Phthalocyanine derivative attenuates TNF-α production in macrophage culture and prevents alveolar bone loss in experimental periodontitis.

AIM: This study investigated the activity and mechanism of action of the iron tetracarboxyphthalocyanine (FeTcPc) on tumor necrosis factor alpha (TNF-α) production and its impact on experimental periodontitis. METHODS: RAW 264.7 macrophages were treated with FeTcPc, activated with lipopolysaccharide (LPS) at 10 ng/mL, and the TNF-α levels were measured, as well as the nuclear factor kappa B (NF-κB) activation. Subsequently, a mouth gel containing 1% FeTcPc was topically administered to the gingival tissue of mice with periodontitis-induced ligatures. Bone loss and the gene expression of Tnfα, p65 (NF-κB), and receptor-activating nuclear factor kappa B ligand (Rankl) were quantified in gingival tissue. Finally, the systemic toxicity of FeTcPc was estimated in Galleria mellonella larvae. RESULTS: In an activated RAW 264.7 macrophage culture, 100 µM FeTcPc reduced TNF-α release and NF-κB activation. Regarding experimental periodontitis, topical application of mouth gel containing 1% FeTcPc blocked alveolar bone loss. Additionally, 1% FeTcPc reduced the expression of Tnfα, p65 (NF-κB), and Rankl in gingival tissue. Finally, administration FeTcPc at doses ranging from 1 to 1000 mg/kg did not cause acute systemic toxicity in G. mellonella. CONCLUSION: Overall, we demonstrated the potential of mouth gel containing FeTcPc as a therapeutic strategy for managing osteolytic inflammatory disorders, such as periodontitis.

Can toothbrushing reduce the intraoral viral load of SARS-CoV-2? A pilot study with a dentifrice containing an antimicrobial phthalocyanine derivative.

The aim of this study was to assess whether toothbrushing with a dentifrice containing an antimicrobial phthalocyanine derivative (APD) can reduce the intraoral viral load of SARS-CoV-2. Twenty COVID-19-positive dentate patients aged ≥18 years were selected instructed to brush their teeth for 2 min with a dentifrice containing APD. Self-collected samples of unstimulated saliva were carried out three times: T0 (baseline), T5 (5 min after toothbrushing), and T30 (30 min after toothbrushing). The analysis of viral RNA was performed by RT-qPCR for detection of three viral genes (ORF1ab, N and S genes). Results were statistically tested using Friedman's test and pairwise comparison with Bonferroni corrections, with a significance level of 5%. There was an increase in the cycle threshold (Ct) value from T0 to T5 in 13 patients (72.2%), and from T0 to T30 in 14 patients (77.8%). In two patients (11.1%) no SARS-CoV-2 was detected at T5 and five patients (27.8%) at T30. The Ct values were statistically significantly higher (p=0.020) at T30 in comparison to T0 and T5. This pilot study suggests that toothbrushing with a dentifrice containing APD could reduce the SARS-CoV-2 viral load in the oral cavity. However, further studies are needed to confirm this possible beneficial effect against SARS-CoV-2.

Use of phthalocyanine-derived mouthwash as a protective factor for COVID-19: a community trial.

Aim: In a population profile corrected for sociodemographic factors, the aim of this study was to examine sociodemographic the protective effect of a phthalocyanine-derived mouthwash (APD) before infection with SARS-CoV-2, in addition to analyzing the survival of the at-risk population and the confirmed diagnosis of COVID-19. Methods: For individuals from the Uru municipality, a structured questionnaire consisting of two parts was completed before the distribution of APD. Subsequently, subjects received two bottles containing 600 mL of APD and were instructed to rinse/gargle with 3 mL of the solution 3 to 5 times per day for 1 min for 2 months. Data were obtained from the electronic system of the municipal health center, organized in a spreadsheet, and analyzed using multiple linear regression and Cox regression analysis. Results: The study included 995 participants with the following sociodemographic data: 98/995 individuals (p<0.002) who did not complete high school used the APD 66.30 times more than did individuals with higher education. The results in terms of survival were meaningful in relation to the duration of APD use. The protective factor for COVID-19 was 14.1%. Conclusion: Daily use of a solution containing phthalocyanine derivatives provided a higher protection factor against COVID-19 infection, predominantly in individuals without a school-completion certificate.

Regenerative and Protective Effects on Dental Tissues of a Fluoride-Silicon-Rich Toothpaste Associated with a Calcium Booster: An In Vitro Study.

Calcium boosters have been used as a supplement for fluoride toothpastes to repair the dental tissues and reduce dentin permeability. This in vitro study aimed to characterize the regenerative and protective effects of the treatment of dental tissues with a fluoride-silicon-rich toothpaste associated with a calcium booster. Bovine enamel and dentin blocks (n = 5) were obtained (4 × 4 × 6 mm). A fluoride-silicon-rich toothpaste and a calcium booster were used to brush the enamel and dentin both immediately and five days afterwards. The outcomes were then compared to those of the untreated control group. After that, the specimens were cross-sectioned. SEM was used to evaluate the micromorphology of the surface and cross-section. Energy-dispersive X-ray spectroscopy (EDS) was used to determine the elemental analyses (weight%). After treatment for 5 days with a booster/silicon-rich toothpaste, EDS analysis demonstrated that it induced a significant mineral change. It was also able to form a protective silicon-enriched mineral layer on both enamel and dentin surfaces. It was demonstrated in vitro that a fluoride-silicon-rich toothpaste associated with a calcium booster regenerates the dental tissues, remineralizing the enamel structure and occluding the dentin tubules.

USE OF MOUTHWASH AND DENTIFRICE CONTAINING AN ANTIMICROBIAL PHTHALOCYANINE DERIVATIVE FOR THE REDUCTION OF CLINICAL SYMPTOMS OF COVID-19: A RANDOMIZED TRIPLE-BLIND CLINICAL TRIAL.

PURPOSE: This clinical trial aimed to evaluate the use of mouthwash and dentifrice containing an antimicrobial phthalocyanine derivative (APD) to reduce the clinical symptoms in patients with COVID-19. METHODS: This randomized, triple-blind clinical trial enrolled 134 patients aged 18 years or older who underwent COVID-19 testing through the use of nasopharyngeal swab RT-qPCR in a reference center for the diagnosis of COVID-19, had no clinical contraindications to mouthwash and gargle, and had access to cell phones with communication applications. According to the use of a mouthwash and dentifrice containing antimicrobial phthalocyanine derivatives (APD), patients were randomly assigned (1:1) to the APD or non-APD (control) group. All participants were instructed to floss twice a day, brush teeth for 2 minutes 3 times a day, and gargle/rinse (5 mL) for 1 min/3 times a day for 7 days. An online questionnaire was sent to collect data on the clinical symptoms of COVID-19 3 times: T0 (baseline before using the oral hygiene products), T3 (3 days after), and T7 (7 days after). The investigators, patients, and outcome assessors were blinded to group assignment. The Mann-Whitney, Chi-Square, Fisher's exact, and Cochran's tests were used according to the nature of the variables studied, with the level of significance set at P < .05. RESULTS: No statistically significant difference was found in the prevalence of symptoms between groups at baseline. A statistically significant reduction in clinical symptoms was found in the control group (fatigue, shortness of breath, hoarse voice, sore throat, nasal congestion, and chest pain) and APD group (cough, fatigue, shortness of breath, hyposmia/anosmia, dysgeusia, hoarse voice, sore throat, nasal congestion, chest pain, diarrhea, and irritability/confusion) during the follow-up period. There were statistically significant differences, with a higher prevalence of symptoms in the control group at T3 and T7. Dysgeusia, sore throat, and irritability/confusion were less prevalent in the APD group at T3, and shortness of breath, hyposmia/anosmia, dysgeusia, hoarse voice, sore throat, diarrhea, and irritability/confusion were more prevalent in the control group at T7. CONCLUSIONS: Based on this methodology, the results demonstrated that the regular use of mouthwash and dentifrice-containing APD had a positive impact on the clinical symptoms, as reported by patients with COVID-19.

Effectiveness of Experimental Whitening Toothpastes Containing Colorants on the Optical Properties of Enamel.

Objective: This in vitro study investigated the whitening potential of experimental toothpastes containing optical colorants in their formulations in comparison with commercial products. The chemical and physical characteristics of the toothpastes, the morphology, and elemental analysis of the enamel surface after treatment were also analyzed. Materials and Methods: One hundred twenty-five bovine incisor teeth were randomly divided into five groups according to the treatment: (i) experimental PHTALOX dental gel (PHT); (ii) experimental blue silica dental gel (SDG); (iii) Sensodyne Whitening Repair & Protect (WRP); (iv) Sensodyne True White (STW); (v) Snow White Toothpaste (SWS). The whiteness index differences (ΔWI D ) and color alteration (CIELab-ΔE, CIEDE2000-ΔE 00) were calculated after color change analysis using a spectrophotometer before and after the enamel treatment (n = 25). The surface and cross-sectional micromorphology were assessed using scanning electron microscopy. The elemental analyses were determined with energy-dispersive X-ray spectroscopy (EDS). The pH, particle size, zeta potential, and polydispersity index of toothpaste were evaluated. Data was statistically analyzed (ANOVA/Tukey, 5%). Results: Whitening toothpastes containing optical colorants were effective for whitening the enamel, as whiter teeth were observed following treatment (higher means of ΔWI D ). In addition, when the parameters ΔE ab and ΔE 00 were evaluated, these toothpastes were graded as very good effectiveness (grade 4). Other toothpastes were graded as 3 (good effectiveness). PHT had a neutral pH and a larger mean particle size (412.8 nm). Elemental analysis demonstrated enamel with a silicon-enriched mineral layer on the enamel surface treated with SDG. The Ca/P ratio after enamel treatment varied from 1.74 (SWS) to 2.04 (SDG and WRP). Conclusions: Experimental whitening toothpastes containing optical colorants are effective at bleaching the enamel. The synergism among the different parameters analyzed seems to positively affect the color change after brushing with whitening toothpastes containing optical colorants.

Beneficial effects of a mouthwash containing an antiviral phthalocyanine derivative on the length of hospital stay for COVID-19: randomised trial.

The risk of contamination and dissemination by SARS-CoV-2 has a strong link with nasal, oral and pharyngeal cavities. Recently, our research group observed the promising performance of an anionic phthalocyanine derivative (APD) used in a mouthwash protocol without photoexcitation; this protocol improved the general clinical condition of patients infected with SARS-CoV-2. The present two-arm study evaluated in vitro the antiviral activity and cytotoxicity of APD. Additionally, a triple-blind randomized controlled trial was conducted with 41 hospitalized patients who tested positive for COVID-19. All the included patients received World Health Organization standard care hospital treatment (non-intensive care) plus active mouthwash (experimental group AM/n = 20) or nonactive mouthwash (control group NAM/n = 21). The adjunct mouthwash intervention protocol used in both groups consisted one-minute gargling/rinsing / 5 times/day until hospital discharge. Groups were compared considering age, number of comorbidities, duration of symptoms prior admission and length of hospital stay (LOS). The associations between group and sex, age range, presence of comorbidities, admission to Intensive care unit (ICU) and death were also evaluated. The in vitro evaluation demonstrated that APD compound was highly effective for reduction of SARS-CoV-2 viral load in the 1.0 mg/mL (99.96%) to 0.125 mg/mL (92.65%) range without causing cytotoxicity. Regarding the clinical trial, the median LOS of the AM group was significantly shortened (4 days) compared with that of the NAM group (7 days) (p = 0.0314). Additionally, gargling/rinsing with APD was very helpful in reducing the severity of symptoms (no ICU care was needed) compared to not gargling/rinsing with APD (28.6% of the patients in the NAM group needed ICU care, and 50% of this ICU subgroup passed way, p = 0.0207). This study indicated that the mechanical action of the protocol involving mouthwash containing a compound with antiviral effects against SARS-CoV-2 may reduce the symptoms of the patients and the spread of infection. The use of APD in a mouthwash as an adjuvant the hospital COVID-19 treatment presented no contraindication and reduced the hospital stay period.Trial registration: The clinical study was registered at REBEC-Brazilian Clinical Trial Register (RBR-58ftdj).

Enamel remineralization and surface roughness after treatment with herbal-containing toothpastes.

BACKGROUND: Oral care products containing bioactive agents obtained from extracts of plant drugs were launched. This in vitro study investigated the effects of herbal-containing toothpastes associated or not with fluoride to remineralize the enamel after cariogenic challenge with pH cycling. The chemical and physical factors of toothpastes and the enamel surface roughness after brushing were also analyzed. MATERIAL AND METHODS: Sixty bovine enamel blocks were obtained and divided into 3 thirds: intact (untreated), demineralized (artificial caries lesion), and treated (caries lesion, pH cycling, and brushing with toothpastes). Toothpastes containing herbal compounds contained no fluoride [Galla chinensis (GCH)], low-F concentration [D'Or (DOR); Herbal Bliss (HBL)], or a different fluoride type [Elmex Anticaries (EAC)]. The results were compared to NaF-containing toothpastes: 1450 and 5000 ppm. Enamel blocks were brushed with the toothpastes using a pH-cycling model (7 days). The Knoop hardness (25g/10s) of the surface and the longitudinal sections were then evaluated. The percentage of surface hardness recovery (%SHR) was calculated. The enamel surface roughness, pH, particle size, zeta potential, and polydispersity index of toothpaste slurries were also evaluated. Data were statistically analyzed (α=5%). RESULTS: No significance was observed when %SHR was compared (p>0.05). DOR, GCH, and HBL were more effective in remineralizing the enamel subsurface. Significantly higher surface roughness was observed when treated with EAC and GCH (p<0.05). CONCLUSIONS: All toothpastes were able to remineralize the enamel, especially the subsurface, with results equal or better than that of standard toothpastes. Key words:Enamel, hardness, roughness, toothpaste, tooth remineralization.

Virucidal Activity of the Antiseptic Mouthwash and Dental Gel Containing Anionic Phthalocyanine Derivative: In vitro Study.

AIM: This research suggested an in vitro virucidal action of a dental gel and a mouthwash with phthalocyanine derivative. PURPOSE: The aim of this study was to report an in vitro study evaluating the virucidal capacity of mouthwash and dental gel containing anionic phthalocyanine derivate (APD). METHODS: The research followed the recommendations of the National Health Surveillance Agency (ANVISA) and adapted methodology, described in the standards EN14776: 2015; ASTM E1053-11 and the Robert Koch Institute - RKI, in addition to good laboratory practices (GLP). The determination of the percentage of inactivation of the SARS-CoV-2 virus particles was carried out by imposing the viral solution in contact with the respective tested products, with intervals of 30 seconds, 1 and 5 minutes, with subsequent submission of the aliquots, recovered in cell culture microplates following virus titration using the TCID50 (50% Median Tissue Culture Infectious Dose). RESULTS: The Mouthwash APD presented 90% of viral inactivation percentage, while the dental gel APD demonstrated 99.99% of viral inactivation. CONCLUSION: In vitro analyses showed that mouthwash and dental gel APD can reduce the viability of SARS-CoV-2 virus particles.

A Phthalocyanine Derivate Mouthwash to Gargling/Rinsing as an Option to Reduce Clinical Symptoms of COVID-19: Case Series.

AIM: This case series demonstrated that phthalocyanine derivate mouthwash is a promising alternative for reducing the viral load of SARS-CoV-2 and for clinical improvement of infected patients who presented mild and moderate symptoms. PURPOSE: The aim of this study was to report a case series of patients diagnosed with COVID-19 that used the phthalocyanine derivate mouthwash to reduce clinical symptoms. PATIENTS AND METHODS: Eight patients used 5mL of phthalocyanine derivate mouthwash gargling/rinsing for one minute, five times daily, over a fourteen day period. Two measurement scales were applied for each patient in different periods to verify sore throat - VAS - Visual Analogue Scale for Pain and the clinical conditions - PS - Performance Status. RESULTS: All patients presented a significant reduction in clinical symptoms with the use of the mouthwash for gargling/rinsing after few days of use, without hospitalization. CONCLUSION: The phthalocyanine derivate mouthwash protocol appears as a potential alternative for clinical improvement of COVID-19 infected patients. Daily use of this mouthwash rapidly reduced clinical symptoms such as sore throats, cough and mouth ulcers. Large, high-quality randomized controlled trials with larger sample size are necessary to confirm the effectiveness of this mouthwash protocol against COVID-19.

Biomimetic Mechanism of Action of Fluoridated Toothpaste Containing Proprietary REFIX Technology on the Remineralization and Repair of Demineralized Dental Tissues: An In Vitro Study.

OBJECTIVES: This in vitro study aimed to characterize the mineral content and surface and cross-sectional morphology of enamel and dentin tissues treated with a 1450 ppm fluoride-containing toothpaste with REFIX technology. MATERIALS AND METHODS: Bovine enamel blocks (n = 5) were obtained (4 × 4 × 6 mm), demineralized (artificial caries lesion), and treated (pH cycling and brushing with the toothpaste). During the pH cycling, which lasted for 7 days (demineralization and remineralization took 6 and 18 hours, respectively), the enamel was brushed for 5 minutes using an electric toothbrush before being immersed in a remineralizing solution. The dentin blocks were acid-etched for 2 minutes (0.05 M citric acid, pH 1.8) to expose the dentinal tubules (n = 5). Morphological analysis of the dentin was performed immediately and after 7 days of brushing with the dentifrice, and compared with the control group. The specimens were then cross-sectioned. The surface and cross-sectional micromorphology were assessed using scanning electron microscopy (SEM). The elemental analyses (weight%) were determined with an energy-dispersive X-ray spectroscopy (EDS). RESULTS: The toothpaste with REFIX technology remineralized and repaired the surface enamel effectively. The elemental analysis also demonstrated that treating the enamel with the toothpaste formed a silicon-enriched mineral layer on the enamel surface. Elemental analysis of the enamel cross-sections showed that the toothpaste induced a mineral change. The results were also consistent in the dentin, where the dentinal tubules were progressively occluded until there was complete occlusion after 7 days. CONCLUSIONS: We prove the biomimetic mechanism of action of fluoridated toothpaste containing proprietary REFIX technology for obtaining silicon-enriched, remineralized and repaired dental tissues.

Silicon-enriched hydroxyapatite formed induced by REFIX-based toothpaste on the enamel surface / Formação de hidroxiapatita enriquecida com silício induzida por dentifrício à base de REFIX na superfície do esmalte

Objetivo: Este estudo in vitro objetivou caracterizar o teor de minerais e a morfologia das partículas de um dentifrício fluoretado contendo tecnologia REFIX e o teor de minerais e a morfologia da superfície do esmalte tratada com este produto. Material e Métodos: Blocos de esmalte bovino (n=5) foram obtidos (4×4×6 mm), desmineralizados (lesão de cárie artificial) e tratados (ciclagem de pH e escovação com dentifrício). Durante a ciclagem de pH de 7 dias (desmineralização e remineralização levaram 6 h e 18 h, respectivamente), o esmalte foi escovado por 5 min com uma escova dental elétrica antes de ser imerso em uma solução de remineralização. Avaliou-se a micromorfologia e os níveis elementares (% em peso) dos íons presentes no dentifrício e na camada formada sobre o esmalte após o tratamento. A morfologia da superfície foi avaliada por meio de microscopia eletrônica de varredura (MEV). As análises elementares foram realizadas usando um espectrômetro de energia dispersiva de raios-X (EDS). Resultados: A análise por MEV do dentifrício demonstrou uma distribuição de tamanho uniforme de partículas de forma regular que foram organizados esparsamente em "nanoclusters". A análise elementar da formulação do dentifrício demonstrou a presença de Si (silício), Na (sódio), P (fósforo) e F (flúor), entre outros. Observou-se também uma camada mineral na superfície de esmalte com espessura uniforme consistente em torno de 14 µm. Conclusão: De acordo com os resultados, conclui-se que a tecnologia REFIX modifica a superfície e subsuperfície da estrutura do esmalte, formando uma camada mineral rica em silício. (AU)

Objective: This in vitro study aimed at characterizing the mineral content and filler particle morphology of a fluoridated toothpaste containing REFIX technology and the mineral content and the morphology of the enamel surface treated with this product. Material and Methods: Bovine enamel blocks (n=5) were obtained (4×4×6 mm), demineralized (artificial caries lesion), and treated (pH cycling and brushing with the toothpaste). During the pH cycling, which lasted for 7 days (demineralization and remineralization took 6 h and 18 h, respectively), the enamel was brushed for 5 min using an electric toothbrush before being immersed in a remineralization solution. We evaluated the micromorphology and elemental levels (wt%) of ions present in the toothpaste and on the layer formed over the enamel following the treatment. The surface morphology was assessed using scanning electron microscopy (SEM). The elemental analyses were performed using an energy-dispersive X-ray spectrometer (EDS). Results: Scanning electron micrographs of the toothpaste demonstrated a uniform size distribution of regular-shaped fillers sparsely organized in nanoclusters. Elemental analysis of the toothpaste's formulation demonstrated the presence of Si (silicon), Na (sodium), P (phosphorus), and F (fluorine), among others. We also detected a mineral layer that had formed on the treated enamel surface; the layer had a consistent uniform thickness of ~14 µm. Conclusion: Based on the results, it can be stated that REFIX technology modifies the surface and subsurface of the enamel structure, forming a Si-rich mineral layer. (AU)

The interaction of sodium trimetaphosphate with collagen I induces conformational change and mineralization that prevents collagenase proteolytic attack.

OBJECTIVES: This study evaluated the cell viability and expression of different major genes involved in mineralization in odontoblast-like cells exposed to sodium trimetaphosphate (STMP). It was also investigated the influence of STMP on the rate of calcium phosphate crystal growth, its anti-proteolytic action against the enzymatic degradation of type I collagen, the binding mechanism of STMP to collagen fibrils, and the potential mechanism to induce collagen stabilization. METHODS: Immortalized rat odontoblast MDPC-23 cells were cultured. Cell viability was assessed by trypan blue staining, and the changes in gene expression balance induced by STMP were assessed by quantitative reverse transcription (qRT) PCR assays. Crystalline particle formation was monitored by light-scattering detectors to estimate pH variation and the radial size of the crystalline particles as a function of reaction time (pH 7.4, 25°C) in the presence of STMP in supersaturated calcium phosphate solution (Ca/P=1.67). Images were obtained under atomic force microscopy (AFM) to measure the particle size in the presence of STMP. A three-point bending test was used to obtain the elastic modulus of fully demineralized dentin beams after immersion in STMP solution. The binding mechanism of STMP to collagen fibrils and potential stabilization mechanism was assessed with circular dichroism spectrometry (CD). The data were analyzed statistically (α=0.05). RESULTS: STMP had no significant influence on the cell viability and gene expression of the MDPC-23 cells. STMP greatly increased the rate of crystal growth, significantly increasing the average radial crystal size. AFM corroborated the significant increase of STPM-treated crystal size. Mineralized collagen I fibrils exhibited less collagenase degradation with lower STMP concentration. CD analysis demonstrated changes in the conformational stability after STMP binding to type I collagen. SIGNIFICANCE: The increased resistance of collagen against the proteolytic activity of collagenases appears to be related to the conformational change induced by STMP binding in collagen I and the STMP capacity for promoting biomimetic mineralization in type I collagen fibrils.