Personal immune profiles: Diversity and prognostic value for oral tongue squamous cell carcinoma evaluated by comprehensive immune parameter analyses with multiplex immunofluorescence.

OBJECTIVES: Understanding the tumor immune microenvironment is becoming increasingly necessary for risk prediction and treatment selection. In particular, oral cancer has various immunosuppressive characteristics in the tumor microenvironment. Therefore, we comprehensively assessed the immune profiles of oral tongue squamous cell carcinoma (OTSCC). MATERIALS AND METHODS: Multiplex immunofluorescence and tissue imaging analyses were performed to evaluate immune profiles at the invasive tumor front of 60 OTSCC surgical specimens. We analyzed 58 immune parameters including the density and proportion (%) of total leukocytes (Leu) and T cells, six subsets of T and myeloid cells, and the expression of programmed cell death-1 (PD-1) and PD-1 ligand 1 (PD-L1). RESULTS: The density, proportion, and location of CD45+ Leu, three T cell subsets (CD8+, Foxp3-CD4+ conventional, and Foxp3+CD4+ regulatory T cells), CD163-CD68+ M1 and CD163+CD68+ M2 macrophages, and neutrophils were highly variable at the individual level. The density and proportion of M2 macrophages were significantly lower in the T1 stage group. Risk prediction analyses for recurrence and/or metastasis (R/M) showed that R/M (+) T1 cases had significantly higher M2 density and percentages. CONCLUSIONS: The immune profiles of OTSCC patients are diverse and cannot be predicted from clinicopathological information alone. The M2 macrophage abundance is a potential candidate biomarker for R/M in the early stage of OTSCC. Personal immune profiling may provide beneficial information for risk prediction and treatment selection.

Morus alba Stem Extract Suppresses Matrix Metalloproteinases (MMP)-1, MMP-9, and Tissue Inhibitors of Metalloproteinase (TIMP)-1 Expression via Inhibition of IκBα Degradation Induced by Porphyromonas gingivalis LPS Signal in THP-1 Cells.

OBJECTIVES: The aim of this study is to evaluate the inhibitory effects of M. alba stem extract (MSE) on the expression of matrix metalloproteinases (MMP)-1, MMP-9, and tissue inhibitors of metalloproteinase (TIMP)-1 in Porphyromonas gingivalis lipopolysaccharide (LPS)-activated-acute monocytic leukemia cell line (THP-1). MATERIALS AND METHODS: THP-1 cells were treated with noncytotoxic concentrations of MSE combined with 1 µg/mL of P. gingivalis LPS. The mRNA levels of MMP-1, MMP-9, and TIMP-1 were evaluated via quantitative real-time polymerase chain reaction. The secreted proteins in the culture media were detected by enzyme-linked immunosorbent assay. The degradation of inhibitor of kappa B-alpha (IκBα) protein was tracked by Western blotting. STATISTICAL ANALYSIS: Comparisons in experiments were analyzed with analysis of variance followed by Tukey honestly significant difference comparison test. RESULTS: Twenty and 40 µg/mL of MSE significantly downregulated MMP-1 and MMP-9 genes and protein expression but upregulated the gene expression of TIMP-1 (p < 0.05). P. gingivalis LPS induced degradation of IκBα, while addition of MSE (20 and 40 µg/mL) increased IκBα cytosolic levels. MSE was able to suppress the P. gingivalis LPS-induced MMPs expression and also increased the gene expression of TIMP-1 via the inhibition of the cytoplasmic IκBα degradation in THP-1 cells. CONCLUSIONS: The present observations suggest that MSE exerted a positive effect on the regulatory mechanism between MMPs and TIMP, which is an important implication for the therapeutic potential of MSE in periodontitis.

Clinical profile of 102 patients with oral lichen planus in Thailand.

BACKGROUND: Oral lichen planus (OLP) is a chronic inflammatory disease of the skin and mucous membrane presented with various clinical appearances. The aim of the present study was to elucidate the clinical profile of patients with OLP. MATERIAL AND METHODS: The dental records of 102 patients who visited Oral Medicine Clinic, Dental Hospital, Naresuan University during 2002-2018 were retrospectively reviewed. RESULTS: There were 75 (73.5%) women and 27 (26.5%) men, giving a female to male ratio of 2.8:1. The age of OLP patients ranged 20-81 years old with the mean age of 56.4 ± 13.2 years old. Seventy-eight patients (76.5%) had the history of systemic diseases and hypertension was the predominantly one. Most patients were non-smokers (98%), non-drinkers (86.3%) and non-betel nut chewers (98%). The atrophic form (93.1%) was the most common OLP. The lesions were mainly symptomatic (92.2%) and involved multiple locations (67.6%) where the buccal mucosa (79.4%) primarily affected. Only 2% were extraoral lesions detected on the skin. Patients had no family history of OLP or malignant transformation. Ninety-one patients (89.2%) were treated with topical steroid and only 4 patients were prescribed a combination of tropical and systemic steroid. CONCLUSIONS: The results of the study indicated that most of characteristics are in accordance with previous studies. Since, OLP is a chronic inflammatory oral mucosal disease with high recurrence rate, early detection, accurately diagnosis, and long-term follow-up are necessary to evaluate the exacerbation and malignant transformation. Key words:Clinical profile, demographic, oral lichen planus, retrospective study.

Cyclodextrin-based oral dissolving films formulation of taste-masked meloxicam.

This work deals with fast-dissolving drug delivery systems of meloxicam (MX) derived from electrospun polyvinylpyrrolidone (PVP)/2-hydroxypropyl-ß-cyclodextrin (HPßCD) nanofiber mats. Electrospinning of solutions with different solvent systems [dimethylformamide (DMF) and ethyl alcohol (EtOH)] was performed. Prepared films were evaluated for morphology, physical, and mechanical properties. MX content, dissolving time, MX release, and cytotoxicity of films were investigated. In vivo studies were also performed in healthy human volunteers. The results showed that MX/HPßCD complexes improved the solubility of MX. PVP also increased MX solubility and the stability of MX/HPßCD complexes. Films were successfully prepared by two solvent systems with fiber in the nanometer range. MX was well incorporated into the films (100% efficiency). The X-ray patterns and DSC experiment indicated an amorphous form of MX. A fast disintegration time and burst release of MX was obtained from EtOH system. Cytotoxicity testing of the films produced by EtOH system proved safer than the DMF system. In vivo studies revealed that films rapidly dissolved in the mouth and had a less bitter taste than MX. These results suggest that electospun films from EtOH system may be a good candidate for fast-dissolving drug delivery systems to increase palatability of dosage forms.

Inhibitory effect of alpha-mangostin on Candida biofilms.

The objective of this study was to determine the inhibitory effect of alpha-mangostin on Candida biofilms. Candida species including Candida albicans, Candida krusei, Candida tropicalis, and Candida glabrata were tested. Candida biofilms were formed in flat-bottomed 96-well microtiter plates. The metabolic activity of cells within biofilms was quantified using the XTT assay. The results demonstrated that alpha-mangostin showed a significant anti-biofilm effect on both developing biofilms and preformed biofilms of Candida species. It may be concluded that alpha-mangostin could be an anti-biofilm agent against Candida species. Further in vivo investigations are needed to uncover the therapeutic values of this medicinal plant.

Fabrication and Evaluation of Nanostructured Herbal Oil/Hydroxypropyl-ß-Cyclodextrin/Polyvinylpyrrolidone Mats for Denture Stomatitis Prevention and Treatment.

This work aims to develop the herbal oil-incorporated nanostructure mats with antifungal activity for the prevention and treatment of Candida-associated denture stomatitis. The nanofiber mats loaded with betel oil or clove oil were fabricated via electrospinning process. The morphologies and physicochemical properties of the herbal oil loaded nanofiber mats were examined using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), and mechanical testing. The release characteristic, antifungal activity, and cytotoxicity were also investigated. The SEM images confirmed the homogeneous and smooth nanoscale fibers. The addition of the herbal oil into the nanofiber mats reduced the fiber diameters. The DSC and FT-IR results confirmed the presence of the oil in the nanofiber mats. The herbal oils can be released from the mats in a very fast manner and inhibit the growth of candida cells within only few minutes after contact. These nanofiber mats may be beneficial for the prevention and treatment of denture stomatitis.

Fabrication of mucoadhesive chitosan coated polyvinylpyrrolidone/cyclodextrin/clotrimazole sandwich patches for oral candidiasis.

This study aims to fabricate clotrimazole (CZ)-composite sandwich nanofibers using electrospinning. The CZ-loaded polyvinylpyrrolidone (PVP)/hydroxypropyl-ß-cyclodextrin (HPßCD) fiber was coated with chitosan-cysteine (CS-SH)/polyvinyl alcohol (PVA) to increase the mucoadhesive properties and to achieve a sustained release of the drug from the nanofibers. The nanofibers were characterized using scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy and X-ray diffractometry (XRD). The nanofibers mechanical and mucoadhesive properties, drug release, antifungal activity and cytotoxicity were also assessed. The fibers were in the nanoscale with good mucoadhesive properties. The XRPD revealed a molecular dispersion of amorphous CZ in the nanofibers. The initial fast release of CZ from the nanofibers was achieved. Moreover, the sandwich nanofibers coated for longer times resulted in slower release rates compared with the shorter coating times. The CZ-loaded nanofibers killed the Candida significantly faster than the commercial CZ lozenges at 5, 15 and 30 min and were safe for a 2-h incubation. Therefore, these nanofibers may be promising candidates for the treatment of oral candidiasis.

Fast releasing oral electrospun PVP/CD nanofiber mats of taste-masked meloxicam.

Fast release and taste masking of meloxicam (MX)-loaded polyvinylpyrrolidone (PVP)/cyclodextrin (CD) nanofiber mats were developed using an electrospinning process. CDs were blended to improve the stability of the mats. The morphology and diameter of the mats were determined using scanning electron microscopy (SEM); physical and mechanical properties were also studied. The MX content, disintegration time, MX release and cytotoxicity of the mats were investigated. In vivo studies were also performed in healthy human volunteers. The results indicated that the mats were successfully prepared with fiber in the nanometer range. MX was well incorporated into the mats, with an amorphous form. The mats showed suitable tensile strength. CDs improved the physical stability by their cage-like supramolecular structure to protect from humidity and moisture, and create bead free nanofiber mats. The nanofiber mats with CDs were physically stable without any hygroscopicity and fusion. A fast disintegration and release of MX was achieved. Moreover, this mat released MX faster than the MX powder and commercial tablets. The cytotoxicity test revealed that mats were safe for a 5-min incubation. The disintegration studies indicated that in vivo disintegration agreed with the in vitro studies; the mat rapidly disintegrated in the mouth. The less bitter of MX was occurred in the mats that incorporated CD, menthol and aspartame. In addition, this mat was physical stable for 6 months. The results suggest that these mats may be a good candidate for fast dissolving drug delivery systems of bitter drugs to increase the palatability of dosage forms.

Fabrication and In Vitro/In Vivo Performance of Mucoadhesive Electrospun Nanofiber Mats Containing α-Mangostin.

This study aimed to fabricate mucoadhesive electrospun nanofiber mats containing α-mangostin for the maintenance of oral hygiene and reduction of the bacterial growth that causes dental caries. Synthesized thiolated chitosan (CS-SH) blended with polyvinyl alcohol (PVA) was selected as the mucoadhesive polymer. α-Mangostin was incorporated into the CS-SH/PVA solution and electrospun to obtain nanofiber mats. Scanning electron microscopy, differential scanning calorimetry, X-ray diffraction, and tensile strength testing were used to characterize the mats. The swelling degree and mucoadhesion were also determined. The nanofiber mats were further evaluated regarding their α-mangostin content, in vitro α-mangostin release, antibacterial activity, cytotoxicity, in vivo performance, and stability. The results indicated that the mats were in the nanometer range. The α-mangostin was well incorporated into the mats, with an amorphous form. The mats showed suitable tensile strength, swelling, and mucoadhesive properties. The loading capacity increased when the initial amount of α-mangostin was increased. Rapid release of α-mangostin from the mats was achieved. Additionally, a fast bacterial killing rate occurred at the lowest concentration of nanofiber mats when α-mangostin was added to the mats. The mats were less cytotoxic after use for 72 h. Moreover, in vivo testing indicated that the mats could reduce the number of oral bacteria, with a good mouth feel. The mats maintained the amount of α-mangostin for 6 months. The results suggest that α-mangostin-loaded mucoadhesive electrospun nanofiber mats may be a promising material for oral care and the prevention of dental caries.

Inhibitory Effect of Alpha-Mangostin on Adhesion of Candida albicans to Denture Acrylic.

OBJECTIVE: Candida-associated denture stomatitis is a very common disease affecting denture wearers. It is characterized by the presence of yeast biofilm on the denture, primarily associated with C. albicans. The investigation of agents that can reduce C. albicans adhesion may represent a significant advancement in the prevention and treatment of this disease. This study aims to investigate the effect of alpha-mangostin on the in vitro adhesion of C. albicans to denture acrylic and germ tube formation by C. albicans and to compare its activity with clotrimazole which is a topical antifungal agent commonly used for the treatment of Candida-associated denture stomatitis. MATERIALS AND METHODOLOGY: Alpha-mangostin was extracted by thin layer chromatography. The effect of alpha-mangostin on adhesion of C. albicans to denture acrylic was determined by using a colorimetric tetrazolium assay and germ tube formation by C. albicans was determined by using the counting chamber. RESULTS: A significant reduction of C. albicans adhesion to denture acrylic was evident after exposure to 2,000 µg/ml of alpha-mangostin for only 15 min. In addition, the 2,000 µg/ml of the alpha-mangostin-treated C. albicans had a reduced ability for germ tube formation. These inhibitory effects of alpha-mangostin were as effective as clotrimazole. CONCLUSION: Alpha-mangostin has antifungal property against C. albicans by inhibiting the adhesion to denture acrylic and germ tube formation in vitro. These results suggest the potential application of alpha-mangostin as a topical medication or a natural oral hygiene product for treatment of Candida-associated denture stomatitis.

Pyoderma gangrenosum-like oral ulcerations in an elderly patient.

OBJECTIVE: To present a case of pyoderma gangrenosum (PG)-like oral ulcerations in an elderly patient. BACKGROUND: PG is an uncommon idiopathic, ulcerative, chronic inflammatory cutaneous disorder of unknown etiology, which is associated with systemic diseases found in more than 50% of patients. Oral lesions of PG are extremely rare and have not been previously reported on chronic leukemia patient. CLINICAL REPORT: This report presents the first case of a 73 year-old man who had PG-like oral ulcerations which offered the possibility of an initial finding of chronic myeloid leukemia. CONCLUSION: Clinicians should always take into consideration that PG in the oral mucosa is a recalcitrant ulcer and can precede the development of underlying clonal malignancy.

Fabrication of a novel scaffold of clotrimazole-microemulsion-containing nanofibers using an electrospinning process for oral candidiasis applications.

Clotrimazole (CZ)-loaded microemulsion-containing nanofiber mats were developed as an alternative for oral candidiasis applications. The microemulsion was composed of oleic acid (O), Tween 80 (T80), and a co-surfactant such as benzyl alcohol (BzOH), ethyl alcohol (EtOH) or isopropyl alcohol (IPA). The nanofiber mats were obtained by electrospinning a blended solution of a CZ-loaded microemulsion and a mixed polymer solution of 2% (w/v) chitosan (CS) and 10% (w/v) polyvinyl alcohol (PVA) at a weight ratio of 30:70. The nanofiber mats were characterized using various analytical techniques. The entrapment efficiency, drug release, antifungal activity and cytotoxicity were investigated. The average diameter of the nanofiber mats was in the range of 105.91-125.56 nm. The differential scanning calorimetry (DSC) and powder X-ray diffractometry (PXRD) results revealed the amorphous state of the CZ-loaded microemulsions incorporated into the nanofiber mats. The entrapment efficiency of CZ in the mats was approximately 72.58-98.10%, depended on the microemulsion formulation. The release experiment demonstrated different CZ release characteristics from nanofiber mats prepared using different CZ-loaded microemulsions. The extent of drug release from the fiber mats at 4h was approximately 64.81-74.15%. The release kinetics appeared to follow Higuchi's model. In comparison with CZ lozenges (10mg), the nanofiber mats exhibited more rapid killing activity. Moreover, the nanofiber mats demonstrated desirable mucoadhesive properties and were safe for 2h. Therefore, the nanofiber mats have the potential to be promising candidates for oral candidiasis applications.

Mucoadhesive electrospun chitosan-based nanofibre mats for dental caries prevention.

The mucoadhesive electrospun nanofibre mats were developed using chitosan (CS) and thiolated chitosan (CS-SH) as mucoadhesive polymers. Garcinia mangostana (GM) extract was incorporated into nanofibre mats. The antibacterial activity in the single and combined agents was evaluated against dental caries pathogens. The morphology of mats was observed using SEM. The mats were evaluated for GM extract amount, mucoadhesion, in vitro release, antibacterial activity and cytotoxicity. The mucoadhesion and antibacterial activity were determined in healthy human volunteers. The prepared mats were in nanoscale with good physical and mucoadhesive properties. The CS-SH caused the higher mucoadhesion. All mats rapidly released active substances, which had the synergistic antibacterial activity. In addition, the reduction of bacteria and good mucoadhesion in the oral cavity occurred without cytotoxicity. The results suggest that mats have the potential to be mucoadhesive dosage forms to maintain oral hygiene by reducing the bacterial growth that causes the dental caries.

Alpha-mangostin suppresses IL-6 and IL-8 expression in P. gingivalis LPS-stimulated human gingival fibroblasts.

This study aims to investigate the effect of alpha-mangostin on interleukin (IL)-6 and IL-8 expression in human gingival fibroblasts (HGFs). HGFs were challenged with Porphyromonas gingivalis LPS and then treated with various concentrations of alpha-mangostin. The cytotoxicity was determined using MTS assay and cytokine expressions were evaluated by Real-time PCR and ELISA. The results showed that 5 µg/ml P. gingivalis LPS and alpha-mangostin at 1 µg/ml or less did not affect the viability of HGFs. Alpha-mangostin reduced IL-6 and IL-8 mRNA and protein in P. gingivalis LPS-stimulated HGFs. These findings suggested that alpha-mangostin might be used as an adjunct to the periodontal therapy.

Fast-acting clotrimazole composited PVP/HPßCD nanofibers for oral candidiasis application.

PURPOSE: This study investigates fabrication of clotrimazole (CZ)-composited electrospun Polyvinylpyrrolidone/Hydroxypropyl-ß-cyclodextrin (PVP/HPßCD) blended nanofiber mats for oral candidiasis applications. METHODS: PVP/HPßCD blended nanofiber mats containing clotrimazole were electrospun and characterized using SEM, DSC and XRPD. The solvent system ethanol: water: benzyl alcohol (EtOH:H2O:BzOH) with a 70:20:10 ratio was optimal for the electrospinning process. Various amounts of CZ were loaded into the nanofiber mats. The nanofiber mats was further investigated for drug release, antifungal activity and cytotoxicity. RESULTS: The fiber diameters in the mats were in the nanometer range. The DSC and XRPD revealed a molecular dispersion of amorphous CZ in the nanofiber mats. The loading capacity increased when CZ content was raised. A fast dissolved and released of CZ from the nanofibers mat was achieved. The ability of the CZ-loaded nanofiber mats to kill the Candida depended on the amount of CZ in the mats; moreover, the CZ-loaded nanofibers killed the Candida significantly faster than the CZ powder and lozenges with low cytotoxicity. CONCLUSIONS: CZ-loaded nanofiber mats were successfully electrospun. They exhibited rapid antifungal activity in vitro relative to CZ powder and lozenges. Further in vivo studies are needed to investigate for their application in oral candidiasis.

Alpha-mangostin suppresses MMP-2 and MMP-9 expression in head and neck squamous carcinoma cells.

The purpose of this study was to investigate the effect of alpha-mangostin on matrix metalloproteinase (MMP)-2 and MMP-9 expression in head and neck squamous cell carcinoma (HNSCC). The human HNSCC cell lines were treated with alpha-mangostin and the cytotoxicity of alpha-mangostin in HNSCC was determined using the MTS assay. To determine the effect of alpha-mangostin on the expression of MMP-2 and MMP-9 in HNSCC, gelatin zymography and RT-PCR were performed. The results showed that alpha-mangostin increased in growth inhibition of HNSCC cell lines in a concentration-dependent manner. Treatment with alpha-mangostin decreased MMP-2 and MMP-9 expression in a concentration-dependent manner in all cell lines. These findings suggested that alpha-mangostin might be a potential therapeutic agent for HNSCC.

Antifungal activity of coronarin D against Candida albicans.

OBJECTIVE: The objective of this study was to investigate the antifungal activity of coronarin D on Candida albicans and its activity was compared with clotrimazole and nystatin. METHODS: Coronarin D was extracted by liquid chromatography and used in antifungal testing. The inhibitory effect of coronarin D on C. albicans was determined by cultures and an applied broth dilution test. The rate of fungicidal activity was evaluated by time-kill curves. Morphologic alterations of fungal cells were investigated using scanning electron microscopy. RESULTS: Coronarin D was effective against C. albicans; the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) were 2 and 4 mg/mL, respectively. The C. albicans killing activity of coronarin D was higher than clotrimazole and nystatin at 2 × MFC and 4 × MFC, respectively. Morphologic alterations of fungal cells consistent with cell membrane damage were observed in the coronarin D-treated cells. CONCLUSIONS: Coronarin D showed promising antifungal activity against C. albicans in vitro.

Apoptotic effect of alpha-mangostin on head and neck squamous carcinoma cells.

OBJECTIVE: The purposes of this study were to measure the cytotoxic effect of alpha-mangostin on head and neck squamous cell carcinoma (HNSCC) cell lines, to evaluate the apoptotic aspect of dead cells, and to identify the molecular mechanism involved in apoptosis. METHODS: The human HNSCC cell lines HN-22, HN-30 and HN-31 were treated with alpha-mangostin. The apoptotic effects of alpha-mangostin on HNSCC cells were determined by observation the morphological changes of cells, immunofluorescence for single-stranded DNA (ssDNA), and DNA fragmentation analysis. The expression of bax, bcl-2, and p53 were detected by RT-PCR and Western blot analysis. RESULTS: Alpha-mangostin showed excellent apoptotic effects on HNSCC cell lines, which induced the down-regulation of bcl-2, but up-regulation of bax and p53 in HN-22, HN-30 and HN-31. CONCLUSION: The present study suggests that the induction of apoptosis by alpha-mangostin seemed to be modulated by bcl-2, bax and p53 level in HNSCC cell lines.