This study aimed to evaluate carbonated hydroxyapatite (CHA)’s ability for mineral induction and its in vitro cytotoxicity with human dental pulp cells.

Precursors for the study include di-ammonium hydrogen phosphate and calcium nitrate tetrahydrate, with sodium hydrogen carbonate added to achieve different levels of carbonate substitution. The synthesized CHA samples are characterized using X-ray diffraction, Fourier transform infrared spectroscopy, and Raman spectroscopy. Scanning electron microscopy (SEM) was used to observe morphology. For 14 days at 37°C, samples were submerged in simulated body fluid to assess their mineral induction capabilities. SEM was used to confirm apatite formation on sample surfaces. The cytotoxicity assay was used to assess the vitality of the cells following their exposure to various concentrations of CHA.

The Joint Committee on Powder Diffraction Standards data for HA aligned well with the results from X-ray diffraction analysis of CHA across 3 different concentrations, indicating strong agreement. Fourier transform infrared spectra indicated the presence of phosphate, hydroxyl, and carbonate groups within the samples. SEM and Energy-dispersive X-ray analysis show agglomerated and flaky nanoparticles. All the samples are bioactive, but the formation of apatite differs from one another. In vitro cytotoxicity assay showed that over 70% of cells maintain viability.

The results of this study may provide insight into the potential use of carbonated HA as a dental pulp-capping material for vital pulp therapy.

This study aimed to investigate the endodontic characteristics of mandibular premolars with dens evaginatus (DE) that require endodontic treatment.

Patients who underwent endodontic treatment were enrolled. The inclusion criteria were patients who underwent root canal treatment in the lower permanent teeth with DE and were followed up for at least 1 year. Preoperative clinical and radiographic variables were obtained. The frequency distribution of the preoperative variables was compared using the χ² or Fisher’s exact tests. The significance of the change in periapical health index (PAI) and root development stages before and after treatment was examined using the Wilcoxon signed-rank test.

A total of 150 teeth of 134 patients with an average age of 15.3 years were included. The percentage distribution comparison of the preoperative variables and obturation techniques revealed significant differences in pulpal and periapical diagnosis, and percussion, and especially regarding age, root development stage, and PAI. Age was the only statistically significant preoperative variable associated with root growth (p < 0.05).

Approximately, 60% of DEs requiring endodontic treatment had immature roots. Age being the most significant predisposing factor, early treatment provides the greatest opportunity for full root development.

This study aimed to investigate the effectiveness of different topical/systemic agents in reducing the damage caused by bleaching gel to pulp tissue or cells.

Electronic searches were performed in July 2023. In vivo and in vitro studies evaluating the effects of different topical or systemic agents on pulp inflammation or cytotoxicity after exposure to bleaching agents were included. The risk of bias was assessed.

Out of 1,112 articles, 27 were included. Nine animal studies evaluated remineralizing/anti-inflammatories agents in rat molars subjected to bleaching with 35%–38% hydrogen peroxide (HP). Five of these studies demonstrated a significant reduction in inflammation caused by HP when combined with bioglass or MI Paste Plus (GC America), or following KF-desensitizing or Otosporin treatment (n = 3). However, orally administered drugs did not reduce pulp inflammation (n = 4). Cytotoxicity (n = 17) was primarily assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on human dental pulp cells and mouse dental papilla Cell-23 cells. Certain substances, including sodium ascorbate, butein, manganese chloride, and peroxidase, were found to reduce cytotoxicity, particularly when applied prior to bleaching. The risk of bias was high in animal studies and low in laboratory studies.

Few in vivo studies have evaluated agents to reduce the damage caused by bleaching gel to pulp tissue. Within the limitations of these studies, it was found that topical agents were effective in reducing pulp inflammation in animals and cytotoxicity. Further analyses with human pulp are required to substantiate these findings.

PROSPERO Identifier: CRD42022337192

This study aimed to determine whether collagen triple helix repeat containing-1 (CTHRC1), which is involved in vascular remodeling and bone formation, can stimulate odontogenic differentiation and angiogenesis when administered to human dental pulp stem cells (hDPSCs).

The viability of hDPSCs upon exposure to CTHRC1 was assessed with the WST-1 assay. CTHRC1 doses of 5, 10, and 20 µg/mL were administered to hDPSCs. Reverse-transcription polymerase reaction was used to detect dentin sialophosphoprotein, dentin matrix protein 1, vascular endothelial growth factor, and fibroblast growth factor 2. The formation of mineralization nodules was evaluated using Alizarin red. A scratch wound assay was conducted to evaluate the effect of CTHRC1 on cell migration. Data were analyzed using 1-way analysis of variance followed by the Tukey post hoc test. The threshold for statistical significance was set at p < 0.05.

CTHRC1 doses of 5, 10, and 20 µg/mL had no significant effect on the viability of hDPSCs. Mineralized nodules were formed and odontogenic markers were upregulated, indicating that CTHRC1 promoted odontogenic differentiation. Scratch wound assays demonstrated that CTHRC1 significantly enhanced the migration of hDPSCs.

CTHRC1 promoted odontogenic differentiation and mineralization in hDPSCs.

This study evaluated 2 nickel-titanium rotary systems and a complementary protocol with an ultrasonic tip and a small-diameter instrument in flattened root canals.

Thirty-two human maxillary second premolars with flattened canals (buccolingual diameter ≥4 times larger than the mesiodistal diameter) at 9 mm from the radiographic apex were selected. The root canals were prepared by ProDesign Logic (PDL) 30/0.01 and 30/0.05 or Hyflex EDM (HEDM) 10/0.05 and 25/0.08 (n = 16), followed by application of the Flatsonic ultrasonic tip in the cervical and middle thirds and a PDL 25/0.03 file in the apical third (FPDL). The teeth were scanned using micro-computed tomography before and after the procedures. The percentage of volume increase, debris, and uninstrumented surface area were analyzed using the Kruskal-Wallis, Dunn, Wilcoxon, analysis of variance/Tukey, and paired and unpaired t-tests (α = 0.05).

No significant difference was found in the volume increase and uninstrumented surface area between PDL and HEDM (p > 0.05). PDL had a higher percentage of debris than HEDM in the middle and apical thirds (p < 0.05). The FPDL protocol resulted in less debris and uninstrumented surface area for PDL and HEDM (p < 0.05). This protocol, with HEDM, reduced debris in the middle and apical thirds and uninstrumented surface area in the apical third (p < 0.05).

High percentages of debris and uninstrumented surface area were observed after preparation of flattened root canals. The HEDM, Flatsonic tip, and 25/0.03 instrument protocol enhanced cleaning in flattened root canals.

This systematic review evaluated the efficacy of the supplementary use of the XP-endo Finisher on bacteria content reduction in the root canal system.

In-vitro studies evaluating the use of the XP-endo Finisher on bacteria content were searched in four databases in July 2020. Two authors independently screened the studies for eligibility. Data were extracted, and risk of bias was assessed. Data were meta-analyzed by using random-effects model to compare the effect of the supplementary use (experimental) or not (control) of the XP-endo Finisher on bacteria counting reduction, and results from different endodontic protocols were combined. Four studies met the inclusion criteria while 1 study was excluded from the meta-analysis due to its high risk of bias and outlier data. The 3 studies that made it to the meta-analysis had an unclear risk of bias for at least one criterion.

No heterogeneity was observed among the results of the studies included in the meta-analysis. The study excluded from the meta-analysis assessing the bacteria counting deep in the dentin demonstrated further bacteria reduction upon the use of the XP-endo Finisher.

This systematic review found no evidence supporting the supplementary use of the XP-endo Finisher on further bacteria counting the reduction in the root canal.

This study aimed to assess the presence of pulp stones through an examination of cone beam computed tomography images and correlate their prevalence with age, sex, dental arch and side, tooth type, and restoration type and depth.

Cone beam computed tomography images obtained from 673 patients and archival data on 11,494 teeth were evaluated. The associations of pulp stones with age, sex, dental arch and side, tooth type, and restoration type and depth were noted. All the measurements were subjected to a χ² test and one sample χ² test (p < 0.05).

In the study group, 163 (24.2%) patients and 379 (3.3%) teeth had at least one pulp stone. The pulp stone frequency in those aged 30–39 years was significantly greater than in those aged 18–29 and ≥ 60 years, and the frequency was higher in females than in males (p < 0.05). The highest prevalence of pulp stones was found in maxillary dental arches and molar teeth (p < 0.05). Pulp stones were significantly more common in medium-depth restorations (p < 0.05).

Maxillary molar teeth, medium-depth restorations, individuals aged 30–39 years and females had a greater percentage of pulp stones.

The aim of the present systematic review was to investigate the cryopreservation process of dental pulp mesenchymal stromal cells and whether cryopreservation is effective in promoting cell viability and recovery.

This systematic review was developed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement and the research question was determined using the population, exposure, comparison, and outcomes strategy. Electronic searches were conducted in the PubMed, Cochrane Library, Science Direct, LILACS, and SciELO databases and in the gray literature (dissertations and thesis databases and Google Scholar) for relevant articles published up to March 2019. Clinical trial studies performed with dental pulp of human permanent or primary teeth, containing concrete information regarding the cryopreservation stages, and with cryopreservation performed for a period of at least 1 week were included in this study.

The search strategy resulted in the retrieval of 185 publications. After the application of the eligibility criteria, 21 articles were selected for a qualitative analysis.

The cryopreservation process must be carried out in 6 stages: tooth disinfection, pulp extraction, cell isolation, cell proliferation, cryopreservation, and thawing. In addition, it can be inferred that the use of dimethyl sulfoxide, programmable freezing, and storage in liquid nitrogen are associated with a high rate of cell viability after thawing and a high rate of cell proliferation in both primary and permanent teeth.

This systematic review aimed to identify mean oxygen saturation values (SpO₂) using pulse oximetry in permanent maxillary anterior teeth.

The MEDLINE, Scientific Electronic Library Online, Cochrane Central Register of Controlled Trials, EMBASE, and Literatura Latino Americana em Ciências da Saúde electronic databases were searched. Combinations and variations of “oximetry” AND “dental pulp test” were used as search terms. Studies reporting means and standard deviations of SpO₂ values were included. Two reviewers independently extracted data following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses checklist. Heterogeneity was assessed using the I² statistic, and all analyses were performed using R software. Study quality was assessed using the Quality Assessment of Diagnostic Accuracy Studies-2 tool and the Newcastle-Ottawa scale.

Of the 251 studies identified, 19 met the eligibility criteria and were included (total sample, 4,541 teeth). In the meta-analysis, the mean SpO₂ values were 84.94% (95% confidence interval [CI], 84.85%–85.04%) for the central incisors, 89.29% (95% CI, 89.22%–89.35%) for the lateral incisors, and 89.20% (95% CI, 89.05%–89.34%) for the canines. The studies were predominantly low-quality due to the high risk of bias associated with the index test, unclear risk regarding patient selection, and concerns about outcome assessment.

Although most studies were low-quality, the oxygen saturation levels in normal pulp could be established (minimum saturation, 77.52%). Despite the risk of bias of the included studies, the reference values reported herein are clinically relevant for assessments of changes in pulp status.

International Prospective Register of Systematic Reviews Identifier: CRD42018085598

This study investigated the indirect effect of calcium-enriched mixture (CEM) cement and mineral trioxide aggregate (MTA), as 2 calcium silicate-based hydraulic cements, on human dental pulp stem cells (hDPSCs) through different dentin thicknesses.

Two-chamber setups were designed to simulate indirect pulp capping (IPC). Human molars were sectioned to obtain 0.1-, 0.3-, and 0.5-mm-thick dentin discs, which were placed between the 2 chambers to simulate an IPC procedure. Then, MTA and CEM were applied on one side of the discs, while hDPSCs were cultured on the other side. After 2 weeks of incubation, the cells were removed, and cell proliferation, morphology, and attachment to the discs were evaluated under scanning electron microscopy (SEM). Energy-dispersive X-ray (EDXA) spectroscopy was performed for elemental analysis. Alkaline phosphatase (ALP) activity was assessed quantitatively. The data were analyzed using the Kruskal-Wallis and Mann-Whitney tests.

SEM micrographs revealed elongated cells, collagen fibers, and calcified nucleations in all samples. EDXA verified that the calcified nucleations consisted of calcium phosphate. The largest calcifications were seen in the 0.1-mm-thick dentin subgroups. There was no significant difference in ALP activity across the CEM subgroups; however, ALP activity was significantly lower in the 0.1-mm-thick dentin subgroup than in the other MTA subgroups (p < 0.05).

The employed capping biomaterials exerted biological activity on hDPSCs, as shown by cell proliferation, morphology, and attachment and calcific precipitations, through 0.1- to 0.5-mm-thick layers of dentin. In IPC, the bioactivity of these endodontic biomaterials is probably beneficial.

To achieve pulp-dentin complex regeneration with tissue engineering, treatment efficacies and safeties should be evaluated using in vivo orthotopic transplantation in a sufficient number of animals. Mice have been a species of choice in which to study stem cell biology in mammals. However, most pulp-dentin complex regeneration studies have used large animals because the mouse tooth is too small. The purpose of this study was to demonstrate the utility of the mouse tooth as a transplantation model for pulp-dentin complex regeneration research.

Experiments were performed using 7-week-old male Institute of Cancer Research (ICR) mice; a total of 35 mice had their pulp exposed, and 5 mice each were sacrificed at 1, 2, 4, 7, 9, 12 and 14 days after pulp exposure. After decalcification in 5% ethylenediaminetetraacetic acid, the samples were embedded and cut with a microtome and then stained with hematoxylin and eosin. Slides were observed under a high-magnification light microscope.

Until 1 week postoperatively, the tissue below the pulp chamber orifice appeared normal. The remaining coronal portion of the pulp tissue was inflammatory and necrotic. After 1 week postoperatively, inflammation and necrosis were apparent in the root canals inferior to the orifices. The specimens obtained after experimental day 14 showed necrosis of all tissue in the root canals.

This study could provide opportunities for researchers performing in vivo orthotopic transplantation experiments with mice.

Direct pulp capping is a treatment for mechanically exposed pulp in which a biocompatible capping material is used to preserve pulpal vitality. Biocompatibility tests in animal studies have used a variety of experimental protocols, particularly with regard to the exposure site. In this study, pulp exposure on the occlusal and mesial surfaces of molar teeth was investigated in a rat model.

A total of 58 maxillary first molars of Wistar rats were used. Forty molars were mechanically exposed and randomly assigned according to 3 factors: 1) the exposure site (occlusal or mesial), 2) the pulp-capping material (ProRoot White MTA or Bio-MA), and 3) 2 follow-up periods (1 day or 7 days) (n = 5 each). The pulp of 6 intact molars served as negative controls. The pulp of 12 molars was exposed without a capping material (n = 3 per exposure site for each period) and served as positive controls. Inflammatory cell infiltration and reparative dentin formation were histologically evaluated at 1 and 7 days using grading scores.

At 1 day, localized mild inflammation was detected in most teeth in all experimental groups. At 7 days, continuous/discontinuous calcified bridges were formed at exposure sites with no or few inflammatory cells. No significant differences in pulpal response according to the exposure site or calcium-silicate cement were observed.

The location of the exposure site had no effect on rat pulpal healing. However, mesial exposures could be performed easily, with more consistent results. The pulpal responses were not significantly different between the 2 capping materials.

The aim of this in vivo study was to assess the accuracy of 2 third-generation electronic apex locators (EALs), Propex II (Dentsply Maillefer) and Root ZX II (J. Morita), and radiographic technique for locating the major foramen (MF).

Thirty-two premolars with single canals that required extraction were included. Following anesthesia, access, and initial canal preparation with size 10 and 15 K-flex files and SX and S1 rotary ProTaper files, the canals were irrigated with 2.5% sodium hypochlorite. The length of the root canal was verified 3 times for each tooth using the 2 apex locators and once using the radiographic technique. Teeth were extracted and the actual WL was determined using size 15 K-files under a × 25 magnification. The Biostat 4.0 program (AnalystSoft Inc.) was used for comparing the direct measurements with those obtained using radiographic technique and the apex locators. Pearson's correlation analysis and analysis of variance (ANOVA) were used for statistical analyses.

The measurements obtained using the visual method exhibited the strongest correlation with Root ZX II (r = 0.94), followed by Propex II (r = 0.90) and Ingle's technique (r = 0.81; p < 0.001). Descriptive statistics using ANOVA (Tukey's post hoc test) revealed significant differences between the radiographic measurements and both EALs measurements (p < 0.05).

Both EALs presented similar accuracy that was higher than that of the radiographic measurements obtained with Ingle's technique. Our results suggest that the use of these EALs for MF location is more accurate than the use of radiographic measurements.

In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications.

HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated. In vitro differentiation capacities of HDPCs into adipogenic, osteogenic, and chondrogenic lineages were compared. Differentiation markers were analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blotting.

Our data indicated that whole HDPCs-ED, HPDCs-OG, and HDPCs-Combined could be differentiated into adipogenic, chrondrogenic, and osteogenic cell types. However, we found that the methods for isolating and culturing HDPCs influence the differentiation capacities of cells. HDPCs-OG and HDPCs-ED were preferably differentiated into adipogenic and osteogenic cells, respectively. Differentiation markers shown by RT-PCR and western blotting analysis were mostly upregulated in the treated groups compared with the control groups.

Our findings confirmed that cell populations formed by two different culture methods and the combined culture method exhibited different properties. The results of this study could provide an insight into regenerative endodontic treatment using HDPCs.

The purpose of this study was to investigate the involvement of TRPA1 in the cinnamaldehyde-induced pulpal blood flow (PBF) change in the feline dental pulp.

Mandibles of eight cats were immobilized and PBF was monitored with a laser Doppler flowmetry at the mandibular canine tooth. To evaluate the effect of cinnamaldehyde on PBF, cinnamaldehyde was injected into the pulp through the lingual artery at a constant rate for 60 seconds. As a control, a mixture of 70% ethanol and 30% dimethyl sulfoxide (DMSO, vehicle) was used. To evaluate the involvement of transient receptor potential ankyrin 1 (TRPA1) in PBF change, AP18, a specific TRPA1 antagonist, was applied into the pulp through the Class V dentinal cavity followed by cinnamaldehyde-administration 3 minutes later. The paired variables of experimental data were statistically analyzed using paired t-test. A p value of less than 0.05 was considered as statistically significant.

Administration of cinnamaldehyde (0.5 mg/kg, intra-arterial [i.a.]) induced significant increases in PBF (p < 0.05). While administration of a TRPA1 antagonist, AP18 (2.5 - 3.0 mM, into the dentinal cavity [i.c.]) caused insignificant change of PBF (p > 0.05), administration of cinnamaldehyde (0.5 mg/kg, i.a.) following the application of AP18 (2.5 - 3.0 mM, i.c.) resulted in an attenuation of PBF increase from the control level (p < 0.05). As a result, a TRPA1 antagonist, AP18 effectively inhibited the vasodilative effect of cinnamaldehyde (p < 0.05).

The result of the present study provided a functional evidence that TRPA1 is involved in the mechanism of cinnamaldehyde-induced vasodilation in the feline dental pulp.

The purpose of this study was to evaluate the histopathological effects of an antioxidant therapy on the pulp tissue of rat teeth exposed to a bleaching gel with 35% hydrogen peroxide.

Forty rats were subjected to oral ingestion by gavage of distilled water (DW) or ascorbic acid (AA) 90 min before the bleaching therapy. For the bleaching treatment, the agent was applied twice for 5 min each to buccal surfaces of the first right mandibular molars. Then, the animals were sacrificed at 6 hr, 24 hr, 3 day, or 7 day post-bleaching, and the teeth were processed for microscopic evaluation of the pulp tissue.

At 6 hr, the pulp tissue showed moderate inflammatory reactions in all teeth of both groups. In the DW and AA groups, 100% and 80% of teeth exhibited pulp tissue with significant necrosis and intense tissue disorganization, respectively. At 24 hr, the AA-treated group demonstrated a greater regenerative capability than the DW group, with less intense inflammatory reaction and new odontoblast layer formation in 60% of the teeth. For up to the 7 day period, the areas of pulpal necrosis were replaced by viable connective tissue, and the dentin was underlined by differentiated odontoblast-like cells in most teeth of both groups.

A slight reduction in initial pulpal damage during post-bleaching was promoted by AA therapy. However, the pulp tissue of AA-treated animals featured faster regenerative potential over time.

Sodium hypochlorite (NaOCl) is an excellent bactericidal agent, but it is detrimental to stem cell survival, whereas intracanal medicaments such as calcium hydroxide (Ca[OH]₂) promote the survival and proliferation of stem cells. This study evaluated the effect of sequential NaOCl and Ca[OH]₂ application on the attachment and differentiation of dental pulp stem cells (DPSCs).

DPSCs were obtained from human third molars. All dentin specimens were treated with 5.25% NaOCl for 30 min. DPSCs were seeded on the dentin specimens and processed with additional 1 mg/mL Ca[OH]₂, 17% ethylenediaminetetraacetic acid (EDTA) treatment, file instrumentation, or a combination of these methods. After 7 day of culture, we examined DPSC morphology using scanning electron microscopy and determined the cell survival rate with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. We measured cell adhesion gene expression levels after 4 day of culture and odontogenic differentiation gene expression levels after 4 wk using quantitative real-time polymerase chain reaction.

DPSCs did not attach to the dentin in the NaOCl-treated group. The gene expression levels of fibronectin-1 and secreted phosphoprotein-1 gene in both the Ca[OH]₂- and the EDTA-treated groups were significantly higher than those in the other groups. All Ca[OH]₂-treated groups showed higher expression levels of dentin matrix protein-1 than that of the control. The dentin sialophosphoprotein level was significantly higher in the groups treated with both Ca[OH]₂ and EDTA.

The application of Ca[OH]₂ and additional treatment such as EDTA or instrumentation promoted the attachment and differentiation of DPSCs after NaOCl treatment.

The aim of this study was to investigate the expression of 7 different sirtuin genes (SIRT1-SIRT7) in human dental pulp cells (HDPCs), and to determine the role of SIRTs in the odontoblastic differentiation potential of HDPCs.

HDPCs were isolated from freshly extracted third molar teeth of healthy patients and cultulred in odontoblastic differentiation inducing media. Osteocalcin (OCN) and dentin sialophosphoprotein (DSPP) expression was analyzed to evaluate the odontoblastic differentiation of HDPCs by reverse transcription-polymerase chain reaction (RT-PCR), while alizarin red staining was used for the mineralization assay. To investigate the expression of SIRTs during odontoblastic differentiation of HDPCs, real time PCR was also performed with RT-PCR.

During the culture of HDPCs in the differentiation inducing media, OCN, and DSPP mRNA expressions were increased. Mineralized nodule formation was also increased in the 14 days culture. All seven SIRT genes were expressed during the odontogenic induction period. SIRT4 expression was increased in a time-dependent manner.

Our study identified the expression of seven different SIRT genes in HDPCs, and revealed that SIRT4 could exert an influence on the odontoblast differentiation process. Further studies are needed to determine the effects of other SIRTs on the odontogenic potential of HDPCs.

We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization.

Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data.

Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated.

Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

The aim was to confirm the stem cell-like properties of the dental pulp stromal cells and to evaluate the morphologic changes during in vitro chondrogenesis.

Stromal cells were outgrown from the dental pulp tissue of the premolars. Surface markers were investigated and cell proliferation rate was compared to other mesenchymal stem cells. Multipotency of the pulp cells was confirmed by inducing osteogenesis, adipogenesis and chondrogenesis. The morphologic changes in the chondrogenic pellet during the 21 day of induction were evaluated under light microscope and transmission electron microscope. TUNEL assay was used to evaluate apoptosis within the chondrogenic pellets.

Pulp cells were CD90, 105 positive and CD31, 34 negative. They showed similar proliferation rate to other stem cells. Pulp cells differentiated to osteogenic, adipogenic and chondrogenic tissues. During chondrogenesis, 3-dimensional pellet was created with multi-layers, hypertrophic chondrocyte-like cells and cartilage-like extracellular matrix. However, cell morphology became irregular and apoptotic cells were increased after 7 day of chondrogenic induction.

Pulp cells indicated mesenchymal stem cell-like characteristics. During the in vitro chondrogenesis, cellular activity was superior during the earlier phase (within 7 day) of differentiation.