CN114533782A - Application of full-spectrum CBD oil in preparation of medicine for preventing or treating periodontitis - Google Patents

Abstract

The invention belongs to the technical field of new application of full-spectrum CBD oil, and provides application of the full-spectrum CBD oil in preparation of a medicine for preventing or treating periodontitis. According to the invention, researches show that the full-spectrum CBD oil reduces the secretion of TNF-alpha and IL-1 beta, so that alveolar bone resorption of an experimental periodontitis model is inhibited. In addition, the full-spectrum CBD oil can effectively relieve Lipopolysaccharide (LPS) induced inflammatory reaction of periodontal ligament cells by inhibiting TLR 4-mediated activation of NF-kB and MAPK signal channels. Therefore, the invention has important significance for preventing or treating periodontitis diseases and improving the life quality of patients.

Description

Application of full-spectrum CBD oil in preparation of medicine for preventing or treating periodontitis

Technical Field

The invention relates to the technical field of new application of full-spectrum CBD oil, in particular to application of full-spectrum CBD oil in preparation of a medicine for preventing or treating periodontitis.

Background

Periodontitis is a chronic inflammatory disease, and its severe inflammatory reaction and destruction of periodontal tissues are major causes of tooth loss. The existing research proves that the periodontal plaque microorganisms are the initiation factor of periodontal disease, directly participate in destroying the periodontal tissues through the periodontal plaque microorganisms and the metabolites thereof, and simultaneously cause the immune response of a host to infection through antigen components, further cause the damage of the periodontal tissues and are the important reason for the destruction of the periodontal tissues. At present, the clinical treatment technology of periodontitis only achieves the purpose of mechanically removing dental plaque to treat periodontitis, does not cover the influence of the immune response of a host on the disease development, and usually needs to be supplemented with local or systemic drug treatment to consolidate the curative effect of basic treatment and block the development of the basic treatment. The periodontal anti-inflammatory drugs commonly used at present mainly include nitroimidazoles, tetracyclines, penicillins, macrolides and other antibiotics. And the long-term use of antibiotics has large side effect and inevitable harm to human bodies. Therefore, there is an urgent need to develop a traditional Chinese patent medicine or natural extract for local or systemic anti-inflammatory treatment.

The Full Spectrum CBD Oil (Full Spectrum CBD Oil) refers to a hemp plant natural component which is removed toxic component Tetrahydrocannabinol (THC) by modern chromatographic technology and contains CBD (Cannabidiol) and other cannabinoids, other terpene compounds and the like. The full spectrum CBD oils are generally more viscous, darker in color, and have the taste of the hemp plant. At present, there are many reports on the anti-inflammatory properties of pure Cannabidiol (CBD) and the full spectrum of CBD oils. However, the protection effect of the full-spectrum CBD oil on human periodontal ligament stem cells and the mechanism of inhibiting periodontitis are not reported yet.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provide the application of full-spectrum CBD oil in preparing a medicament for preventing or treating periodontitis.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides application of full-spectrum CBD oil in preparation of a medicine for preventing or treating periodontitis.

Preferably, the periodontitis refers to alveolar bone resorption and LPS-induced inflammation of periodontal ligament cells.

Preferably, the alveolar bone resorption is due to inflammatory mediators TNF- α and IL-1 β, and the LPS-induced inflammation of periodontal ligament cells is due to TLR 4-mediated activation of NF-kB and MAPK signaling pathways.

Preferably, the medicament is injection, powder, granules, powder, pills, oral liquid or tablets.

Preferably, the medicine also contains medically acceptable auxiliary materials.

The invention also provides application of the full-spectrum CBD oil in preparation of mouthwash for preventing or treating periodontitis.

The invention also provides application of the full-spectrum CBD oil in preparing toothpaste for preventing or treating periodontitis.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a new medical application of full-spectrum CBD oil. The inventor finds that the full-spectrum CBD oil reduces the secretion of TNF-alpha and IL-1 beta, thereby inhibiting the alveolar bone resorption of an experimental periodontitis model. In addition, the full-spectrum CBD oil can effectively relieve Lipopolysaccharide (LPS) induced inflammatory reaction of periodontal ligament cells by inhibiting TLR 4-mediated activation of NF-kB and MAPK signal channels. Therefore, the invention has important significance for preventing or treating periodontitis diseases and improving the life quality of patients.

Drawings

FIG. 1 is a Micro-CT scan reconstruction diagram of the absorption degree of the maxillary first molar lingual side and the section alveolar bone of rats in each group (note: blank control group (CON group); control group (L group); experimental group (CBD group)) from top to bottom;

FIG. 2 is a graph of the extent of alveolar bone height loss in rats of each group (note: data all represent mean. + -. standard deviation, n-9);

FIG. 3 is a graph of bone volume loss under the abutment surface of the first and second molars in each group of rats (note: data are mean. + -. standard deviation, n is 9);

FIG. 4 is a graph showing the variation of TNF-. alpha.in gingival tissues of rats in each group measured by ELISA (note: data all represent mean. + -. standard deviation, n-9);

FIG. 5 is a graph showing the variation of IL-1. beta. in gingival tissues of rats in each group measured by ELISA (note: data all represent mean. + -. standard deviation, and n is 9);

FIG. 6 is a graph showing the variation of TLR4 in the gingival tissue of rats in each group measured by ELISA (note: data all represent mean. + -. standard deviation, n-9);

FIG. 7 is a graph showing the variation of Cat-k in gingival tissues of rats in each group measured by ELISA (note: data all represent mean. + -. standard deviation, n is 9);

FIG. 8 is a graph of the effect of a full spectrum of CBD oils at different concentrations on human periodontal ligament cell proliferation;

FIG. 9 is a graph of the effect of different concentrations of the full spectrum of CBD oil on periodontal ligament cell activity;

FIG. 10 shows the RNA transcription levels of TNF-. alpha.in periodontal ligament cells of rats in each group (note: data each indicates mean. + -. standard deviation, and n. sup.3);

fig. 11 shows the transcription level of TLR4 RNA in periodontal ligament cells of rats in each group (note: data all represent mean ± sd, n ═ 3);

FIG. 12 shows the transcript levels of RNA of P38 in periodontal ligament cells of rats in each group (note: data each indicates mean. + -. standard deviation, and n is 3);

FIG. 13 shows the transcription levels of Erk1 RNA in periodontal ligament cells of rats in each group (note: data all indicate mean. + -. standard deviation, n-3);

FIG. 14 shows the transcription levels of Erk2 RNA in periodontal ligament cells of rats in each group (note: data all indicate mean. + -. standard deviation, n-3);

FIG. 15 shows the transcription levels of RNA of NF-kB (p65) in periodontal ligament cells of rats in each group (note: data each indicates mean. + -. standard deviation, and n is 3);

figure 16 is a graph of the effect of pan-CBD oil on ERK, NF-kB protein expression (note: data all mean ± sd, n ═ 3).

Detailed Description

The technical solutions provided by the present invention will be described in detail with reference to the following experimental examples, but they should not be construed as limiting the scope of the present invention.

The invention is characterized in that full-spectrum CBD oil (selected from full-spectrum cannabidiol oil containing 60% CBD and produced by Yunnan Hansen biotechnology and Limited liability company) is externally coated on an experimental rat periodontitis model, upper jaw samples are collected, Micro-CT scanning and reconstruction are carried out, the distance (CEJ-AC) between a second molar glaze cementum boundary of each sample and a crest top of an alveolar ridge and the bone density bone body integral ratio are used as bone morphology measurement indexes, and meanwhile, the expression of inflammation-related proteins TNF-alpha, IL-1 beta, TLR4 and Cat-k in rat periodontal tissues is detected by an enzyme-linked immunosorbent assay. Lipopolysaccharide (LPS) is used for inducing inflammatory reaction of human periodontal ligament stem cells and simulating inflammatory process of periodontitis. The activity of human periodontal ligament stem cells was examined by the cck-8 method. The expression of TNF-alpha, TLR4, Erk1, Erk2, P38 and NF-kB genes in cells is detected by real-time fluorescent quantitative PCR, and the expression of Erk and NF-kB proteins in the cells is detected by immunoblot analysis.

1. Experimental rat periodontitis model establishment and material drawing

In the experiment, 30 male SD rats with the weight of 200g +/-20 g and the weight of 30 male SD rats with the age of 8 weeks are selected and bred in cages of polypropylene synthetic plastic rats, each 5 rats are bred in one cage, and standard feed is provided by the experimental animal center of Sichuan university and is free to drink water and diet.

Rats were randomly divided into 3 groups of 10 rats each, two groups were used for the establishment of periodontitis models, and the remaining group was used as a normal control group. Two sets of rows for modeling 7% chloral hydrate (500mg/kg) were injected intraperitoneally for anesthesia. The neck parts of first molar teeth of the upper jaw at the two sides of a rat are ligated by using 3-0 sterile nylon threads, and the ligation threads should enter the gingival sulcus and are knotted at the middle part, so that whether the threads fall off or not can be conveniently observed in the experimental process.

And (3) drug treatment: blank control group (CON group), ligature the neck of the tooth with sterile nylon thread, ligature the silk thread into the gingival sulcus; in the control group (group L), the neck of the tooth is ligated with sterile nylon thread, the ligation thread enters the gingival sulcus, and edible vinegar is added; experimental group (CBD group), the neck of the tooth was ligated with sterile nylon thread, the silk thread was ligated into the gingival sulcus, and edible vinegar was added, along with the full spectrum CBD oil added. Experimental rats in the CBD group were treated under ether inhalation anesthesia and the drug was applied evenly around the maxillary periodontal tissues of the rats once a day. After feeding for four weeks, cervical vertebra is deprived of white under anesthesia, and the maxilla on both sides is separated, the tissue of the maxilla on one side is placed in 0.5% of a poly formic acid solution for Micro-CT scanning, and the gum on the other side, 2mm below the gingival margin around the first molar of the maxilla on the other side, is separated for protein extraction.

Micro-CT analysis of alveolar bone resorption

Micro-CT scanning and reconstruction, measuring the distance (CEJ-AC) from the first molar enamel cementum boundary to the crest of the alveolar ridge of each specimen, namely measuring along the long axis of the root on the buccolingual side of the first molar, 3 measured values (mum) per tooth, and taking the average value as the CEJ-AC distance. Bone volume was calculated using the CT machine's own analysis software. The area adjacent the first and second molars was selected as a selection area on the scan cross-section and the percentage of bone volume to total volume (BV/TV) was calculated.

3. Enzyme Linked Immunosorbent (ELISA) kit for detecting expression of inflammatory protein

Collecting the rat gingival tissue, and extracting total protein in the tissue. And (3) performing determination according to the instruction of an enzyme-linked immunosorbent kit, reading the absorbance of each sample at 450nm, and calculating the contents of inflammatory proteins TNF-alpha, IL-1 beta, TLR4 and Cat-k according to a standard curve.

4. Cell culture

Periodontal ligament cells were obtained from premolar teeth of 16-20 year old volunteers with healthy periodontal tissue extracted by orthodontic subtraction (patients and families informed consent), after tooth washing with PBS, periodontal ligament tissue of 1/3 in roots was scraped under aseptic conditions, primary culture of hPDLCs was performed by tissue block method, and tissue culture medium (DMEM supplemented with 0.1% penicin-Streptomycin Solution and 10% FBS) was replaced every 2 days. The first passage was performed when the cells had climbed out of the tissue mass and spread out of the culture plate to 80%. The present invention employs passage 3 to passage 5 cells.

CCK-8 method for detecting cell proliferation

hPDLCs were seeded in 96-well plates (5000/well, 100. mu.L). Cells were treated with different concentrations (0.5, 1, 2, 4, 8. mu. mol/L) of CBD. Cell concentrations were measured by the CCK-8 method at 1, 3, 5, 7d after the start of the experiment, and the survival of periodontal ligament cells in the medium was evaluated in DMEM blank control to generate cell proliferation curves according to the formula: cell viability ═ a (dosed) -a (blank) ]/[ a (control) -a (blank) ] cell viability was calculated and plotted with time as x-axis and cell viability as y-axis.

6. Real-time fluorescent quantitative PCR (RT-qPCR) technology for determining expression level of inflammatory marker

Cells were plated at 1 x 10⁶Is seeded in 6-well cell culture plates. After 2 days in DMEM with 10% FBS, the medium was changed to DMEM without FBS starved for 1 day, 10mg/L LPS was added, as well as various concentrations of CBD (0, 1, 2, 4 and 8. mu. mol/L; full spectrum CBD oil contains 67% CBD, all concentrations are measured as pure CBD). Total cellular RNA from each group was extracted after 24 hours using Trizol reagent and 2. mu.g of total RNA was used for cDNA synthesis using Prime script RT kit (Takara, DRR037A) according to the manufacturer's instructions. The primer is designed as follows: two primers of GAPDH are respectively shown as SEQ ID NO.1 and SEQ ID NO.2, two primers of TNF-alpha are respectively shown as SEQ ID NO.3 and SEQ ID NO.4, two primers of TLR4 are respectively shown as SEQ ID NO.5 and SEQ ID NO.6, two primers of Erk1 are respectively shown as SEQ ID NO.7 and SEQ ID NO.8, and two primers of Erk2 are respectively shown as SEQ ID NO.9 and SEQ ID NO.10 (also shown in the following table). The conditions for RT-qPCR were as follows: reaction at 95 deg.C for 3 min. ② 9 ℃ for 10 seconds; 55 ℃ for 30 seconds, 39 cycles of reaction. Repeated measurement is carried out for 3 times, and relative quantitative calculation is carried out on target genes by the following formula: Δ CT (x) -CT (gapdh), Y ═ 2^-ΔΔCT。

Primer design sheet

Westernblot analysis of inflammatory protein expression

hPDSCs at 1 x 10⁶Is seeded in 6-well cell culture plates. Cultivation in DMEM containing 10% FBSAfter 2 days of culture, the medium was changed to DMEM without FBS and starved for 1 day, and 10mg/L LPS and 8. mu. mol/L CBD were added. After 24 hours, the hPDLSCs were washed with cold PBS, the cells were lysed extensively with RIPA lysis buffer, and centrifuged at 13000g for 10 min. Protein concentration was determined using the BCA protein assay kit. Total proteins were separated on SDS-PAGE gels and transferred to PVDF membranes. PVDF membranes were blocked with 5% skim milk and incubated overnight at 4 ℃ with primary anti-NF-kB (1: 800), ERK (1:1000), beta-actin (1: 1000). PVDF membrane was washed 4 times (5 min/time) with TBST and then placed in the corresponding secondary antibody (1:8000) and incubated for 1h at room temperature in a shaker. Finally, the film was washed 4 times (5 min/time), irradiated, developed, and fixed. Quantitative analysis was performed using Image-J software.

8. Statistical analysis

Statistical analysis SPSS 22.0 and GraphPadPrism 8.0 software were used. Experimental results values are presented as mean ± standard deviation, and multiple comparisons between groups were performed using the Tukey test using univariate analysis of variance (ANOVA). It is statistically significant to have p < 0.05.

Experimental example 1

The effect of the full spectrum of CBD oil on the experimental rat periodontitis model was studied and the results are shown in fig. 1 to 7.

FIG. 1 is a Micro-CT scanning reconstruction diagram showing the first molar lingual side of the upper jaw and the degree of alveolar bone resorption of the section of each group of rats.

The results of the three sets of CEJ-AC measurements are: 0.89 + -0.13 mm for CON group, 1.18 + -0.08 mm for L group, and 1.00 + -0.08 mm for CDB group. Micro-CT reconstruction showed that silk ligation resulted in significant alveolar bone resorption around the first molars. As can be seen from FIG. 2, the CEJ-AC distance was significantly smaller in the CBD-treated group than in the L group, indicating that CBD inhibited the loss of alveolar bone height to some extent.

The extent of bone volume loss under the abutment surface of the first and second molars, as indicated by the percentage of bone volume in the measured area to total volume (BV/TV), was calculated as follows: 0.80 plus or minus 0.09, 0.53 plus or minus 0.04 and 0.61 plus or minus 0.02. As can be seen from FIG. 3, the decrease in BV/TV was less in the CBD group than in the L group, indicating that the CBD-treated group was able to significantly reduce alveolar bone loss.

In conclusion, compared with the CON group, the L group and the CBD group of rats with experimental periodontitis induced by silk thread ligation both show the increase of CEJ-AC distance, and the difference has statistical significance (# # # # p <0.0001, compared with the control group); the increase in CEJ-AC distance was significantly greater in the L group than in the CBD group (×) p <0.0001 compared to the L group. Both the L and CBD groups showed a reduction in BV/TV compared to the CON group, and the difference was statistically significant (# # p <0.01, compared to the control group); BV/TV decreased significantly more in the L groups than in the CBD group (× p <0.0001 compared to the L groups). Data are presented as mean ± standard deviation (n ═ 9).

ELISA measured TNF- α, IL-1 β, TLR4, Cat-k in gingival tissue. The results are shown in FIGS. 4 to 7. As can be seen from fig. 4-7, both the L and CBD groups showed an increase in the detection of inflammatory proteins, and the differences were statistically significant (# p <0.05, # # # # p <0.0001, compared to the CON group). The levels of inflammatory cytokines in gingival tissues of animals in the CBD group were significantly reduced compared to those in the L group of ligated animals, with statistical significance of the differences (. p <0.01,. p <0.0001 compared to the L group). Data represent mean ± sd (n-9). This indicates that the full spectrum of CBD oils can inhibit the inflammatory response in the experimental periodontitis model in rats.

Experimental example 2

The effect of the full spectrum of CBD oils on human periodontal ligament cell proliferation was studied and the results are shown in fig. 8 to 9.

As can be seen from fig. 8 and 9, after the periodontal ligament cells were incubated for 1, 3, 5, and 7 days with CBD (0.5, 1, 2, 4, and 8 μmol/L) at different concentrations, the absorbance values measured by the CCK-8 method were not significantly different among the groups, and the cell viability of each group was not significantly different. This indicates that the individual concentrations of CBD used in the experiment did not affect the activity of periodontal ligament fibroblasts. CBD did not produce cytotoxicity to periodontal ligament fibroblasts during the longest observation period of the experiment.

Experimental example 3

The effect of the full spectrum of CBD oil on LPS-stimulated human pericellular membranes was studied and the results are shown in fig. 10-16.

Lipopolysaccharide is a key toxic factor in the pathogenesis of periodontitis. Therefore, LPS can increase the transcription level of TNF-alpha, TLR4, P38, Erk1, Erk2 and NF-kB (P65) RNA in human periodontal ligament cells after being stimulated for 24 hours. As can be seen from fig. 10 to 15, the RT-qPCR results showed that after LPS stimulation, TNF- α, TLR4, P38, Erk1, Erk2, NF-kB expression was up-regulated, and the LPS control group was statistically different from the blank control group (# P <0.05, # P <0.01, # P <0.001, compared to the control group). With the addition of CBD, TNF- α, TLR4, P38, Erk1, Erk2, NF-kB expression were all down-regulated with increasing CBD concentration, with the more pronounced the down-regulation was with greater CBD concentration and the LPS + CBD group was statistically different from the LPS group (P <0.05, > P <0.01, > P <0.001, > P <0.0001 compared to the LPS group). This indicates that CBD can inhibit increase of transcription levels of TNF-alpha, TLR4, Erk1, Erk2, P38 and NF-KB caused by LPS stimulation in a concentration-dependent manner, and the inhibition effect is more obvious when the concentration is higher.

ERK is analyzed through Westernblot, and the expression of NF-kB protein is 24h after being stimulated by LPS. As shown in fig. 16, after LPS stimulation, ERK and NF-kB protein expression was up-regulated, and the LPS stimulated group was statistically different from the blank control group ([ p <0.01, [ p <0.001, compared to the control group), and after CBD addition, ERK and NF-kB protein expression was down-regulated, and the LPS + CBD group was statistically different from the LPS group (# p <0.05, # p <0.01, compared to the LPS group). Data are expressed as mean soil standard deviation (n-3). Consistent with the qPCR results, CBD inhibited the production of ERK as well as NF-kB proteins. This suggests that LPS, upon stimulation of human periodontal ligament cells, by binding to TLR4, may initiate MAPK and NF-kB signaling pathways, which are associated with an increase in the proinflammatory cytokine TNF- α. And CBD inhibits the activation of MAPK and NF-kB signal channels, thereby reducing the expression of TNF-alpha and further inhibiting the amplification of inflammation.

As can be seen from the above, in order to evaluate the effect of CBD on inflammation and alveolar bone loss, the present invention measured TNF-. alpha.IL-1. beta., TLR4 and Cat-k levels in the gums of experimental rats and Micro-CT scan reconstruction was performed on the maxillary samples. Micro-CT analysis showed that the distance of CEJ-AC in the L group was significantly higher than that in the CBD group, and BV/TV in the CBD group was reduced less than that in the L group. The above results all show that the CBD treatment group significantly inhibited alveolar bone resorption. It has been shown that the expression of the inflammatory mediators TNF- α and IL-1 β play an important role in the pathogenesis of periodontitis. The results of the invention show that the CDB treatment group remarkably inhibits the expression of TNF-alpha and IL-1 beta caused by periodontitis, and the CBD has certain anti-inflammatory effect. TLR4 was the first receptor to respond to LPS and recognition of LPS by TLR4 induced activation of NF-kB and MAPK. The result of the invention shows that the expression of TLR4 is inhibited in the CBD group, so that the invention starts with TLR4 and discusses the action mechanism of CBD in periodontitis host inflammation and periodontal bone metabolism regulation.

The cytotoxicity of CBD was first studied. The present data show that the CBD treatment used at the highest concentration of 8. mu. mol/L had no effect on periodontal ligament cell survival. TNF-alpha is a critical inflammatory cytokine in the process of periodontium pathology, has the effect of initiating and amplifying inflammatory response and plays an important role in periodontium destruction, and the research of the invention finds that CBD can inhibit the production of TNF-alpha in LPS-induced periodontal ligament cells in vitro and can also inhibit the increase of TNF-alpha caused by filament ligation-induced periodontitis in vivo, which is probably one of the mechanisms of CBD in preventing periodontal destruction.

Previous studies have demonstrated that LPS and bacteria promote the production of TNF- α, IL-1 β by mediating the expression of TLRs, suggesting that the release of inflammatory mediators can be controlled by the inhibition of TLRs. Activation of TLRs usually activates NF-kB and MAPK-ERK1/2 signaling pathways, leading to upregulation of inflammatory gene expression, which is critical to the innate immune response to inflammation. The invention stimulates hPDLSCs through LPS, induces the TLR4/NF-kB signal channel to be up-regulated, and inhibits the TLR4/NF-kB signal channel after CBD treatment. LPS-stimulated cells induced phosphorylation and activation of ERK1/2 and p38 MAPK. The present invention discusses the effect of CBD on LPS-induced activation of ERK1/2 and p38 MAPK. The results show that CBD has a dose-dependent inhibition effect on NF-kB and MAPK activation induced by LPS.

In summary, the full spectrum of CBD oils reduced the secretion of TNF- α and IL-1 β, thereby inhibiting alveolar bone resorption in the experimental periodontitis model. And the LPS-induced inflammatory response of periodontal ligament cells is relieved by inhibiting TLR 4-mediated activation of NF-kB and MAPK. Thus, the full spectrum of CBD oils can be considered as a potential therapeutic agent for periodontal disease.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

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Claims (7)

1. Application of full-spectrum CBD oil in preparation of medicine for preventing or treating periodontitis.

2. Use of the full spectrum CBD oil according to claim 1, in the manufacture of a medicament for the prevention or treatment of periodontitis, wherein periodontitis is alveolar bone resorption and LPS-induced inflammation of periodontal ligament cells.

3. Use of the full spectrum CBD oil in the manufacture of a medicament for the prevention or treatment of periodontitis according to claim 1 or 2, wherein alveolar bone resorption is caused by inflammatory mediators TNF- α and IL-1 β, and LPS-induced inflammation of periodontal ligament cells is caused by TLR4 mediated activation of NF-kB and MAPK signaling pathways.

4. Use of the full spectrum CBD oil according to claim 3, in the preparation of a medicament for the prevention or treatment of periodontitis, wherein the medicament is an injection, a powder, a granule, a powder, a pill, an oral liquid or a tablet.

5. The use of the full-spectrum CBD oil according to claim 4, for the preparation of a medicament for the prevention or treatment of periodontitis, wherein the medicament further comprises pharmaceutically acceptable excipients.

6. Use of full spectrum CBD oil in the preparation of a mouthwash for the prevention or treatment of periodontitis.

7. Use of full spectrum CBD oil in the manufacture of a toothpaste for the prevention or treatment of periodontitis.

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