Neuropeptides Regulate Expression of Angiogenic Growth Factors in Human Dental Pulp Fibroblasts

Abstract
INTRODUCTION:
Neuropeptides play an important role in inflammation and repair and have been implicated in mediating angiogenesis. Pulp fibroblasts express neuropeptide receptors, and the aim of this research was to investigate whether neuropeptides could regulate angiogenic growth factor expression in vitro
METHODS:
An angiogenic array was used to determine the levels of 10 angiogenic growth factors expressed by human pulp fibroblasts.
RESULTS:
Pulp fibroblasts were shown to express angiogenin, angiopoietin-2, epidermal growth factor, basic fibroblast growth factor, heparin-binding epidermal growth factor, hepatocyte growth factor, leptin, platelet-derived growth factor, placental growth factor, and vascular endothelial growth factor. Furthermore, the neuropeptides substance P, calcitonin gene-related peptide, vasoactive intestinal polypeptide, and neuropeptide Y altered angiogenic growth factor expression in vitro.
CONCLUSIONS:
The regulation of angiogenic growth factor expression by neuropeptides suggests a novel role for neuropeptides in pulpal inflammation and repair.

Re-treatment profiles during long-term maintenance therapy in a periodontal practice in Norway

Background: Periodontal therapy coupled with active maintenance has been shown to be effective in maintaining periodontal health, however, the question of re-treatment is rarely alluded to in the literature.

Interviewing children with acquired brain injury (ABI).

Research into the lives of children with acquired brain injury (ABI) often neglects to incorporate children as participants, preferring to obtain the opinions of the adult carer (e.g. McKinlay et al., 2002). There has been a concerted attempt to move away from this position by those working in children’s research with current etiquette highlighting the inclusion of children and the use of a child-friendly methodology (Chappell, 2000). Children with disabilities can represent a challenge to the qualitative researcher due to the combination of maintaining the child’s attention and the demands placed on them by their disability. The focus of this article is to discuss possible impediments to interviewing children with acquired brain injury (ABI) and provide an insight into how the qualitative researcher may address these.

PAR-2 Regulates Dental Pulp Inflammation Associated with Caries

Protease-activated receptors (PARs) are G-protein-coupled receptors that are activated enzymatically by proteolysis of an N-terminal domain. The cleavage and activation of PARs by serine proteases represent a novel mechanism by which such enzymes could influence the host inflammatory response. The aim of this study was to determine whether PAR-2 expression and activation were increased in dental caries. Using immunohistochemistry, we showed PAR-2 to be localized to pulp cells subjacent to caries lesions, but minimally expressed by healthy pulp tissue. Trypsin and the PAR-2 agonist (PAR2-AP) activated PAR-2 in an in vitro functional assay. Endogenous molecules present in pulp cell lysates from carious teeth specifically activated PAR-2, but those from healthy teeth failed to do so. The activation of PAR-2 in vitro was shown to increase the expression of the pro-inflammatory mediator cyclo-oxygenase-2 (COX-2), providing a mechanism whereby PAR-2 could modulate pulpal inflammation.

Healthcare professionals’ attitudes towards traumatic brain injury (TBI): The influence of profession, experience, aetiology and blame on prejudice towards survivors of brain injury

Primary objective: To investigate the attitudes of healthcare professionals towards individuals with traumatic brain injury (TBI) and their relationship to intended healthcare behaviour.

Research design: An independent groups design utilized four independent variables; aetiology, group, blame and gender to explore attitudes towards survivors of brain injury. The dependent variables were measured using the Prejudicial Evaluation and Social Interaction Scale (PESIS) and Helping Behaviour Scale (HBS).

Methods and procedures: A hypothetical vignette based methodology was used. Four hundred and sixty participants (131 trainee nurses, 94 qualified nurses, 174 trainee doctors, 61 qualified doctors) were randomly allocated to one of six possible conditions.

Main outcomes and results: Regardless of aetiology, if an individual is to blame for their injury, qualified healthcare professionals have more prejudicial attitudes than those entering the profession. There is a significant negative relationship between prejudice and helping behaviour for qualified healthcare professionals.

Conclusions: Increased prejudicial attitudes of qualified staff are related to a decrease in intended helping behaviour, which has the potential to impact negatively on an individual's recovery post-injury.

"To be accepted as normal": Public understanding and misconceptions concerning survivors of brain injury.

PRIMARY OBJECTIVE: To determine the views held by the general public in Northern Ireland towards survivors of brain injury. RESEARCH DESIGN: Qualitative semi-structured interviews. METHODS AND PROCEDURES: Interviews were conducted with 16 members of the general public. Ten questions addressed issues such as the role of survivors of brain injury in society, the challenges they face and the characteristics ascribed to them. MAIN OUTCOMES AND RESULTS: When asked to describe someone with a brain injury participants typically used negative labels and identified the most common problems as relating to physical, cognitive, emotional and social functioning. There was a general failure to recognize that brain injury was a 'hidden' disability, with most participants expecting some outward manifestation. Relatively few previous studies have employed a qualitative approach to explore how the public perceives survivors of brain injury. CONCLUSION: Members of the public have an increasing awareness of the challenges faced by survivors of brain injury. However, in spite of this, perceptions of aggressiveness, dependency and unhappiness were still evident, suggesting potential problems in reintegrating survivors of brain injury with their communities.

Long-term outcomes for cross-arch stabilizing bridges in periodontal maintenance patients - a retrospective study

Background: Cross-arch bridges are used to stabilize teeth for patients with reduced periodontal support. Little is known about technical or biological complications, whether teeth and implants can be combined in this type of bridge and the long-term effects on tooth loss.

A comparative study of nursing attitudes towards young male survivors of brain injury: A questionnaire survey.

Background The attitudes members of the nursing profession hold towards survivors of brain injury may impact on the level of help, and degree of involvement they are willing to have. Given that the manner in which an individual receives their brain injury has been shown to impact on public prejudices, the importance of exploring nursing attitudes to this vulnerable group, and the subsequent impact this may have on the caring role, requires investigation. Objective To investigate the attitudes held by members of the nursing profession towards young male survivors of brain injury whose behaviour either contributed, or did not contribute, to their injury. Design Independent groups design. Setting and participants Ninety trainee and sixty-nine qualified nurses respectively drawn from a university in the south west of England and the emergency, orthopaedic and paediatric Departments of the Royal Devon and Exeter Hospital, UK. Methods Participants were randomly assigned to one of four fictional brain injury scenarios. A young male character was portrayed as sustaining a brain injury as a result of either an aneurysm, or through drug taking, with their behaviour being either a contributory or non-contributory factor. On reading these, participants were asked to complete the prejudicial evaluation scale, the social interaction scale and the helping behaviour scale. Results Analysis of variance showed that qualified nurses held more prejudicial attitudes than student nurses towards survivors of brain injury. Mean scores indicated that individuals seen as contributing towards their injury were likely to experience more prejudice (blame total = 42.35 vs. no blame total = 38.34), less social interaction (blame total = 37.54 vs. no blame total = 41.10), and less helping behaviour (blame total = 21.49 vs. no blame total = 22.34) by both groups. Conclusions Qualified nurses should be mindful of the impact their attitudes and judgements of survivors of brain injury may have on the subsequent care they provide. Greater emphasis on the effects of negative attitudes on patient interactions during training may provide nurses with the understanding to recognise and avoid challenges to their caring role in the future.

Attitudes of qualified versus student mental health nurses towards an individual diagnosed with schizophrenia

Aims: This paper is the report of a study which sought to compare the attitudes held by student and qualified mental health nurses towards individuals with schizophrenia in the Republic of Ireland. Background: Media portrayals of individuals with schizophrenia often include images of aggression and violence. With global initiatives aimed at reducing the stigma and exclusion associated with mental illness, the attitudes of those who care for people with schizophrenia are of particular interest. Methods: A survey was administered to 66 student mental health nurses, and 121 qualified mental health nurses. Participants completed the community attitudes to mental illness scale (CAMI) and the social interaction scale (SIS) in 2009. Multivariate analysis of variance was used to test for the effects of qualification, work setting, years of experience and education on the measures. Results: A statistically significant difference was found between community mental health nurses and those employed in an inpatient setting on the social restrictiveness and community mental health ideology subscales of the CAMI and on the SIS. Findings also showed a statistically significant difference between nurses in the 10-14 years of experience group and the 5-9 years of experience group on the SIS. Conclusions: Mental health nurses employed in an inpatient setting are often confronted with patients who have challenging behavioural presentations which may explain their socially restrictive attitudes. However, nurses must be alerted to the fact that such negative attitudes may adversely affect the therapeutic relationship and ultimately lead to stigmatisation and its negative consequences.

Human odontoblasts express functional thermo-sensitive TRP channels: implications for dentin sensitivity

Odontoblasts form the outermost cellular layer of the dental pulp where they have been proposed to act as sensory receptor cells. Despite this suggestion, evidence supporting their direct role in mediating thermo-sensation and nociception is lacking. Transient receptor potential (TRP) ion channels directly mediate nociceptive functions, but their functional expression in human odontoblasts has yet to be elucidated. In the present study, we have examined the molecular and functional expression of thermo-sensitive TRP channels in cultured odontoblast-like cells and in native human odontoblasts obtained from healthy wisdom teeth. PCR and western blotting confirmed gene and protein expression of TRPV1, TRPA1 and TRPM8 channels. Immunohistochemistry revealed that these channels were localised to odontoblast-like cells as determined by double staining with dentin sialoprotein (DSP) antibody. In functional assays, agonists of TRPV1, TRPA1 and TRPM8 channels elicited [Ca2+]i transients that could be blocked by relevant antagonists. Application of hot and cold stimuli to the cells also evoked rises in [Ca2+]i which could be blocked by TRP-channel antagonists. Using a gene silencing approached we further confirmed a role for TRPA1 in mediating noxious cold responses in odontoblasts. We conclude that human odontoblasts express functional TRP channels that may play a crucial role in mediating thermal sensation in teeth. Cultured and native human odontoblasts express functional TRP channels that may play a crucial role in mediating thermal sensation in teeth.

Human Dental Pulp Fibroblasts Express the “Cold-sensing” Transient Receptor Potential Channels TRPA1 and TRPM8

Introduction: Transient receptor potential (TRP) channels comprise a group of nonselective calcium-permeable cationic channels, which are polymodal sensors of environmental stimuli such as thermal changes and chemicals. TRPM8 and TRPA1 are cold-sensing TRP channels activated by moderate cooling and noxious cold temperatures, respectively. Both receptors have been identified in trigeminal ganglion neurones, and their expression in nonneuronal cells is now the focus of much interest. The aim of this study was to investigate the molecular and functional expression of TRPA1 and TRPM8 in dental pulp fibroblasts.
Methods: Human dental pulp fibroblasts were derived from healthy molar teeth. Gene and protein expression was determined by polymerase chain reaction and Western blotting. Cellular localization was investigated by immunohistochemistry, and TRP functionality was determined by Ca2+ microfluorimetry.
Results: Polymerase chain reaction and Western blotting showed gene and protein expression of both TRPA1 and TRPM8 in fibroblast cells in culture. Immunohistochemistry studies showed that TRPA1 and TRPM8 immunoreactivity co-localized with the human fibroblast surface protein. In Ca2+ microfluorimetry studies designed to determine the functionality of TRPA1 and TRPM8 in pulp fibroblasts, we showed increased intracellular calcium ([Ca2+]i) in response to the TRPM8 agonist menthol, the TRPA1 agonist cinnamaldehyde, and to cool and noxious cold stimuli, respectively. The responses to agonists and thermal stimuli were blocked in the presence of specific TRPA1 and TRPM8 antagonists.
Conclusions: Human dental pulp fibroblasts express TRPA1 and TRPM8 at the molecular, protein, and functional levels, indicating a possible role for fibroblasts in mediating cold responses in human teeth.

A comparison of four extraction methods for substance P, neurokinin A and calcitonin gene-related peptide from human dental pulp tissue

Measuring neuropeptides in biological tissues by radioimmunoassay requires efficient extraction that maintains their immunoreactivity. Many different methods for extraction have been described, but there is little information on optimal extraction methods for individual neuropeptides from human dental pulp tissue. The aim was therefore to identify an effective extraction procedure for three pulpal neuropeptides: substance P. neurokinin A and calcitonin gene-related peptide. Tissue was obtained from 20 pulps taken from teeth freshly extracted for orthodontic reasons. The pulp samples were divided into four equal groups and different extraction methods were used for each group. Boiling whole pulp in acetic acid gave the highest overall yield and, in addition, offered an easy and rapid means of pulp tissue processing. The use of protease inhibitors did not increase the recovery of the immunoreactive neuropeptides but did provide the best combination of maximal recoveries and minimal variability. These results should be useful for planning the extraction of these neuropeptides from human pulp tissue in future studies. (C) 1999 Elsevier Science Ltd. All rights reserved.