Paediatric endodontics Part 2: Pulp regeneration: current approaches.

Dental pulp regeneration: An overview of the current approaches. Regenerative Endodontic Procedures (REPs) are biologically based procedures aimed at restoring the damaged structures and physiological functions of the pulp-dentine complex. Clinically, two strategies have been proposed so far to induce REP: cell transplantation and cell homing. REPs success relies primarily on the clinical and biological conditions of the tooth; therefore, cell homing strategies will not be consistently successful in every condition. Root canal treatment remains the standard of care for mature teeth with necrotic pulps and closed apex.

Dental pulp regeneration via cell homing.

The typical treatment for irreversibly inflamed/necrotic pulp tissue is root canal treatment. As an alternative approach, regenerative endodontics aims to regenerate dental pulp-like tissues using two possible strategies: cell transplantation and cell homing. The former requires exogenously transplanted stem cells, complex procedures and high costs; the latter employs the host's endogenous cells to achieve tissue repair/regeneration, which is more clinically translatable. This systematic review examines cell homing for dental pulp regeneration, selecting articles on in vitro experiments, in vivo ectopic transplantation models and in situ pulp revascularization. MEDLINE/PubMed and Scopus databases were electronically searched for articles without limits in publication date. Two reviewers independently screened and included papers according to the predefined selection criteria. The electronic searches identified 46 studies. After title, abstract and full-text examination, 10 articles met the inclusion criteria. In vitro data highlighted that multiple cytokines have the capacity to induce migration, proliferation and differentiation of dental pulp stem/progenitor cells. The majority of the in vivo studies obtained regenerated connective pulp-like tissues with neovascularization. In some cases, the samples showed new innervation and new dentine deposition. The in situ pulp revascularization regenerated intracanal pulp-like tissues with neovascularization, innervation and dentine formation. Cell homing strategies for pulp regeneration need further understanding and improvement if they are to become a reliable and effective approach in endodontics. Nevertheless, cell homing currently represents the most clinically viable pathway for dental pulp regeneration.

Styrene enhances the noise induced oxidative stress in the cochlea and affects differently mechanosensory and supporting cells.

Experimental and human investigations have raised the level of concern about the potential ototoxicity of organic solvents and their interaction with noise. The main objective of this study was to characterize the effects of the combined noise and styrene exposure on hearing focusing on the mechanism of damage on the sensorineural cells and supporting cells of the organ of Corti and neurons of the ganglion of Corti. The impact of single and combined exposures on hearing was evaluated by auditory functional testing and histological analyses of cochlear specimens. The mechanism of damage was studied by analyzing superoxide anion and lipid peroxidation expression and by computational analyses of immunofluorescence data to evaluate and compare the oxidative stress pattern in outer hair cells versus the supporting epithelial cells of the organ of Corti. The oxidative stress hypothesis was further analyzed by evaluating the protective effect of a Coenzyme Q10 analogue, the water soluble Qter, molecule known to have protective antioxidant properties against noise induced hearing loss and by the analysis of the expression of the endogenous defense enzymes. This study provides evidence of a reciprocal noise-styrene synergism based on a redox imbalance mechanism affecting, although with a different intensity of damage, the outer hair cell (OHC) sensory epithelium. Moreover, these two damaging agents address preferentially different cochlear targets: noise mainly the sensory epithelium, styrene the supporting epithelial cells. Namely, the increase pattern of lipid peroxidation in the organ of Corti matched the cell damage distribution, involving predominantly OHC layer in noise exposed cochleae and both OHC and Deiters' cell layers in the styrene or combined exposed cochleae. The antioxidant treatment reduced the lipid peroxidation increase, potentiated the endogenous antioxidant defense system at OHC level in both exposures but it failed to ameliorate the oxidative imbalance and cell death of Deiters' cells in the styrene and combined exposures. Current antioxidant therapeutic approaches to preventing sensory loss focus on hair cells alone. It remains to be seen whether targeting supporting cells, in addition to hair cells, might be an effective approach to protecting exposed subjects.

The redox protein p66(shc) mediates cochlear vascular dysfunction and transient noise-induced hearing loss.

p66(shc), a member of the ShcA protein family, is essential for cellular response to oxidative stress, and elicits the formation of mitochondrial Reactive Oxygen Species (ROS), thus promoting vasomotor dysfunction and inflammation. Accordingly, mice lacking the p66 isoform display increased resistance to oxidative tissue damage and to cardiovascular disorders. Oxidative stress also contributes to noise-induced hearing loss (NIHL); we found that p66(shc) expression and serine phosphorylation were induced following noise exposure in the rat cochlea, together with markers of oxidative stress, inflammation and ischemia as indicated by the levels of the hypoxic inducible factor (HIF) and the vascular endothelial growth factor (VEGF) in the highly vascularised cochlear lateral region and spiral ganglion. Importantly, p66(shc) knock-out (p66 KO) 126 SvEv adult mice were less vulnerable to acoustic trauma with respect to wild type controls, as shown by preserved auditory function and by remarkably lower levels of oxidative stress and ischemia markers. Of note, decline of auditory function observed in 12 month old WT controls was markedly attenuated in p66KO mice consistent with delayed inner ear senescence. Collectively, we have identified a pivotal role for p66(shc) -induced vascular dysfunction in a common pathogenic cascade shared by noise-induced and age-related hearing loss.

Molecular targets for anticancer redox chemotherapy and cisplatin-induced ototoxicity: the role of curcumin on pSTAT3 and Nrf-2 signalling.

BACKGROUND: In oncology, an emerging paradigm emphasises molecularly targeted approaches for cancer prevention and therapy and the use of adjuvant chemotherapeutics to overcome cisplatin limitations. Owing to their safe use, some polyphenols, such as curcumin, modulate important pathways or molecular targets in cancers. This paper focuses on curcumin as an adjuvant molecule to cisplatin by analysing its potential implications on the molecular targets, signal transducer and activator of transcription 3 (STAT3) and NF-E2 p45-related factor 2 (Nrf-2), in tumour progression and cisplatin resistance in vitro and the adverse effect ototoxicity in vivo. METHODS: The effects of curcumin and/or cisplatin treatment have been evaluated in head and neck squamous cell carcinoma as well as in a rat model of cisplatin-induced ototoxicity by using immunofluorescence, western blot, and functional and morphological analysis. RESULTS: This study demonstrates that curcumin attenuates all stages of tumour progression (survival, proliferation) and, by targeting pSTAT3 and Nrf-2 signalling pathways, provides chemosensitisation to cisplatin in vitro and protection from its ototoxic adverse effects in vivo. CONCLUSIONS: These results indicate that curcumin can be used as an efficient adjuvant to cisplatin cancer therapy. This treatment strategy in head and neck cancer could mediate cisplatin chemoresistance by modulating therapeutic targets (STAT3 and Nrf2) and, at the same time, reduce cisplatin-related ototoxic adverse effects.

Fine-tuning of Th17 Cytokines in Periodontal Disease by IL-10.

Periodontitis (PD) is a chronic disease caused by the host inflammatory response to bacteria colonizing the oral cavity. In addition to tolerance to oral microbiome, a fine-tuned balance of IL-10 levels is critical to efficiently mount antimicrobial resistance without causing immunopathology. Clinical and animal studies support that adaptive T-helper (Th) cytokines are involved in the pathogenesis of alveolar bone destruction in PD. However, it remains unclear what type of Th response is related to human PD progression and what role IL-10 has on this process. We addressed the contribution of IL-10 in limiting Th1 and Th17 inflammatory response in murine and human PD. Through a combination of basic and translational approaches involving selected cytokine-deficient mice as well as human genetic epidemiology, our results demonstrate the requirement for IL-10 in fine-tuning the levels of Th17 (IL-17A and IL-17F) cytokines in experimental and human PD. Of novelty, we found that IL-17F correlated with protection in murine and human PD and was positively regulated by IL-10. To our knowledge, this is the first demonstration of the protective role for IL-17F in PD, its positive regulation by IL-10, and the potential differential role for IL-17A and IL-17F in periodontal disease.

Rosmarinic acid up-regulates the noise-activated Nrf2/HO-1 pathway and protects against noise-induced injury in rat cochlea.

Noise-induced hearing loss depends on progressive increase of reactive oxygen species and lipoperoxidative damage in conjunction with the imbalance of antioxidant defenses. The redox-sensitive transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) plays a critical role in the regulation of cellular defenses against oxidative stress, including heme oxygenase-1 (HO-1) activation. In this work we describe a link between cochlear oxidative stress damage, induced by noise exposure, and the activation of the Nrf2/HO-1 pathway. In our model, noise induces superoxide production and overexpression of the lipid peroxidation marker 4-hydroxy-nonenals (4-HNE). To face the oxidative stress, the endogenous defense system is activated as well, as shown by the slight activation of superoxide dismutases (SODs). In addition, we observed the activation of the Nrf2/HO-1 pathway after noise exposure. Nrf2 appears to promote the maintenance of cellular homeostasis under stress conditions. However, in this model the endogenous antioxidant system fails to counteract noise-induced cell damage and its activation is not effective enough in preventing cochlear damage. The herb-derived phenol rosmarinic acid (RA) attenuates noise-induced hearing loss, reducing threshold shift, and promotes hair cell survival. In fact, RA enhances the endogenous antioxidant defenses, as shown by decreased superoxide production, reduced expression of 4-HNE, and up-regulation of SODs. Interestingly, RA potentiates the Nrf2/HO-1 signaling pathway, as shown by immunohistochemical and Western blot analyses. Thus, protective effects of RA are associated with the induction/activation of the Nrf2-ARE signaling pathway in addition to RA direct scavenging capability.

Carious affected dentine: its behaviour in adhesive bonding.

BACKGROUND: Carious affected dentine (CAD) represents a very common substrate in adhesive dentistry. Despite its ability to interact with adhesive systems, the intrinsic character of CAD leads to lower bonding compared with sound dentine, regardless of the adhesive systems used. This low bonding may be more susceptible to leakage and hydrolysis of the interface by matrix metalloproteinases (MMPs). This systematic review aimed to determine current knowledge of CAD bonding, together with bond strength and MMP inhibitors' ability to prevent hybrid layer instability. METHODS: MEDLINE/Pubmed, Scopus and The Cochrane Library databases were electronically searched for articles published from 1 January 1960 to 31 August 2014. Two reviewers independently screened and included papers according to predefined selection criteria. RESULTS: The electronic searches identified 320 studies. After title, abstract and full-text examinations, 139 articles met the inclusion criteria. Data highlighted that a poor resin saturation of the already demineralized collagen matrix in CAD is strictly related to nanoleakage in interdiffusion and is the basis of the progressive decrease in strength with hydrolysis by MMPs. The use of mild self-etching systems seems to be the more accredited method to establish bonding in CAD. Inhibitors of MMPs may ensure better performance of CAD bonding, allowing undisturbed remineralization of the affected matrix. CONCLUSIONS: CAD bonding needs further understanding and improvement, particularly to enhance the strength and durability of the hybrid layer.

The effect of the NMDA channel blocker memantine on salicylate-induced tinnitus in rats.

Short-term tinnitus develops shortly after the administration of a high dose of salicylate. Since salicylate selectively potentiates N-methyl- D-aspartate (NMDA) currents in spiral ganglion neurons, it may play a vital role in tinnitus by amplifying NMDA-mediated neurotransmission. The aim of this study was to determine whether systemic treatment with a NMDA channel blocker, memantine, could prevent salicylate-induced tinnitus in animals. Additional experiments were performed to evaluate the effect of memantine on the auditory brainstem response (ABR) and distortion product otoacoustic emissions (DPOAE) to test for changes in hearing function. Thirty-six rats were divided into 3 groups and treated daily for four consecutive days. One group (n = 12) was injected with salicylate (300 mg/kg/d, IP), the second (n = 12) was treated with memantine (5 mg/kg/d, IP) and the third group (n = 12) was injected with salicylate and memantine. All rats were tested for tinnitus and hearing loss at 2, 24, 48 and 72 h after the first drug administration and 24 h post treatment; tinnituslike behaviour was assessed with gap prepulse inhibition of acoustic startle (GPIAS), and hearing function was measured with DPOAE, ABR and noise burst prepulse inhibition of acoustic startle (NBPIAS). Rats in the salicylate group showed impaired GPIAS indicative of transient tinnitus-like behaviour near 16 kHz that recovered 24 h after the last salicylate treatment. Memantine did not cause a significant change in GPIAS. Combined injection of salicylate and memantine significantly attenuated GPIAS tinnitus-like behaviour at 48 hours after the first injection. None of the treatments induced permanent threshold shifts in the ABR and DPOAE, which recovered completely within one day post treatment. Animals treated with salicylate plus memantine showed results comparable to animals treated with salicylate alone, confirming that there is no effect of memantine on DPOAE which reflects OHC function. The present study confirms the role of cochlear NMDA receptors in the induction of salicylate-induced tinnitus.

Antioxidant treatment with coenzyme Q-ter in prevention of gentamycin ototoxicity in an animal model.

Aminoglycosides, such as gentamycin, are well known ototoxic agents. Toxicity occurs via an activation process involving the formation of an iron-gentamycin complex with free radical production. Antioxidants like Q-ter (a soluble formulation of coenzyme Q(10), CoQ(10)), can limit or prevent cellular ototoxic damage. The present study was designed to investigate the possible protective effects of Q-ter on gentamycin ototoxicity in albino guinea pigs (250-300 g). Animals were divided into five experimental groups: I, a sham control group given an intra-peritoneal (I.P.) injection of 0.5 ml saline (SHAM); II, gentamycin group (GM), treated with an injection of gentamycin (100 mg/ kg); III, gentamycin + Q-ter group (GM+Q-ter), treated with gentamycin (same dose as group II) and an I.P. injection of coenzyme Q(10) terclatrate (Q-ter) at 100 mg/kg body weight; IV, injected with gentamycin (100 mg/kg) plus saline; V, treated with Q-ter alone (100 mg/ kg). All animals were treated for 14 consecutive days. Auditory function was evaluated by recording auditory brainstem responses (ABR) at 15 and 30 days from the beginning of treatment. Morphological changes were analyzed by rhodamine-phalloidine staining. Gentamycin-induced progressive high-frequency hearing loss of 45-55 dB SPL. Q-ter therapy slowed and attenuated the progression of hearing loss, yielding a threshold shift of 20 dB. The significant loss of outer hair cells (OHCs) in the cochlear medio-basal turn in gentamycin-treated animals was not observed in the cochleae of animals protected with Q-ter. This study supports the hypothesis that Q-ter interferes with gentamycin-induced free radical formation, and suggests that it may be useful in protecting OHC function from aminoglycoside ototoxicity, thus reducing hearing loss.

Pit and fissure sealants: review of literature and application technique.

Molars and premolars are the most vulnerable teeth to caries attack. The high susceptibility of these teeth to caries is directly related to morphology of their occlusal surface that prevents both chemical cleaning by saliva and mechanical cleaning by toothbrush. Pit and fissures are therefore the most prone areas to caries and need special protection to prevent carious lesions. Fluoride is the only chemical element used for caries prevention. In fact, it favors the remineralisation of initial lesions, prevents the production of polysaccharides essential for the development and sustainment of bacterial plaque, and the absorption of salivary glycoprotein. Fluoride also reinforces enamel, making it less susceptible to caries. Two methods of fluoroprophylaxis have been proposed: the first is the systemic fluoroprophylaxis which is particularly efficient in preventing interproximal caries, but it does not form an adequate protective barrier on the occlusal surfaces; the other is the topical application of a fluoride gel to the tooth surface, although this second method does not significantly reduce the incidence of caries. The efficacy of the sealing procedures depends on the correct application technique. Observing an operative protocol will ensure a longer lasting retention of the sealant on the occlusal surface and subsequently prolongs the protection against caries.

In vivo protective effect of ferulic acid against noise-induced hearing loss in the guinea-pig.

Ferulic acid (FA) is a phenolic compound whose neuroprotective activity was extensively studied in vitro. In this study, we provided functional in vivo evidence that FA limits noise-induced hearing loss. Guinea-pigs exposed to acoustic trauma for 1 h exhibited a significant impairment in auditory function; this injury was evident as early as 1 day from noise exposure and persisted over 21 days. Ferulic acid (150 mg/kg i.p. for 4 days) counteracted noise-induced hearing loss at days 1, 3, 7 and 21 from noise exposure. The improvement of auditory function by FA was paralleled by a significant reduction in oxidative stress, apoptosis and increase in hair cell viability in the organ of Corti. Interestingly in the guinea-pig cochleae, the neuroprotective effect of FA was functionally related not only to its scavenging ability in the peri-traumatic period but also to the up-regulation of the cytoprotective enzyme heme oxygenase-1 (HO-1); in fact, FA-induced improvement of auditory function was counteracted by the HO inhibitor zinc-protoporphyrin-IX and paralleled the time-course of HO-1 induction over 3-7 days. These results confirm the antioxidant properties of FA as free-radical scavenger and suggest a role of HO-1 as an additional mediator against noise-induced ototoxicity.

N-acetyl-beta-D-glucosaminidase activity in urine of dental personnel.

BACKGROUND: Dental personnel is exposed to several potential nephrotoxic agents. Urinary N-acetyl-beta-d-glucosaminidase (U-NAG) activity has emerged as a sensitive marker of early nephrotoxicity. METHODS: U-NAG was evaluated, by fluorimetric assay, in urine from 30 healthy subjects and 30 dental personnels. RESULTS: The median value of U-NAG activity (133.5 U/mmol urinary creatinine (U-Cr) in urines of dental personnel was not statistically different (P>0.05) from activity (100.7 U/mmol U-Cr) of control urines. CONCLUSIONS: The results suggest that, for dental personnel, exposure to potential nephrotoxic agents is not usually high enough to increase U-NAG activity.

[A SEM analysis of in vivo adhesive interface between a bonding agent and normal or sclerotic dentin]. / Analisi in vivo al SEM dell'interfaccia tra un agente adesivo e dentina sana sclerotica.

AIM: Although dentine is a dynamic sub-stratum of the tooth, it seems correct to say that as far as etching and bonding to the sclerotic dentine is concerned, there are different micro-morphological aspects to be analysed in comparison to those on the healthy dentin. METHODS: This paper seeks to analysis the interface between the dentin and the adhesive bonding material resulting from in vivo placement of a 5(th) generation dentin bonding system on the base of cavities in 8 teeth. The teeth were removed for periodontal or orthodontic reasons and then examined by scanning electron microscopy. RESULTS: The results confirm the notable complexity of the dentin-adhesive interface that is developed or formed in-vivo and is due to the different state in which the dentine is found (healthy or sclerotic). CONCLUSION: It is underlined that the passage from the state of healthy to sclerotic dentin is very fast.

[Histopathological and therapeutical aspects of cervical lesions]. / Aspetti istopatologici e terapeutici delle lesioni cervicali.

The term "cervical lesions" definies all the alterations consisting in a loss of hard dental tissues located along the cement-enamel junction, and may be more specifically defined as carious lesions (LCC) and non-carious lesions (NCCL). The non-carious cervical lesions are characterized by the loss of mineralised dental tissue in the absence of a carious process. This definition includes three different lesion categories: abrasion, erosion and the cervical lesions caused by stress (abfractions). In this paper the authors explain the histological, clinical, preventive findings of NCCL and try to give some guidelines to choose the appropriate material to restore different types of non carious cervical lesions.

[Fluoride release properties of GRACs. A survey of the literature]. / Meccanismi del rilascio di fluoro dai CVI. Una revisione della letteratura.

While numerous publications have documented the anticariogenic properties of GPACs through fluoride release measurements and artificial caries studies, considerable debate still exists concerning the fluoride release properties of these materials in terms of dissolution and diffusion mechanisms. This paper will review some of the more important aspects of GPAC fluoride release. Numerous studies have reported on the fluoride release properties of GPACs. The first point to note is that all the studies were performed on commercial materials. This approach has inherent difficulties as the main factor governing fluoride release is the GPAC composition and formulation. Traditional GPACs are two component systems, consisting essentially of a ion leachable fluoroaluminosilicate glass and a polymeric acid, especially a polycarboxylic acid; fluoride ions are of prime importance in this reaction, in that they have a major influence on the structure of the glass, its reactivity, cement formation and bioactivity. GPACs may be viewed as particle reinforced polymeric composites, in which the degraded residual glass particles with their silicious layer reinforce the polysalt matrix, improving mechanical properties.

Fluoride release from model glass ionomer cements.

Glass ionomer cements (GICs) are an important class of biomedical material used extensively for color matched mercury free, dental restorations. GICs can release clinically beneficial amounts of fluoride and have acceptable handling properties which make them suitable as dental restoratives. The fluoride release of model GICs produced from specially synthesized fluoro-alumino-silicate glasses was studied. Nine glasses of varying fluoride content based on 4.5SiO(2)-3Al(2)O(3)-1.5P(2)O(5)-(5-Z)CaO-ZCaF(2) were synthesized and cement disks were prepared from them. The glass transition temperature reduced with increasing fluorine content of the glass. Fluoride ion release was measured into distilled water as a function of time for up to 140 days using a fluoride ion selective electrode. The quantity of fluoride released was found to be proportional to the fluorine content of the glass at all intervals time. The cumulative fluoride release was proportional to square root time. Substituting strontium for calcium in the glass had little influence on the fluoride release behavior of the cements.

[ELISA analysis of salivary cotinine in smokers]. / Determinazioni ELISA della cotinina salivare in pazienti fumatori.

BACKGROUND: The presence in saliva of cotinine, the main and inactive metabolite of nicotine, reflects the extent of systemic distribution of nicotine and explains the increased susceptibility to periodontal disease in smokers. The aim of this study was to investigate the comparative amount of cotinine in the saliva of habitual cigarette smokers, non-smokers and passive smokers. METHODS: Saliva sample were obtained from 14 cigarette smokers and 13 non-smokers (8 passive-smokers), all without periodontal disease, and analyzed by Microplate EIA (a variation of ELISA based on cross-reactivity of cotinine with anti-cotinine antibody revealed by absorbance in spectrophotometry) to determine the presence and the amount of cotinine. RESULTS: Cotinine was detected in the saliva of smokers with a mean of 92.3 +/- 4.15 ng/ml and, unexpectedly, there was evidence of cotinine also in the saliva of non-smokers (mean 5.4 +/- 1.22 ng/ml), particularly, in passive-smokers (mean 12.9 +/- 6.67 ng/ml). CONCLUSIONS: The salivary concentration of cotinine can be used to estimate nicotine intake and its possible role in the pathogenesis of periodontal disease also in passive-smokers.

Dentin dysplasia type I. Report of case and ultrastructural study.

Radicular dentin dysplasia (DD-I) is a rare hereditary dental alteration. It is characterized clinically by almost normal looking crowns and severe hypermobility of the teeth. The radiographic analysis, on the other hand, discloses the obliteration of all pulp chambers, the short, malformed roots and plenty of periapical bone radiolucencies on noncarious teeth. A case of radicular dentin dysplasia is presented. In this 43-year-old woman the diagnosis was supported, besides the clinical and radiographic analysis, by the pedigree of the proband, which showed the autosomal dominant pattern of feature transmission. Further-more, the electron microscopic analysis of one extracted molar revealed the atubular structure of the secondary dentin, and its globular organization.

[Tests on ionic release from glass-ionomer cements]. / Prove sperimentali sul rilascio ionico da parte di sistemi ionomerovetrosi.

BACKGROUND: Dissolution process in oral liquids by the presence of glass-ionomer systems (due to surface corrosion, to diffusion through solutions and through mass) make an ionic release (particularly F, Al, Pb, As) which is a non secondary problem, due to the usual utilization of these materials in pedodontic and restorative dentistry. METHODS: In this work, considering the high toxicity of low quantity of Arsenic ion, a comparative research has been made in order to determine, by using high level liquid Cromatography (HPCL), the quantity in ppm of As hydro- and acid soluble given by five ionomeric products, in water and in nitric acid concentrated solution. RESULTS AND CONCLUSIONS: The results show that in some products arsenical concentrations are higher then the quantity accepted by ISO-FDI; therefore, a better control in the production of these products is needed as well as a limited use in dentistry. It is suggested to use glass-ionomer systems in patients with dental dike and varnish on the surfaces that are in contact with oral liquids action.