Melbourne Dental School - Research Publications
Now showing items 1-12 of 218
The immune phenotype of tongue squamous cell carcinoma predicts early relapse and poor prognosis
BACKGROUND: In patients with squamous cell carcinoma of the oral tongue (OTSCC), current tumor node metastasis staging system fails to identify at-risk patients associated with early relapse and poor prognosis despite complete surgical resection. Given the importance of tumor-infiltrating lymphocytes (TILs) in the development of cancers, here we investigated the prognostic significance of the immune phenotype in OTSCC. METHODS: Hematoxylin-eosin stained sections of OTSCCs from 211 patients were evaluated. Cancers were classified as (a) immune-inflamed when TILs were found next to tumor cell nests; (b) immune-excluded when TILs were found in the stroma, outside the tumor; and (c) immune-desert for tumors lacking lymphocyte infiltrate. The prognostic significance of these immune phenotypes classes was investigated. Data were further validated on an independent cohort from The Cancer Genome Atlas database. RESULTS: Immune-desert phenotype was the least represented group of OTSCCs in our cohort (11.8%) and served as an independent prognostic factor. Patients with immune-desert tumors exhibited worse disease-specific survival (HR = 2.673; [CI: 95% 1.497-4.773]; P = .001), overall survival (HR = 2.591; [CI: 95% 1.468-4.572]; P = .001), and disease-free survival (HR = 2.313; [CI: 95% 1.118-4.786]; P = .024) at multivariate analysis. CONCLUSIONS: We identified a specific subgroup of OTSCCs with poor prognosis. Tumor-infiltrating lymphocytes density and localization could serve as an integrative parameter to the current staging system and inform the selection of most appropriate treatments. In particular, the tumor immune phenotype could improve the stratification of patients with more aggressive disease.
Knowledge, attitudes, and beliefs regarding molar incisor hypomineralization (MIH) amongst German dental students
BACKGROUND: Knowledge of molar incisor hypomineralization (MIH) has relevance for paediatric dentists. AIM: To assess final-year German dental students' knowledge, attitudes, and beliefs regarding MIH. MATERIALS AND METHODS: A previously validated questionnaire was posted to the 31 German dental schools. Demographic covariates as well as knowledge regarding diagnosis and prevalence, and attitudes and beliefs around aetiology and management were collected. RESULTS: Twenty-two (71%) dental schools responded and a total of 877 students participated. Most (97%) were familiar with MIH and 88% were aware of the diagnostic criteria for MIH; however, only 42% knew how to implement them. One-third were able to identify MIH and 16% reported diagnostic confidence when doing so; 90% assumed the MIH prevalence to be <10%. Two-thirds of the respondents implicated genetic components as the main aetiological factor of MIH. Resin composite (60%) and preformed metal crowns (46%) were the dental materials most often suggested for restorative management. Almost all (98%) respondents were interested in receiving more clinical training. CONCLUSION: German students were familiar with MIH; however, they reported low levels of knowledge and confidence regarding its prevalence and diagnosis. Standardized nationwide, up-to-date curricula should be implemented to educate future dentists in Germany.
The 9-Fluorenylmethoxycarbonyl (Fmoc) Group in Chemical Peptide Synthesis - Its Past, Present, and Future
(CSIRO PUBLISHING, 2020-01-01)
<jats:p> The chemical formation of the peptide bond has long fascinated and challenged organic chemists. It requires not only the activation of the carboxyl group of an amino acid but also the protection of the Nα-amino group. The more than a century of continuous development of ever-improved protecting group chemistry has been married to dramatic advances in the chemical synthesis of peptides that, itself, was substantially enhanced by the development of solid-phase peptide synthesis by R. B. Merrifield in the 1960s. While the latter technology has continued to undergo further refinement and improvement in both its chemistry and automation, the development of the base-labile 9-fluorenylmethoxycarbonyl (Fmoc) group and its integration into current synthesis methods is considered a major landmark in the history of the chemical synthesis of peptides. The many beneficial attributes of the Fmoc group, which have yet to be surpassed by any other Nα-protecting group, allow very rapid and highly efficient synthesis of peptides, including ones of significant size and complexity, making it an even more valuable resource for research in the post-genomic world. This review charts the development and use of this Nα-protecting group and its adaptation to address the need for more green chemical peptide synthesis processes. </jats:p>
How to intervene in the caries process in adults: proximal and secondary caries? An EFCD-ORCA-DGZ expert Delphi consensus statement.
(Springer Science and Business Media LLC, 2020-09)
OBJECTIVES: To provide consensus recommendations on how to intervene in the caries process in adults, specifically proximal and secondary carious lesions. METHODS: Based on two systematic reviews, a consensus conference and followed by an e-Delphi consensus process were held with EFCD/ORCA/DGZ delegates. RESULTS: Managing an individual's caries risk using non-invasive means (oral hygiene measures including flossing/interdental brushes, fluoride application) is recommended, as both proximal and secondary carious lesions may be prevented or their activity reduced. For proximal lesions, only cavitated lesions (confirmed by visual-tactile, or radiographically extending into the middle/inner dentine third) should be treated invasively/restoratively. Non-cavitated lesions may be successfully arrested using non-invasive measures in low-risk individuals or if radiographically confined to the enamel. In high-risk individuals or if radiographically extended into dentine, for these lesions, additional micro-invasive (lesion sealing and infiltration) treatment should be considered. For restoring proximal lesions, adhesive direct restorations allow minimally invasive, tooth-preserving preparations. Amalgams come with a lower risk of secondary lesions and may be preferable in more clinically complex scenarios, dependent on specific national guidelines. In structurally compromised (especially endodontically treated) teeth, indirect cuspal coverage restorations may be indicated. Detection methods for secondary lesions should be tailored according to the individual's caries risk. Avoiding false positive detection and over-treatment is a priority. Bitewing radiographs should be combined with visual-tactile assessment to confirm secondary caries detections. Review/refurbishing/resealing/repairing instead of replacing partially defective restorations should be considered for managing secondary caries, if possible. CONCLUSIONS: An individualized and lesion-specific approach is recommended for intervening in the caries process in adults. CLINICAL SIGNIFICANCE: Dental clinicians have an increasing number of interventions available for the management of dental caries. Many of them are grounded in the growing understanding of the disease. The best evidence, patients' expectations, clinicians' expertise, and the individual clinical scenario all need to be considered during the decision-making process.
Study of Mothers' and Infants' Life Events Affecting Oral Health (SMILE) birth cohort study: cohort profile
(BMJ PUBLISHING GROUP, 2020-01-01)
PURPOSE: The long-term goal of the Study of Mothers' and Infants' Life Events Affecting Oral Health (SMILE) birth cohort study is to identify and evaluate the relative importance and timing of critical factors that shape the oral health of young children. It will then evaluate those factors in their inter-relationship with socioeconomic influences. PARTICIPANTS: SMILE is a single-centre study conducted in Adelaide, Australia. All newborns at the main three public hospitals between July 2013 and August 2014 were eligible for inclusion. The final recruited sample at birth was 2181 mother/infant dyads. Participants were followed up with questionnaires when the child was 3 and 6 months of age, and 1, 2 and 5 years of age. Oral epidemiological examinations and anthropometric assessments were conducted at age 2 and 5 years. FINDINGS TO DATE: SMILE has contributed comprehensive data on dietary patterns of young children. Intakes of free sugars, core and discretionary foods and drinks have been detailed. There was a sharp increase in free sugars intake with age. Determinants of dietary patterns, oral health status and body weight during the first 5 years of life have been evaluated. Socioeconomic characteristics such as maternal education and household income and area-level socioeconomic profile influenced dietary patterns and oral health behaviours and status. FUTURE PLAN: Funding has been obtained to conduct oral epidemiological examinations and anthropometric assessments at age 7-8 years. Plans are being developed to follow the cohort into adolescent years.
Pathogenesis of Molar Hypomineralisation: Aged Albumin Demarcates Chalky Regions of Hypomineralised Enamel
(FRONTIERS MEDIA SA, 2020-09-30)
Molar hypomineralisation (MH) is becoming globally recognised as a significant public health problem linked to childhood tooth decay. However, with causation and pathogenesis unclear after 100 years of investigation, better pathological understanding is needed if MH is to become preventable. Our studies have implicated serum albumin in an extracellular pathomechanism for chalky enamel, opposing longheld dogma about systemic injury to enamel-forming cells. Hypothesising that chalky enamel arises through developmental exposure to serum albumin, this study used biochemical approaches to characterise demarcated opacities from 6-year molars. Addressing contradictory literature, normal enamel was found to completely lack albumin subject to removal of surface contamination. Querying surface permeability, intact opacities were found to lack salivary amylase, indicating that "enamel albumin" had become entrapped before tooth eruption. Thirdly, comparative profiling of chalky and hard-white enamel supported a dose-response relationship between albumin and clinical hardness of opacities. Moreover, albumin abundance delineated chalky enamel from white transitional enamel at opacity borders. Finally, addressing the corollary that enamel albumin had been entrapped for several years, clear signs of molecular ageing (oxidative aggregation and fragmentation) were identified. By establishing aged albumin as a biomarker for chalky enamel, these findings hold methodological, clinical, and aetiological significance. Foremost, direct inhibition of enamel-crystal growth by albumin (here termed "mineralisation poisoning") at last provides a cogent explanation for the clinical presentation of demarcated opacities. Together, these findings justify pursuit of an extracellular paradigm for the pathogenesis of MH and offer exciting new prospects for alleviating childhood tooth decay through medical prevention of MH.
Kava constituents exert selective anticancer effects in oral squamous cell carcinoma cells in vitro
(NATURE RESEARCH, 2020-09-28)
Kava is a beverage made from the ground roots of the plant Piper Methysticum. Active compounds of Kava have previously been demonstrated to exert an antiproliferative effect through cell cycle arrest and promotion of apoptosis. Our aim was to investigate the in vitro effects of the main constituents derived from Kava on oral squamous cell carcinoma (OSCC) activity. Gas chromatography mass spectrometry (GCMS) was used to characterise the main constituents of two Kava preparations. Cell proliferation was assessed in two human OSCC cell lines (H400 and BICR56) and in normal oral keratinocytes (OKF6) treated with the identified Kava constituents, namely Flavokawain A (FKA), Flavokawain B (FKB), yangonin, kavain and methysticin using an MTS in vitro assay. Cell migration at 16 h was assessed using a Transwell migration assay. Cell invasion was measured at 22 h using a Matrigel assay. Cell adhesion was assessed at 90 min with a Cytoselect Adhesion assay. The two Kava preparations contained substantially different concentrations of the main chemical constituents. Treatment of malignant and normal oral keratinocyte cell lines with three of the identified constituents, 10 μg/ml FKA, 2.5 μg/ml FKB and 10 μg/ml yangonin, showed a significant reduction in cell proliferation in both H400 and BICR56 cancer cell lines but not in normal OKF6 cells. Remarkably, the same Kava constituents induced a significant reduction of OSCC cell migration and invasion. We have demonstrated, for the first time, that Kava constituents, FKA, FKB and yangonin have potential anticancer effects on OSCC. This highlights an avenue for further research of Kava constituents in the development of future cancer therapies to prevent and treat OSCC.
Computational analysis of TP53 mutational landscape unveils key prognostic signatures and distinct pathobiological pathways in head and neck squamous cell cancer
BACKGROUND: Mutations of the tumour-suppressor gene TP53 are the most frequent somatic genomic alterations in head and neck squamous cell carcinoma (HNSCC). However, it is not yet clear whether specific TP53 mutations bear distinct clinical and pathophysiological significance in different HNSCC subgroups. METHODS: A systematic bioinformatics appraisal of TP53 mutations was performed on 415 HNSCC cases available on The Cancer Genome Atlas (TCGA). The following features were analysed and correlated with known clinicopathological variables: mutational profile of TP53, location (within secondary structure and predicted domains of p53 protein) and well-known hotspot mutations. Interactome-genome-transcriptome network analysis highlighted different gene networks. An algorithm was generated to develop a new prognostic classification system based on patients' overall survival. RESULTS: TP53 mutations in HNSCCs exhibited distinct differences in different anatomical sites. The mutational profile of TP53 was an independent prognostic factor in HNSCC. High risk of death mutations, identified by our novel classification algorithm, was an independent prognostic factor in TCGA HNSCC database. Finally, network analysis suggested that distinct p53 molecular pathways exist in a site- and mutation-specific manner. CONCLUSIONS: The mutational profile of TP53 may serve as an independent prognostic factor in HNSCC patients, and is associated with distinctive site-specific biological networks.
A biophysically-defined hyaluronic acid-based compound accelerates migration and stimulates the production of keratinocyte-derived neuromodulators
(TAYLOR & FRANCIS INC, 2019-01-01)
Hyaluronic acid (HA) preparations are widely used in clinical practice and recent data suggest that commercially available HA-based compounds promote ulcer re-epithelialization and induce pain relief. However, the pathophysiological basis of these effects remains poorly understood. In the present study, we investigated the biophysical, biomolecular and functional properties of a HA preparation combined with a pool of collagen precursor synthetic aminoacids, namely l-proline, l-leucine, l-lysine and glycine (Aminogam®). Hydrodynamic characterization of Aminogam® by size exclusion chromatography-triple detector array (SEC-TDA) revealed an average molecular weight in the range of 700-1700 kDa. Rheological measurements of the 1700kDa Mw lot showed a pseoudoplastic behaviour with a zero-shear viscosity (η0) equal to 90 ± 9 Pa∙s at 25°C and 55 ± 6 Pa∙s at 37°C. Automated time-lapse videomicroscopy studies in a fibroblast-free system demonstrated that 1% (v/v) Aminogam® significantly reduced the healing time of wounded keratinocyte monolayers. In AKGOS assays, Aminogam® stimulated cellular locomotion (chemokinesis) and directional migration (chemotaxis) of keratinocytes. Analysis of microarray data suggested that keratinocytes had a functional neuroendocrine machinery, and this was confirmed by testing the secretion of six neuroactive molecules by ELISA, namely α-MSH, β-endorphins, melatonin, substance P, cortisol, and neurotensin. Interestingly, Aminogam® regulated the production of several neuropeptides, including β-endorphins. In conclusion, our data shed light on the epithelial-dependent mechanisms that underlie the efficacy of Aminogam®, particularly in reference to wound healing and nociception.
Self-assembly of dental surface nanofilaments and remineralisation by SnF2 and CPP-ACP nanocomplexes
(NATURE PUBLISHING GROUP, 2019-02-04)
Dental caries, erosion and hypersensitivity are major public health problems. SnF2 is used widely in oral care products to help prevent/treat these conditions. Casein phosphopeptide-stabilised amorphous calcium phosphate nanocomplexes (CPP-ACP) are a biomimetic nanotechnology of salivary phosphopeptide-ACP complexes that deliver bioavailable calcium and phosphate ions to promote dental remineralisation (repair). We show here using in vitro studies and a double-blind, randomised controlled, cross-over design in situ clinical trial that SnF2 and CPP-ACP interact to form a nanofilament coating on the tooth surface and that together they are superior in their ability to promote dental remineralisation. Sn(II) by cross-linking the CPP-ACP helps to stabilise the complexes which improves delivery to the tooth surface and enhances binding and ion incorporation into tooth mineral. The combination of SnF2 and CPP-ACP in oral care products may significantly improve their efficacy in prevention/treatment of dental caries/erosion and hypersensitivity.