Sex estimation of Malaysian sub-adults using craniometrics: A computed tomography study.

Sex estimation is crucial in biological profiling of skeletal human remains. Methods used for sex estimation in adults are less effective for sub-adults due to varied cranium patterns during the growth period. Hence, this study aimed to develop a sex estimation model for Malaysian sub-adults using craniometric measurements obtained through multi-slice computed tomography (MSCT). A total of 521 cranial MSCT dataset of sub-adult Malaysians (279 males, 242 females; 0-20 years old) were collected. Mimics software version 21.0 (Materialise, Leuven, Belgium) was used to construct three-dimensional (3D) models. A plane-to-plane (PTP) protocol was utilised to measure 14 selected craniometric parameters. Discriminant function analysis (DFA) and binary logistic regression (BLR) were used to statistically analyze the data. In this study, low level of sexual dimorphism was observed in cranium below 6 years old. The level was then increased with age. For sample validation data, the accuracy of DFA and BLR in estimating sex improved with age from 61.6% to 90.3%. All age groups except 0-2 and 3-6 showed high accuracy percentage (≥75%) when tested using DFA and BLR. DFA and BLR can be utilised to estimate sex for Malaysian sub-adult using MSCT craniometric measurements. However, BLR showed higher accuracy than DFA in sex estimation of sub-adults.

Is microbial adhesion affected by the build orientation of a 3-dimensionally printed denture base resin?

STATEMENT OF PROBLEM: How the build orientation of a 3-dimensionally (3D) printed denture affects microbial adhesion is unclear. PURPOSE: The purpose of this in vitro study was to compare the adherence of Streptococcus spp. and Candida spp. on 3D-printed denture bases prepared at different build orientations with conventional heat-polymerized resin. MATERIAL AND METHODS: Resin specimens (n=5) with standardized 28.3 mm2 surface area were 3D printed at 0 and 60 degrees, and heat-polymerized (3DP-0, 3DP-60, and HP, respectively). The specimens were placed in a Nordini artificial mouth (NAM) model and exposed to 2 mL of clarified whole saliva to create a pellicle-coated substratum. Suspensions of Streptococcus mitis and Streptococcus sanguinis, Candida albicans and Candida glabrata, and a mixed species, each at 108 cfu/mL were pumped separately into the model for 24 hours to promote microbial adhesion. The resin specimens were then removed, placed in fresh media, and sonicated to dislodge attached microbes. Each suspension (100 µL) was aliquoted and spread on agar plates for colony counting. The resin specimens were also examined under a scanning electron microscope. The interaction between types of specimen and groups of microbes was examined with 2-way ANOVA and then further analysis with Tukey honest significant test and Kruskal-Wallis post hoc tests (α=.05). RESULTS: A significant interaction was observed between the 3DP-0, 3DP-60, and HP specimen types and the groups of microbes adhering to the corresponding denture resin specimens (P<.05). The difference was statistically significant among 3DP-0, 3DP-60, and HP specimens (P<.05). The adherence of candida was 3.98-times lower on the 3DP-0 than that of HP (P<.05). However, adherence of the mixed-species microbes and streptococci on the 3DP-60 were 1.75 times and 2-fold higher, respectively (P<.05). The scanning electron micrographs showed that 3DP-0 exhibited the lowest microbial adherence compared with HP and 3DP-60. CONCLUSIONS: Adherence affinity of denture base resin is affected by the build orientation rather than by the group of different microbes. Three-dimensionally printed denture base resin fabricated at a 0-degree build orientation exhibited low affinity for microbial adhesion. Three-dimensionally printed dentures may reduce microbial adhesion when printed at a 0-degree build orientation.

The efficacy of okra fruit extract on the expression of transforming growth factor beta 1 in the tooth socket of diabetic Wistar rats.

BACKGROUND: Patients with diabetes mellitus suffer from an additional macrophage dysfunction in the secretion of growth factor, which later decreases transforming growth factor beta 1 (TGF-ß1). This condition disrupts proliferation and angiogenesis. Extract of okra fruit (Abelmoschus esculentus) contains flavonoid, an active substance which acts as antioxidant, anti-inflammation, and antidiabetes. The purpose of this study is to analyze the difference in TGF-ß1 expression in wound-healing process after tooth extraction of diabetic Wistar rats. MATERIALS AND METHODS: This is a laboratory experimental study using pretest and posttest on 24 Wistar rats which are divided into two groups: control group (treated with streptozotocin induction but without administration of okra fruit extract) and treatment group (treated with streptozotocin induction and oral administration of 250 mg/kg okra fruit extract once a day). Extractions of the rats' mandibular left incisors were performed using a pair of modified forceps and an elevator. The tooth sockets were then irrigated using saline solution. Four rats in each group were sacrificed on day 3 (KO1, PO1), 5 (KO2, PO2), and 7 (KO3, PO3). The socket tissues from the rats were then immunohistochemically analyzed. Data were analyzed at level significance of 0.05. RESULTS: The average level of TGF-ß1 expression in the treatment groups was higher compared to the control group: PO1 (11.59 ± 0.58), PO2 (15.15 ± 1.07), and PO3 (18.75 ± 2.73) as compared to KO1 (5.32 ± 1.69), KO2 (8.47 ± 0.60), and KO3 (9.28 ± 1.16) with P = 0.001. CONCLUSION: The administration of okra fruit extract can increase the level of TGF-ß1 in wounds after tooth extraction of diabetic Wistar rats.

Impact of non-surgical periodontal therapy on serum Resistin and periodontal pathogen in periodontitis patients with obesity.

BACKGROUND: Non-surgical periodontal therapy (NSPT) known as gold standard treatment in managing periodontitis. The aim of this study was to investigate the response of NSPT in periodontitis subjects who were obese. Clinical parameters of periodontitis, changes in serum resistin and periodontal pathogens in subgingival plaque were compared before and after NSPT in periodontitis subjects who were obese and with normal weight. METHODS: A total of 48 periodontitis subjects (obese, n = 18; normal weight, n = 30) were recruited (hereafter will be referred as participants) to participate into a prospective, before and after clinical trial. Obesity status is defined by body mass index (BMI) criteria (obese: ≥30 kg/ m2; normal weight < 25 kg/m2). Visible Plaque Index (VPI), Gingival Bleeding Index (GBI), Probing Pocket Depth (PPD) and Clinical Attachment Loss (CAL) were recorded; and serum and plaque were collected at baseline and following 12 weeks post-NSPT. Serum resistin level was analyzed using enzyme-linked immune-sorbant assay (ELISA), while detection of periodontal pathogens in dental plaque were carried out using real time PCR (qPCR). RESULTS: Following NSPT, means VPI and GBI showed significant improvement between obese and normal weight groups (p <  0.05), but no difference in means PPD and CAL was observed between groups. Obesity remained as a predictor for VPI and GBI after adjusting for smoking habit. No significant difference was observed in serum resistin level and mean counts for P. gingivalis, T. forsythia, and P. intermedia between obese and normal weight groups following NSPT. CONCLUSIONS: Regardless of obesity status, NSPT has a significant impact on VPI and GBI in periodontitis subjects. However, the impact of NSPT towards serum resistin and periodontal pathogens was non-significant in those with periodontitis. TRIAL REGISTRATION: This study followed the Consolidation Standards of Reporting Trials Statement and retrospectively registered on 26/11/2015 at clinicaltrials.gov (No. NCT02618486).

Oral Health Interventions Using Chlorhexidine-Effects on the Prevalence of Oral Opportunistic Pathogens in Stroke Survivors: A Randomized Clinical Trial.

OBJECTIVE: To evaluate the presence of oral opportunistic pathogens among stroke survivors, both before and after oral health care interventions. METHODS: A multicenter randomized clinical trial was conducted on hospitalized stroke survivors. Those in the control group were given standard care of oral hygiene (a manual toothbrush and toothpaste), whereas those in the test group were given intense care of oral hygiene (a powered toothbrush and 1% chlorhexidine oral gel). Oral clinical assessments were carried out, and microbiological samples were collected, using concentrated oral rinse samples at 3 time points: baseline, 3 months, and 6 months. RESULTS: The prevalence of oral yeast was significantly reduced in the test group at 6 months (P < .05), but no significant difference was observed over time. A significant reduction was observed in the prevalence of Staphylococcus aureus (P < .01) and aerobic and facultative gram-negative bacilli over time (P < .05), but there were no significant differences noted between groups at 6 months. Candida albicans and Klebsiella pneumoniae were the prominent pathogens determined throughout the trial. Kluyvera strains have also been isolated from this cohort. CONCLUSION: Oral hygiene intervention using a powered tooth brush and 1% chlorhexidine oral gel was effective in reducing the prevalence of oral opportunistic pathogens.

Vitamin E (α-Tocopherol) Exhibits Antitumour Activity on Oral Squamous Carcinoma Cells ORL-48.

Cancers involving the oral cavity, head, and neck regions are often treated with cisplatin. In cancer therapy, the main target is to eliminate unwanted cancerous cells. However, reports on the nonselective nature of this drug have raised few concerns. Incorrect nutritional habits and lifestyle practices have been directly linked to cancer incidence. Nutrients with antioxidant activity inhibit cancer cells development, destroying them through oxidative stress and apoptosis. α-tocopherol, the potent antioxidant form of vitamin E is a known scavenger of free radicals. In vitro study exhibited effective antitumor activity of α-tocopherol on ORL-48 at 2.5 ± 0.42 µg/mL. Cisplatin exhibited stronger activity at 1.0 ± 0.15 µg/mL, but unlike α-tocopherol it exhibited cytotoxicity on normal human epidermal keratinocytes at very low concentration (<0.1 µg/mL). Despite the lower potency of α-tocopherol, signs of apoptosis such as the shrinkage of cells and appearance of apoptotic bodies were observed much earlier than cisplatin in time lapse microscopy. No apoptotic vesicles were formed with cisplatin, instead an increased population of cells in the holoclone form which may suggest different induction mechanisms between both agents. High accumulation of cells in the G0/G1 phase were observed through TUNEL and annexin V-biotin assays, while the exhibition of ultrastructural changes of the cellular structures verified the apoptotic mode of cell death by both agents. Both cisplatin and α-tocopherol displayed cell cycle arrest at the Sub G0 phase. α-tocopherol thus, showed potential as an antitumour agent for the treatment of oral cancer and merits further research.

Alternative sweeteners influence the biomass of oral biofilm.

OBJECTIVE: Compact-structured oral biofilm accumulates acids that upon prolonged exposure to tooth surface, causes demineralisation of enamel. This study aimed to assess the effect of alternative sweeteners Equal Stevia®, Tropicana Slim®, Pal Sweet® and xylitol on the matrix-forming activity of plaque biofilm at both the early and established stages of formation. METHODS: Saliva-coated glass beads (sGB) were used as substratum for the adhesion of a mixed-bacterial suspension of Streptococcus mutans, Streptococcus sanguinis and Streptococcus mitis. Biofilms formed on sGB at 3h and 24h represented the early and established-plaque models. The biofilms were exposed to three doses of the sweeteners (10%), introduced at three intervals to simulate the exposure of dental plaque to sugar during three consecutive food intakes. The treated sGB were (i) examined under the SEM and (ii) collected for turbidity reading. The absorbance indicated the amount of plaque mass produced. Analysis was performed comparative to sucrose as control. RESULTS: Higher rate of bacterial adherence was determined during the early compared to established phases of formation. Comparative to the sweeteners, sucrose showed a 40% increase in bacterial adherence and produced 70% more plaque-mass. Bacterial counts and SEM micrographs exhibited absence of matrix in all the sweetener-treated biofilms at the early phase of formation. At the established phase, presence of matrix was detected but at significantly lower degree compared to sucrose (p<0.05). CONCLUSION: Alternatives sweeteners promoted the formation of oral biofilm with lighter mass and lower bacterial adherence. Hence, suggesting alternative sweeteners as potential antiplaque agents.

Assessment of Antifungal Activity of Bakuchiol on Oral-Associated Candida spp.

Bakuchiol is an active component of Psoralea glandulosa and Psoralea corylifolia, used in traditional Chinese medicine. The study aimed at investigating the antifungal activity of bakuchiol on planktonic and biofilm forms of orally associated Candida species. The antifungal susceptibility testing was determined by the broth micro dilution technique. Growth kinetics and cell surface hydrophobicity (CSH) of Candida were measured to assess the inhibitory effect of bakuchiol on Candida planktonic cells. Biofilm biomass and cellular metabolic activity were quantitatively estimated by the crystal violet (CV) and the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) assays. All Candida strains have been shown to be susceptible to bakuchiol with the MIC ranges from 12.5 to 100 µg/mL. Significant decrease in specific growth rates and viable counts demonstrates the inhibitory effect of bakuchiol on Candida planktonic cells. A brief exposure to bakuchiol also reduced CSH of Candida (P < 0.05), indicating altered surface properties of yeast cells towards hydrophobic interfaces. Biofilm biomass and cell metabolic activity were mostly decreased, except for C. glabrata (P = 0.29). The antifungal properties of bakuchiol on Candida species in this in vitro study may give insights into the application in therapeutic strategy against Candida infections.

Evaluation of Wound Closure Activity of Nigella sativa, Melastoma malabathricum, Pluchea indica, and Piper sarmentosum Extracts on Scratched Monolayer of Human Gingival Fibroblasts.

Nigella sativa, Melastoma malabathricum, Pluchea indica, and Piper sarmentosum are common Asian traditional medicines to treat minor wounds. This study aimed to investigate the in vitro wound healing properties of aqueous extracts of these plants using human gingival fibroblast (HGF) monolayer as study model. DPPH scavenging activity of the extracts was evaluated and effect on HGF proliferation was determined. Their effect on HGF's function to synthesize collagen was indicated by the level of hydroxyproline produced and effect on wound healing activity was assessed using an in vitro scratch assay. The influence of the extracts on expression of bFGF and TGF-ß was also determined. Results revealed all four extracts to exhibit low free radical scavenging activity. The extract from N. sativa (NSSE) compared to the others showed favourable enhancement of HGF proliferation with EC50 of 22.67 ± 3.06 µg/mL (P < 0.05) with accelerated wound closure activity despite its nonsignificant effect on collagen synthesis. In addition to the elevated level of bFGF by up to 15% at 100 µg/mL of NSSE, a slightly better effect was observed on the expression of TGF-ß. NSSE thus showed that promising wound healing properties and data obtained may contribute towards validation of its traditional use for the healing of oral wounds.

Is synthetic mouthwash the final choice to treat oral malodour?

The aim of the review was to critically appraise the various pros and cons of the synthetic and herbal agents used in mouthwashes against halitosis and facilitate users to choose appropriate mouthwashes according to their need. Oral Malodour (OMO) or halitosis is a global epidemic with social and psychological impact. Use of mouthwash has been adopted worldwide to control halitosis within a past few decades. Alcohol and Chlorhexidine are common agents in synthetic mouthwashes, while Tannins and Eugenol are derived traditional herbal extracts. Each agent signifies some unique properties distinguishing them from others. Herbal ingredients are gaining the attention of the profession due to its mild side effects and competitive results. Herbal mouthwashes can be a safer choice in combating OMO, as an alternate to synthetic mouthwashes.

Growth inhibitory response and ultrastructural modification of oral-associated candidal reference strains (ATCC) by Piper betle L. extract.

Candida species have been associated with the emergence of strains resistant to selected antifungal agents. Plant products have been used traditionally as alternative medicine to ease mucosal fungal infections. This study aimed to investigate the effects of Piper betle extract on the growth profile and the ultrastructure of commonly isolated oral candidal cells. The major component of P. betle was identified using liquid chromatography-mass spectrophotometry (LC-MS/MS). Seven ATCC control strains of Candida species were cultured in yeast peptone dextrose broth under four different growth environments: (i) in the absence of P. betle extract; and in the presence of P. betle extract at respective concentrations of (ii) 1 mg⋅mL(-1); (iii) 3 mg⋅mL(-1); and (iv) 6 mg⋅mL(-1). The growth inhibitory responses of the candidal cells were determined based on changes in the specific growth rates (µ). Scanning electron microscopy (SEM) was used to observe any ultrastructural alterations in the candida colonies. LC-MS/MS was performed to validate the presence of bioactive compounds in the extract. Following treatment, it was observed that the µ-values of the treated cells were significantly different than those of the untreated cells (P<0.05), indicating the fungistatic properties of the P. betle extract. The candidal population was also reduced from an average of 13.44×10(6) to 1.78×10(6) viable cell counts (CFU)⋅mL(-1). SEM examination exhibited physical damage and considerable morphological alterations of the treated cells. The compound profile from LC-MS/MS indicated the presence of hydroxybenzoic acid, chavibetol and hydroxychavicol in P. betle extract. The effects of P. betle on candida cells could potentiate its antifungal activity.

Antifungal susceptibility and growth inhibitory response of oral Candida species to Brucea javanica Linn. extract.

BACKGROUND: Candida species have been associated with the emergence of resistant strains towards selected antifungal agents. Plant products have been used traditionally as alternative medicine to ease candidal infections. The present study was undertaken to investigate the antifungal susceptibility patterns and growth inhibiting effect of Brucea javanica seeds extract against Candida species. METHODS: A total of seven Candida strains that includes Candida albicans ATCC14053, Candida dubliniensis ATCCMYA-2975, Candida glabrata ATCC90030, Candida krusei ATCC14243, Candida lusitaniae ATCC64125, Candida parapsilosis ATCC22019 and Candida tropicalis ATCC13803 were used in this study. The antifungal activity, minimum inhibitory concentration and minimum fungicidal concentration of B. javanica extract were evaluated. Each strain was cultured in Yeast Peptone Dextrose broth under four different growth environments; (i) in the absence and presence of B. javanica extract at respective concentrations of (ii) 1 mg/ml (iii) 3 mg/ml and (iv) 6 mg/ml. The growth inhibitory responses of the candidal cells were determined based on changes in the specific-growth rates (µ) and doubling time (g). The values in the presence of extract were computed as percentage in the optical density relative to that of the total cells suspension in the absence of extract. RESULTS: B. javanica seeds extract exhibited antifungal properties. C. tropicalis showed the highest growth rate; 0.319 ± 0.002 h(-1), while others were in the range of 0.141 ± 0.001 to 0.265 ± 0.005 h(-1). In the presence of extract, the lag and log phases were extended and deviated the µ- and g-values. B. javanica extract had significantly reduced the µ-values of C. dubliniensis, C. krusei and C. parapsilosis at more than 80% (ρ < 0.05), while others were reduced within the range of 2.28% to 57.05%. The g-values of most candidal strains were extended and significantly reduced (ρ < 0.05) in relative to the untreated. The candidal population was reduced from an average of 10 x 106 to 6 x 106 CFU/ml. CONCLUSIONS: B. javanica extract exhibited in vitro antifungal activity against seven oral Candida species. The fungistatic and growth inhibiting effects of B. javanica extract have shown that it has potential to be considered as a promising candidate for the development of antifungal agent in oral health products.

An in vitro study on the anti-adherence effect of Brucea javanica and Piper betle extracts towards oral Candida.

OBJECTIVE: The adherence of Candida to mucosal surfaces is the initial step for successful invasive process of the oral cavity. The study aimed to investigate the effect of two plant extracts on the non-specific and specific bindings of oral candida. METHODS: In the former, adsorption to hexadecane was used to measure the hydrophobic interaction of the candida cells. In the later, glass beads coated with saliva represented the experimental pellicles in specific adhesion of oral candida to hard tissue surface. RESULTS: Candida krusei, Candida dubliniensis and Candida tropicalis showed the highest adsorption to hexadecane at 30.23%, 26.19% and 19.70%, respectively, while the others within the range of 7-10%. All candidal species were significantly affected by the extracts (P<0.05) with Brucea javanica exhibited more than 60% reduction of CSH than Piper betle. Candida parapsilosis showed the highest affinity in specific-bindings to pellicle with 18.72±0.71×10(5)CFU/ml. Exposing to P. betle-treated pellicle has drastically reduced the adherence of C. tropicalis, Candida albicans and C. krusei by 86.01%, 61.41% and 56.34%, respectively. B. javanica exhibited similar effect on C. tropicalis (89.86%), Candida lusitaniae (88.95%), C. albicans (79.74%), Candida glabrata (76.85%) and C. krusei (67.61%). CONCLUSION: The extracts demonstrated anti-adherence activities by modifying the CSH and the characteristics of the experimental pellicle.

Effect of phenotypic switching on the biological properties and susceptibility to chlorhexidine in Candida krusei ATCC 14243.

Phenotypic switching is characterized as a virulence factor of Candida spp. This study was carried out to evaluate the phenotypic switching ability of C. krusei ATCC 14243 and to determine its effect on the biological properties, adherence capacity and susceptibility towards chlorhexidine digluconate (CHX). To induce switched generations C. krusei was cultured under nitrogen-depleted growth conditions by adding phloxine B. These phenotypically switched colonies were designated as the 1st generation. Subsequent sub-culturing was performed to produce the 2nd, 3rd and 4th switched generations. The recovery of the 3rd generation was the highest at 85.7% while that of the 4th generation was lower at 70.8%, and the recovery of the 1st and 2nd generations gradually reduced to 46.6% and 36.4%, respectively. All generations of C. krusei were susceptible towards CHX. The unswitched C. krusei was the most susceptible but the least adherent to coated hard surfaces. The 2nd generation was the least susceptible, but with the highest adherent ability. The minimum inhibition concentration and minimal fungicidal concentration of C. krusei of all generations were determined at 0.4 mg mL(-1) . These observations suggest that the switching activity of C. krusei induces changes to its biological properties and susceptibility towards CHX.