DOI: 10.5061/dryad.8gtht76p0
title: A pilot study on the degree of tooth staining caused by pollen water and Chlorhexidine: in vitro study

description: Background: Chlorhexidine oral rinse has been used as an adjuvant in the
 treatment of periodontal disease. However, there are draw backs of using
 chlorhexidine i.e. tooth staining and other side effects including allergy
 reaction. In light of proven therapeutic properties of pollen water as
 well as its relatively cheap cost in the market, pollen water has a
 potential to be an effective alternative to chlorhexidine oral rinse. The
 aim of this study is to compare the degree of tooth staining influenced by
 pollen water-based mouthwash to the standard Chlorhexidine mouthwash using
 spectrophotometer. Materials and Methods: 24 Specimens from extracted
 intact human teeth were soaked into the three different solutions, Pollen
 water, and normal water. Color measurements were carried out by a
 spectrophotometer devise and recorded at 5 different time intervals. Color
 change (ΔE), Chroma (C*) and Hue (H*) were analyzed and compared among the
 three solutions. Results: Overall mean ΔE was similar in all groups,
 significant difference between all time points was found only in Pollen
 water. The change in C* was higher in pollen water as compare to other
 solutions. There was a subtle increase in H* in the Chlorhexidine samples
 after week 3. The H* values in pollen water was stable but sudden decrease
 was observed in week 6. The difference in H* among the three solutions was
 significant after 3 weeks. Conclusion: Within the limitation of our study,
 it can be concluded that Pollen water stained teeth to a lesser extent
 than did Chlorhexidine. It might be beneficial to use Pollen water as
 mouthwash however further investigation is needed regarding efficacy of
 plaque control.
description: This study was
 conducted at the laboratory of Imam Abdurahman bin Faisal University
 College of Dentistry, after gaining ethical approval (EA #2018029) from
 the Scientific Research Unit. It consisted of the following
 steps. 1) Preparation of three
 solutions: Chlorhexidine 0.12 % licensed for
 use as a mouthwash. (positive control)
 Pollen water
 licensed for internal/external use (100% pure, cold-pressed). (test
 group) Normal water. (negative
 control) 2) Teeth preparation: 24 extracted
 intact human teeth were used in this study. Inclusion criteria were:
 caries-free teeth with no cracks, no previous root canal therapy, and no
 crowns, which were recently extracted for orthodontic treatment purposes.
 The teeth surfaces were cleaned using a scaler to dislodge debris and
 periodontal instruments to remove calculus. This was followed by brushing
 using a rubber cup. Subsequently, each sample was prepared to be at a size
 of 8 mm ×11 mm × 2.5 mm using a diamond bur (Iwanson, Ustomed, Tuttlingen,
 Germany). The total sample size was 24 (8 samples for each solution).
 3) Color Measurements: Each
 prepared sample was placed onto the chamber of a benchtop digital imaging
 spectrophotometer (X-rite color-Eye 7000A; Gmbh, Regensdorf, Switzerland),
 which was previously calibrated according to the manufacturer’s
 specifications. Color measurements were obtained in triplicate, and the
 mean value was considered as the final measurement.
 4) Each specimen
 was soaked in Chlorhexidine, pollen water, or normal water, and staining
 was measured weekly for 6 weeks. 5) The following parameters were
 analyzed: ΔE: indicates the change in color
 from baseline to that at the measurement time. 
 Chroma (C*):
 indicates the color intensity or saturation
 Hue (H*):
 describes the perception of an object’s color (e.g., red, orange, green,
 blue). <b><i>2.1
 Statistical Analysis</i></b><i> </i>
 The color change (ΔE) was calculated using the following Equation
 1:                           ΔEab* =
 [(ΔL*)<sup> 2</sup> + (Δa*)<sup> 2</sup> +
 (Δb*)<sup> 2</sup>]<sup>
 ½</sup>                                (1)
 Where: Δ represents the difference between the
 object being measured and its reference L* represents
 lightness a* and b* are the chromaticity
 coordinates Data normality was checked using the
 Shapiro-Wilk test with p &lt; 0.05
 indicating non-normally distributed data. The color differences for L, a,
 b, C*, and H* were calculated and analyzed using the Friedman test
 (non-parametric method of repeated-measure analysis of variance [ANOVA]).
 Repeated-measure ANOVA was applied separately for each solution. For
 significant findings, the paired sample t-test was performed to assess for
 individual significant results. We considered statistically significant at
 P &lt; 0.05. All analysis was done using SPSS version 22 software
 (IBM, Armonk, NY).