Effect of glycosaminoglycans on growth factor-stimulated trophoblast invasion

Objectives: To determine the effect of glycosaminoglycans and a series of growth factors on the viability and invasion of the extravillous trophoblast cell line SGHPL4. Methods: Cells were cultured in Hams F10 media supplemented with fetal bovine serum and L-glutamine. For viability studies cells were seeded into 96-well culture plates (104 cells/well), maintained in serum free medium for 24h and then incubated with glycosaminoglycans (heparin, heparin sulphate and hyaluronic acid; each 100ng/ml) ± growth factors (VEGF, FGFand HB-EGF). Cell viability was measured in cells using the MTS assay. Cellular invasion was assessed using the FluoroBlok invasion assay. Cells were serum-starved for 24 h, incubated with the fluorescent dye DiIC12(3) (10mg/ml) for 1 hour prior to seeding onto an artificial extracellular matrix coated 8 mm FluoroBlok porous membrane inserts (2.5 x 105 cells per insert). Growth factors ± GAGs were added to the cell suspension and the inserts were lowered into a 96-well plate containing 10% fetal calf serum. Plates were incubated at 37°C for 24h. Invasion was determined by measurement of fluorescence of invaded cells using a fluorescent plate reader (Ex549/Em565 nm). Results: Cell numbers were significantly increased following incubation with VEGF, FGF and HB-EGF. Cell number was also increased after incubation with each of the glycosaminoglycans tested. The largest increase was observed following incubation with heparin sulphate. Cell numbers were further increased when the GFs were incubated with HS and heparin, but not with hyaluronic acid. Invasion was increased following incubation with VEGF, HBEGF and HGF. Heparan sulphate and heparin increased invasiveness in a dose-dependent manner. In contrast, hyaluronic acid had no significant effect. Conclusion: This study demonstrates a role for glycosaminoglycans in key features of trophoblast function

The impact of dental caries on children and young people: What they have to say?

Background: Dental caries affects 60-90% of children across the world and is associated with a variety of negative impacts. Despite its ubiquity, there has been surprisingly little exploration of these impacts from the child's perspective. Aim: The aim was to allow children to describe the impact of dental caries on their daily lives and to describe the terminology they used. Design: Children, aged 5-15 years, with caries experience were purposively sampled from primary and secondary care dental clinics. Focus groups (n = 5) and in-depth interviews (n = 16) were recorded and transcribed verbatim. Data analysis took a narrative approach, and themes were derived from the data using framework analysis. Results: Pain was the main theme to emerge. Within this, three subthemes were identified: impacts related to pain, strategies adopted to reduce pain, and emotional aspects resulting from pain. A second theme was also identified relating to the aesthetic aspects of caries. Conclusion: Children as young as 5 years of age were able to competently discuss their experiences of dental caries. Participants reported a number of impacts affecting various aspects of their lives. These will be incorporated into the future development of a caries-specific measure of oral health-related quality of life

In-vivo validity of proximal caries detection in primary teeth, with histological validation.

BACKGROUND: Detection and diagnosis of proximal caries in primary molars is challenging. AIM: The aim of this in-vivo study was to assess the validity and reproducibility of four methods of proximal caries detection in primary molar teeth. DESIGN: Eighty-two children (5-10 yrs) were recruited. Initially 1030 proximal surfaces were examined using meticulous visual examination (ICDAS) (VE1), bitewing radiographs (RE), and a laser fluorescence pen device (LF1). Temporary tooth separation (TTS) was achieved for 447 surfaces and these were re-examined visually (VE2) and using the LF-pen (LF2). Three hundred and fifty-six teeth (542 surfaces) were subsequently extracted and provided histological validation. RESULTS: At D1 (enamel and dentine caries) diagnostic threshold, the sensitivity of VE1, RE, VE2, LF1 and LF2 examination were 0.52, 0.14, 0.75, 0.58, 0.60 and the specificity values were 0.89, 0.97, 0.88, 0.85, 0.77 respectively. At D3 (dentine caries) threshold, the sensitivity values were 0.42, 0.71, 0.49, 0.63, 0.65 respectively, while specificity was 0.93 for VE1 and VE2, and 0.98, 0.87 and 0.88 for RE, LF1 and LF2 examinations respectively. ROC analysis showed radiographic examination to be superior at D3 . CONCLUSION: Meticulous caries diagnosis (ICDAS) should be supported by radiographs for detection of dentinal proximal caries in primary molars

Selection and validation of a classification system for a child-centred caries-specific utility measure

Background: Caries Impacts and Experiences Questionnaire for Children (CARIES-QC) is a child-centred caries-specific quality of life measure. This study aimed to select, and validate with children, a classification system for a paediatric condition-specific preference-based measure, based on CARIES-QC. Methods: First, a provisional classification system for a preference-based measure based on CARIES-QC was developed using Rasch analysis, psychometric testing, involvement of children and parents, and the developer of CARIES-QC. Second, qualitative, semi-structured ‘think aloud’ validation interviews were undertaken with a purposive sample of children with dental caries. The interviewer aimed to identify whether items were considered important and easily understood, whether any were overlapping and if any excluded items should be reintroduced. Interview recordings were transcribed verbatim and thematic analysis conducted. Results: Rasch analysis identified poor item spread for the items ‘cross’ and ‘school’. Items relating to eating were correlated and the better performing items were considered for selection. Children expressed some confusion regarding the items ‘school’ and ‘food stuck’. Parent representatives thought that impacts surrounding toothbrushing (‘brushing’) were encompassed by the item ‘hurt’. Five items were selected from CARIES-QC for inclusion in the provisional classification system; ‘hurt’, ‘annoy’, ‘carefully’, ‘kept awake’ and ‘cried’. Validation interviews were conducted with 20 children aged 5-16 years old. Participants thought the questionnaire was straightforward and covered a range of impacts. Children thought an item about certain foods being ‘hard to eat’ was more relevant than one about having to eat more carefully because of their teeth and so the ‘carefully’ item was replaced with ‘hard to eat’. Conclusion: Following child-centred modification, the preliminary five-item classification system is considered valid and suitable for use in a valuation survey. The innovative child-centred methods used to both identify and validate the classification system can be applied in the development of other preference-based measures

Adolescent valuation of CARIES-QC-U: a child-centred preference-based measure of dental caries

Objectives: This study develops an adolescent value set for a child-centred dental caries-specific measure of oral health-related quality of life (OHRQoL) based upon CARIES-QC (Caries Impacts and Experiences Questionnaire for Children). This study develops a new approach to valuing child health by eliciting adolescent preferences and anchoring these onto the 1-0 full health-dead QALY (quality adjusted life year) scale using ordinal adult preferences.Methods: Two online surveys were created to elicit preferences for the CARIES-QC classification system. The first comprised best-worst scaling (BWS) tasks for completion by adolescents aged 11-16 years. The second comprised discrete choice experiment tasks with a duration attribute (DCETTO) for completion by adults aged over 18 years. Preferences were modelled using the conditional logit model. Mapping regressions anchored the adolescent BWS data onto the QALY scale using adult DCETTO values, since the BWS survey data alone cannot generate anchored values.Results: 723 adolescents completed the BWS survey and 626 adults completed the DCE(TTO )survey. The samples were representative of UK adolescent and adult populations. Fully consistent and robust models were produced for both BWS and DCETTO data. BWS preferences were mapped onto DCETTO values, resulting utility estimates for each health state defined by the classification system.Conclusion: This is the first measure with predetermined scoring based on preferences to be developed specifically for use in child oral health research, and uses a novel technique to generate a value set using adolescent preferences. The estimates can be used to generate QALYs in economic evaluations of interventions to improve children's oral health

‘I would rather be having my leg cut off than a little needle’: A supplementary qualitative analysis of dentally anxious children’s experiences of needle fear

Fear of needles is common in childhood, with up to 50% being affected to some degree. In individuals who are dentally anxious, the prevalence may be as high as 91%. Fear of needles, and therefore intra-oral injections can have negative impacts on children’s quality of life and healthcare experiences, including a requirement for pharmacological methods to facilitate dental treatment. The aim of this study is to identify whether dentally anxious children report fear of injections and explore how these children experience a fear of needles in a dental setting. A supplementary analysis of interviews collected as part of two previous studies relating to children with dental anxiety. Five main themes were identified: feelings about needles; the nature of needle fear; the context of the fear, its consequences and how children tried to control the process. Children showed a desire to have control of their healthcare interventions, and wanted to trust the healthcare professionals giving the injections. There is evidence that children with dental anxiety also experience fear of needles, including intra-oral injections. Further primary qualitative research is needed to explore this topic in more depth and to design appropriate child centred interventions to reduce needle fear

‘Your teeth you are in control’

The use of cognitive behavioural therapy in reducing dental anxiety for paediatric patients </jats:p

Cognitive behavioural therapy and the adolescent patient

Dental anxiety is common in adolescents and is associated with poor oral health status and worse oral health-related quality of life. As these patients move into adulthood, they may continue to be reliant on pharmacological approaches in order to cope with dental treatment and only attend when they have a problem. Managing anxious adolescents is also a source of stress for dental practitioners. This article describes how cognitive behavioural therapy (CBT) can be used to reduce dental anxiety in this group of patients

Discrete choice experiments or best-worst scaling? A qualitative study to determine the suitability of preference elicitation tasks in research with children and young people

Background Ordinal tasks are increasingly used to explore preferences for health states. This study aimed to determine the suitability of two ordinal preference elicitation tasks (discrete choice experiments (DCE) and best-worst scaling (BWS)) for use with children and young people to generate health state utility values. The study explored children’s understanding, the relationship between their age and level of understanding, and how many tasks they felt they could complete. Methods Children aged 11–16 years were recruited from a secondary school in South Yorkshire, UK. Participants were asked to ‘think aloud’ as they completed a computer-based survey that contained both DCE and BWS tasks relating to dental caries (tooth decay) health states. Health states involved descriptions of the impact of tooth decay on children’s daily lives. One-to-one semi-structured interviews were then held with participants, with use of a topic guide. Qualitative data were transcribed verbatim and analysed thematically. Results A total of 33 children (12 male, 21 female) participated, comprising 5–6 children from each school year group. Children expressed a preference for BWS and demonstrated a better understanding of these tasks than DCE. There was no clear relationship between children’s level of understanding and age. Children felt they could manage between 8 and 10 BWS tasks comfortably. Conclusion This study suggests that BWS tasks are the most appropriate type of preference elicitation task to value health states for children and young people aged 11–16 years to complete

The impact of dental caries and its treatment under general anaesthetic on children and their families

Objective To assess the impact of dental caries and treatment under general anaesthetic (GA) on the everyday lives of children and their families, using child-reported measures of quality of life (QoL) and oral health-related quality of life (OHRQoL). Method Participants, aged 5–16 years old having treatment for dental caries under GA, were recruited from new patient clinics at Charles Clifford Dental Hospital, Sheffield. OHRQoL was measured before and 3-months after treatment using the Caries Impacts and Experiences Questionnaire for Children (CARIES-QC). Overall QoL was measured using the Child Health Utility 9D (CHU9D). Parents/caregivers completed the Family Impact Scale (FIS). Results Eighty five parent–child dyads completed the study. There was statistically significant improvement in OHRQoL (mean interval score difference in CARIES-QC = 4.43, p < 0.001) and QoL (mean score difference in CHU9D = 2.48, p < 0.001) following treatment, with moderate to large effect sizes. There was statistically significant improvement in FIS scores (mean score difference = 5.48, p = 0.03). Conclusions Treatment under GA was associated with improvement in QoL and OHRQoL as reported by children, and reduced impacts on the family. This work highlights the importance of GA services in reducing the caries-related impacts experienced by children. Further work is needed investigate the impact of clinical, environmental and individual factors