3D-printed nanocomposite denture base resin: The effect of incorporating TiO2 nanoparticles on the growth of Candida albicans.

PURPOSE: To develop a biocompatible denture base resin/TiO2 nanocomposite material with antifungal characteristics that is suitable for 3D-printing denture bases. MATERIALS AND METHODS: TiO2 nanoparticles (NPs) with a 0.10, 0.25, 0.50, and 0.75 weight percent (wt.%) were incorporated into a commercially available 3D-printed resin material. The resulting nanocomposite material was analyzed using Lactate dehydrogenase (LDH) and AlamarBlue (AB) assays for biocompatibility testing with human gingival fibroblasts (HGF). The composite material was also tested for its antifungal efficacy against Candida albicans. Fourier transform infrared (FTIR) and Energy Dispersive X-ray Spectroscopy (EDX) mapping were conducted to assess the surface coating and the dispersion of the NPs. RESULTS: LDH and AB assays confirmed the biocompatibility of the material showing cell proliferation at a rate of nearly 100% at day 10, with a cytotoxicity of less than 13% of the cells at day 10. The concentrations of 0.10, 0.25, and 0.50 wt.% caused a significant reduction (p < 0.05) in the number of candida cells attached to the surface of the specimens (p < 0.05), while 0.75 wt.% did not show any significant difference compared to the control (no TiO2 NPs) (p > 0.05). FTIR and EDX analysis confirmed the presence of TiO2 NPs within the nanocomposite material with a homogenous dispersion for 0.10 and 0.25 wt.% groups and an aggregation of the NPs within the material at higher concentrations. CONCLUSION: The addition of TiO2 NPs into 3D-printed denture base resin proved to have an antifungal effect against Candida albicans. The resultant nanocomposite material was a biocompatible material with HGFs and was successfully used for 3D printing.

Improved method for skin reflectance reconstruction from camera images.

A improved spectral reflectance reconstruction method is developed to transform camera RGB to spectral reflectance for skin images. Rather than using conventional direct or two-step processes, we transform camera RGB to skin reflectance directly using a principal component analysis (PCA) approach. The novelty in our direct method (RGB to spectra) is the use of a skin-specific colour characterisation chart with spectra closer to human skin spectra, and a new database of skin reflectances to derive the PCA bases. The experimental results using the facial images of 17 subjects demonstrate that our new direct method gives a significantly better performance than conventional, two-step methods and direct methods with traditional characterization charts. This new spectral reconstruction algorithm is sufficiently precise to reconstruct spectral properites relating to chromophores and its performance is within the acceptable range for maxillofacial soft tissue prostheses (error < 3 ΔE*ab units).

Interventions for iatrogenic inferior alveolar and lingual nerve injury.

BACKGROUND: Iatrogenic injury of the inferior alveolar or lingual nerve or both is a known complication of oral and maxillofacial surgery procedures. Injury to these two branches of the mandibular division of the trigeminal nerve may result in altered sensation associated with the ipsilateral lower lip or tongue or both and may include anaesthesia, paraesthesia, dysaesthesia, hyperalgesia, allodynia, hypoaesthesia and hyperaesthesia. Injury to the lingual nerve may also affect taste perception on the affected side of the tongue. The vast majority (approximately 90%) of these injuries are temporary in nature and resolve within eight weeks. However, if the injury persists beyond six months it is deemed to be permanent. Surgical, medical and psychological techniques have been used as a treatment for such injuries, though at present there is no consensus on the preferred intervention, or the timing of the intervention. OBJECTIVES: To evaluate the effects of different interventions and timings of interventions to treat iatrogenic injury of the inferior alveolar or lingual nerves. SEARCH METHODS: We searched the following electronic databases: the Cochrane Oral Health Group's Trial Register (to 9 October 2013), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2013, Issue 9), MEDLINE via OVID (1946 to 9 October 2013) and EMBASE via OVID (1980 to 9 October 2013). No language restrictions were placed on the language or date of publication when searching the electronic databases. SELECTION CRITERIA: Randomised controlled trials (RCTs) involving interventions to treat patients with neurosensory defect of the inferior alveolar or lingual nerve or both as a sequela of iatrogenic injury. DATA COLLECTION AND ANALYSIS: We used the standard methodological procedures expected by The Cochrane Collaboration. We performed data extraction and assessment of the risk of bias independently and in duplicate. We contacted authors to clarify the inclusion criteria of the studies. MAIN RESULTS: Two studies assessed as at high risk of bias, reporting data from 26 analysed participants were included in this review. The age range of participants was from 17 to 55 years. Both trials investigated the effectiveness of low-level laser treatment compared to placebo laser therapy on inferior alveolar sensory deficit as a result of iatrogenic injury.Patient-reported altered sensation was partially reported in one study and fully reported in another. Following treatment with laser therapy, there was some evidence of an improvement in the subjective assessment of neurosensory deficit in the lip and chin areas compared to placebo, though the estimates were imprecise: a difference in mean change in neurosensory deficit of the chin of 8.40 cm (95% confidence interval (CI) 3.67 to 13.13) and a difference in mean change in neurosensory deficit of the lip of 21.79 cm (95% CI 5.29 to 38.29). The overall quality of the evidence for this outcome was very low; the outcome data were fully reported in one small study of 13 patients, with differential drop-out in the control group, and patients suffered only partial loss of sensation. No studies reported on the effects of the intervention on the remaining primary outcomes of pain, difficulty eating or speaking or taste. No studies reported on quality of life or adverse events.The overall quality of the evidence was very low as a result of limitations in the conduct and reporting of the studies, indirectness of the evidence and the imprecision of the results. AUTHORS' CONCLUSIONS: There is clearly a need for randomised controlled clinical trials to investigate the effectiveness of surgical, medical and psychological interventions for iatrogenic inferior alveolar and lingual nerve injuries. Primary outcomes of this research should include: patient-focused morbidity measures including altered sensation and pain, pain, quantitative sensory testing and the effects of delayed treatment.

Ibuprofen and/or paracetamol (acetaminophen) for pain relief after surgical removal of lower wisdom teeth.

BACKGROUND: Both paracetamol and ibuprofen are commonly used analgesics for the relief of pain following the surgical removal of lower wisdom teeth (third molars). In 2010, a novel analgesic (marketed as Nuromol) containing both paracetamol and ibuprofen in the same tablet was launched in the United Kingdom, this drug has shown promising results to date and we have chosen to also compare the combined drug with the single drugs using this model. In this review we investigated the optimal doses of both paracetamol and ibuprofen via comparison of both and via comparison with the novel combined drug. We have taken into account the side effect profile of the study drugs. This review will help oral surgeons to decide on which analgesic to prescribe following wisdom tooth removal. OBJECTIVES: To compare the beneficial and harmful effects of paracetamol, ibuprofen and the novel combination of both in a single tablet for pain relief following the surgical removal of lower wisdom teeth, at different doses and administered postoperatively. SEARCH METHODS: We searched the Cochrane Oral Health Group'sTrials Register (to 20 May 2013); the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2013, Issue 4); MEDLINE via OVID (1946 to 20 May 2013); EMBASE via OVID (1980 to 20 May 2013) and the metaRegister of Controlled Trials (to 20 May 2013). We checked the bibliographies of relevant clinical trials and review articles for further studies. We wrote to authors of the identified randomised controlled trials (RCTs), and searched personal references in an attempt to identify unpublished or ongoing RCTs. No language restriction was applied to the searches of the electronic databases. SELECTION CRITERIA: Only randomised controlled double-blinded clinical trials were included. Cross-over studies were included provided there was a wash out period of at least 14 days. There had to be a direct comparison in the trial of two or more of the trial drugs at any dosage. All trials used the third molar pain model. DATA COLLECTION AND ANALYSIS: All trials identified were scanned independently and in duplicate by two review authors, any disagreements were resolved by discussion, or if necessary a third review author was consulted. The proportion of patients with at least 50% pain relief (based on total pain relief (TOTPAR) and summed pain intensity difference (SPID) data) was calculated for all three drugs at both two and six hours postdosing and meta-analysed for comparison. The proportion of participants using rescue medication over both six and eight hours was also collated and compared. The number of patients experiencing adverse events or the total number of adverse events reported or both were analysed for comparison. MAIN RESULTS: Seven studies were included, they were all parallel-group studies, two studies were assessed as at low risk of bias and three at high risk of bias; two were considered to have unclear bias in their methodology. A total of 2241 participants were enrolled in these trials.Ibuprofen was found to be a superior analgesic to paracetamol at several doses with high quality evidence suggesting that ibuprofen 400 mg is superior to 1000 mg paracetamol based on pain relief (estimated from TOTPAR data) and the use of rescue medication meta-analyses. The risk ratio for at least 50% pain relief (based on TOTPAR) at six hours was 1.47 (95% confidence interval (CI) 1.28 to 1.69; five trials) favouring 400 mg ibuprofen over 1000 mg paracetamol, and the risk ratio for not using rescue medication (also favouring ibuprofen) was 1.50 (95% CI 1.25 to 1.79; four trials).The combined drug showed promising results, with a risk ratio for at least 50% of the maximum pain relief over six hours of 1.77 (95% CI 1.32 to 2.39) (paracetamol 1000 mg and ibuprofen 400 mg) (one trial; moderate quality evidence), and risk ratio not using rescue medication 1.60 (95% CI 1.36 to 1.88) (two trials; moderate quality evidence).The information available regarding adverse events from the studies (including nausea, vomiting, headaches and dizziness) indicated that they were comparable between the treatment groups. However, we could not formally analyse the data as it was not possible to work out how many adverse events there were in total. AUTHORS' CONCLUSIONS: There is high quality evidence that ibuprofen is superior to paracetamol at doses of 200 mg to 512 mg and 600 mg to 1000 mg respectively based on pain relief and use of rescue medication data collected at six hours postoperatively. The majority of this evidence (five out of six trials) compared ibuprofen 400 mg with paracetamol 1000 mg, these are the most frequently prescribed doses in clinical practice. The novel combination drug is showing encouraging results based on the outcomes from two trials when compared to the single drugs.

Systemic interventions for recurrent aphthous stomatitis (mouth ulcers).

BACKGROUND: Recurrent aphthous stomatitis (RAS) is the most frequent form of oral ulceration, characterised by recurrent oral mucosal ulceration in an otherwise healthy individual. At its worst RAS can cause significant difficulties in eating and drinking. Treatment is primarily aimed at pain relief and the promotion of healing to reduce the duration of the disease or reduce the rate of recurrence. A variety of topical and systemic therapies have been utilised. OBJECTIVES: To determine the clinical effect of systemic interventions in the reduction of pain associated with RAS, a reduction in episode duration or frequency. SEARCH METHODS: We undertook electronic searches of: Cochrane Oral Health Group and PaPaS Trials Registers (to 6 June 2012); CENTRAL via The Cochrane Library (to Issue 4, 2012); MEDLINE via OVID (1950 to 6 June 2012); EMBASE via OVID (1980 to 6 June 2012); CINAHL via EBSCO (1980 to 6 June 2012); and AMED via PubMed (1950 to 6 June 2012). We searched reference lists from relevant articles and contacted the authors of eligible trials to identify further trials and obtain additional information. SELECTION CRITERIA: We included randomised controlled trials (RCTs) in which the primary outcome measures assess a reduction of pain associated with RAS, a reduction in episode duration or a reduction in episode frequency. Trials were not restricted by outcome alone. We also included RCTs of a cross-over design. DATA COLLECTION AND ANALYSIS: Two review authors independently extracted data in duplicate. We contacted trial authors for details of randomisation, blindness and withdrawals. We carried out risk of bias assessment on six domains. We followed The Cochrane Collaboration statistical guidelines and risk ratio (RR) values were to be calculated using fixed-effect models (if two or three trials in each meta-analysis) or random-effects models (if four or more trials in each meta-analysis). MAIN RESULTS: A total of 25 trials were included, 22 of which were placebo controlled and eight made head-to-head comparisons (five trials had more than two treatment arms). Twenty-one different interventions were assessed. The interventions were grouped into two categories: immunomodulatory/anti-inflammatory and uncertain. Only one study was assessed as being at low risk of bias. There was insufficient evidence to support or refute the use of any intervention. AUTHORS' CONCLUSIONS: No single treatment was found to be effective and therefore the results remain inconclusive in regard to the best systemic intervention for RAS. This is likely to reflect the poor methodological rigour of trials, and lack of studies for certain drugs, rather than the true effect of the intervention. It is also recognised that in clinical practice, individual drugs appear to work for individual patients and so the interventions are likely to be complex in nature. In addition, it is acknowledged that systemic interventions are often reserved for those patients who have been unresponsive to topical treatments, and therefore may represent a select group of patients.

In-Vitro Phenotypic Response of Human Osteoblasts to Different Degrees of Titanium Surface Roughness.

Objectives: This study aimed to investigate human osteoblast (HOB) responses towards different degrees of titanium (Ti) implant surface roughness. Methods: Four degrees of Ti surface roughness were investigated on a micrometer roughness scale: smooth (S: 0.08−0.1 µm), minimally rough (MM: 0.3−0.5 µm), moderately rough (MR: 1.2−1.4 µm), and rough (R: 3.3−3.7 µm). HOB cells were cultured, expanded, and maintained according to the supplier's protocol. Cell proliferation and cytotoxicity were assessed at day 1, 3, 5, and 10 using alamarBlue and lactate dehydrogenase colorimetric assays. Data were analyzed with one-way ANOVA, two-way ANOVA, and Tukey's post hoc test (p = 0.05 for all tests). Results: There was no significant difference in the cell proliferation or cytotoxicity of the HOB cells in contact with the different degrees of Ti surface roughness. There was, however, a significant time effect on cell proliferation (p < 0.0001) with different exposure durations for each roughness degree. Furthermore, a positive correlation (non-significant) between proliferation and cytotoxicity was observed for all investigated degrees of surface roughness. Conclusion: All investigated roughness degrees showed comparable HOB proliferation, with the MR surface presenting the highest percentage, followed by the R, MM, ad S, surfaces, respectively. The S surface showed the highest cytotoxic effect on HOBs; however, it did not reach the cytotoxic level suggested by the ISO for any medical device to be considered cytotoxic.

Human osteoblasts response to different dental implant abutment materials: An in-vitro study.

OBJECTIVES: This study aimed to investigate human osteoblasts (HOB) response towards different dental implant abutment materials. METHODS: Five dental implant abutment materials were investigated: (1) titanium (Ti), (2) titanium coated nitride (TiN), (3) cobalt chromium (CoCr), (4) zirconia (ZrO2), and (5) modified polyether ether ketone (m-PEEK). HOBs were cultured, expanded, and seeded according to the supplier's protocol (PromoCell, UK). Cell proliferation and cytotoxicity were evaluated at days 1, 3, 5, and 10 using Alamar Blue (alamarBlue) and lactate dehydrogenase (LDH) colorimetric assays. Data were analysed via two-way ANOVA, one-way ANOVA and Tukey's post hoc test (significance was determined as p < 0.05 for all tests). RESULTS: All the investigated materials showed high and comparable initial proliferation activities apart from ZrO2 (46.92%), with P% of 79.91%, 68.77%, 73.20%, and 65.46% for Ti, TiN, CoCr, and m-PEEK, respectively. At day 10, all materials exhibited comparable and lower P% than day 1 apart from TiN (70.90%) with P% of 30.22%, 40.64%, 37.27%, and 50.65% for Ti, CoCr, ZrO2, and m-PEEK, respectively. The cytotoxic effect of the investigated materials was generally low throughout the whole experiment. At day 10, the cytotoxicity % was 7.63%, 0.21%, 13.30%, 5.32%, 8.60% for Ti, TiN, CoCr, ZrO2, and m-PEEK. The Two-way ANOVA and Tukey's Multiple Comparison Method highlighted significant material and time effects on cell proliferation and cytotoxicity, and a significant interaction (p < 0.0001) between the tested materials. Notably, TiN and m-PEEK showed improved HOB proliferation activity and cytotoxic levels than the other investigated materials. In addition, a non-significant negative correlation between viability and cytotoxicity was found for all tested materials. Ti (p = 0.07), TiN (p = 0.28), CoCr (p = 0.15), ZrO2 (p = 0.17), and m-PEEK (p = 0.12). SIGNIFICANCE: All the investigated materials showed excellent biocompatibility properties with more promising results for the newly introduced TiN and m-PEEK as alternatives to the traditionally used dental implant and abutment materials.

Two Gingival Cell Lines Response to Different Dental Implant Abutment Materials: An In Vitro Study.

Objectives: This study aimed to investigate the response of human gingival fibroblasts (HGFB) and human gingival keratinocytes (HGKC) towards different dental implant abutment materials. Methods: Five materials were investigated: (1) titanium (Ti), (2) titanium nitride (TiN), (3) cobalt-chromium (CoCr), (4) zirconia (ZrO2), and (5) modified polyether ether ketone (m-PEEK). Both cell lines were cultured, expanded, and seeded in accordance with the protocol of their supplier. Cell proliferation and cytotoxicity were evaluated at days 1, 3, 5, and 10 using colourimetric viability and cytotoxicity assays. Data were analysed via two-way ANOVA, one-way ANOVA, and Tukey's post hoc test (p < 0.05 for all tests). Results: There was a statistically significant difference in cell proliferation of HGKC and HGFB cells in contact with different abutment materials at different time points, with no significant interaction between different materials. There was a significant effect on cell proliferation and cytotoxicity with different exposure times (p < 0.0001) for each material. Cell proliferation rates were comparable for both cell lines at the beginning of the study, however, HGFB showed higher proliferation rates for all materials at day 10 with better proliferation activities with ZrO and m-PEEK (40.27%) and (48.38%) respectively. HGKC showed significant interactions (p < 0.0001) in cytotoxicity between different materials. Conclusion: The present in vitro assessment investigated the biocompatibility of different abutment materials with soft tissue cells (HGFB and HGKC). The findings suggest that m-PEEK and TiN are biologically compatible materials with human cells that represent the soft tissue and can be considered as alternative implant abutment materials to Ti and ZrO2, especially when the aesthetic is of concern.

Biological response of epithelial and connective tissue cells to titanium surfaces with different ranges of roughness: An in-vitro study.

OBJECTIVES: This study aimed to investigate how titanium (Ti) surface with different range roughness created by industrial machining influence the biological response of primary human gingival fibroblasts (HGFB) and keratinocytes (HGKC) in terms of cell proliferation and cytotoxicity. METHODS: Four Ti surfaces of different roughness ranges were investigated: smooth (S: 0.08-0.1 µm), minimally rough (MM: 0.3-0.5 µm), moderately rough (MR: 1.2-1.4 µm) and rough (R: 3.3-3.7 µm). Discs topography and surface roughness were evaluated by scanning electron microscopy (SEM) and non-contact profilometer. Both cell lines were cultured, expanded, and maintained according to their supplier's protocols. Cell proliferation and cytotoxicity were evaluated at days 1, 3, 5, and 10 using cell viability and cytotoxicity colorimetric assays. Data were analysed via two-way ANOVA, one-way ANOVA and Tukey's post hoc test (p = 0.05 for all tests). RESULTS: Both cell lines showed comparable initial proliferation activity of 70-86% for all the investigated roughnesses. HGKC showed better and higher proliferation % with S surface at all time points than all the other investigated surfaces which was significantly higher than MM at day 3 and higher than all the other investigated surfaces at day 5 and 10. On the other hand, HGFB exhibited the best proliferation with both MM and R surfaces with no significant differences from the other two surfaces (S and MR). Different surface roughnesses and exposure times showed significant effect on cell proliferation in both cell lines. Cytotoxicity for both cell lines was generally the highest on day 3, with the following order from highest to lowest: S (19.86%)> R> MR> MM for HGKC and MM (39.48%)> MR> S> R for HGFB. Different exposure times showed a significant effect on cell cytotoxicity in both cell lines and a significant effect of surface roughness in HGFB. SIGNIFICANCE: All investigated roughness levels were sufficiently biologically compatible with cells representative of the major population of the soft tissue surrounding dental implants. However, the S surface was most cytotoxic to HGKC, while the MM surface was most cytotoxic to HGFB cells.

Schwannoma presenting in the soft palate of a nine-year-old boy.

Swellings and tumours within the oral cavity are a common finding, however, benign intra-oral schwannoma or neurolemma is relatively uncommon, especially in younger patients. Involvement of the palate is a rare presentation although there have been a few reported cases relating to the lingual and other tissues. This paper reviews intra-oral schwannomas and presents a case of such a tumour of the soft palate in a paediatric patient and discusses the presenting features, differential diagnoses, along with the management of the condition.

Clinically applicable artificial intelligence system for dental diagnosis with CBCT.

In this study, a novel AI system based on deep learning methods was evaluated to determine its real-time performance of CBCT imaging diagnosis of anatomical landmarks, pathologies, clinical effectiveness, and safety when used by dentists in a clinical setting. The system consists of 5 modules: ROI-localization-module (segmentation of teeth and jaws), tooth-localization and numeration-module, periodontitis-module, caries-localization-module, and periapical-lesion-localization-module. These modules use CNN based on state-of-the-art architectures. In total, 1346 CBCT scans were used to train the modules. After annotation and model development, the AI system was tested for diagnostic capabilities of the Diagnocat AI system. 24 dentists participated in the clinical evaluation of the system. 30 CBCT scans were examined by two groups of dentists, where one group was aided by Diagnocat and the other was unaided. The results for the overall sensitivity and specificity for aided and unaided groups were calculated as an aggregate of all conditions. The sensitivity values for aided and unaided groups were 0.8537 and 0.7672 while specificity was 0.9672 and 0.9616 respectively. There was a statistically significant difference between the groups (p = 0.032). This study showed that the proposed AI system significantly improved the diagnostic capabilities of dentists.

Response of two gingival cell lines to CAD/CAM composite blocks.

OBJECTIVE: This study aimed to investigate the influence of CAD/CAM composite materials on human gingival fibroblasts (HGF) and gingival keratinocytes (HGK). METHODS: Four materials were investigated: two resin-composite blocks (RCB), Grandio Blocs (GR) and Block HC (HC); one polymer-infiltrated ceramic network (PICN) (Enamic, EN); and one conventional resin-composite, Grandioso (GND). HGF and HGK were cultured as per the supplier's protocol (ATCC, UK). Cell proliferation and cytotoxicity were evaluated at 1, 3, 5 and 10 days using LDH and Alamar Blue assays. Indirect immunostaining was used to assess the Caspase-3 activity. Data were analysed via two-way ANOVA, one-way ANOVA and Tukey's post hoc test (α = 0.05 for all tests). RESULTS: There was significant difference in cell proliferation of the HGK and HGF cells in contact with different composite materials but no significant differences in their cytotoxicity. There was a significant effect on cell proliferation and cytotoxicity with different exposure times, for each type of resin-composite. HGF cell proliferation was higher than HGK with almost all investigated materials and at all time points. No Caspase-3 activity was detected in either cell lines. SIGNIFICANCE: HGK proliferation and cytotoxicity appeared to be more influenced by composite materials compared to HGF, demonstrating EN cytotoxic effects in HGK. Different manufacturing techniques of resin-composites (photo curing versus heat/pressure curing) had no significant effect on their biocompatibility.

P2X(3) expression is not altered by lingual nerve injury.

We have investigated a possible role for the ATP receptor subunit P2X(3), in the development of neuropathic pain following injury to a peripheral branch of the trigeminal nerve. In nine anaesthetised adult ferrets the left lingual nerve was sectioned and recovery permitted for 3 days, 3 weeks or 3 months (3 ferrets per group). A retrograde tracer, fluorogold, was applied to the nerve to allow identification of cell bodies in the trigeminal ganglion with axons in the injured nerve. Indirect immunofluorescence for P2X(3) and image analysis was used to quantify the percentage area of staining at the site of injury. Additionally, the proportion of fluorogold-positive cells that expressed P2X(3) was determined and compared with expression in non-fluorogold containing cells in another part of the ganglion. Comparisons were made with results from control animals that only received the tracer injection. After lingual nerve injury there was no significant change in P2X(3) expression at the site of nerve injury or within cell bodies linked to either injured (lingual) or uninjured (ophthalmic) axons, at any of the time periods investigated. Overall, this study suggests that P2X(3) expression at these sites is not involved in the development of neuropathic pain following lingual nerve injury.

Effect of SB-750364, a specific TRPV1 receptor antagonist, on injury-induced ectopic discharge in the lingual nerve.

Abnormal neural activity generated at a site of nerve injury is thought to contribute to the development of dysaesthesia. Vanilloid receptor 1 (TRPV1), a transducer of noxious stimuli, may be involved in the initiation of this abnormal activity and could provide a useful therapeutic target. We investigated the effect of a specific TRPV1 antagonist (SB-750364) on injury-induced discharge in the lingual nerve. In 12 anaesthetised adult ferrets the left lingual nerve was sectioned and animals were allowed to recover for 3-7 days. In terminal experiments under general anaesthesia, the nerve was re-exposed and electrophysiological recordings made from spontaneously active axons in fine filaments dissected from the nerve central to both the injury site and the junction with the chorda tympani. SB-750364 was infused via the cephalic vein in order to achieve three increasing but stable systemic blood levels of the compound (0.3, 1.0 and 3.0 microM). Twenty-eight spontaneously active units were studied, with discharge frequencies ranging from 0.02 to 4.9 Hz. There was a significant reduction in spontaneous activity in 17 units (61%) at 1.0 microM or less of SB-750364 (p<0.01; Friedman test with Dunn's multiple comparisons). A further 4 units (14%) showed a significant reduction in activity at 3.0 microM (p<0.01). In the remaining 7 units (25%) the discharge was unaffected (p>0.05). These data show that the TRPV1 antagonist SB-750364 can reduce the level of spontaneous activity initiated in some axons following lingual nerve injury.

A simple rocker-induced mechanical stimulus upregulates mineralization by human osteoprogenitor cells in fibrous scaffolds.

Biodegradable electrospun polycaprolactone scaffolds can be used to support bone-forming cells and could fill a thin bony defect, such as in cleft palate. Oscillatory fluid flow has been shown to stimulate bone production in human progenitor cells in monolayer culture. The aim of this study was to examine whether bone matrix production by primary human mesenchymal stem cells from bone marrow or jaw periosteal tissue could be stimulated using oscillatory fluid flow supplied by a standard see-saw rocker. This was investigated for cells in two-dimensional culture and within electrospun polycaprolactone scaffolds. From day 4 of culture onwards, samples were rocked at 45 cycles/min for 1 h/day, 5 days/week (rocking group). Cell viability, calcium deposition, collagen production, alkaline phosphatase activity and vascular endothelial growth factor secretion were evaluated to assess the ability of the cells to undergo bone differentiation and induce vascularisation. Both cell types produced more mineralized tissue when subjected to rocking and supplemented with dexamethasone. Mesenchymal progenitors and primary human mesenchymal stem cells from bone marrow in three-dimensional scaffolds upregulated mineral deposition after rocking culture as assessed by micro-computed tomography and alizarin red staining. Interestingly, vascular endothelial growth factor secretion, which has previously been shown to be mechanically sensitive, was not altered by rocking in this system and was inhibited by dexamethasone. Rocker culture may be a cost effective, simple pretreatment for bone tissue engineering for small defects such as cleft palate.

Vanilloid receptor 1 (TRPV1) expression in lingual nerve neuromas from patients with or without symptoms of burning pain.

The lingual nerve, a peripheral branch of the trigeminal nerve, can be damaged during the surgical removal of lower third molar teeth. This damage can lead to the development of dysaesthesia, with some patients complaining of burning pain. We investigated the hypothesis that vanilloid receptor 1 (TRPV1), a transducer of noxious heat stimuli, was involved in the development of this burning pain. Neuroma specimens were obtained from patients undergoing microsurgical repair of a damaged lingual nerve. Repair was undertaken where there was little evidence of spontaneous recovery, 7-41 months after the initial injury. Preoperatively the incidence of dysaesthesia was determined by reported symptoms and using visual analogue scales (VAS) for pain, tingling and discomfort. Nine neuromas were studied from patients with burning dysaesthesia and six from patients with a sensory deficit but no dysaesthesia. Indirect immunofluorescence for protein gene product (PGP) 9.5 and TRPV1 was used to quantify the percentage area of PGP 9.5 positive neuronal tissue that also expressed TRPV1. The results showed no significant difference between the mean percentage area of TRPV1 expression in neuromas from patients with or without burning dysaesthesia. Furthermore, there was no correlation between TRPV1 expression and the VAS scores for pain, tingling or discomfort. However, if data from all patients was pooled, there was a negative correlation between the level of TRPV1 expression and the time after initial injury. These data do not rule out involvement of TRPV1 in the aetiology of burning dysaesthesia following lingual nerve injury but suggest that TRPV1 at the injury site does not play a primary role.

Changes in vanilloid receptor 1 (TRPV1) expression following lingual nerve injury.

We have investigated a possible role for vanilloid receptor 1 (TRPV1), a transducer of noxious stimuli, in the development of neuropathic pain following injury to a peripheral branch of the trigeminal nerve. In nine adult ferrets the left lingual nerve was sectioned and recovery permitted for 3 days, 3 weeks or 3 months (3 ferrets per group). A retrograde tracer, fluorogold, was injected into the damaged nerve to identify associated cell bodies in the trigeminal ganglion. Three further ferrets, receiving only tracer injection, served as uninjured controls. Indirect immunofluorescence for TRPV1 and image analysis was used to quantify the percentage area of staining (PAS) of TRPV1 in the left and right lingual nerves. Additionally, the proportion of fluorogold positive and fluorogold negative cells expressing TRPV1 in the ganglion was determined. TRPV1 expression increased significantly at the injury site of damaged nerves 3 days after injury and this was matched by a reduction in the proportion of fluorogold positive cells expressing TRPV1 in the ganglion. At 3 weeks TRPV1 expression at the injury site was still high, while in the ganglion was significantly greater than in the controls. In the 3-month recovery group TRPV1 expression in both nerve fibres and ganglion cells, was not significantly different from controls and there were no changes in expression in the fluorogold negative cells in the ganglion at any time point studied. These data suggest that after injury there is an increase in the axonal transport of TRPV1 from the cell bodies to the damaged axons and this is followed by an increase in synthesis in the ganglion. These changes in expression may be involved in development of sensory disturbances or dysaesthesia after injury.

Regeneration of Corneal Epithelium With Dental Pulp Stem Cells Using a Contact Lens Delivery System.

PURPOSE: The corneal epithelium is sloughed off surface of the eye by the action of blinking and is continually replaced by division and maturation of the limbal stem cells (LSCs). In the case of injury or disease, LSCs can be lost or damaged to a point at which the corneal epithelial layer is no longer maintained. leading to LSC deficiencies (LSCDs). When this occurs, the opaque conjunctiva overgrows the anterior surface of the eye, leading to vision impairment or loss. Dental pulp stem cells (DPSCs) are promising candidates as autologous LSC substitutes. In this study, contact lenses (CLs) are used as a novel medical device to deliver DPSCs onto corneal surface to enhance corneal epithelium regeneration. METHODS: Dental pulp stem cells labeled with green fluorescent Qtracker 525 were seeded onto the pretreated CLs, allowed to adhere, then delivered to debrided human corneas. Expression of KRT3, 12, 13, and 19 was investigated by immunostaining, then standard and confocal microscopy. RESULTS: Dental pulp stem cells were successfully isolated, labeled, and delivered to the corneal surface using CLs. Following removal of CLs, confocal microscopy showed that the DPSCs had migrated onto the cornea. Coexpression of KRT12 and green fluorescent Qtracker 525 confirmed that the DPSCs had transdifferentiated into corneal epithelial progenitors. Delimitation of KRT 19 and green fluorescence provides evidence that Qtracker 525-labeled DPSCs establish a barrier to the invasion of the cornea by conjunctiva. CONCLUSIONS: In this study we show that DPSCs, delivered using CLs, can be used to enhance repair and regeneration of the human corneal epithelium.

The effect of carbamazepine on injury-induced ectopic discharge in the lingual nerve.

Previous studies have shown that the development of ectopic activity from damaged axons following nerve injury may contribute to the aetiology of sensory disturbances, including dysaesthesia. Pharmacological manipulation of this activity could provide a method of treatment for this intractable condition. In this study we have investigated the effect of carbamazepine, an anti-convulsant, as it is known to have membrane stabilising properties. In eight anaesthetised adult ferrets the left lingual nerve was sectioned and the animals allowed to recover for 3 days. Then, in terminal experiments under general anaesthesia, the nerve was re-exposed and electrophysiological recordings were made from spontaneously active units in fine filaments dissected from the nerve proximal to the injury site. Carbamazepine in a modified cyclodextrin (hydroxypropyl-beta-cyclodextrin) was administered intravenously in increments, in order to achieve a progressively increasing systemic concentration, and serum levels were determined at the point that activity ceased. Twenty-one spontaneously active units were studied, with conduction velocities of 2.1-28.9 m s(-1) and discharge frequencies of 0.25-15.3 Hz. Spontaneous activity ceased in 13 units with a serum concentration of carbamazepine ranging from 3.5 to 8.4 mg/l, which was within the normal therapeutic range (4-12 mg/l). Four units ceased activity with carbamazepine levels above the therapeutic range (15.4-17.2 mg/ml), but the remaining four continued to discharge throughout the recording period. These data suggest that systemic carbamazepine can reduce the level of spontaneous activity initiated in some axons following lingual nerve injury.