Characterization of Odontoblast-like Cell Phenotype and Reparative Dentin Formation In Vivo: A Comprehensive Literature Review.

INTRODUCTION: The primary aim was to explore the criteria used in characterization of reparative cells and mineralized matrices formed after treatment of pulp exposures, and the sequence of relative events. The secondary aim was to evaluate whether the reparative events depend on the experimental model species, age, and therapeutic intervention. METHODS: A literature search of databases using different combinations of the key words was undertaken. Data analysis was based only on studies having histological or histochemical assessment of the pulp tissue responses. The search yielded 86 studies, 47 capping material-based and 39 bioactive application-based experiments, which provided data on morphological or functional characterization of the mineralized matrices and the associated cells. RESULTS: In 64% of capping material-based and 72% of bioactive application-based experiments, a 2-zone mineralized matrix formation (atubular followed by tubular) was detected, whereas characterization of odontoblastic differentiation is provided in only 25.5% and 46.1% of the studies, respectively. In 93.3% of the studies showing odontoblast-like cells, differentiated cells were in association with tubular mineralized matrix formation. Analyses further showed that cell- and matrix-related outcomes do not depend on experimental model species, age, and therapeutic intervention. CONCLUSIONS: The evidence of the reviewed scientific literature is that dental pulp cells secrete a dentin-like matrix of tubular morphology in relation to primitive forms of atubular or osteotypic mineralized matrix. Furthermore, data analysis showed that dental pulp cells express in vivo the odontoblastic phenotype, and secrete matrix in a predentin-like pattern, regardless of the model species, age, and therapeutic intervention used.

Preparation Prerequisites for Effective Irrigation of Apical Root Canal: A Critical Review.

BACKGROUND: It is well recognized that disinfection of the complex root canal system at the apical root canal remains the most critical therapeutic measure to treat apical periodontitis. MATERIAL AND METHODS: Observational and experimental data in relation to the anatomy of the apical root canal in different tooth types and the cross sectional diameters of the apical part of the most commonly used hand and rotary files are critically reviewed. RESULTS: The present data analysis confirm that the challenging issue of antibacterial efficacy of modern preparation protocols in non-surgical endodontics requires more attention to apical root canal irrigation as a balance between safety and effectiveness. Ex vivo investigations clearly indicate that a specific design of the chemo-mechanical preparation is needed at the onset of RCT, more particularly in infected teeth. Design should be based on specific anatomical parameters, and must determine the appropriate size and taper of preparation as pre-requirements for effective and safe apical irrigation. CONCLUSIONS: The optimal irrigation protocols might be designed on the basis of technical specifications of the preparations procedures, such as the penetration depth, the type of the needle, the required time for continuous irrigant flow, the concentration of NaOCl, and the activation parameters. Key words:Endodontics, root canal treatment, instrumentation, irrigation, apical root canal.

Dentine-pulp tissue engineering in miniature swine teeth by set calcium silicate containing bioactive molecules.

OBJECTIVE: The present study aims to investigate whether reparative dentinogenesis could be guided at central pulpal sites or at a distance from the amputated pulp of miniature pig teeth, by using set calcium silicate-based carriers containing human recombinant bioactive molecules. DESIGN: Pulp exposures were performed in 72 permanent teeth of 4 healthy miniature swine. The teeth were capped with pre-manufactured implants of set calcium silicate-based material containing BMP-7, TGFß1 or WnT-1, for 3 weeks. Conical-shaped intrapulpal implants were exposed in the central pulp core, while disc-shaped extrapulpal implants were placed at a distance from the amputated pulp. Implants without bioactive molecules were used as controls. Thickness and forms of new matrix mineralized deposition were assessed histologically at post-operative periods of 3 weeks by light microscopy. RESULTS: Intrapulpal applications: Calcified structures composed of osteodentine were found in contact with the BMP-7 implants. An inhomogeneous calcified tissue matrix was found around the WnT-1 carriers. A two-zone calcified structure composed of osteodentine and a thicker tubular matrix zone was seen at the TGFß1 carrier-pulp interface. Extrapulpal applications: The space between WnT-1 implants and pulp periphery had been invaded by soft tissue with traces of calcified foci. Thick calcified structures composed of osteodentine were found surrounding pulp exposure sites in response to application of BMP-7. Spindle-shaped cells associated with atubular calcified matrix or elongated polarized cells associated with tubular dentine-like matrix were found along the cut dentinal walls of the TGFß1 group. CONCLUSION: The present experiments indicated that set calcium silicate could be used as carrier for biologically active molecules. TGFß1 was shown to be an effective bioactive molecule in guiding tertiary dentine formation.

Designing and testing regenerative pulp treatment strategies: modeling the transdentinal transport mechanisms.

The need for simulation models to thoroughly test the inflammatory effects of dental materials and dentinogenic effects of specific signaling molecules has been well recognized in current dental research. The development of a model that simulates the transdentinal flow and the mass transfer mechanisms is of prime importance in terms of achieving the objectives of developing more effective treatment modalities in restorative dentistry. The present protocol study is part of an ongoing investigation on the development of a methodology that can calculate the transport rate of selected molecules inside a typical dentinal tubule. The transport rate of biological molecules has been investigated using a validated CFD code. In that framework we propose a simple algorithm that, given the type of molecules of the therapeutic agent and the maximum acceptable time for the drug concentration to attain a required value at the pulpal side of the tubules, can estimate the initial concentration to be imposed.

Dentinogenic Activity of Biodentine in Deep Cavities of Miniature Swine Teeth.

INTRODUCTION: The aim of the present study was to evaluate comparatively the bioactivity potential of a calcium silicate-based material (Biodentine; Septodont, Saint-Maur-des-fosses Cedex, France) after the restoration of deep dentinal cavities of miniature swine teeth with or without the application of a calcium hydroxide-containing pulp protective base (Dycal; Caulk Lab, Milford, DE). METHODS: Thirty-three permanent teeth (premolars, canines, and incisors) of 3 miniature swine were used. Class V cavities were prepared on the buccal surface of teeth. The cavities were restored with Biodentine in the presence (control group) or absence (experimental group) of a Dycal protective base. The pulpal tissue responses were histologically and histomorphometrically assessed at postoperative periods of 3 and 8 weeks. Three specimens were further evaluated with scanning electron microscopy. The maximum thickness of the postoperatively formed mineralized matrix beneath the cavity floor was measured. Data were statistically analyzed by the Kruskal-Wallis and the Mann-Whitney U tests. RESULTS: A bacterial staining reaction along the cavity walls or intense inflammatory infiltration in the pulp was not detected in any of the specimens. A continuous zone of the postoperatively formed mineralized matrix mostly of atubular structure with scattered defects and cellular inclusions and occasionally followed by a thin zone with tubular morphology was detected in all specimens of the control group and 13 of 18 experimental group teeth. In the remaining teeth of the experimental group, a separate zone composed of the osteotypic mineralized matrix and soft tissues was noted between the circumpulpal and the newly formed matrix. Scanning electron microscopy confirmed the fibrous structural morphology of the tertiary dentin. A significantly higher rate of the postoperatively formed mineralized matrix had been formed in the teeth of the experimental group in both periods of 3 and 8 weeks (P < .01). CONCLUSIONS: The present investigation indicates that under the present experimental conditions tertiary dentin with occasional intermediate formation of osteodentin is observed after the application of Biodentine in the presence or absence of a Dycal protective base. The thickness of the tertiary dentine zone was significantly higher in the absence of Dycal.

Dentinogenic Specificity in the Preclinical Evaluation of Vital Pulp Treatment Strategies: A Critical Review.

Reviews on the clinical performance of vital pulp treatment strategies and capping materials repeatedly showed an insufficient grade of evidence concerning their therapeutic validity. The biological mechanisms underlying the regenerative potential of pulp-dentin complex have attracted much attention during the last two decades, since new pulp treatment modalities have been designed and tested at the preclinical level. It has been recognized that evaluation should be based on the specific ability of therapeutic interventions to signal recruitment and differentiation of odontoblast-like cells forming a matrix in a predentin-like pattern, rather than uncontrolled hard tissue deposition in a scar-like form. The aim of the present article was to critically review data from histological experimental studies on pulp capping, published during the last 7 decades. A comprehensive literature search covering the period from 1949 to 2015 was done using the Medline/Pubmed database. Inclusion of a study was dependent on having sufficient data regarding the type of capping material used and the unit of observation (human permanent tooth in vivo or animal permanent dentition; primary teeth were excluded). The post-operatively deposited matrix was categorized into three types: unspecified, osteotypic, or dentin-like matrix. One hundred fifty-two studies were included in the final evaluation. Data from the present systematic review have shown that only 30.2% of the 152 experimental histological pulp capping studies described the heterogenic nature of the hard tissue bridge formation, including osteotypic and tubular mineralized tissue. Structural characteristics of the new matrix and the associated formative cells were not provided by the remaining 106 studies. Analysis showed that more careful preclinical evaluation with emphasis on the evidence regarding the dentinogenic specificity of pulp therapies is required. It seems that selection of appropriate vital pulp treatment strategies and pulp capping materials would be further facilitated in terms of their therapeutic validity if international consensus could be reached on a select number of mandatory criteria for tissue-specific dentinogenic events.

Dentinogenic responses after direct pulp capping of miniature swine teeth with Biodentine.

INTRODUCTION: The aim of this study was to evaluate pulpal responses after experimental direct pulp capping of mechanically exposed teeth with a new calcium silicate-based dentin replacement material. METHODS: Thirty-four anterior and posterior teeth of 3 miniature swine were used. Class V or I cavities were prepared on the buccal or occlusal surfaces, respectively. Pulpal exposures were further performed using a round carbide bur 0.8 mm in diameter. Exposures were treated with white MTA Angelus (Angelus, Londrina, PR, Brazil) or Biodentine (Septodont, Saint Maur des Fosses, France), and the cavities were further restored with Biodentine. The pulpal tissue responses were histologically assessed at postoperative periods of 3 and 8 weeks. Data were statistically analyzed using the Kruskal Wallis and the Mann-Whitney U tests. RESULTS: Inflammatory infiltration or pulp tissue necrosis was not found in any of the specimens. All teeth showed mineralized matrix formation in the form of a complete hard tissue bridge composed of osteodentin or osteodentin followed by a discontinuous or continuous reparative dentin zone. A significantly higher thickness of the hard tissue bridge was found in the group of teeth treated with Biodentine at both 3 and 8 weeks. A number of teeth, which were under root development at the onset of the experimental procedures, exhibited ectopic pulp calcification. CONCLUSIONS: The application of both calcium silicate-based materials in direct contact with the mechanically exposed pulp of healthy miniature swine teeth led to pulp repair with complete hard tissue bridge formation. The thickness of hard tissue bridges was significantly higher after pulp capping with Biodentine.

Pulp response to dentine adhesives: A study on mature human pulps.

OBJECTIVE: To clinically investigate the antibacterial effects of a commercially available self-etch 12-methacryloyloxy- dodecylpyridinium bromide (MDPB)-containing adhesive system in comparison with its respective non-MDPB-containing adhesive and to evaluate the pulp responses when in use on human teeth. MATERIALS AND METHODS: Sixty-two viable human teeth scheduled for extraction were used. Class V cavities were prepared on the buccal surfaces of the teeth and filled with the tested materials (Protect Bond/Clearfil AP-X, SE Bond/Clearfil AP-X and Dycal/Ketac Fill Plus) as a control group, according to manufacturer's recommendations. Randomly divided to two groups teeth remained intra-orally for 4 and 8 weeks. After extraction, teeth were decalcified, sectioned and stained using the Mayer's hematoxylin and eosin, and modified Brown-Brenn's technique. Pulp responses were evaluated microscopically under a microscope and remaining dentine thickness measured under a stereomicroscope. RESULTS: No statistically significant differences regarding pulp inflammation or bacterial infiltration were found either for the materials tested or for periods of post-operative evaluation. CONCLUSIONS: The results suggested that for a short period of evaluation there are no quantitative differences, regardless to restoring material used.

Experimental formation of dentin-like structure in the root canal implant model using cryopreserved swine dental pulp progenitor cells.

OBJECTIVES: The purpose of the present study was to present histological and immunohistochemical evidence showing the regenerative capacity of swine dental pulp stem cells (S-DPSCs) seeded on organic or synthetic scaffolds and implanted as hybrid root implants in the jaw bone of minipigs. METHODS: Immature permanent incisor teeth and unerupted premolars at the early root-forming stage were extracted from three 7-month-old minipigs, and mesenchymal stem/progenitor cells were isolated from dental pulp. Cells were cryopreserved in liquid nitrogen. A year later, new permanent incisor and premolar teeth were extracted; pulp tissue was removed; and pieces of root canals of the extracted teeth, containing collagen or Poly(lactic-co-glycolic acid) scaffolds seeded with the autologous cryopreserved DPSCs, were implanted into the fresh post-extraction socket of the mini pig jaw. The resulting constructs were harvested after 6 and 10 weeks and evaluated by histological and immunohistochemical analyses. RESULTS: Six weeks postoperatively, the central canal space of the root implants showed degrading scaffold material. New extracellular matrix had been deposited in a polar predentin-like pattern on the canal dentinal walls by cuboidal nonpolarized cells. Ten weeks postoperatively, newly formed organic matrix had been consistently deposited on the canal walls. The presence of a continuous layer of polarized cells showing typical columnar morphology adjacent to the newly deposited organic matrix was evident. CONCLUSIONS: The interactions of S-DPSCs with the dentin matrix of roots implanted in the jawbone of minipigs constitute a model to study in vivo organization and differentiation potential of DPSCs.

Periradicular Tissue Responses to Biologically Active Molecules or MTA When Applied in Furcal Perforation of Dogs' Teeth.

The aim of this study was the comparative evaluation of inflammatory reactions and tissue responses to four growth factors, or mineral trioxide aggregate (MTA), or a zinc-oxide-eugenol-based cement (IRM) as controls, when used for the repair of furcal perforations in dogs' teeth. Results showed significantly higher inflammatory cell response in the transforming growth factorß1 (TGFß1) and zinc-oxide-eugenol-based cement (IRM) groups and higher rates of epithelial proliferation in the TGFß1, basic fibroblast growth factor (bFGF), and insulin growth factor-I (IGF-I) groups compared to the MTA. Significantly higher rates of bone formation were found in the control groups compared to the osteogenic protein-1 (OP-1). Significantly higher rates of cementum formation were observed in the IGF-I and bFGF groups compared to the IRM. None of the biologically active molecules can be suggested for repairing furcal perforations, despite the fact that growth factors exerted a clear stimulatory effect on cementum formation and inhibited collagen capsule formation. MTA exhibited better results than the growth factors.

Differentiation potential of dental papilla, dental pulp, and apical papilla progenitor cells.

INTRODUCTION: Regenerative endodontic procedures use the differentiation potential of embryonic and adult pulp progenitor cell populations to reconstitute dental structures. METHODS: An in-depth search of the literature was accomplished to review biologic knowledge from basic research on tooth morphogenesis and differentiation, root development, dentin-pulp regeneration, pulp revascularization and apexification, experimental and clinical studies on the dentinogenic differentiation potential of progenitor cells in the embryonic dental papilla, dental pulp, and associated mesenchymal tissues of the developing root. RESULTS: Odontogenic potential is determined during early tooth morphogenesis in the odontogenic mesenchyme. Progenitor cells from the odontogenic mesenchyme give rise to primary dentin-forming cells (odontoblasts) in the presence of stage-specific enamel epithelium and/or basement membrane and tertiary dentin-forming cells (odontoblast-like cells) in experimental conditions. The specificity of odontogenic mesenchymal cells to form tertiary dentin might be related to the repertoire of signaling pathways operated by the temporospatial pattern of epithelial-mesenchymal interactions during tooth formation. Dental papilla cells isolated from tooth germs before the onset of odontoblast differentiation have not shown any competence to become odontoblasts in the absence of enamel epithelium. On the other hand, the specificity of progenitor cells in the mesenchymal cell populations of the developing root apex remains to be determined. CONCLUSIONS: It seems evident that the dental pulp might be only used as a source of progenitor cells with dentinogenic competence for the regeneration of the dentin-pulp complex. The nature of dental or apical papilla progenitor cells in terms of their specificity for dentin regeneration has to be first characterized.

Pulpal responses after application of current adhesive systems to deep cavities.

The aim of the present study was to evaluate comparatively the pulpal tissue reactions of four adhesive systems placed in experimental cavities of healthy dog's teeth. Class V cavities with a mean value of remaining dentin thickness (RDT) ranging between 0.55 +/- 0.30-0.68 +/- 0.38 mm were prepared. The cavities were treated with the following adhesive systems: Etch and Prime 3.0 (EP), Single Bond (SB), Clearfil SE Bond (CSE), and Prompt L-Pop (PLP). The pulpal tissue responses to dentin adhesives were assessed histopathologically at postoperative periods of 7, 21, and 65 days, and the results were subjected to statistical analysis. A significantly greater adverse inflammatory response was observed with the materials EP and PLP, while a significantly lesser degree of disorganization in the odontoblastic zone was found with the materials SB and CSE, in the postoperative period of 65 days. In addition, a thicker predentin zone was observed where SB material was applied. Application of the selected adhesive systems to non-exposed cavities, with an RDT, which ranged between the above-mentioned rates, was correlated with slight to moderate inflammation and odontoblast reduction depending on the materials used as well as upon the RDT.