Tertiary Dentin Formation Research Articles

Bortezomib Facilitates Reparative Dentin Formation after Pulp Access Cavity Preparation in Mouse Molar

IntroductionThe aim of this study was to evaluate in vitro and ex vivo roles of bortezomib, a proteasome inhibitor that binds to the active site of the 26S proteasome, in tertiary dentin formation. MethodsWe established pulpal access cavity preparation that was treated with or without bortezomib before direct pulp capping with a calcium hydroxide–based material. We also analyzed bone morphogenetic protein (Bmp)- and Wnt-related signaling molecules using quantitative real-time polymerase chain reaction. ResultsIn the short-term observation period, the bortezomib-treated pulp specimens showed the period-altered immunolocalization patterns of nestin, CD31, and myeloperoxidase, whereas the control specimens did not. The bortezomib-treated group showed a complete dentin bridge with very few irregular tubules after 42 days. The micro–computed tomographic images showed more apparent dentin bridge structures in the treated specimens than were in the controls. Quantitative real-time polymerase chain reaction analysis showed up-regulated Bmp and Wnt. ConclusionsThese findings revealed that treatment with 1 μmol/L bortezomib induced reparative dentin formation that facilitated the maintenance of the integrity of the remaining pulpal tissue via early vascularization and regulation of Bmp and Wnt signaling.

IGF-1 Mediates EphrinB1 Activation in Regulating Tertiary Dentin Formation

Eph receptors belong to a subfamily of receptor tyrosine kinases that are activated by membrane-spanning ligands called ephrins. Previously, we demonstrated that the ephrinB1-EphB2 interaction regulates odontogenic/osteogenic differentiation from dental pulp cells (DPCs) in vitro. The goal of this study was to identify the molecular mechanisms regulated by the EphB2/ephrinB1 system that govern tertiary dentin formation in vitro and in vivo. During tooth development, ephrinB1, and EphB2 were expressed in preodontoblast and odontoblasts at postnatal day 4. EphrinB1 was continuously expressed in odontoblasts and odontoblastic processes until the completion of tooth eruption. In addition, ephrinB1 was expressed in odontoblastic processes 2 wk following tooth injury without pulp exposure, whereas EphB2 was expressed in the center of pulp niches but not odontoblasts. In a model of tooth injury with pulp exposure, ephrinB1 was strongly expressed in odontoblasts 4 wk postinjury. In vitro studies with human and mouse DPCs treated with calcium hydroxide (CH) or mineral trioxide aggregate (MTA) showed an increased expression of insulin-like growth factor 1 (IGF-1). Experiments using several inhibitors of IGF-1 receptor signaling revealed that inhibiting the Ras/Raf-1/MAPK pathway inhibited EphB2 expression, and inhibiting the PI3K/Akt/mTOR pathway specifically inhibited ephrinB1 gene expression. Tooth injury in mice with odontoblast-specific IGF-1 receptor ablation exhibited a reduced tertiary dentin volume, mineral density, and ephrinB1 expression 4 wk following injury. We conclude that the IGF-1/ephrinB1 axis plays significant roles in the early stages of tooth injury. Further research is needed to fully understand the potential of targeting ephrinB1 as a regenerative pulp therapy.

Could the application of bioactive molecules improve vital pulp therapy success? A systematic review.

This study aimed to systematically review the literature of animal studies to evaluate whether bioactive dentin proteins could improve vital pulp therapy success. The review is reported in accordance with the PRISMA Statement. Two reviewers independently conducted a literature search of seven databases: PubMed (Medline), Lilacs, IBECS, BBO, Web of Science, Scopus, and SciELO. Animal experiments in which bioactive dentin proteins were applied directly or indirectly to the pulp tissue were included. Data regarding the characteristics of the proteins evaluated, the delivery systems used and the main findings from each study were tabulated to assess the outcomes of interest (tertiary dentin formation, inflammatory response, intratubular mineralization). After screening, 32 papers were subjected to qualitative analysis. In 75% of the studies, direct pulp capping was performed. Additionally, the most studied proteins were BMP-7, TGF-β1, and extracted soluble dentin matrix proteins. In conclusion, there is evidence in the literature suggesting that bioactive dentin molecules could enhance tertiary dentin formation with fewer initial inflammatory responses in direct and indirect pulp therapy in animal models. There are potential areas to be explored for novel therapeutic approaches for dental tissue repair and regeneration with bioactive materials. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 941-956, 2017.

Evaluation of a new mouse model for studying dental pulpal responses to GaAlAs laser irradiation

ObjectivesThe molecular mechanisms regulating pulpal responses to GaAlAs laser irradiation remain to be clarified. This study aimed to assess the feasibility of a mouse model for studying pulpal responses to GaAlAs laser irradiation. MethodsMaxillary first molars of 5-week-old ICR mice were irradiated at an output power of 1.0W for 180s, and samples were collected at intervals of 0, 1, 3, 5, 7, 10, and 14 days. The demineralized paraffin sections were processed for hematoxylin and eosin staining, immunohistochemistry for nestin (a marker for odontoblast differentiation) and Ki67 (a marker for cell proliferation), and a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. ResultsThe intense nestin immunoreactivity in the odontoblast layer of the mesial pulp was weakened immediately after irradiation and was almost lost on Days 1–3, although the extent of pulpal damage was variable among individual animals. At around Day 1, numerous TUNEL-positive cells appeared in the degenerative zone and gradually decreased in number by Day 14. Active cell proliferation occurred in the mesial pulp during Days 5–10. Nestin-positive odontoblast-like cells appeared along the pulp-dentin border by Day 10, resulting in tertiary dentin formation on Day 14. ConclusionsThe current output energy induced apoptosis in the affected dental pulp, followed by active cell proliferation, resulting in tertiary dentin formation. This is the first report regarding laser irradiation of teeth in an in vivo mouse model. This model could enable further understanding of the function of certain proteins, including transcriptional factors.

Effect of adhesive system application for cavities prepared with erbium, chromium: yttrium scandium gallium garnet laser on rat dental pulp tissue.

We examined the effects of adhesive systems under study applied for a laser-cut cavity using an Er,Cr:YSGG laser on rat dental pulp at 24h and 14days postoperatively. Group 1, laser-cut cavities were treated with a self-etching-primer and bonding agent; group 2, pretreated with a phosphoric-acid, and then treated with a self-etching-primer and bonding agent; group 3, pretreated with a phosphoric-acid and sodium-hypochlorite, and then treated with a self-etching-primer and bonding agent; and group 4, treated with an all-in-one adhesive. A flowable resin composite was used as filling material for each cavity treated with each group. A glass-ionomer-cement was used as a control. The following items were evaluated: pulp-tissue-disorganization (PTD), inflammatory-cell-infiltration (ICI), tertiary-dentin-formation (TDF), and bacterial-penetration (BP). The results were statistically analyzed using the Kruskal-Wallis test and Mann-Whitney U test. No significant differences were observed among the experimental groups for all parameters after 24h and 14days (P>0.05). The majority of the specimens showed PTD with edema formation after 24h; however, all the specimens demonstrated pulpal healing with TDF after 14days. On the parameter of TDF, all groups showed significant differences between the two postoperative periods (P<0.01). On the parameter of ICI, a significant difference was found between the two postoperative periods in group 4 (P<0.05). No specimens showed BP. The pretreatment on the cavity prepared with the laser using phosphoric-acid or sodium-hypochlorite did not affect the dental pulp healing of rat tooth.

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. DesignPulp 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, TGFβ1 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. ResultsIntrapulpal 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 TGFβ1 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 TGFβ1 group. ConclusionThe present experiments indicated that set calcium silicate could be used as carrier for biologically active molecules. TGFβ1 was shown to be an effective bioactive molecule in guiding tertiary dentine formation.

Pulp and apical tissue response to deep caries in immature teeth: A histologic and histobacteriologic study

Descriptions of the pathologic changes in the pulp and associated apical structures of human immature teeth in response to deep caries are lacking in the literature. ObjectivesThis article describes the histologic events associated with the radicular pulp and the apical tissues of human immature teeth following pulp inflammation and necrosis. MethodsTwelve immature teeth with destructive caries lesions were obtained from 8 patients. Two intact immature teeth served as controls. Teeth were extracted for reasons not related to this study and immediately processed for histopathologic and histobacteriologic analyses. Serial sections were examined for the pulp conditions and classified as reversible or irreversible pulp inflammation, or pulp necrosis. Other histologic parameters were also evaluated. ResultsIn the 3 cases with reversible pulp inflammation, tissue in the pulp chamber showed mild to moderate inflammation and tertiary dentin formation related to tubules involved in the caries process. Overall, the radicular pulp tissue, apical papilla and Hertwig’s epithelial root sheath (HERS) exhibited characteristics of normality. In the 3 cases with irreversible pulp inflammation, the pulps were exposed and severe inflammation occurred in the pulp chamber, with minor areas of necrosis and infection. Large areas of the canal walls were free from odontoblasts and lined by an atubular mineralized tissue. The apical papilla showed extremely reduced cellularity or lack of cells and HERS was discontinuous or absent. In the 6 cases with pulp necrosis, the coronal and radicular pulp tissue was necrotic and colonized by bacterial biofilms. The apical papilla could not be discerned, except for one case. HERS was absent in the necrotic cases. ConclusionWhile immature teeth with reversible pulpitis showed histologic features almost similar to normal teeth in the canal and in the apical region, those with irreversible pulpitis and necrosis exhibited significant alterations not only in the radicular pulp but also in the apical tissues, including the apical papilla and HERS. Clinical significanceAlterations in the radicular pulp and apical tissues help explain the outcome of current regenerative/reparative therapies and should be taken into account when devising more predictable therapeutic protocols for teeth with incomplete root formation.

Histologic Evaluation of Tertiary Dentine after Indirect Pulp Capping Procedures

Early carious lesions induce odontoblast stimulation in tertiary dentine formation even before the lesion reaches the dentine. The most used material for these procedures was calcium hydroxide, which was recently replaced by Mineral Trioxide Aggregate (MTA).The purpose of our study was to assess the histological characteristics of tertiary dentine induced by these materials in permanent molar teeth.We used 23 molars scheduled for extraction due to orthodontic reasons, in patients of 17-24 years of age. In a time interval of 4-8 weeks prior to extraction, occlusal cavities were prepared and filled with MTA or calcium hydroxide and glass ionomer cement.In teeth filled with MTA we noticed an early development of tertiary dentin layer, with a tubular structure, similar to secondary dentine. In the case of calcium hydroxide, the process of new dentine deposition was delayed and diffuse calcification, with formation of pulp stones was noticed.MTA proved to be superior to calcium hydroxide in inducing tertiary dentine formation, which appeared early after treatment. In time the differences in the amount of tertiary dentine between these materials are reduced, but there is a tendency of diffuse mineralization induced by calcium hydroxide.

Tertiary Dentin Formation after Indirect Pulp Capping Using Protein CPNE7

If there is a partial loss of dentin, the exposed dentinal surface should be protected by an indirect pulp capping (IPC) procedure to preserve pulp vitality and prevent symptoms of dentin hypersensitivity. In our previous study, copine7 (CPNE7) induced odontoblast differentiation in vitro and promoted dentin formation in vivo. The aim of this study was to investigate the possibility of IPC therapy using the CPNE7 protein at the exposed dentinal surface and the resulting effects on tertiary dentin formation in a beagle model. CPNE7 promoted mineralization of odontoblasts and had high calcium ion-binding capacity. The in vivo IPC model with canine teeth showed that regeneration of physiologic reactionary dentin with dentinal tubule structures was clearly observed beneath the remaining dentin in the CPNE7 group, whereas irregular features of reparative dentin were generated in the mineral trioxide aggregate (MTA) group. The CPNE7+MTA group also showed typical reactionary dentin without reparative dentin, showing synergistic effects of CPNE7 with MTA. A scanning electron microscopy analysis showed that dentinal tubules beneath the original dentin were occluded by the deposition of peritubular dentin in the CPNE7 and CPNE7+MTA groups, whereas those in the control group were opened. Therefore, CPNE7 may be able to serve as a novel IPC material and improve symptoms of dentin hypersensitivity.

Effects of human vascular endothelial growth factor on reparative dentin formation

It is a challenge for dentists to save dental pulp in patients with pulp disease without resorting to root canal therapy. Formation of tertiary dentin to maintain pulp vitality is a key odontoblast response to dental pulp injury. Vascular endothelial growth factor (VEGF) is the most potent angiogenic and vasculogenic factor involved in tertiary dentin formation. It was hypothesized that VEGF may be used to treat pulp diseases such as pulpitis. To explore this hypothesis, the first step was to assess whether VEGF affects dental pulp cells to promote reparative dentin formation. In the current study, an AdCMV-hVEGF vector was constructed to deliver hVEGF into dental pulp cells of exfoliated deciduous teeth (hDPCs) in vitro and dental pulp cells in a rat model in vivo. The collected data clearly demonstrated that hVEGF increased alkaline phosphatase and mineralization by enzymatic activity. RT-qPCR data demonstrated that hVEGF significantly increased the expression levels of genes commonly involved in osteogenesis/odontogenesis. Data from the in vivo assays indicated that hVEGF enhanced pulp cell proliferation and neovascularization, and markedly increased formation of reparative dentin in dental pulp. The in vitro and in vivo data suggest that hVEGF may have potential clinical applications, thus may aid in the development of novel treatment strategies for dental pulpitis.

Restored teeth can be used as samples for genotyping?

The aim of this study is to warn that such samples should not be overlooked for a successful genotyping.Six restored dental pieces were randomly selected from extreme forensic situations. For each tooth two portions have been selected: the dentin of adjacent portions of the restoration area (DS) and the remaining root dentin (DR). DNA extraction was executed using the PrepFiler​TM BTA kit according to the manufacturer’s instructions and its quantification was realized with the Quantifiler​TM Duo kit (both from Applied Biosystems). DS quantification values were comparatively higher than those of DR portion.These results may be related to a tertiary dentin formation, restorative and disorganized. The use of a sample with dentin tissue adjacent to a restoration area, covering the entire portion up to the pulp cavity, proved to be a good sample for genetic analysis, optimizing the quality of the sample volume.

Responses of pulp vasculature after cavity preparation in rat molars

ObjectivesOur recent study showed that cavity preparation increases the number of proliferative cells in the dental pulp during postoperative days 2–5. This study aimed to clarify pulp vascular changes following cavity preparation. MethodsGroove-shaped cavities were prepared on the medial surfaces of the upper first molars of 100-day-old Wistar rats. The animals were perfusion-fixed and tissues were collected during postoperative days 1–30, with subsequent India ink perfusion, immunohistochemistry for type IV collagen, CD31, and protein gene product (PGP) 9.5 and scanning electron microscopy with KOH digestion. The untreated upper first molars were used as controls. ResultsIn the controls, blood vessels with large diameters were located in the center of the pulp tissue and ramified to make capillary networks and PGP9.5-positive nerves were extensively arborized to form the subodontoblastic nerve plexus beneath the odontoblast layer. Cavity preparation induced disturbance in injured odontoblasts and subodontoblastic capillaries and nerves. Blood vessel density and thickness subsequently increased in the center and periphery of the pulp tissues with the exception of the subodontoblastic capillary network during postoperative days 3–5. PGP9.5-positive nerves overlapped with CD31-positive blood vessels in the mesial coronal pulp. Until day 30, when the tertiary dentin formation was completed, the pulp blood vasculature showed the same distribution and morphological features as that of the controls. ConclusionsThese results suggest that increased vascular flow under neuronal regulation plays an important role in cell proliferation in the dental pulp following cavity preparation.

Sympathetic Regulation of Tertiary Dentinogenesis via Beta-2 Adrenergic Receptor on Rat Odontoblasts

IntroductionBeta-2 adrenergic receptor has been found within the osteoblast membrane meditating bone remodeling. Propranolol is a sympatholytic beta antagonist commonly used as long-term medication for the management of many common diseases such as hypertension. This study was performed to verify the presence of this receptor on odontoblasts in rats and, if present, to investigate its possible association with tertiary dentinogenesis. MethodsTwenty male Sprague-Dawley rats (9 weeks old) were randomly assigned to 4 groups: CP0.8 group, cavity preparation + propranolol treatment (0.8 mg/day, n = 5); CP4 group, cavity preparation + propranolol treatment (4.0 mg/day, n = 7); CON group, cavity preparation + saline treatment (0.2 mL/day, n = 5); and NT group, no treatment (n = 3). Cavity preparation was performed on the mesial aspect of the maxillary first molars bilaterally. After 2 weeks, the tertiary dentinogenesis (CP0.8, CP4, and CON) was examined by hematoxylin-eosin staining, and the localization of beta-2 adrenergic receptor (NT) was examined by immunohistochemistry. ResultsThe beta-2 adrenergic receptor immunoreactivity was observed in the odontoblastic layer in normal rat molar dental pulp. The tertiary dentinogenesis beneath the prepared cavity was significantly higher in the rats receiving 2-week systemic administration of propranolol than in those without the propranolol treatment. The higher-dose treatment of propranolol (P < .001) presented more effective up-regulation of tertiary dentinogenesis than the lower-dose treatment (P < .01). ConclusionsThese results indicate that the sympathetic nervous system decreases tertiary dentin formation via beta-2 adrenergic receptors located on rat odontoblasts. It suggests that adrenergic beta antagonist is expected to use in the treatment of inducing tertiary dentin formation to protect dental pulp.

Wnt Acts as a Prosurvival Signal to Enhance Dentin Regeneration.

Wnt proteins are lipid-modified, short-range signals that control stem cell self-renewal and tissue regeneration. We identified a population of Wnt responsive cells in the pulp cavity, characterized their function, and then created a pulp injury. The repair response was evaluated over time using molecular, cellular, and quantitative assays. We tested how healing was impacted by wound environments in which Wnt signaling was amplified. We found that a Wnt-amplified environment was associated with superior pulp healing. Although cell death was still rampant, the number of cells undergoing apoptosis was significantly reduced. This resulted in significantly better survival of injured pulp cells, and resulted in the formation of more tertiary dentin. We engineered a liposome-reconstituted form of WNT3A then tested whether this biomimetic compound could activate cells in the injured tooth pulp and stimulate dentin regeneration. Pulp cells responded to the elevated Wnt stimulus by differentiating into secretory odontoblasts. Thus, transiently amplifying the body's natural Wnt response resulted in improved pulp vitality. These data have direct clinical implications for treating dental caries, the most prevalent disease affecting mankind.

Gypsum-based biomaterials: Evaluation of physical and mechanical properties, cellular effects and its potential as a pulp liner.

This in vitro study aimed to evaluate setting time and compressive strength of gypsum-based chitosan biomaterials and its effect on proliferation of stem cells from human exfoliated deciduous teeth (SHED) and alkaline phosphatase (ALP) activity. Pure-GYP was mixed with water (2.5 g: 1.9 mL); Gyp-CHT was prepared with gypsum, chitosan, and water (2.5 g: 0.285 g: 1.9 mL). Cell viability and ALP activity were assessed at different periods. Data were analyzed using SPSS (p<0.05). The setting times were 2.7 min and 2.8 min for pure-GYP and Gyp-CHT, respectively. Significantly higher compressive strength was observed with Gyp-CHT. SHED treatments with both materials were not cytotoxic. ALP was consistently higher in the treated groups compared with the control. Cellular attachments were evident with SEM. Excellent cellular viability with pure-GYP and Gyp-CHT, as well as increased ALP activities, suggested the possibility of tertiary dentin formation. Further studies are necessary to evaluate the biomaterials for its pulp protective potentialities.

Iloprost Induces Tertiary Dentin Formation

IntroductionProstacyclin (PGI2), a member of the prostaglandin family, can promote angiogenesis and cell proliferation. MethodsIn this study, the effect of the application of a PGI2 analog (iloprost) on dentin repair was examined in vitro and in vivo. ResultsIloprost significantly stimulated the expression of vascular endothelial growth factor and osteo-/odontogenic marker messenger RNA in human dental pulp cells (HDPCs) under osteoinductive conditions in vitro. In addition, iloprost enhanced HDPC alkaline phosphatase enzymatic activity and mineral deposition. An in vivo study was performed using a rat molar mechanical pulp exposure model. After 30 days, histologic analysis revealed that there was a dramatic tertiary dentin formation in the iloprost-treated group compared with the calcium hydroxide and the untreated control groups. Furthermore, vascular endothelial growth factor protein expression in dental pulp tissue was increased in the iloprost-treated group as determined by immunohistochemical staining. ConclusionsTaken together, the present study, for the first time, shows that iloprost induces the expression of osteo-/odontogenic markers in vitro and promotes angiogenic factor expression and enhances tertiary dentin formation in vivo. This implies the potential clinical usefulness of iloprost in vital pulp therapy.

Effects of a Triple Antibiotic Solution on Pulpal Dynamics after Intentionally Delayed Tooth Replantation in Mice

IntroductionThis study analyzed the detailed biological events underlying pulpal dynamics evoked by 3Mix (the mixture of ciprofloxacin, metronidazole, and minocycline) solution after intentionally delayed tooth replantation because 3Mix improves pulpal healing after tooth injuries. MethodsThe maxillary first molars of 3-week-old mice were extracted and immersed in 3Mix solution for 30 minutes in comparison with phosphate buffered saline (PBS) alone. Cell proliferation, apoptosis, and differentiation were assessed in extracted/replanted teeth during days 0–14 using immunohistochemistry, apoptosis assay, and reverse-transcriptase polymerase chain reaction. Results3Mix solution accelerated odontoblast differentiation in the coronal pulp on day 7 and tertiary dentin formation on day 14, whereas the regenerative process was delayed in the PBS group. Cell proliferation and apoptosis occurred in the pulp of the 3Mix group during days 5–7 and subsequently decreased from days 7–14. On day 5, dentin sialophosphoprotein and nestin were first recovered in the 3Mix group, whereas expression levels for alkaline phosphatase, osteopontin, and osteocalcin increased in the PBS group. The expression levels for octamer-binding factor 3/4A and 3/4B reached the maximum level on day 1 and were sharply decreased on day 3 in both groups. High expression levels of Cd11c were first observed in the 3Mix group on day 1 and later at days 5 and 7. ConclusionsThe results suggest that the application of 3Mix may suppress osteoblast differentiation by the migration of dendritic cells to the injury site and via the activation of stem/progenitor cells, resulting in the acceleration of odontoblastlike cell differentiation.

Minimally invasive clinical approach in indirect pulp therapy and healing of deep carious lesions.

Indirect pulp treatment is a conservative vital pulp procedure performed in deep carious lesion approximating the pulp, but without signs or symptoms of pulp degeneration. Removing the carious biomass along with sealing the residual caries from extrinsic substrate and oral bacteria makes residual caries after the first excavation less active. This allows time for pulpo dentinal complex to form tertiary dentine so that at the second excavation, there is less likelihood of pulpal exposure. It has also been suggested that by changing the cavity environment from an active lesion into a more slowly progressing lesion, will be accompanied by more regular tubular tertiary dentin formation. The success of this approach has been demonstrated by various randomized controlled studies comparing conventional treatment of such lesions with stepwise excavation. These results are echoed at clinical, radiographic, macroscopic, microscopic and ultrastructural level during follow up visits. This study reviews promising concepts and rationale of minimally invasive indirect pulp therapy technique where conventional wisdom of caries removal is challenged

Odontogenic Effect of a Fast-setting Pozzolan-based Pulp Capping Material

IntroductionMineral trioxide aggregate (MTA) is widely used as a pulp capping material. Recently, a MTA-derived fast-setting pozzolan cement (Endocem; Maruchi, Wonju, Korea) was introduced in the endodontic field. Our aim in this study was to investigate the odontogenic effects of this cement in vitro and in vivo. MethodsHuman dental pulp cells (hDPCs) were cultured, and the effects of Endocem and a previously marketed MTA (ProRoot; Dentsply, Tulsa, OK) on biocompatibility were evaluated by assessing cell morphology and performing a cell viability test. Chemical composition of each material was analyzed by energy-dispersive X-ray spectroscopic analysis. Odontoblastic differentiation was analyzed by alkaline phosphatase activity and alizarin red S staining. The expression of odontogenic-related markers, namely dentin sialophosphoprotein, dentin matrix protein 1, and osteonectin, was evaluated by real-time polymerase chain reaction, Western blotting, and immunofluorescence analysis. Pinpoint pulp exposures were made on rat teeth and then capped with ProRoot or Endocem. After 4 weeks, reparative tertiary dentin formation and inflammatory responses were investigated histologically. ResultsThe biocompatibility of Endocem was similar to that of ProRoot. Energy-dispersive X-ray spectroscopic analysis showed that ProRoot and Endocem contained similar elemental constituents such as calcium, oxygen, and silicon. Alkaline phosphatase activity and mineralized nodule formation increased in ProRoot- and Endocem-treated cells compared with medium only–treated cells in the control group (P < .05). The expression of odontogenic-related markers was significantly higher in the ProRoot- and Endocem-treated groups than the control group (P < .05), but there was no significant difference in the expression of these markers between the 2 experimental groups (P > .05). Four weeks after the pulp capping procedure, continuous tertiary dentin had formed directly underneath the capping materials and the pulp exposure area in all samples in the 2 treated groups. Furthermore, most specimens either had no inflammation or minor pulpal inflammation. ConclusionsOur results indicate that ProRoot and Endocem have similar biocompatibility and odontogenic effects. Therefore, Endocem is as effective a pulp capping material as ProRoot.

Evaluation of antibacterial and remineralizing nanocomposite and adhesive in rat tooth cavity model

Antibacterial and remineralizing dental composites and adhesives were recently developed to inhibit biofilm acids and combat secondary caries. It is not clear what effect these materials will have on dental pulps in vivo. The objectives of this study were to investigate the antibacterial and remineralizing restorations in a rat tooth cavity model, and determine pulpal inflammatory response and tertiary dentin formation. Nanoparticles of amorphous calcium phosphate (NACP) and antibacterial dimethylaminododecyl methacrylate (DMADDM) were synthesized and incorporated into a composite and an adhesive. Occlusal cavities were prepared in the first molars of rats and restored with four types of restoration: control composite and adhesive; control plus DMADDM; control plus NACP; and control plus both DMADDM and NACP. At 8 or 30days, rat molars were harvested for histological analysis. For inflammatory cell response, regardless of time periods, the NACP group and the DMADDM+NACP group showed lower scores (better biocompatibility) than the control group (p=0.014 for 8days, p=0.018 for 30days). For tissue disorganization, NACP and DMADDM+NACP had better scores than the control (p=0.027) at 30days. At 8days, restorations containing NACP had a tertiary dentin thickness (TDT) that was five- to six-fold that of the control. At 30days, restorations containing NACP had a TDT that was four- to six-fold that of the control. In conclusion, novel antibacterial and remineralizing restorations were tested in rat teeth in vivo for the first time. Composite and adhesive containing NACP and DMADDM exhibited milder pulpal inflammation and much greater tertiary dentin formation than the control adhesive and composite. Therefore, the novel composite and adhesive containing NACP and DMADDM are promising as a new therapeutic restorative system to not only combat oral pathogens and biofilm acids as shown previously, but also facilitate the healing of the dentin–pulp complex.