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The basic premise of gene therapy is that genes can be used to produce in situ therapeutic proteins. The controlled delivery of DNA complexes from biomaterials offers the potential to enhance gene transfer by maintaining an elevated concentration of DNA within the cellular microenvironment. Immobilization of the DNA to the substrate to which cells adhere maintains the DNA in the cell microenvironment for subsequent cellular internalization. Here, layer-by-layer (LBL) films made from poly(L-glutamic acid) (PLGA) and poly(L-lysine) (PLL) containing DNA were built in the presence of charged cyclodextrins. The biological activities of these polyelectrolyte films were tested by means of induced production of a specific protein in the nucleus or in the cytoplasm by cells in contact with the films. This type of coating offers the possibility for either simultaneous or sequential interfacial delivery of different DNA molecules aimed at cell transfection. These results open the route to numerous potential applications in patch vaccination, for example.

The implantation of cultured dental cell-cell re-associations allows for the reproduction of fully formed teeth, crown morphogenesis, epithelial histogenesis, mineralized dentin and enamel deposition, and root-periodontium development. Since vascularization is critical for organogenesis and tissue engineering, this work aimed to study: (a) blood vessel formation during tooth development, (b) the fate of blood vessels in cultured teeth and re-associations, and (c) vascularization after in vivo implantation. Ex vivo, blood vessels developed in the dental mesenchyme from the cap to bell stages and in the enamel organ, shortly before ameloblast differentiation. In cultured teeth and re-associations, blood-vessel-like structures remained in the peridental mesenchyme, but never developed into dental tissues. After implantation, both teeth and re-associations became revascularized, although later in the case of the re-associations. In implanted re-associations, newly formed blood vessels originated from the host, allowing for their survival, and affording conditions organ growth, mineralization, and enamel secretion.

Post-eruptive loss of ameloblasts requires identification of alternative sources for these cells to realize tooth-tissue-engineering strategies. Recent reports showed that bone-marrow-derived cells can give rise to different types of epithelial cells, suggesting their potential to serve as a source for ameloblasts. To investigate this potential, we mixed c-Kit+-enriched bone marrow cells with embryonic dental epithelial cells and cultured them in re-association with dental mesenchyme. Non-dividing, polarized, and secretory ameloblast-like cells were achieved without cell fusion. Before basement membrane reconstitution, some bone marrow cells migrated to the mesenchyme, where they exhibited morphological, molecular, and functional characteristics of odontoblasts. These results show, for the first time, that bone-marrow-derived cells can be reprogrammed to give rise to ameloblast-like cells, offering novel possibilities for tooth-tissue engineering and the study of the simultaneous differentiation of one bone marrow cell subpopulation into cells of two different embryonic lineages.

The biological mechanisms of tooth movement result from the cellular responses of connective tissues to exogenous mechanical forces. Among these responses, the degradation of the extracellular matrix takes place, but the identification of the molecular basis as well as the components implicated in this degradation are poorly understood. To contribute to this identification, we subjected human fibroblasts obtained from the periodontal ligament (PDLs) and from the gingiva (HGFs) to a continuous stretch to quantify the mRNAs encoding for various metalloproteinases (MMPs), their tissue inhibitors (TIMPs), and a and β integrin subunits. Both cell lines reacted by inducing the expression of the mRNAs encoding for MMP-1, MMP-2, TIMP-1, and TIMP-2, while other mRNAs did not vary (MTI-MMP, TIMP-3) or were not expressed (MMP-9). PDLs expressed selectively the mRNAs encoding for a4 and av, with no difference measurable under stretching, while the mRNAs encoding for a6 and β1 were increased and the one encoding for a5 was decreased. HGFs increased the mRNAs encoding for α2, α6, β1, and β3 and decreased the one encoding for a3. Analysis of our data indicated that stretched HGFs and PDLs induced the same pattern of mRNAs encoding for MMPs and TIMPs but differed for those encoding various integrin subunits, known to act as protein receptors in mechanotransduction.

A new type of coating involving a layer-by-layer technique has been recently reported. This coating is composed of a polyelectrolyte multilayer film that confers specific properties on surfaces to which it is applied. Here, we studied the applicability of such a technique to the coating of oral prostheses, by first testing the construction of polyelectrolyte multilayer films on several polymers used in oral prosthesis bases, and, subsequently, by studying the stability of these coatings in vitro, in human saliva, and in vivo in a rat model. We demonstrated that the multilayered films are able to coat the surfaces of all tested polymers completely, thus increasing their wettability. We also showed that saliva does not degrade the film after 7 days in vitro and after 4 days in vivo. Taken together, our results establish that the layer-by-layer technique is suitable for the coating of oral devices.

Objectives The aim of this study was to assess the filling quality of five obturation techniques in oval-shaped root canals. Materials and methods A total of 212 mandibular first molars with one distal oval canal were selected. Distal canals, shaped with WaveOne Gold Primary, were randomly divided in five groups ( n  = 40) for obturation: continuous wave condensation, GuttaCore, Thermafil, single cone with AH plus, and single cone with BioRoot RCS. The proportions of gutta-percha-filled areas (GPFA), sealer-filled areas (SFA), void areas (VA), and the sealer/gutta tags into dentinal tubules at 4 mm and 2 mm from the apex were analyzed by an optical numeric microscope, SEM, and energy-dispersive X-ray (EDX). Data were then compared by Kruskal-Wallis one-way ANOVA on ranks ( α  = 0.05). Results At 4 mm, a statistically significant higher GPFA and lower SFA were observed in the GuttaCore and Thermafil groups compared with the 3 other groups. A statistically significant lower VA was observed in the continuous wave condensation, GuttaCore, and Thermafil groups than in the two single-cone groups. At 2 mm, there were a statistically significant higher GPFA and lower SFA and VA in GuttaCore and Thermafil groups than in the 3 other groups. At the two levels investigated, the presence of gutta-percha tags was clearly demonstrated for GuttaCore and Thermafil groups; no tags were observed in the 3 other groups. Conclusions Obturation quality was overall improved in GuttaCore and Thermafil groups. Clinical relevance Carrier-based techniques may significantly improve the filling quality compared to continuous wave condensation and single-cone technique. The single-cone technique might have inherent limitations especially in oval root canals regardless of the sealer used.

The aim of this study was to investigate and compare, radiographically and clinically, the impacts of calcium-silicate based-cement (CSBC), nano-hydroxyapatite and platelet-rich fibrin (PRF) as pulpotomy agents in permanent immature molars with incomplete root development. Sixty-three participants (63 permanent immature molars) were included in this study. The patients were randomly divided into three equal groups. Fast setting MTA (MM-MTA), nano-hydroxyapatite and platelet-rich fibrin were used as pulpotomy agents. The teeth were evaluated clinically and radiographically after 6 and 12 months by two blinded examiners. Apical closure and pulp canal obliteration percentages were recorded. The in vitro reaction of the tested materials after a 7-day immersion period of the different materials in phosphate-buffered solution was analyzed using scanning electron microscopy to associate the in vitro mineralization with in vivo pulp canal obliteration percentages. Data were analyzed using Chi-square and ANOVA tests (α = 0.05). No significant difference was found between the three tested groups in terms of clinical and radiographic success (p > 0.05). All cases demonstrated evidence of root growth, including complete apical closure or continued apical closure. At 12 months, complete apical closure was found among the MM-MTA group (50%), nano-hydroxyapatite group (55%) and platelet-rich fibrin group (60%) (p > 0.05). After 12 months, pulp canal obliteration was more observed in the MM-MTA and nano-hydroxyapatite groups than in the PRF group (p < 0.05). MM-MTA (auto-mixed), NHA (hand-mixed) and PRF (autologous) could be used as pulpotomy agents since they exhibit comparable high clinical and radiographic success rates. However, the fact that the groups managed with MM-MTA and NHA have a higher tendency to canal obliteration might indicate that PRF should be considered the first choice material as pulpotomy agent, as it would make retreatment considerably easier.

The present study aimed to evaluate the effectiveness of customized handle toothbrush in dental plaque removal in children with Down syndrome in comparison to children with no special needs. A randomized clinical trial was performed on 48 children aged 6–9 years old. Participants were divided into two groups (n = 24, children with no special needs or with Down syndrome). Each group was divided into two subgroups (customized and conventional toothbrush). Plaque accumulation was evaluated using Turesky modification of the Quigley–Hein plaque index (TMQHPI) at four times (pre-brushing (T0), post-brushing at baseline (T1), 1 week (T2) and 3 weeks (T3)), and the modified gingival index (MGI) was used to evaluate the gingivitis at three times (T0, T2 and T3). Data were statistically analyzed. Plaque accumulation and gingivitis decreased significantly for all groups between T0 and T3 (p < 0.05). Both customized groups demonstrated significant lower plaque accumulation compared to conventional groups (p < 0.05) at T1, T2 and T3 and significant lower gingivitis at T3. Customizing the toothbrush handle improved child’s ability for plaque control, especially in children with Down syndrome.

Despite the availability of melanoma treatment at the primary site, the recurrence of local melanoma can metastasize to any distant organ. Currently, the available therapies for the treatment of metastatic melanoma are of limited benefit. Thus, the functional analysis of conventional therapies may help to improve their efficiency in the treatment of metastatic melanoma. In the present study, the exposure of melanoma cells to vinblastine was found to trigger apoptosis as evidenced by the loss of mitochondrial membrane potential, the release of both cytochrome c and apoptosis inducing factor, activation of caspase-9 and 3, and cleavage of Poly (ADP-ribose)-Polymerase. Also, vinblastine enhances the phosphorylation of Ras homologous protein A, the accumulation of reactive oxygen species, the release of intracellular Ca 2+ , as well as the activation of apoptosis signal-regulating kinase 1, c-jun-N-terminal kinase, p38, inhibitor of kappaBα (IκBα) kinase, and inositol requiring enzyme 1α. In addition, vinblastine induces the DNA-binding activities of the transcription factor NF-κB, HSF1, AP-1, and ATF-2, together with the expression of HSP70 and Bax proteins. Moreover, inhibitory experiments addressed a central role for Rho A in the regulation of vinblastine-induced apoptosis of melanoma cells via mitochondrial and non-mitochondrial-dependent mechanisms. In conclusion, the present study addresses for the first time a central role for Rho A in the modulation of vinblastine-induced apoptosis of melanoma cells and thereby provides an insight into the molecular action of vinblastine in melanoma treatment.

The formation of bacterial biofilms on the surface of implanted materials is a critical factor that may lead to chronic microbial infection and tissue necrosis. In the present study we analysed the stability of polyelectrolyte multilayer (ML) films on suture materials and the antibacterial effect obtained with chlorhexidine (CHX)-functionalized films built on different types of suture materials such as silk, polyester and copolymer of glycolide and l -lactide. The comparison of Escherichia coli culture on glass coverslips and glass coverslips with ML and CHX showed at 24 h an inhibition of the bacterial relative luminescence (40.68%, P < 0.5) and at 48 h (99.46%, P < 0.001). In another way, simple soaking of suture material overnight in CHX digluconate 20% without polyelectrolyte films did not at all protect sutures from bacterial colonization but CHX-functionalized polyelectrolyte films, made from poly- l -glutamic acid and poly- l -lysine, inhibited Escherichia coli proliferation.