A comparative evaluation of efficacy of biomin F and propolis containing toothpastes on dentinal tubule occlusion with and without use of an adjunct 810 nm diode laser: An In vitro scanning electron microscope study

Komal Khare; Prashant Bhusari; Abhishek Soni; Sachin K Malagi; Dennis Abraham; Lynn Johnson

doi:10.4103/jmau.jmau_133_20

ORIGINAL ARTICLE

Year : 2023 | Volume : 11 | Issue : 1 | Page : 41-46

A comparative evaluation of efficacy of biomin F and propolis containing toothpastes on dentinal tubule occlusion with and without use of an adjunct 810 nm diode laser: An In vitro scanning electron microscope study

Komal Khare, Prashant Bhusari, Abhishek Soni, Sachin K Malagi, Dennis Abraham, Lynn Johnson
Department of Periodontics, Maitri College of Dentistry and Research Centre, Chhattisgarh, India

Date of Submission	10-Dec-2020
Date of Decision	27-May-2021
Date of Acceptance	19-Jun-2021
Date of Web Publication	17-Jan-2022

Correspondence Address:
Dr. Komal Khare
MDS Periodontology, Maitri College of Dentistry and Research Centre, Anjora, Durg, Chhattisgarh
India

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jmau.jmau_133_20

Abstract

Introduction: Dentinal hypersensitivity (DH) is a common chronic condition that affects a high proportion of the adult population and is one of the main reasons why patients seek dental opinion and treatment. Despite considerable success in reducing DH, unfortunately, most of the present methods can provide only temporary and unpredictable desensitization. Therefore, new treatment methods that are effective for long periods and at the same time do not have the complications are needed. Recently, two new desensitizing agents BioMin F and Propolis have been introduced for the treatment of DH. Aim: The aim of the study was to compare and evaluate the efficacy of BioMin F and Propolis containing toothpastes on dentinal tubule occlusion with and without the use of an adjunct 810 nm Diode Laser. Materials and Methods: Forty-five freshly extracted teeth were taken out of which thirty were sectioned into halves and divided into four test groups BioMin F, Propolis, BioMin F + Laser, and Propolis + Laser and control group. All the specimens were treated twice a day for 7 days and then evaluated under scanning electron microscope for partial and complete dentinal tubule occlusion. Results: A significantly higher number of completely occluded tubules were seen in BioMin F + laser group followed by Propolis + laser, Biomin F, and Propolis. Conclusion: Combination approach of desensitizing agent and laser provided a better result than the desensitizing agent alone and when compared individually Biomin F was more effective desensitizer as compared to Propolis.

Keywords: BioMin F, dentinal hypersensitivity, dentinal tubule occlusion, dentinal tubules, desensitizing agent, diode laser, propolis

How to cite this article:
Khare K, Bhusari P, Soni A, Malagi SK, Abraham D, Johnson L. A comparative evaluation of efficacy of biomin F and propolis containing toothpastes on dentinal tubule occlusion with and without use of an adjunct 810 nm diode laser: An In vitro scanning electron microscope study. J Microsc Ultrastruct 2023;11:41-6

How to cite this URL:
Khare K, Bhusari P, Soni A, Malagi SK, Abraham D, Johnson L. A comparative evaluation of efficacy of biomin F and propolis containing toothpastes on dentinal tubule occlusion with and without use of an adjunct 810 nm diode laser: An In vitro scanning electron microscope study. J Microsc Ultrastruct [serial online] 2023 [cited 2023 Mar 20];11:41-6. Available from: https://www.jmau.org/text.asp?2023/11/1/41/335865

Introduction

Dentinal hypersensitivity (DH) is defined as a pain arising from exposed dentin typically in response to chemical, thermal, tactile, or osmotic stimuli, which cannot be explained as arising from any other form of defect or pathology.^[1] The reported prevalence of DH varies from 4% to 57%.^[2],[3] DH can present from early to old age, with majority of sufferers aged between 20 and 40 years.^[4] The discomfort of DH experienced by the patients is highly subjective and can vary substantially between patients. For some patients, the discomfort may present as a disturbing, severe pain which can affect their quality of life.

Some dentifrices are available in the market containing occluding agents, especially one of either strontium or stannous salts for the treatment of DH. One disadvantage of strontium chloride is fluoride incompatibility, so the strontium salts are not ideal for a daily use toothpaste. Stannous salts on the other hand have the disadvantage of poor taste and tooth staining.^[5] Recently, two new desensitizing agents have been introduced for the treatment of DH. First is BioMin F^[6] which is a novel multicomponent bioactive glass of varying fluoride content. They have the combined effect of apatite formation and ion release which make BioMin F an attractive component for use in remineralizing dentifrices. Second is Propolis^[7] which is a natural resinous substance from plant buds or exudate by Apis mellifera bees. Almas et al.^[8] and Sales-Peres et al.^[7] had reported in their studies that Propolis could provide rapid, long-lasting tubule occlusion, and resistance to the challenges in the oral environment, the mechanism being unknown.

Despite considerable success in reducing DH, unfortunately, most of the present methods can provide only temporary and unpredictable desensitization.^[9] Studies have listed some advantages in using laser desensitizing treatment in comparison with non laser ones such as higher rate of patient response, early and greater durability and predictability of laser desensitization effects, as well as ease and time efficiency of application.^[10],[11] Lasers can result in either an occluding effect on dentinal tubules or a desensitizing effect by reducing the pulpal nerve's pain threshold.^[11] Hence, to increase the effectiveness of treatment, researchers postulated combination treatments such as lasers and chemical agents.^[12]

Since the literature was lacking the information regarding the effectiveness of BioMin F and Propolis alone as desensitizing agents and their combination with laser, the present scanning electron microscope (SEM) in vitro study was carried out aiming to evaluate the in vitro effects of BioMin F and Propolis with and without the use of adjunct 810 nm diode laser by using SEM analysis.

We started the study with the null hypothesis that there is no significant differences in the number or percentages of open, closed, or partially closed dentinal tubules in BioMin F alone, Propolis alone, and combined treatment group (BioMin F + laser and Propolis + laser).

Materials and Methods

We calculated the sample size by the formula

n = 2 (Zα + Z_1−ß)² σ²/Δ²

Where, Zα, Z = Constant set by convention according to accepted α error.

Z_1−ß = Constant set by convention according to power of study (Power – 80% =0.84)

Σ = Standard deviation estimated (from previous studies) ~ 0.6

Δ = Estimated effect size (effect size = 70%)

n = 2 (1.96 + 0.84)² 0.6²/0.7 × 0.7 = 11.5

□ Minimum sample = 12–15

Hence, 45 teeth freshly extracted for orthodontic purpose were taken and cleaned with distilled water. The ethical clearance was given by IRB of Maitri College of Dentistry and Research Centre (MCDRC) (MCDRC/2018/343) on 8.12.2018

Thirty teeth were split into two halves by the means of a diamond disc and micromotor which resulted in sixty samples for the study. The remaining 15 teeth were used as control group in order to calculate the mean number of dentinal tubules present in the given magnification. After cleaning the extracted teeth with distilled water, enamel layer was removed from the cervical region with the help of a round diamond bur so as to expose the underlying relatively unaltered dentin (dentin disc model). After exposing the dentin, 17% ethylenediaminetetraacetic acid (Endo-L, MAARC, Thane, Maharashtra, India) was applied within the exposed dentin for 1 min to remove the smear layer, and then the particular treatment was done in the different groups after rinsing it with distilled water. Then, these were divided into five groups containing 15 specimens each. These were:

Control group: Specimens were not treated by any of the toothpaste or laser to calculate the average number of dentinal tubules in dentin disc
Group 1: Specimens treated with BioMin F toothpaste (Bf) (Elsenz toothpaste, Group Pharmaceuticals ltd. Bengaluru, Karnataka, India) brushed for 2 min twice daily for 7 days
Group 2: Specimens treated with Propolis toothpaste (Pp) (Superbee Propolis toothpaste, Hi-Tech Natural products, Delhi, India) brushed for 2 min twice daily for 7 days
Group 3: Specimens treated with BioMin F and Diode Laser (Bf + L) brushed for 2 min twice daily for 7 days with Biomin F toothpaste followed by laser (NV Pro3 microlaser, DenMat holdings llc, Lompoc, USA) irradiation in non contact mode for 60 s
Group 4: Specimens treated with Propolis toothpaste and Diode Laser (Pp + L) brushed for 2 min twice daily for 7 days with Propolis toothpaste followed by laser irradiation in non contact mode for 60 s.

Artificial saliva was used as a substitute for saliva in the present study. All specimens from each group were kept in the artificial saliva for 7 days which was replaced every 24 h. SEM photomicrographs were obtained from each specimen surface examined at ×3000 magnifications. All specimens were viewed using SEM, and a single blind assessment was performed by one investigator. Surface characteristics studied for each specimen include: The total number of tubules and number of occluded tubules (both partially occluded [PO] and completely occluded tubules CO]) counted manually in each photograph of all the specimens. The tubules that showed reduction of the tubule by more than 50% or circumferential closure of the tubule with the presence of a central opening in the canal were considered PO. All data obtained were analyzed using Student's unpaired-”t” test to determine the significance of the result.

Results

The mean number of tubules seen in control group was 16. The individual scores for CO and PO tubules are given in [Table 1] and [Graph 1] and [Graph 2]. The number of open tubules seen in control group can be seen in [Figure 1]. The number of PO and CO tubules are represented in BioMin F is represented in [Figure 2], Propolis in [Figure 3], BioMin F + Laser in [Figure 4], and Propolis + laser in [Figure 5].

Table 1: Number of partially and completely occluded tubules by desensitizing agents

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Figure 1: Open dentinal tubules in control group scanning electron microscope view (×3000 magnification)

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Figure 2: Scanning electron microscope view of specimens treated with BioMin F toothpaste showing partially and completely occluded dentinal tubules (×3000 magnification)

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Figure 3: Scanning electron microscope view of specimens treated with Propolis toothpaste showing partially and completely occluded dentinal tubules (×3000 magnification)

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Figure 4: Scanning electron microscope view of specimens treated with BioMin F + 810 nm diode laser showing partially and completely occluded dentinal tubules (×3000 magnification)

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Figure 5: Scanning electron microscope view of specimens treated with Propolis + 810 nm diode laser showing partially and completely occluded dentinal tubules (×3000 magnification)

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The mean number of PO tubules seen in the Propolis group was 16.6 ± 2.67 (98.81 ± 15. 88%), whereas in the BioMin F group, it was found to be 7.87 ± 0.35 (46.83% ± 2.09%). The mean difference of 8.73 was found to be statistically highly significant (P<0.0001). The mean number of CO tubules seen in the BioMin F group was 9.13 ± 0.35 (54.37% ± 2.09%), whereas Propolis group showed a mean number of 0.73 ± 1.62 (4.37% ±9.67%) CO tubules. The mean difference of 8.4 was found to be statistically highly significant (P < 0.0001).

BioMin F used along with an 810 nm diode laser showed statistically highly significant outcome than the use of BioMin F alone. The mean number of tubules that were PO by BioMin F group was 7.87 ± 0.35 (46.83% ± 2.09%) and that of BioMin F along with laser group was 4.27 ± 3.01 (25.4% ± 17.92%). The mean difference of 3.6 was found to be statistically highly significant (P < 0.0001). The mean number of CO tubules in the BioMin F alone group was 9.13 ± 0.35 (54.37% ± 2.09%) and that of BioMin F and laser group was found to be 12.73 ± 3.01 (75.79% ± 17.92%). The mean difference of 3.6 was found to be statistically highly significant (P < 0.0001).

Number of PO tubules by Propolis alone was 16.6 ± 2.67 (98.81% ±15.4%) and that of Propolis + laser group was found to be 6.47 ± 2.59 (38.49% ±15.4%). The mean difference of 10.13 was found to be statistically highly significant (P < 0.0001). The mean number of CO tubules in the Propolis alone group was 0.73 ± 1.62 (4.37% ± 9.67%) and in Propolis + laser group was found to be 10.53 ± 2.59 (62.7% ±15.4%). The mean difference of 9.8 was found to be statistically highly significant (P < 0.0001).

Higher number of PO tubule was seen in propolis + laser group compared to BioMin F + laser. The result was statistically significant. Number of PO tubules in the Propolis + laser group was 6.47 ± 2.59 (38.49% ± 15.4%) and that of BioMin F + laser group was 4.27 ± 3.01 (25.4% ± 17.92%). The mean difference of 2.27 was found to be statistically highly significant (P < 0.0001). A higher number of CO tubules were seen in the BioMin F + laser group compared to Propolis + Laser group, and the mean difference was found to be statistically significant. The mean number of CO tubules in BioMin F + laser group was found to be 12.73 ± 3.01 (75.79% ± 17.92%) and that of propolis + laser group was found to be 10.53 ± 2.59 (62.7% ±15.4%). The mean difference of 2.2 was found to be statistically highly significant (P < 0.0001).

Discussion

DH is described clinically as an exaggerated response to non- noxious stimuli and satisfies all the criteria to be classified as a true pain syndrome.^[13]

The most widely accepted theory for DH is the hydrodynamic theory proposed by Brännström and Aström.^[14] Based on this theory, open dentinal tubules permit fluid flow through the tubules, which results in the stimulation of nerve endings in the dental pulp. Scanning electron microscopy has shown that tubules in clinically characterized, “sensitive,” exfoliated teeth are eight times more numerous, two times wider in diameter and more permeable, whereas tubules in “non sensitive” teeth are fewer, smaller, and usually blocked as seen in the study by Brännström and Aström^[14] According to the laws of hydrodynamics, decline in dentinal fluid movement causes a reduction in DH.^[15]

Mechanism of action of BioMin F includes the combined action of apatite formation and ion release makes these glasses attractive components for use in remineralizing dentifrices, particularly for treating dentine hypersensitivity.^[6] Research on the properties of Propolis for oral applications has shown that it has an anti-inflammatory action and stimulates reparative dentin formation, which could be the possible mechanism to reduce the dentin permeability.^[7] Following diode laser application, dentin structure was changed because of the thermal effects caused by laser energy being absorbed by dentin's mineral content which includes carbonate and phosphate, resulting in melting (photo-biomodulation) and tubule occlusion.^[16]

To our knowledge, this the first study using BioMin F and Propolis along with diode laser. Hence, comparison is done with studies using other materials. The results of PO tubules in Group 1 are comparable to the results reported by Shah et al.^[6] where they found 18 ± 7.88 mean number of PO tubules after the treatment of hypersensitive dentin by self-applied dentifrices containing BioMin F. On the contrary in the same study, Novamin and herbal dentifrice (Hiora-K) resulted in 30.5 ± 10.65 mean number of PO tubules and 21.5 ± 11.06 mean number of PO, respectively. Results were also not at par with the results reported by Rajeshwari et al.,^[17] who found 32.27 ± 1.21 mean number of PO tubules at 1 week after application of Novamin once a week. The CO tubules are comparable to results reported by Rajeshwari et al.,^[17] who found only 39.88% of CO tubules after the application of Sensodyne Repair. This might be due to very less frequency of application of desensitizer (once a week).

Since no similar studies have been reported in the literature, the results of PO dentinal tubules in the present study by Propolis could not be compared. However, Sales Peres et al.^[7] evaluated the effectiveness of Propolis gel on the in vitro reduction of dentin permeability and found that 10% Propolis gel and 30% Propolis gel resulted in minimal reduction of dentin hydraulic conductance compared to smear layer. The result of CO tubules in Group 2 is comparable with the results reported by Chen et al.,^[18] who observed the mean percentage of 2.88% CO tubules after the application of red Propolis extract on dentine disc.

The results of Group 3 can be compared with the results of other investigators who used other material and laser such as Novamin and laser for the treatment of DH; Rajeshwari et al.^[17] reported 20.22% of mean percentage of PO tubule after the application of combination of 980 nm GaAlAs diode laser at 1 week. The result of CO tubules is again comparable with the result of Rajeshwari et al.^[17] after the application of Novamin and 980 nm GaAlA diode laser at 1 week (mean 73.81% of CO tubules at 1 week). Results are not comparable with the results reported by Ali et al.,^[19] Han et al.,^[20] and Reddy et al.,^[21] who found mean percentage of 99.8% ± 3.3% (after the use of Nano-fluor-hydroxyapatite and neodymium-doped yttrium aluminum garnet [Nd-YAG] laser), 52% (after the use of Nano-carbonate apatite dentifrice and erbium-doped yttrium aluminum garnet [Er: YAG] laser), and 97.33% (after the use of ProArgin and laser) CO tubules at 1 week. The differences in the results of various studies and variety of alterations in the morphology of dentin irradiation with 980 nm diode laser could be attributed mainly to various parameters of the lasers. These include the output power, wavelength, frequency, and application mode because these parameters are directly related to energy transmitted to the dentin.

In Group 4, the result is not at par with the results reported by Rajeshwari et al.,^[17] who reported only 20.22% of PO tubules after the application of Novamin and laser irradiation at 1 week. The mean number of CO tubules are not at par with the results reported by Han et al.,^[19] Ali et al.,^[20] and Reddy et al.,^[21] who used nano-carbonate Apatite dentifrice and Er: YAG laser (52%), Nano-Fluor-Hydroxyapatite, and Nd: YAG laser (99.8% ±3.3%) and Novamin and 980 nm diode laser (97.33%), ProArgin + 980 nm diode laser (96.92%) respectively. Better results found in above studies could be attributed to the use of nano-sized particles along with laser.

When the effectiveness of BioMin F and Propolis in completely occluding the dentinal tubules was compared, BioMin F was found to be more effective (mean difference 8.4) (P < 0.0001 HS) and Propolis was more effective in partially occluding the tubules. Studies on dentin bridge formation by Propolis showed that the reparative dentin formation requires 4 weeks to be completed, and our study duration was 1 week which could be the possible reason of this outcome.^[22] When the effectiveness of BioMin F and BioMin F + laser in occluding dentinal tubules was compared BioMin F + laser was found to be significantly more effective in completely occluding dentinal tubules than BioMin F alone (mean difference 3.6; P < 0.0001 HS). The results of present approach (BioMin F + Laser) is at par with the results reported by Reddy et al.^[21] where Novamin formulated toothpaste followed by diode laser irradiation were evaluated for the effectiveness in occluding dentinal tubules. When the effectiveness of Propolis and Propolis + laser in CO dentinal tubules was compared, Propolis + laser was found to be significantly more effective than Propolis alone (mean difference 9.8) (P < 0.0001). This result is comparable with the result of the other study where another combination of desensitizing agents and laser was used. Ali et al.^[19] reported that the combination of Nano-Fluor-hydroxyapatite and diode laser was significantly more effective in occluding dentinal tubules than Nano-Fluor-Hydroxyapatite alone (mean difference 17.7). However, when the effectiveness of BioMin F + laser was compared with Propolis + laser in CO dentinal tubules, BioMin F + laser was found to be significantly better than Propolis + laser (mean difference 2.2). This was attributed to the combined hastened effect of BioMin F + laser as both BioMin F and laser are significantly effective in completely occluding the dentinal tubules. Manual counting of the dentinal tubules for calculating the percentage of tubule occlusion is prone to human error, and in vitro dentin permeability test in human dentin section is also required to supplement the results from tubular occlusion aspect of this study. Further studies to investigate the durability of occlusion, hydraulic conductance, and morphologic changes in the dentinal tubules with different concentrations of test materials are needed.

Conclusion

After analyzing the results, the following conclusions were drawn:

BioMin F and Propolis as desensitizing agent was shown to partially and completely occlude the dentinal tubules with statistically highly significant result
BioMin F was found to be more effective in completely occluding dentinal tubules than Propolis with statistically highly significant result
The combined approach of BioMin F + laser and Proplis + laser was found to be more effective in completely occluding dentinal tubules than BioMin F and Propolis alone with statistically highly significant result
Out of two combined approach, BioMin F + laser was found to be more effective in completely occluding dentinal tubules than Propolis + laser with statistically highly significant result.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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