Unique role for glycogen synthase kinase 3 beta (GSK3B) in transforming growth factor beta1 (TGFB1) regulation of connective tissue growth factor (CCN2CTGF) in human gingival fibroblasts

PLEASE NOTE: This work is protected by copyright. Downloading is restricted to the BU community: please click Download and log in with a valid BU account to access. If you are the author of this work and would like to make it publicly available, please contact [email protected] (DScD) --Boston University, Henry M. Goldman School of Dental Medicine, 2009 (Department of Periodontology and Oral Biology).Includes bibliography: leaves 148-182

Evaluation and Comparison of Genomic DNA Extraction Methods and PCR Optimization on Archival Formalin-Fixed and Paraffin-Embedded Tissues of Oral Squamous Cell Carcinoma

Recovery and amplification of nucleic acids from archived formalin-fixed tissue samples is the most developing field in retrospective genetic studies. We compared different deparaffinization methods and DNA isolation techniques, and intergroup comparisons were performed to evaluate the effectiveness of different storing methods for archival OSCC samples based on obtained mean DNA quantity, quality, and PCR amplification of the P53 gene. The study comprised 75 archival histologically diagnosed OSCC samples which were divided into Group I: Formalin-fixed paraffin-embedded tissue blocks and Group II: Long-term formalin-fixed tissue. A comparison of different deparaffinization methods showed that xylene deparaffinization is an efficient method to obtain suitable DNA. Comparing different DNA isolation techniques illustrated that the conventional phenol–chloroform method gives better integrity to DNA in contrast with the kit method. Comparison between FFPET and long-term FFT samples demonstrated that samples fixed in formalin overnight and embedded in wax yield better quality and quantity DNA in comparison with long-term samples fixed in formalin. To obtain suitable integrity of DNA, tissue samples should be stored by fixing in formalin overnight followed by preparation of paraffin tissue blocks, deparaffinization by xylene, and subjecting them to the conventional phenol–chloroform DNA isolation protocol

Alternate Special Stains for the Detection of Mycotic Organisms in Oral Cyto-Smears—A Histomorphometric Study

In the wake of the COVID-19 pandemic, fungal infections of the maxillofacial region have become prevalent, making their accurate diagnosis vital. Histopathological staining remains a simple, cost-effective technique for differentiation and diagnosis of the causative fungal organisms. The present study aims to evaluate the staining efficacy of Periodic Acid-Schiff (PAS), Alcian Blue, Safranin-O and Gomori’s Methenamine Silver (GMS) on fungal smears. This research work also attempts to study the morphometric characteristics of Candida albicans, Aspergillus flavus, Rhizopus oryzae. Candida albicans, Aspergillus flavus and Rhizopus oryzae, 10 smears each, were stained using PAS, Alcian Blue, Safranin-O and GMS. The morphological characteristics and staining efficacy were examined, and semi-quantitative scoring was performed. Candida albicans, Aspergillus flavus and Rhizopus oryzae were stained for the first time with Safranin-O. The morphometric traits were then analyzed using an image analysis software. Safranin-O provided the most reliable staining efficacy amongst the stains and optimum morphological definition for all three organisms. Safranin-O was found to be superior to PAS and GMS, ensuring detection of even the most minute mycotic colonies. The hyphae of Aspergillus flavus to be the largest, and the spores and fruiting body of Rhizopus oryzae were found to be the largest amongst the three organisms compared. Early and accurate diagnosis of fungal infections can significantly reduce morbidity in orofacial fungal infections

Evaluating the Effect of Tideglusib-Loaded Bioactive Glass Nanoparticles as a Potential Dentine Regenerative Material

Dental pulp treatment is the least intrusive procedure currently available for preserving the vitality of the pulp. Several studies are underway to improve the bioactivity of pulp capping materials. Tideglusib isa potent anti-inflammatory, antioxidant, and a regenerative drug developed against Alzheimer’s disease and has been shown to be effective in the treatment of dental cavities. However, its bioactive properties encapsulated within the nanoparticles as a component of pulp capping material are largely unknown. In this study, tideglusib-loaded bioactive glass nanoparticles were synthesized (tideglusib-BgNPs) and mixed at various concentrations into the calcium silicate cement to testits physiomechanical and bioactivitiescompared with biodentine (control). The calcium silicate cement with 10wgt% tideglusib-BgNPs showed comparable physiomechanical properties to that of biodentine. Additionally, the assessment of cytotoxicity and bioactivity (cell proliferation, wound healing, and cell migration assays) showed increased bioactivity in terms of better wound healing, increased proliferation, and better migration of human dental pulp stem cells than biodentine. These findings suggest new opportunities to use tideglusib-BgNPs in pulp therapy

Red-Complex Bacterial Levels in Pregnant Women With Preeclampsia and Chronic Periodontitis

ABSTRACT: Objectives: The presence of red-complex bacteria (RCB) has been associated with the occurrence of not only periodontal disease but also other systemic diseases, thereby potentiating an oral–systemic relationship. This study aimed to explore the levels of periodontal RCB in pregnant preeclamptic women with periodontal disease, potentiating the pathophysiology of a bacterial association in both periodontitis and preeclampsia. Methods: Six hundred pregnant women (aged 20 to 35 years) in their third trimester were selected and divided into 3 groups comprising a control group (systemically and periodontally healthy pregnant women), group A (preeclamptic women with chronic periodontitis), and group B (preeclamptic women without chronic periodontitis). Demographic variables and periodontal parameters were recorded. Other variables such as systolic blood pressure (SBP) and diastolic blood pressure (DBP) and the urine protein content were also estimated. RCB, namely Porphyromonas gingivalis (Pg), Treponema denticola (Td), and Tenerella forsythia (Tf), were assessed in both subgingival plaque samples and placental samples of all the pregnant participants in the study. Results: The mean weight, mean body mass index, mean SBP, mean DBP, and mean urine protein content were significantly elevated in group A and group B when compared to the control group. Preeclamptic women with chronic periodontitis (group A) had higher levels of plaque index, probing pocket depth, bleeding on probing, and clinical attachment level as compared with the other groups. The numbers of the RCB were found to be elevated in group A, out of which Pg was found to be more prevalent. Conclusions: Our findings indicate that the periodontal indices, blood pressure, and urine protein content were found to be higher in preeclamptic women with periodontitis. Hence, reducing the periodontal bacterial load in pregnant women via good oral hygiene maintenance would further reduce the putative risk of adverse pregnancy outcomes including preeclampsia in these women

Evaluating the efficacy of Centella asiatica on enhancement of oral health status in hyperglycemic patients - A randomized clinical trial

Aim: The aim of the study was to evaluate the oral secretagogue, anti-microbial and immunomodulatory properties of Centella asiatica in patients with type II diabetes mellitus. Materials and methods: In-vivo and In- vitro assessment was conducted for a period of 3 months. A before and after trial for a period of 3 months of intervention was performed involving 20 participants. Unstimulated Saliva and gingival crevicular fluid (GCF) samples was collected from all the participants before and after the intervention period. The effectiveness of Centella asiatica was estimated by evaluating the pre and post intervention values of salivary flow rate, salivary pH, S.mutans count, GCF flow rate, neutrophils count and alpha 1 defensin levels. Results: This clinico-interventional study inferred that after the usage of aqueous extract of Centella asiatica mouth wash, there was a high statistically significant improvement in the GCF flow rate, GCF neutrophil count and alpha 1 defensin levels from baseline to 90thday (after 3 months) (P = 0.001). The salivary flow rate, salivary pH and S.mutans colony count showed no statistically significant difference. Conclusion: The outcome of the current study enlightens the utility of ethnomedicinal plants and their usage, it is also helpful in enhancement of oral health status in hyperglycemic conditions. The current study concluded that rinsing twice with aqueous extract of Centella asiatica as mouth rinse for a period of 3 months has an immunomodulatory action. Clinical significance: Centella asiatica mouthwash can be used as an immunomodulatory agent

The Efficacy of Low-Level Laser Therapy Combined with Single Flap Periodontal Surgery in the Management of Intrabony Periodontal Defects: A Randomized Controlled Trial

This study aimed at assessing the clinical outcomes of the Single Flap Approach (SFA) with the additional use of Low-level laser therapy (LLLT). The defects were treated as per the principles of SFA, whereby 20 defects received only SFA (control group) and 20 defects received additional LLLT for bio stimulation/bio modulation (test group). Stable primary closure of the flaps was obtained with vertical internal mattress sutures. Plaque indices (FMPS), clinical attachment levels (CAL), probing pocket depth (PPD), and gingival bleeding scores (FMBS) were calculated at baseline, and at the 3rd and 6th months in both groups. An EHI score of 1 was observed at all sites except for two, where a score of 2 in the control group at week 2 was observed. In the test group, the PPD reduction at 6 months was 3.60 ± 0.95 and in the control group it was 3.75 ± 0.91 mm. CAL gain at 6 months was 2.70 ± 1.36 mm and 3.45 ± 1.2 mm in the test group and showed no statistical significance. These data suggested the positive effect of LLLT over CAL gain; thus, LLLT may be combined with SFA to potentially enhance the early wound healing and higher clinical outcomes in terms of increase in CAL and decrease in PPD

Comparative Evaluation of Microleakage of Flowable Composite Resin Using Etch and Rinse, Self-Etch Adhesive Systems, and Self-Adhesive Flowable Composite Resin in Class V Cavities: Confocal Laser Microscopic Study

The essential factor in determining the preservation of restoration is the marginal seal. Restoring cervical lesions with a resin composite has always been a challenge. Composite resins with various viscosities and different bonding systems are being researched to reduce the microleakage. Confocal laser scanning microscopy (CLSM) is the latest non-destructive technique for visualizing the microleakage. Objectives: To evaluate and compare the microleakage of Universal Flo composite resin (G-aenial) using etch and rinse adhesive system ER-2 steps (Adper Single Bond 2), self-etch adhesive system SE-1 step (G-Bond), and self-adhesive flowable composite resin (Constic) in Class V cavities using a confocal laser scanning microscope. Materials and Method: Class V cavities were prepared on 27 caries-free human extracted premolar teeth on the buccal and lingual surfaces with standardized dimensions of 2 mm height, width 4 mm, and a depth of 2 mm. After the cavity preparation, all teeth were randomly divided into three groups, namely Group-I: G-aenial Universal Flo with Single Bond 2 (n = 9 teeth); Group-II: G- aenial Universal Flo with G-Bond (n = 9 teeth), and Group-III: Constic (n = 9 teeth). The prepared and restored specimens were then subjected to thermocycling for 500 cycles in a water bath at 5 °C and 55 °C with a dwelling time of 30 s. The specimens were placed in 0.6% aqueous rhodamine dye for 48 h. Sectioning was carried out bucco-lingually and specimens were evaluated for microleakage under a confocal laser scanning microscope. Results: There was a significant difference (p = 0.009) in microleakage when comparing total etch and rinse, specifically between Adper Single Bond 2 ER-2 steps (fifth generation) and self-adhesive flowable composite resin, which is Constic. There was more microleakage in the self-etch bonding agent, particularly G-Bond, SE-1 step (seventh generation), when compared to ER-2 steps (fifth generation bonding agent); however, the results were not statistically significant (p = 0.468). The self-adhesive flowable composite resin showed more microleakage than SE-1 step and ER-2 steps. Conclusions: None of the adhesive systems tested were free from microleakage. However, less microleakage was observed in the total etch and rinse, especially Adper Single Bond 2 (ER-2 steps), than the self-etch adhesive system SE-1 step and self-adhesive flowable composite resin. Clinical significance: Constant research and technological advancements are taking place in dentin adhesives to improve the marginal seal. This has led to the evolution of total acid-etching dentin bonding agents termed as etch and rinse (ER)-2 steps (fifth generation dentin bonding agents) and self-etching (SE) 2 steps, and SE-1 step dentin bonding agents termed as the sixth and seventh generation bonding agents, respectively

Role of Toothbrushes as Gene Expression Profiling Tool for Oral Cancer Screening in Tobacco and Alcohol Users

Aim: The use of toothbrushes was investigated as a potential RNA source and gene expression profiling tool for oral cancer screening in tobacco and alcohol users. Methodology: A total of 20 subjects were selected on the basis of inclusion and exclusion criteria. They were divided into two groups: group I—healthy controls (n = 6); group II—individuals who consume tobacco and alcohol (n = 14). After the volunteers brushed their teeth using a soft-bristle toothbrush with ~0.5 gm of toothpaste, the toothbrushes were collected, and the gene expression of BAX, BCL2, CDK4, CKDN2A, GNB3, and TCF7L2 was assessed. Results: The gene expression of BAX decreased significantly in alcoholics and smokers (0.13867 ± 0.12014), while the gene expression of BCL2 increased in alcoholics and smokers (1.91001 ± 0.90425) in comparison with healthy controls (p = 0.0054 and p = 0.0055). Although there was increased expression of CDK4, CKDN2A, and TCF7L2 and decreased expression of GNB3 in smokers and alcoholics, the results were not significant. Conclusions: A toothbrush is a good source of RNA, and gene expression analysis can be performed using the genetic material retrieved from toothbrushes, which can aid in the early diagnosis of oral squamous cell carcinoma among tobacco and alcohol users. Further studies with a larger sample size and different durations of toothbrush use should be conducted to explore the role of toothbrushes as a noninvasive tool for disease diagnosis

Transcriptional regulatory signatures of systemic diseases in periodontitis with dyslipidemia

Background: Periodontitis is a chronic bacterial infection of tooth that increases the risk of systemic diseases like diabetes, cancer, and cardiovascular diseases. Although it has been have found altered lipids in periodontitis patients, its gene regulation is largely unknown. This study aimed to examine the lipid meditated regulatory network in periodontitis that may helpful in early detection of periodontal mediated systemic diseases. Methods: We employed a high-throughput gene expression data of 1) patients with periodontitis (n = 6); 2) the periodontitis patients with dyslipidemia (n = 6); and 3) healthy control group (n = 6). The over represented (DEGs) genes in SET-A (control vs. periodontitis) and SET-B (control vs. periodontitis with dyslipidemia) was identified. The protein interaction network was generated for the over represented genes in both the conditions. The constructed network was dissected into multiple regulatory clusters, containing over expressed transcription factors with its interacting proteins. Further the behavior of the clusters was determined through gene ontology and molecular pathways. Results: On expression analysis, 751 in SET-A and 561 in SET-B were over expressed compared to healthy control. Using over expressed genes, protein interaction networks were constructed for SET-A and SET-B, respectively. Topological analysis revel the difference in the complexity of both the network. Four regulatory clusters (ESR1, FOS, RUNX2, and SP1) from SET-A and six (ESR1, ESR2, FOS, JDP2, PBX1, and TAL1) from the SET-B network was extracted. Each cluster displayed a variety of molecular mechanism associated with immune system, cell cycle, and signal processing. Clusters from SET-B showed diverse regulatory pattern in associated with cancer, neurological, psychiatric and metabolic diseases. Conclusion: Our findings demonstrate difference in regulatory patterns between periodontitis and periodontitis with dyslipidemia. These finding may provide evidence for dyslipidemia mediated periodontitis contribute to progressive systemic diseases. Further experiments are required to validate these regulators as biomarkers and drug targets