Titania nanotube-based protein delivery system to inhibit cranial bone regeneration in Crouzon model of craniosynostosis.

BACKGROUND: Craniosynostosis is a developmental disorder characterized by the premature fusion of skull sutures, necessitating repetitive, high-risk neurosurgical interventions throughout infancy. This study used protein-releasing Titania nanotubular implant (TNT/Ti) loaded with glypican 3 (GPC3) in the cranial critical-sized defects (CSDs) in Crouzon murine model (Fgfr2c342y/+ knock-in mutation) to address a key challenge of delaying post-operative bone regeneration in craniosynostosis. MATERIALS AND METHODS: A 3 mm wide circular CSD was created in two murine models of Crouzon syndrome: (i) surgical control (CSDs without TNT/Ti or any protein, n=6) and (ii) experimental groups with TNT/Ti loaded with GPC3, further subdivided into the presence or absence of chitosan coating (on nanotubes) (n=12 in each group). The bone volume percentage in CSDs was assessed 90 days post-implantation using micro-computed tomography (micro-CT) and histological analysis. RESULTS: Nano-implants retrieved after 90 days post-operatively depicted well-adhered, hexagonally arranged, and densely packed nanotubes with average diameter of 120±10 nm. The nanotubular architecture was generally well-preserved. Compared with the control bone volume percentage data (without GPC3), GPC3-loaded TNT/Ti without chitosan coating displayed a significantly lower volume percent in cranial CSDs (P<0.001). Histological assessment showed relatively less bone regeneration (healing) in GPC3-loaded CSDs than control CSDs. CONCLUSION: The finding of inhibition of cranial bone regeneration by GPC3-loaded TNT/Ti in vivo is an important advance in the novel field of minimally-invasive craniosynostosis therapy and holds the prospect of altering the whole paradigm of treatment for affected children. Future animal studies on a larger sample are indicated to refine the dosage and duration of drug delivery across different ages and both sexes with the view to undertake human clinical trials.

Glypican-based drug releasing titania implants to regulate BMP2 bioactivity as a potential approach for craniosynostosis therapy.

Advances in molecular biology and nanomedicine based therapies hold promise to obviate the need of multiple surgical interventions (associated with current management) in craniosynostosis by preventing bone re-ossification. One such adjunctive therapy involves application of glypicans 1 and 3 (GPC1 and GPC3) that are BMP inhibitors implicated in downregulating the BMP2 activity in prematurely fusing sutures. Electrochemically anodized Titania nanotube (TNT) arrays have been recognized as a promising localized, long-term drug delivery platform for bone-related therapies. This study presents the application of nanoengineered TNT/Ti implants loaded with recombinant glypicans for craniosynostosis therapy. By using Dual luciferase Reporter assay, we tested the biofunctionality of eluted glypicans from the TNT/Ti implants for BMP2 bioactivity regulation in C2C12 murine myoblast cell line. BMP2 activity was inhibited significantly for up to 15days by the glypicans released from polymer-coated TNT/Ti implants, indicating their potential application in adjunctive craniosynostosis treatment.

Biological response of human suture mesenchymal cells to Titania nanotube-based implants for advanced craniosynostosis therapy.

Titania nanotubes (TNTs) engineered on titanium (Ti) surfaces (i.e. TNT/Ti) and loaded with specific drugs have been recognised as a promising solution for localised therapeutic delivery to address several medical problems not feasible with conventional drug administration. We propose the use of TNT/Ti protein-releasing implants to treat paediatric craniofacial abnormality in craniosynostosis caused by premature fusion of cranial sutures. In this study, we have analysed the biological response of human suture mesenchymal cells (SMCs), extracted from two different patients undergoing craniofacial reconstruction surgery, at the TNT/Ti implant surface. The experimental groups included large-diameter TNT/Ti implants, with and without biopolymer surface coating (Chitosan and Pluronic-F127) while the controls comprised of flat Ti disc and tissue culture plastic. The non-loaded implant surfaces and the cellular interactions at the implant-cell interface were characterised using scanning electron microscopy (SEM). The SMC adhesion, viability and proliferation were determined by MTT assay and manual cell counting at day 1 and day 3 of cell incubation. SEM showed significant reduction in initial attachment and adhesion of SMCs at TNT-cell biointerface compared with the control Ti discs. Subsequent cell proliferation results also revealed a decrease in the number of viable cells on the TNT surfaces. The nanotopography and structural features along with the surface chemistry dictated the cellular response, with nanotubular surfaces (with and without polymer coating) impeding cell adhesion and proliferation. Our findings hold promise for the use of TNT-based cranial implants as a delivery system to prevent sutural bone growth for advanced craniosynostosis therapy.

Three-dimensional profilometric assessment of early enamel erosion simulating gastric regurgitation.

OBJECTIVES: A priority research area in minimal intervention dentistry is the characterization of the early stages of dental erosion. The aim of this in vitro study was to assess the effect of short, repetitive erosive challenges to human enamel over 2 min at pH 1.5 and 3.0 under conditions simulating gastric regurgitation. METHODS: Enamel surfaces were subjected to erosive challenges at pH 1.5 (Group 1, n=10) and pH 3.0 (Group 2, n=9) for periods of 30s (stage 1), 60s (stage 2) and 120 s (stage 3). Quantitative changes were assessed longitudinally by measuring the 3D average surface roughness (Sa) values using 3D confocal microscopy. Qualitative micrographic assessment of surface changes was also conducted by using environmental scanning electron microscopy. RESULTS: Linear mixed model analysis showed significant effects of the pH values (p<0.001) and the stages (p<0.001) on the observed Sa values. Post hoc tests showed significant increases in the Sa values between baseline and other stages in both groups (p<0.01). The mean Sa values also increased significantly from stage 1 to stage 2 in Group 1 (p<0.05). Micrographic analysis displayed severely etched enamel rods in Group 1, but only subtle changes in Group 2. CONCLUSIONS: The complexity of the enamel surface is influenced by both acid concentration (pH value) and duration of acid exposure during early stages of erosion. Erosion occurring under conditions simulating GORD can be detected in its initial stages, opening up the possibilities of early diagnosis and management of this condition. CLINICAL SIGNIFICANCE: Erosive tooth wear occurs progressively and insidiously, often creating complex treatment challenges. This emphasizes the need for early diagnosis and management in accordance with minimal intervention philosophy. Our findings provide a foundation for further research that could lead to the development of highly-sensitive clinical diagnostic tools and preventive strategies.

Oral manifestations of gastroesophageal reflux disease.

Numerous case-control and other studies involving confirmation of gastroesophageal reflux disease (GERD) by esophageal pH-metry and the assessment of dental erosions have shown significant associations between the two conditions in both adults and children. By contrast, when asked to vote on whether GERD may cause dental erosions, only 42% of physicians strongly agreed that such an association existed in adults, and just 12.5% strongly agreed for children, respectively in two global consensus reports. Part of this divergence between the perceptions of physicians and the findings of research publications may reflect a general lack of oral health education during medical training, and cursory oral examinations being made under less-than-ideal conditions. Adequate salivary secretions are essential for the protection of the teeth and the oropharyngeal and esophageal mucosa. The quantity and quality of the saliva require monitoring as many drugs, including several of the proton pump inhibitors (PPIs), can cause hyposalivation. In addition, PPIs do not always result in adequate acid suppression. Therefore, collaboration between physicians and dentists is strongly advocated to prevent or ameliorate possible adverse oral effects from both endogenous and exogenous acids, and to promote adequate saliva production in patients with GERD.

Gastroesophageal reflux disease and tooth erosion.

The increasing prevalence of gastroesophageal reflux disease (GERD) in children and adults, and of "silent refluxers" in particular, increases the responsibility of dentists to be alert to this potentially severe condition when observing unexplained instances of tooth erosion. Although gastroesophageal reflux is a normal physiologic occurrence, excessive gastric and duodenal regurgitation combined with a decrease in normal protective mechanisms, including an adequate production of saliva, may result in many esophageal and extraesophageal adverse conditions. Sleep-related GERD is particularly insidious as the supine position enhances the proximal migration of gastric contents, and normal saliva production is much reduced. Gastric acid will displace saliva easily from tooth surfaces, and proteolytic pepsin will remove protective dental pellicle. Though increasing evidence of associations between GERD and tooth erosion has been shown in both animal and human studies, relatively few clinical studies have been carried out under controlled trial conditions. Suspicion of an endogenous source of acid being associated with observed tooth erosion requires medical referral and management of the patient as the primary method for its prevention and control.

An anthropological perspective: another dimension to modern dental wear concepts.

For many years, research on tooth wear by dental academics has been diametrically opposite to that of anthropological research, with each discipline having a different understanding as to the nature of the wear processes. Dental focus revolved around preventive and restorative considerations while the anthropological focus was a biological understanding related to human evolution, diet, environment, form, and function and included all the craniofacial structures. Introducing the anthropological perspective into modern dentistry gives an insight into the "bigger picture" of the nature and extent of tooth wear. By combining anthropological evidence with clinical knowledge and experience, it is most likely to provide the best-informed and biologically based approach to the management of tooth wear in modern societies.

The effect of CPP-ACP on enamel wear under severe erosive conditions.

OBJECTIVE: In addition to its role as a remineralizing agent in preventing dental caries, recent evidence has shown that casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) can protect teeth against erosion. The aim of this study was to determine whether CPP-ACP could reduce enamel wear rates under severe erosive conditions simulating heavy attrition and gastric regurgitation. DESIGN: Enamel specimens were subjected to 10,000 wear cycles at a load of 100 N and pH 1.2 in a tooth wear machine. The machine was stopped every 2 min (160 cycles), and CPP-ACP in the form of a paste was applied for 5 min in experimental group 1. A paste with the same formulation but without CPP-ACP was applied in experimental group 2. No paste was applied in the control group. RESULTS: A linear mixed model analysis indicated that the mean wear rates in experimental group 1 (0.44+/-0.05 mm(3) per 1000 cycles) and in experimental group 2 (0.63+/-0.06 mm(3) per 1000 cycles) were significantly lower than that in the control group (0.92+/-0.11 mm(3) per 1000 cycles) (p<0.05). The mean wear rate in experimental group 1 was also lower than that in experimental group 2 (p<0.05). Wear facets in experimental groups 1 and 2 were noted to be smoother and more polished than those in the control group. CONCLUSIONS: Both remineralizing and lubricating properties of the paste containing CPP-ACP appear to contribute to wear reduction in enamel. These findings may lead to new strategies for the clinical management of tooth wear.

An in vitro assessment of the effect of load and pH on wear between opposing enamel and dentine surfaces.

OBJECTIVE: Previous in vitro studies have described the wear characteristics of specimens in which enamel has been opposed to enamel and dentine opposed to dentine. The aim of this study was to assess the characteristics of wear between specimens in which enamel was opposed to dentine at loads simulating attrition and at pH values simulating different erosive environments. It was hypothesized that enamel would wear more slowly than dentine under all conditions. DESIGN: Opposing enamel and dentine specimens from 57 human third molar teeth were worn in electromechanical machines with various loads (32, 62 and 100 N) and lubricants (pH 1.2, 3.0 and 6.1). Tooth wear was quantified by measuring reduction in dentine volume over time using a 3D profilometer. Qualitative assessment was also carried out using scanning electron microscopy. RESULTS: Dentine wear increased with increasing load, and dentine wear was faster at pH 1.2 than at pH 3.0 or 6.1 for all loads tested. Interestingly, enamel wore more rapidly than dentine at pH 1.2 under all loads. At pH values of 3.0 and 6.1, enamel wear rates were not measurably different from zero and they were less than wear rates for opposing dentine specimens at all loads. Micrographic assessment showed extensive surface destruction of dentine wear facets due to erosion at pH 1.2. Dentine wear facets were smoother at pH 3.0 that at pH 6.1. CONCLUSIONS: When enamel wears against dentine in an acidic environment enamel will wear more rapidly at very low pH, while under less acid conditions dentine will wear faster than enamel.

Tooth wear: the view of the anthropologist.

Anthropologists have for many years considered human tooth wear a normal physiological phenomenon where teeth, although worn, remain functional throughout life. Wear was considered pathological only if pulpal exposure or premature tooth loss occurred. In addition, adaptive changes to the stomatognathic system in response to wear have been reported including continual eruption, the widening of the masticatory cycle, remodelling of the temporomandibular joint and the shortening of the dental arches from tooth migration. Comparative studies of many different species have also documented these physiological processes supporting the idea of perpetual change over time. In particular, differential wear between enamel and dentine was considered a physiological process relating to the evolution of the form and function of teeth. Although evidence of attrition and abrasion has been known to exist among hunter-gatherer populations for many thousands of years, the prevalence of erosion in such early populations seems insignificant. In particular, non-carious cervical lesions to date have not been observed within these populations and therefore should be viewed as 'modern-day' pathology. Extrapolating this anthropological perspective to the clinical setting has merits, particularly in the prevention of pre-mature unnecessary treatment.

Differential wear of teeth and restorative materials: clinical implications.

PURPOSE: This study reviewed the wear of commonly used dental restorative materials and their effects on the opposing dentition. MATERIALS AND METHODS: Key words were used with PubMed to retrieve pertinent references to publications on tooth and restoration wear. RESULTS: The wear resistance of newer esthetic restorative materials has generally improved, and the damage caused by several materials to the opposing dentition has been reduced. However, the different structures and physical properties of tooth substance and restorative materials will eventually lead to varying degrees of differential wear. The extent and rate of wear are influenced by many intraoral factors. CONCLUSION: Selection of restorative materials must be based on knowledge of their wear behavior and the individual needs of each patient. The lowest wear rates for restorations and the opposing dentition occur with metal alloys, machined ceramics, and microfilled and microfine hybrid resin composites.

Case report: management of tooth tissue loss from intrinsic acid erosion.

Acid erosive tooth wear is increasingly being reported in many western countries, and is also being recognised as a significant cause of tooth destruction in persons with xerostomia. The primary aetiology is related to a high consumption of dietary sources of acids and also to an increasing awareness of gastric acid as a significant factor in both children and adults. Recommended preventive dental treatments aim to neutralise the effects of acids, reduce the severity of xerostomia, stimulate salivary flow and buffering capacity, and increase the acid resistance of tooth substance. Initial restorative treatments should be conservative, using adhesive dentistry techniques. This treatment approach is illustrated by the oral rehabilitation of a severe erosion case using adhesive onlays, veneers and all ceramic crowns.

The diagnosis and control of extrinsic acid erosion of tooth substance.

As modern societies increase their acidic food and beverage consumption, erosive tooth wear is an increasing cause of tooth destruction among young people. The primary causes can be exacerbated by xerostomia, which can be induced by dehydration, several systemic diseases, or any of a number of drugs. Initial preventive treatments are directed at eliminating extrinsic acids, reducing xerostomia and other co-factors, and increasing teeth's resistance to acid. Initial restorative treatments should be conservative, using adhesive materials. Treatment of advanced tooth tissue loss is difficult; preventive management is emphasized.

Dahl appliances used for the restorative management of localized anterior tooth erosion.

Dahl partial bite-raising appliances have been used over the past 30 years for the management of occlusal tooth tissue loss, particularly in patients who have lost normal occlusal vertical dimension. The more recent use of Dahl-type appliances for individual teeth has changed the restorative management of patients with localized anterior tooth erosion. The Dahl treatment principle can be applied to those patients with and without the loss of occlusal vertical dimension. Palatal bite-raising platforms can be used to re-establish anterior tooth guidance for disocclusion of the posterior teeth without occlusal interferences. The treatment principle is illustrated with clinical cases.

Management of tooth tissue loss from erosion.

Erosive tooth wear appears to be no less frequent in the United States than in the United Kingdom and Europe and is of increasing concern as a significant cause of tooth destruction in younger persons. Consumption of numerous dietary sources of acids is increasing in modern societies. In addition, involuntary regurgitation may be a significant cause of tooth erosion. These primary causes can be exacerbated by xerostomia, which is induced by many drugs. Initial preventive treatments are directed at neutralizing the effects of the acids, and initial restorative treatments should be conservative, using adhesive materials. Treatment of advanced tooth tissue loss is difficult and expensive, and preventive management is emphasized.