Anticandidal Efficacy of Erythrosine with Nano-TiO2 and Blue LED-Mediated Photodynamic Therapy against Candida albicans Biofilms on Acrylic Resin: A Preliminary Study.

OBJECTIVE: Incorporating an enhancer such as nano-titanium dioxide into antimicrobial photodynamic therapy can improve treatment outcome.This study aimed to compare the anticandidal efficacy of photodynamic therapy by erythrosine with nano-titanium dioxide (nano-TiO2) stimulated by a blue light emitting diode with three standard dental antifungal agents. MATERIALS AND METHODS: Candida albicans biofilms on acrylic resin plates were treated for 15 minutes with either nystatin, fluconazole, Polident, 220µM erythrosine + 1% (w/w) nano-TiO2 + 15 J/cm2 blue light photodynamic therapy (Ery PDT), or distilled water. For the Ery PDT group, blue light was applied for 1 minute after incubation. After 1, 3, and 6 hours, the colony forming units in log10 (log10CFU/mL) were compared. The ultrastructure of C. albicans on the acrylic resin plates treated with erythrosine + nano-TiO2 + blue light was examined using transmission electron microscopy at magnification of 30,000x. RESULTS: After 1 hour, nystatin, Polident, and Ery PDT indifferently inhibited C. albicans. At 6 hours, Ery PDT reduced the number of viable C. albicans in biofilms by 0.28log10 CFU/mL, which was equal to the effect of fluconazole and Polident. Transmission electron microscopy demonstrated that Ery PDT altered the C. albicans cell morphology by inducing cell wall/membrane rupture. CONCLUSION: Photodynamic therapy with erythrosine + nano-TiO2 + blue light at low light power density (15 J/cm2) was as effective at inhibiting C. albicans biofilm on acrylic resin as fluconazole and Polident.

Combined bisdemethoxycurcumin and potassium iodide-mediated antimicrobial photodynamic therapy.

Antimicrobial photodynamic therapy is emerging as a promising way to treat infections with minimal side effects. Typically, a single photosensitizer used in photodynamic therapy is capable of generating only one type of reactive oxygen species, which may have inadequate capability to eradicate certain types of microbes, especially Candida species. Thus, the use of combined photosensitizers is examined as a means of achieving superior antimicrobial results. We postulate that bisdemethoxycurcumin, a type I reactive oxygen species generator, combined with potassium iodide, an antimicrobial iodide molecule, might exhibit superior antimicrobial effects compared to a single photosensitizer-mediated photodynamic therapy. The effects of bisdemethoxycurcumin + potassium iodide + dental blue light on Candida albicans reduction were examined. Candida biofilms were treated with 20, 40 or 80 µM bisdemethoxycurcumin, 100 mM potassium iodide or a combination of these species for 20 min before irradiation with a dental blue light (90 J/cm2). The negative and positive controls were phosphate buffer saline and nystatin at 1 : 100,000 units/ml, respectively. Candidal numbers were quantified at 0, 1, 6 and 24 h. Hydroxyl radicals were spectrophotometrically measured using 2-[6-(4'amino phynoxyl-3H-xanthen-3-on-9-yl)] benzoic acid or APF probe-mediated fluorescence intensity (Varioskan) at 490/515 nm (excitation/emission). Candidal counts and hydroxyl radical comparisons were performed using the Kruskal-Wallis test and one-way ANOVA, respectively. Correlations between candidal numbers and hydroxyl radical levels were done with a Pearson correlation test. Forty µM bisdemethoxycurcumin+100 mM KI could provide a 3.5 log10 CFU/ml reduction after 6 h. Bisdemethoxycurcumin alone generated OH levels that were strongly correlated with candidal reduction. In conclusion, 40 µM bisdemethoxycurcumin+100 mM KI could reduce C. albicans biofilm.

An in vitro anti-inflammatory effect of Thai propolis in human dental pulp cells.

OBJECTIVE: To explore the potential for development of Thai propolis extract as a pulp capping agent to suppress pulpal inflammation from dental pulp infections. This study aimed to examine the anti-inflammatory effect of the propolis extract on the arachidonic acid pathway, activated by interleukin (IL)-1ß, in cultured human dental pulp cells. METHODOLOGY: Dental pulp cells, isolated from three freshly extracted third molars, were first characterized for their mesenchymal origin and treated with 10 ng/ml of IL-1ß in the presence or absence of non-toxic concentrations of the extract from 0.08 to 1.25 mg/ml, as determined by the PrestoBlue cytotoxic assay. Total RNA was harvested and analyzed for mRNA expressions of 5-lipoxygenase (5-LOX) and cyclooxygenase-2 (COX-2). Western blot hybridization was performed to investigate COX-2 protein expression. Culture supernatants were assayed for released prostaglandin E2 levels. Immunofluorescence was conducted to determine involvement of nuclear factor-kappaB (NF-kB) in the inhibitory effect of the extract. RESULTS: Stimulation of the pulp cells with IL-1ß resulted in the activation of arachidonic acid metabolism via COX-2, but not 5-LOX. Incubation with various non-toxic concentrations of the propolis extract significantly inhibited upregulated COX-2 mRNA and protein expressions upon treatment with IL-1ß (p<0.05), resulting in a significant decrease in elevated PGE2 levels (p<0.05). Nuclear translocation of the p50 and the p65 subunits of NF-kB upon treatment with IL-1ß was also blocked by incubation with the extract. CONCLUSIONS: Upregulated COX-2 expression and enhanced PGE2 synthesis upon treatment with IL-1ß in human dental pulp cells were suppressed by incubation with non-toxic doses of Thai propolis extract via involvement of the NF-kB activation. This extract could be therapeutically used as a pulp capping material due to its anti-inflammatory properties.

An in vitro anti-inflammatory effect of Thai propolis in human dental pulp cells

Abstract Objective To explore the potential for development of Thai propolis extract as a pulp capping agent to suppress pulpal inflammation from dental pulp infections. This study aimed to examine the anti-inflammatory effect of the propolis extract on the arachidonic acid pathway, activated by interleukin (IL)-1β, in cultured human dental pulp cells. Methodology Dental pulp cells, isolated from three freshly extracted third molars, were first characterized for their mesenchymal origin and treated with 10 ng/ml of IL-1β in the presence or absence of non-toxic concentrations of the extract from 0.08 to 1.25 mg/ml, as determined by the PrestoBlue cytotoxic assay. Total RNA was harvested and analyzed for mRNA expressions of 5-lipoxygenase (5-LOX) and cyclooxygenase-2 (COX-2). Western blot hybridization was performed to investigate COX-2 protein expression. Culture supernatants were assayed for released prostaglandin E2 levels. Immunofluorescence was conducted to determine involvement of nuclear factor-kappaB (NF-kB) in the inhibitory effect of the extract. Results Stimulation of the pulp cells with IL-1β resulted in the activation of arachidonic acid metabolism via COX-2, but not 5-LOX. Incubation with various non-toxic concentrations of the propolis extract significantly inhibited upregulated COX-2 mRNA and protein expressions upon treatment with IL-1β (p<0.05), resulting in a significant decrease in elevated PGE2 levels (p<0.05). Nuclear translocation of the p50 and the p65 subunits of NF-kB upon treatment with IL-1β was also blocked by incubation with the extract. Conclusions Upregulated COX-2 expression and enhanced PGE2 synthesis upon treatment with IL-1β in human dental pulp cells were suppressed by incubation with non-toxic doses of Thai propolis extract via involvement of the NF-kB activation. This extract could be therapeutically used as a pulp capping material due to its anti-inflammatory properties.

Effect of intermittency factor on singlet oxygen and PGE2 formation in azulene-mediated photodynamic therapy: A preliminary study.

In photodynamic therapy, intermittent irradiation modes that incorporate an interval between pulses are believed to decrease the effect of hypoxia by permitting an interval of re-oxygenation. The effect of the irradiation intermittency factor (the ratio of the irradiation pulse time to the total irradiation time) on singlet oxygen formation and inflammatory cytokine production was examined using azulene as a photosensitizer. Effects of difference intermittency factor on singlet oxygen formation and inflammatory cytokine were examined. Azulene solutions (1/10 µM) were irradiated with a 638-nm 500 mW diode laser in fractionation (intermittency factor of 5 or 9) or continuous mode using 50 mW/cm2 at 4 or 8 J/cm2. Singlet oxygen measurement was performed using a dimethyl anthracene probe. Peripheral blood mononuclear cells (PBMC) were stimulated by 10 ng/ml rhTNF-α for 6 h, before addition of 1 and 10 µM azulene solutions and irradiation. PGE2 measurement was undertaken using a human PGE2 ELISA kit. Kruskal-Wallis with Dunn Bonferroni test was used for statistical analyses at p < 0.05.Irradiation of 1 µM azulene+4 J/cm2+intermittency factor of 9 increased singlet oxygen 3-fold (p < 0.0001). Irradiation of 10 µM azulene at either 4 J/cm2+intermittency of 9 or 8 J/cm2+intermittency factor of 5 reduced PGE2 expression in PBMCs to non-inflamed levels. Thus, at 50 mW/cm2, 10 µM azulene-mediated photodynamic therapy with a high intermittency factor and a low energy density generated sufficient singlet oxygen to suppress PGE2 in Inflamed PBMCs.

Formulation development and evaluation of lidocaine hydrochloride loaded in chitosan-pectin-hyaluronic acid polyelectrolyte complex for dry socket treatment.

The main purpose of this study was to assess a lidocaine hydrochloride-loaded chitosan-pectin-hyaluronic polyelectrolyte complex for rapid onset and sustained release in dry socket wound treatment. Nine formulations (LCs) of lidocaine hydrochloride (LH) loaded into a chitosan-pectin-hyaluronic polyelectrolyte complex (PEC) were assessed using full factorial design (two factors × three levels). The formulations ranged between 4 and 10% w/w LH and 0.5-1.5% w/w HA. The following physicochemical properties of LCs were characterized: size, zeta potential, % entrapment efficiency, viscosity, mucoadhesiveness, % drug release, morphology, storage stability, and cytotoxicity. The particle size, zeta potential, % EE, viscosity, and % mucoadhesion increased with increasing LH and HA concentrations. Rapid release of LH followed a zero-order model, and a steady-state percentage of the drug was released over 4 h. LCs were found to be non-cytotoxic compared to LH solution. LH loaded into PEC demonstrated appropriate characteristics-including suitable rate of release-and fit a zero-order model. Furthermore, it was not cytotoxic and showed good stability in a high-HA formula, making it a promising candidate for future topical oral formulations.

Anthocyanin complex niosome gel accelerates oral wound healing: In vitro and clinical studies.

An anthocyanin complex (AC), composed of extracts of purple waxy corn and blue butterfly pea petals, and AC niosomes, bilayered vesicles of non-ionic surfactants, were compared in in vitro and clinical studies. Cultured fibroblasts subjected to a scratch wound were monitored for cell viability, cell migration, nuclear morphology and protein expression. Scratched cells showed accelerated wound healing activity, returning to normal 24 h after treatment with AC niosomes (0.002 mg/mL). Western blots and immunocytochemistry indicated upregulation of type I, III and IV collagens, fibronectin and laminins in AC niosome-treated scratched cells. A randomized block placebo-controlled double-blind clinical trial in 60 volunteers (18-60 years old) with oral wounds indicated that AC niosome gel accelerated wound closure, reduced pain due to the oral wounds and improved participants' quality of life more than AC gel, triamcinolone gel and placebo gel. These data are consistent with enhanced delivery of AC to fibroblasts by use of niosomes. AC niosomes activated fibroblasts within wounded regions and accelerated wound healing, indicating that AC niosomes have therapeutic potential.

Anti-Inflammatory Comparison of Melatonin and Its Bromobenzoylamide Derivatives in Lipopolysaccharide (LPS)-Induced RAW 264.7 Cells and Croton Oil-Induced Mice Ear Edema.

The pineal gland is a neuroendocrine organ that plays an important role in anti-inflammation through the hormone melatonin. The anti-inflammatory effects of melatonin and its derivatives have been reported in both in vitro and in vivo models. Our previous study reported the potent antioxidant and neuroprotective activities of bromobenzoylamide substituted melatonin. In silico analysis successfully predicted that melatonin bromobenzoylamid derivatives were protected from metabolism by CYP2A1, which is a key enzyme of the melatonin metabolism process. Therefore, the anti-inflammatory activities of melatonin and its bromobenzoylamide derivatives BBM and EBM were investigated in LPS-induced RAW 264.7 macrophages and croton oil-induced ear edema in mice. The experiments showed that BBM and EBM significantly reduced production of the inflammatory mediators interleukin-6 (IL-6), prostaglandin E2 (PGE2), and nitric oxide (NO) in a dose-dependent manner, but only slightly affected TNF-α in LPS-induced RAW 264.7 macrophages. This suggests that modifying melatonin at either the N1-position or the N-acetyl side chain affected production of NO, PGE2 and IL-6 in in vitro model. In the croton oil-induced mouse ear edema model, BBM, significantly decreased ear edema thickness at 2-4 h. It leads to conclude that bromobenzoylamide derivatives of melatonin may be one of the potential candidates for a new type of anti-inflammatory agent.

Inhibitory Effects of Erythrosine/Curcumin Derivatives/Nano-Titanium Dioxide-Mediated Photodynamic Therapy on Candida albicans.

This study focuses on the role of photosensitizers in photodynamic therapy. The photosensitizers were prepared in combinations of 110/220 µM erythrosine and/or 10/20 µM demethoxy/bisdemethoxy curcumin with/without 10% (w/w) nano-titanium dioxide. Irradiation was performed with a dental blue light in the 395-480 nm wavelength range, with a power density of 3200 mW/cm2 and yield of 72 J/cm2. The production of ROS and hydroxyl radical was investigated using an electron paramagnetic resonance spectrometer for each individual photosensitizer or in photosensitizer combinations. Subsequently, a PrestoBlue® toxicity test of the gingival fibroblast cells was performed at 6 and 24 h on the eight highest ROS-generating photosensitizers containing curcumin derivatives and erythrosine 220 µM. Finally, the antifungal ability of 22 test photosensitizers, Candida albicans (ATCC 10231), were cultured in biofilm form at 37 °C for 48 h, then the colonies were counted in colony-forming units (CFU/mL) via the drop plate technique, and then the log reduction was calculated. The results showed that at 48 h the test photosensitizers could simultaneously produce both ROS types. All test photosensitizers demonstrated no toxicity on the fibroblast cells. In total, 18 test photosensitizers were able to inhibit Candida albicans similarly to nystatin. Conclusively, 20 µM bisdemethoxy curcumin + 220 µM erythrosine + 10% (w/w) nano-titanium dioxide exerted the highest inhibitory effect on Candida albicans.

Effect of Different Irradiation Modes of Azulene-mediated Photodynamic Therapy on Singlet Oxygen and PGE2 Formation.

Azulene samples in ethanol/distilled water (1, 10 and 100 µm) were irradiated with a 638 nm red laser (0.5 watts, light-to-target distance 2 cm, energy density 4 or 40 J cm-2 ) by either continuous, fractionation or pulse mode. Singlet oxygen in the samples was measured using 10 µm 9,10-dimethyl anthracene (positive control 10 µm erythrosine) and relative fluorescence intensities were measured at 375/436 nm excitation/emission. Peripheral blood mononuclear cells (PBMCs, 1 × 105 cells/well) preincubated with 0.01 µg mL-1 rhTNF-α for 6 h were cultured with irradiated azulene samples in RPMI-1640 under standard conditions. PGE2 was quantified by rhPGE2 ELISA kit using a Varioscan® microplate reader at an excitation wavelength of 420 nm. Kruskal Wallis with Dunn`s test was performed at a significance level of P < 0.05. The highest singlet oxygen amount was found in 10 µm azulene samples irradiated at 40 J cm-2 under continuous mode (P = 0.001 when compared with 10 µm erythrosine). PGE2 expression in rhTNF-α-induced PBMCs was reduced to 45% of control by 1 µm azulene irradiated at 40 J cm-2 under fractionation mode. Fractionation mode with intermediate laser energy density in the presence of low concentration of azulene could increase singlet oxygen and tend to reduce PGE2 .

Preservation of the viability and gene expression of human periodontal ligament cells by Thai propolis extract.

BACKGROUND/AIM: Success of tooth replantation depends on the quality and quantity of periodontal ligament (PDL) cells. The aims of this study were to evaluate Thai propolis extract as a storage medium for maintaining PDL cell viability and preserving gene expressions in PDL tissues. MATERIALS AND METHODS: PDL cells from human premolars were tested for cytotoxicity of the extract by PrestoBlue assay to determine a non-toxic concentration. Subsequently, 96 freshly extracted premolars were allocated into different treatment groups. Control groups were freshly extracted premolars or they had been stored dry for 12 hours. Experimental avulsed teeth were created by leaving them air-dried for 30 minutes immediately after extraction, then they were immersed in Thai propolis extract, HBSS or milk for 3, 6 and 12 hours. After tooth storage, the remaining PDL cells were determined for their cell viability. RNA isolated from PDL tissues of three premolars treated similarly was analysed for periostin and S100A4 expressions using RT-qPCR. RESULTS: Thai propolis extract at 0.625 mg mL-1 promoted the greatest PDL cell viability. Tooth storage in 0.625 mg mL-1 Thai propolis extract, HBSS or milk showed no difference in maintaining cell viability. Periostin mRNA level was preserved by Thai propolis extract. Expression of S100A4 mRNA in PDL tissues stored in all tested media was dampened. CONCLUSIONS: PDL cells from mock avulsed teeth stored in 0.625 mg mL-1 Thai propolis extract for 3, 6 and 12 hours remained viable and the expression of periostin was preserved. This study suggests this extract as an alternative for a tooth storage medium for up to 12 hours. However, transporting an avulsed tooth in a storage medium for extended extra-oral time might affect the PDL cell phenotypes.

Anti-inflammatory effect of photodynamic therapy using guaiazulene and red lasers on peripheral blood mononuclear cells.

INTRODUCTION: Photodynamic therapy improves oral mucositis treatment. The reactive oxygen species (ROS) generated from this reaction could contribute to an anti-inflammatory effect by suppressing inflammatory cells. OBJECTIVE: To evaluate the anti-inflammatory effect of photodynamic therapy using guaiazulene and a red laser in peripheral blood mononuclear cells (PBMCs). METHODS: Guaiazulene solutions (1, 2, 5, 25, 35, and 100 µM in 99.8 % methanol) were irradiated with red laser light (625 nm, 146.2 mW/cm2) in continuous mode at 0, 4, and 8 J/cm2 in black 96-well plates. ROS were measured using spin trapping technique with electron spin resonance (ESR) spectroscopy and fluorescence. The two highest concentrations were tested using cell viability (PrestoBlue®) and anti-inflammation (RANTES and PGE2 ELISA) assay kits. Kruskal-Wallis and Dunn Bonferroni tests were used for statistical analyses with significant differences at p-value < 0.05. RESULTS: Guaiazulene solutions between 2 and 5 µM exposed to red laser light at 4-8 J/cm2 generated significantly more singlet oxygen compared to the no guaiazulene group (p < 0.01) and reduced RANTES and PGE2 levels in TNF-α-inflamed peripheral blood mononuclear cells without affecting cell viability. CONCLUSION: Photodynamic activation of guaiazulene generated singlet oxygen and suppressed inflammatory markers in PBMCs.

Glutaryl Melatonin Niosome Gel for Topical Oral Mucositis: Anti- Inflammatory and Anticandidiasis.

BACKGROUND: Glutaryl melatonin, which is synthesized from melatonin and is a pineal glandderived neurohormone with anti-inflammatory and anti-oxidant properties, was comparatively investigated for its potential use as a topical anti-inflammatory agent. OBJECTIVE: Glutaryl melatonin, synthesized and screened for in vitro anti-candidiasis and in vitro and in vivo anti-inflammatory activities, was formulated as a niosome gel for topical oral evaluation in 5- fluorouracil-induced oral mucositis in mice. METHODS: In vitro anti-fungal activity in Candida albicans, in vitro anti-inflammatory activity in Escherichia coli liposaccharide-induced RAW cells and in vivo anti-inflammatory activity using a croton oilinduced ear edema model in ICR mice were investigated. Mucositis in mice (n= 6/group, 10-week-old mice) was induced by intraperitoneal injections of 5-fluorouracil, and the mice were subjected to a topical oral application of niosome gel containing melatonin (2% w/w) or glutaryl melatonin (2% w/w) and were compared with mice subjected to blank, fluocinolone acetonide (0.5% w/w) and control conditions. RESULTS: Glutaryl melatonin, at a 14.2 mM concentration, showed the highest fungicidal effect on C. albicans using the broth dilution method, indicating a nonsignificant difference from 1 µM of nystatin (p = 0.05). Nitric oxide, interleukin-6 and tumor necrosis factors were analyzed by ELISA. Liposaccharide-induced RAW cells were significantly reduced by glutaryl melatonin (p < 0.01). Ear edema inhibition of glutaryl melatonin was significant 1 h after application compared with that of melatonin (p = 0.03). Food consumption and body weight of the 5-fluorouracil-treated mice were significantly lower than those of the normal mice before all treatments (p < 0.05). Differences in the amount of licking behavior, which were observed in the control group for 5 min, were noticeable in the 5- fluorouracil-treated mice but not in the mice treated with the glutaryl melatonin niosome gel. CONCLUSION: Glutaryl melatonin exhibited mild anti-candidiasis and anti-inflammatory properties. The incorporation of glutaryl melatonin in a niosome gel formulation, demonstrated the potential for topical oral applications to reduce oral discomfort caused by 5-fluorouracil treatment in mice.

Effects of photodynamic therapy with azulene on peripheral blood mononuclear cell viability and singlet oxygen formation.

BACKGROUND: We aimed to investigate the effect of azulene on peripheral blood mononuclear cells (PBMCs) viability and singlet oxygen formation. METHODS: 1 × 105 PBMCs were cultured in a 96-well plate in RPMI-1640 supplemented with 10% FBS (37 °C, 5% CO2) for 24 h. Each well was treated for 30 min with each azulene concentration between 0-500 µM and activated by a 625 ± 5 nm light emitting diode (power 20-23 mW) at energy densities of 0-200 J/cm2. MTT cell viability was recorded using a spectrophotometer at a 570 nm. 9,10-Dimethylanthracene (DMA) was utilized for the measurement of singlet oxygen, using a fluorescence spectrophotometer at 375 and 436 nm as the excitation and emission wavelengths, respectively. Optical density,　relative fluorescence units were compared using one-way ANOVA and a post-hoc test. The correlation between the cell number and singlet oxygen amount was analyzed by the Spearman correlation test. RESUTS: Azulene at all concentrations with 4.2 J/cm2 light significantly induced singlet oxygen formation. 15 µM azulene with 4.2, 100, or 200 J/cm2 light significantly reduced PBMC viability. The inverse relationship between the cell viability and singlet oxygen amount was observed. CONCLUSIONS: An optimum azulene concentration + red light energy density decreased PBMC viability via singlet oxygen formation.

Topical oral wound healing potential of anthocyanin complex: animal and clinical studies.

AIM: An anthocyanin complex (AC), combined Zea mays and Clitoria ternatea extracts, was evaluated for topical oral wound healing in rats and a clinical trial in orthodontic patients. METHODS/RESULTS: AC enhanced anthocyanin permeation in vitro. In rats, 10% w/w of AC in a mucoadhesive gel (AG) reduced erythema and sizes of oral wounds after topical applications at higher extent than its placebo gel. Acute orthodontic wounds in 68 volunteers were randomly assigned to topically receive either AG or placebo gel and double-blind assessed. Wound size reduction and wound closure enhancement were obvious in AG-treated group on day 3 (p < 0.05). CONCLUSION: At 10% w/w, AC promoted wound closure and possessed a potential in healing stimulation of acute oral wounds.

Topical Niosome Gel Containing an Anthocyanin Complex: a Potential Oral Wound Healing in Rats.

Anthocyanins from dietary sources showing potential benefits as anti-inflammatory in oral lesions were developed as an anthocyanin complex (AC), comprised of extracts of Zea mays (CC) and Clitoria ternatea (CT), and formulated into a niosome gel to prove its topical oral wound healing in vitro and in vivo investigations. The AC formed nano-sized clusters of crystalline-like aggregates, occurring through both intra- and inter-molecular interactions, resulting in delivery depots of anthocyanins, following encapsulation in niosomes and incorporation into a mucoadhesive gel. In vitro permeation of anthocyanins was improved by complexation and further enhanced by encapsulation in niosomes. Collagen production in human gingival fibroblasts was promoted by AC and AC niosomes, but not CC or CT. The in vivo wound healing properties of AC gel (1 and 10%), AC niosome gel (1 and 10%), fluocinolone acetonide gel, and placebo gel were investigated for incisional wounds in the buccal cavities of Wistar rats. AC gel and AC niosome gel both reduced wound sizes after 3 days. AC niosome gel (10%) gave the highest reduction in wound sizes after day 3 (compared to fluocinolone acetonide gel, p < 0.05), and resulted in 100% wound healing by day 5. Histological observations of cross-sectioned wound tissues revealed the adverse effects of fluocinolone gel and wound healing potential of AC niosome gel. Topical application of AC niosome gel exhibited an anti-inflammatory effect and promoted oral wound closure in rats, possibly due to the improved mucosal permeability and presence of delivery depots of AC in the niosome gel.

Efficacy of erythrosine and cyanidin-3-glucoside mediated photodynamic therapy on Porphyromonas gingivalis biofilms using green light laser.

BACKGROUND: The purpose of this in vitro study was to evaluate the efficacy of erythrosine and cyanidin-3-glucoside as photosensitizers in PDT for the elimination of Porphyromonas gingivalis (P. gingivalis) biofilms. METHODS: P. gingivalis biofilms were prepared from a chronic periodontitis subject. Erythrosine and cyanidin-3-glucoside were prepared and randomly allocated as follows: 110, 220, 330, and 440µM erythrosine; 101, 202, 303, and 404µM anthocyanin; and 440µM erythrosine+404µM cyanidin-3-glucoside. There were 18 PDT experimental groups (non-irradiated/irradiated with a 532-nm green light diode laser at 1.29J/cm2 for 60s). The 3 controls were grouped as follows: biofilms exposed to the photosensitizers alone, biofilms exposed to the laser alone, and biofilms exposed to 0.12% chlorhexidine. All sample groups were cultured at 1, 3 and 6h after PDT and incubated in an anaerobic chamber at 37°C for 4days. The surviving fraction was calculated from the log10 CFU/ml. The 330 and 440µM erythrosine and the 440µM erythrosine+404µM cyanidin-3-glucoside were mixed with spin traps (TEMPO, DMPO), and the electron spin resonance spectra were evaluated. RESULTS: The log10 CFU/ml measurements showed that the PDT groups with 330µM or 440µM erythrosine and 440µM erythrosine+404µM cyanidin-3-glucoside had statistically significant differences from the other groups (one-way ANOVA and Bonferroni's multiple comparison test, p- value≤0.05). CONCLUSIONS: PDT using 330µM erythrosine, 440µM erythrosine or 440µM erythrosine+404µM cyanidin-3-glucoside irradiated with the laser more effectively inhibited P. gingivalis in biofilms.

Silk fibroin/gelatin-chondroitin sulfate-hyaluronic acid effectively enhances in vitro chondrogenesis of bone marrow mesenchymal stem cells.

Tissue engineering is becoming promising for cartilage repair due to the limited self-repair capacity of cartilage tissue. We previously fabricated and characterized a three-dimensional silk fibroin/gelatin-chondroitin sulfate-hyaluronic acid (SF-GCH) scaffold and showed that it could promote proliferation of human bone marrow mesenchymal stem cells (BM-MSCs). This study aimed to evaluate its biological performance as a new biomimetic material for chondrogenic induction of BM-MSCs in comparison to an SF scaffold and conventional pellet culture. We found that the SF-GCH scaffold significantly enhanced the proliferation and chondrogenic differentiation of BM-MSCs compared to the SF scaffold and pellet culture in which the production of sulfated glycoaminoglycan was increased in concordance with the up-regulation of chondrogenic-specific gene markers. Our findings indicate the significant role of SF-GCH by providing a supportive structure and the mimetic cartilage environment for chondrogenesis which enables cartilage regeneration. Thus, our fabricated SF-GCH scaffold may serve as a potential biomimetic material for cartilage tissue engineering.

Correlation between SNP genotypes and periodontitis in Japanese type II diabetic patients: a preliminary study.

The present study aims to investigate the correlation between SNP genotype patterns and periodontitis severity in Japanese type II diabetic patients. A cross-sectional study in 43 Japanese diabetic patients with periodontitis was performed. Blood samples were drawn for single nucleotide polymorphism (SNP) analyses and periodontal index (probing pocket depth and clinical attachment level) was subsequently recorded. Twelve functional genes with SNPs that had been shown to be associated with diabetes and/or inflammation were genotyped using a nuclease-mediated SNP-specific ligation method. Subjects with two or more sites with clinical attachment level ≥6 mm and who additionally had one or more sites with pocket depth ≥5 mm were classified as having severe periodontitis. Proportions of risk genotypes/non-risk genotypes between severe and non-severe periodontitis were subsequently compared. A high frequency (21/43 participants, 49%) of adiponectin gene polymorphism (ADIPOQ 45T > G) homozygous risk genotype (TT genotype) was observed in the participants. The frequency of TGF-ß1 SNP (29C > T) risk genotype (TT genotype) in severe periodontitis (34%, n = 11) was significantly higher than in non-severe periodontitis (0%, n = 0) (p = 0.04). Our study suggests that TGF-ß1 SNPs (29C > T) may be used as one of the risk indicators for severe periodontitis in Japanese diabetic patients.

Effect of periodontal treatment on adipokines in type 2 diabetes.

The association between adipokines and inflammatory periodontal diseases has been studied over the last two decades. This review was intended to explore the observation that periodontal therapy may lead to an improvement of adipokines in diabetic patients. In summary, substantial evidence suggests that diabetes is associated with increased prevalence, extent and severity of periodontitis. Numerous mechanisms have been elucidated to explain the impact of diabetes on the periodontium. However, current knowledge concerning the role of major adipokines indicates only some of their associations with the pathogenesis of periodontitis in type 2 diabetes. Conversely, treatment of periodontal disease and reduction of oral inflammation may have positive effects on the diabetic condition, although evidence for this remains somewhat equivocal.