org/0000-0002-6136-3230 "ORCID Registry") ⨯ # Short-term effects of different toothpaste formulations on total salivary calcium: A randomized crossover study * Lina Aminy, * Mozhgan Bizhang, * Claudia Klinger, * Stefan Zimmer, * Andreas Savelsbergh ![PLOS](https://journals.plos.org/resource/img/logo-plos-full-color.svg) x * Published: September 17, 2026 * * [Article](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715) * [Authors](https://journals.plos.org/plosone/article/authors?id=10.1371/journal.pone.0358715) * [Metrics](https://journals.plos.org/plosone/article/metrics?id=10.1371/journal.pone.0358715) * [Comments](https://journals.plos.org/plosone/article/comments?id=10.1371/journal.pone.0358715) * [Media Coverage](http://plos.altmetric.com/details/doi/10.1371/journal.pone.0358715) * [Peer Review](https://journals.plos.org/plosone/article/peerReview?id=10.1371/journal.pone.0358715) * [Abstract](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#abstract0) * [Introduction](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#sec005) * [Materials and methods](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#sec006) * [Results](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#sec016) * [Discussion](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#sec021) * [Conclusion](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#sec022) * [Supporting information](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#sec024) * [References](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#references) * [Reader Comments](https://journals.plos.org/plosone/article/comments?id=10.1371/journal.pone.0358715) * [Figures](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715) ## Abstract ### Objectives Dental calculus formation can be inhibited by toothpastes containing pyrophosphate, which chelates calcium ions and prevents hydroxyapatite crystal formation. Although pyrophosphate may also influence salivary calcium availability, the short-term effects of commercially available toothpaste formulations on salivary calcium concentrations remain insufficiently characterized. This study investigated and compared the short-term effects of three fluoride toothpastes on salivary calcium concentration. ### Methods A randomized, three-treatment, three-period crossover trial was conducted in 48 healthy volunteers. The participants used three commercially available sodium fluoride toothpastes in randomized order: a formulation containing methyl vinyl ether/maleic acid copolymer, PVM/MA copolymer (Sensodyne), a formulation containing pyrophosphate and zinc (Dentalux), and a formulation containing sodium bicarbonate (Parodontax). Treatment periods were separated by a 2-day washout. Unstimulated saliva was collected at baseline (T0), immediately after brushing (T1), 30 min (T2), and 60 min (T3). Salivary calcium concentrations were quantified using a colorimetric assay. Data were analysed using linear mixed-effects models with toothpaste formulation, measurement time, treatment sequence, and the toothpaste-by-time interaction as fixed effects and participant as a random effect. ### Results Significant effects were observed for toothpaste formulation (F(2,103.35)=42.31, P < 0.001), measurement time (F(3,141)=39.78, P < 0.001), and the toothpaste-by-time interaction (F(6,141)=23.91, P < 0.001), whereas treatment sequence had no significant effect (P = 0.927). Compared with Dentalux® and Sensodyne®, Parodontax® resulted in significantly lower salivary calcium concentrations (both Bonferroni-adjusted P < 0.001), immediately after brushing (T1). The differences decreased after 30 min and were no longer significant after 60 min. No significant differences were observed between Dentalux® and Sensodyne® at any time point. ### Conclusions Commercially available fluoride toothpastes produced different short-term effects on the salivary calcium concentration, with the greatest differences occurring immediately after brushing. Because the products differed in terms of multiple formulation ingredients, the findings reflect product-level effects rather than ingredient-specific effects. Further multicentre studies are warranted to confirm these findings and evaluate their clinical relevance. ## Figures ![Fig 2](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.g002) ![Table 1](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t001) ![Fig 1](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.g001) ![Table 2](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t002) ![Table 3](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t003) ![Table 4](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t004) ![Table 5](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t005) ![Table 6](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t006) ![Table 7](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t007) ![Table 8](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t008) ![Fig 2](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.g002) ![Table 1](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t001) ![Fig 1](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.g001) ![Table 2](https://journals.plos.org/plosone/article/figure/image?size=inline&id=10.1371/journal.pone.0358715.t002) **Citation:** Aminy L, Bizhang M, Klinger C, Zimmer S, Savelsbergh A (2026) Short-term effects of different toothpaste formulations on total salivary calcium: A randomized crossover study. PLoS One 21(9): e0358715. https://doi.org/10.1371/journal.pone.0358715 **Editor:** Esra Cengiz Yanardag, Mersin University: Mersin Universitesi, TÜRKIYE **Received:** April 13, 2026; **Accepted:** August 31, 2026; **Published:** September 17, 2026 **Copyright:** © 2026 Aminy et al. This is an open access article distributed under the terms of the [Creative Commons Attribution License](http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. **Data Availability:** All relevant data are within the paper and its [Supporting Information](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#sec024) files. **Funding:** This study was supported by the Fördergemeinschaft of Witten/Herdecke University. The funding source had no involvement in the study design; collection, analysis, or interpretation of data; writing of the manuscript; or the decision to submit the article for publication. **Competing interests:** The authors have declared that no competing interests exist. ## Introduction Toothpaste provides multiple oral health benefits, including improved oral freshness, anticariogenic efficacy, inhibition of biofilm regrowth, tooth whitening, and enhanced patient compliance, largely due to the incorporation of chemotherapeutic agents [[1](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref001)]. Dental calculus is a mineralized biofilm composed of an inorganic mineral phase embedded within an organic matrix. Its mineral components consist primarily of brushite (dicalcium phosphate dihydrate), octacalcium phosphate (OCP), hydroxyapatite (HAP), and whitlockite. Calculus formation begins with the acquired pellicle, a protein-rich salivary layer that rapidly forms on the enamel surface and facilitates bacterial adhesion and subsequent biofilm development. As the biofilm matures, calcium and phosphate ions precipitate within the extracellular matrix, leading to progressive mineralization and calculus formation [[2](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref002)–[4](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref004)]. Saliva plays a central role in oral homeostasis and is essential for both calculus formation and enamel remineralization. Its ionic composition resembles that of systemic body fluids, while its buffering capacity helps maintain a stable oral pH. In addition, saliva contains calcium, phosphate, and other inorganic ions that promote the remineralization of early carious lesions and previously demineralized enamel, thereby constituting an intrinsic defence mechanism against mineral loss [[5](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref005)]. Because calculus formation depends on calcium phosphate precipitation, the availability of free calcium ions in saliva is a key determinant of mineral deposition and may influence remineralization processes. Although mechanical plaque removal remains the cornerstone of oral hygiene, chemotherapeutic agents are widely incorporated into toothpastes to inhibit plaque mineralization and calculus formation. Chelating agents reduce calcium availability by forming soluble complexes, thereby interfering with crystal nucleation and growth. Soluble pyrophosphates are among the most extensively studied anti-calculus agents and have consistently been shown to inhibit mineral deposition, delay hydroxyapatite formation, and retard the phase conversion from brushite to hydroxyapatite [[6](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref006)–[10](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref010)]. Zinc salts further inhibit plaque mineralization without compromising the anticaries activity of fluoride and are therefore frequently combined with pyrophosphates [[10](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref010)–[12](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref012)] Another widely used anti-calculus agent is methyl vinyl ether/maleic acid copolymer (PVM/MA, Gantrez®), which reduces bacterial adhesion by modifying enamel surface properties and decreasing calcium availability while also exerting modest inhibitory effects on crystal growth [[13](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref013)–[15](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref015)] Whether anticalculus formulations influence fluoride-mediated remineralization remains controversial. Several in vitro and experimental studies have reported no significant effects of pyrophosphate-containing toothpastes on fluoride uptake or enamel remineralization [[15](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref015)–[18](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref018)]. In contrast, long-term clinical trials have consistently demonstrated greater reductions in caries progression with conventional sodium fluoride toothpastes than with randomized stannous fluoride/pyrophosphate formulations [[19](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref019)–[21](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref021)], whereas randomized controlled trials directly comparing fluoride toothpastes with and without pyrophosphates remain scarce and have yielded inconsistent findings [[12](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref012)]. Consequently, the current evidence does not conclusively exclude the possibility that pyrophosphate-containing formulations may reduce anticaries efficacy. From a mechanistic perspective, pyrophosphates inhibit calcium phosphate nucleation and hydroxyapatite crystal growth, providing a biologically plausible explanation for the potential reduction in remineralization. Moreover, the reported inverse association between calculus accumulation and caries incidence offers indirect evidence for the hypothesis that strategies designed to inhibit calculus formation may unintentionally influence caries development [[22](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref022)]. Although research on toothpaste formulations has increasingly shifted towards other active ingredients, these unresolved questions warrant renewed investigation [[23](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref023)]. To date, little is known about the immediate effects of anticalculus toothpastes on salivary calcium availability. Since pyrophosphate acts primarily through calcium complexation, measuring salivary calcium concentrations may provide valuable insight into the biological availability of calcium following toothpaste use and its potential implications for remineralization. Therefore, the present randomized three-period crossover study was designed to evaluate the effects of a single application of three toothpastes—two containing anticalculus agents (pyrophosphate/zinc and PVM/MA) and one without anticalculus agents—on salivary calcium concentrations. The primary objective was to determine whether changes in salivary calcium concentration over time differed between toothpaste formulations. The secondary objectives were to compare salivary calcium concentrations across toothpaste formulations, measurement times, and treatment sequences. ## Materials and methods ### Ethics Ethical approval was obtained from the Ethics Committee of Witten/Herdecke University (S-324/2023). The study was conducted in accordance with the principles of the Declaration of Helsinki. Before participation, all participants received detailed information about the study and provided written informed consent before enrolment. No minors were included in the study. Trial registration: This study was originally designed as an exploratory physiological crossover study investigating salivary biomarkers as the primary outcome, without evaluating clinical efficacy endpoints. Hence prospective trial registration was not performed before participant enrolment. To ensure transparency, the study was retrospectively registered in the German Clinical Trials Register (DRKS) under the identifier DRKS00039385. ### Sample size Power analysis was conducted using G*Power (version 3.1) [[24](https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0358715#pone.0358715.ref024)]. An a priori sample size calculation for a repeated-measures design indicated that a total sample size of 48 participants would be required to detect a medium effect size (f = 0.25) with 80% statistical power at α = 0.05, assuming four repeated measurements and a correlation of 0.50 among repeated observations (achieved power = 0.804). Accordingly, 48 participants were enrolled in this randomized crossover trial. ### Study design The investigation was designed as a randomized, three-treatment, three-period crossover clinical trial. Each participant received all three toothpaste formulations according to one of three predefined treatment seque
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