Immunohistochemical analysis of stem cells from human exfoliated deciduous teeth seeded in carbonate apatite scaffold for the alveolar bone defect in Wistar rats ( Rattus novergicus).

Background: Stem cells from human exfoliated deciduous teeth (SHED) seeded in carbonate apatite scaffold (CAS) may have multiple functions that could be used to regenerate the alveolar bone defects. The purpose of this study is to examine the ability of SHED and CAS in alveolar bone defects using an immunohistochemical analysis. Methods: ten three-month-old healthy male Wistar rats (R. novergicus) that weighed between 150-250 grams (g) were used as animal models. A simple blind random sampling method was used to select the sample that was assigned to the study group for CAS and SHED seeded in CAS (n=5). The animal study model of the alveolar bone was established by extracting the anterior mandible teeth. Rodent anesthesia was applied to relieve the pain during the procedure for all test animals. Immunohistochemistry was performed after seven days to facilitate the examination of the receptor activator of NF-κß ligand (RANKL), osteoprotegrin (OPG), transforming growth factor-ß (TGF-ß), vascular endothelial growth factor (VEGF), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin, and osteopontin expression. The data was analyzed using the unpaired t-test (p<0.01) and Pearson's correlation test (p<0.05). Results: The OPG, RUNX2, TGF-ß, VEGF, ALP, osteocalcin, and ostepontin expressions were higher in SHED seeded in CAS than CAS only with a significant difference between the groups (p<0.01). Furthermore, the RANKL expression was lower in SHED seeded in CAS compared to CAS only. There was a strong reverse significant correlation between OPG and RANKL expression (p<0.05). Conclusions: The number of osteogenic marker expressing cells, such as OPG, RUNX2, TGF-ß, VEGF, ALP, osteocalcin, and ostepontin, increased. However, RANKL expression in the alveolar bone defects that were implanted with SHED seeded in CAS did not increase after seven days.

Proliferation and osteogenic differentiation of bone marrow-derived mesenchymal stem cell after exposure to red flesh dragon fruit extract.

BACKGROUND: Induction of the proliferation and differentiation of stem cells could represent a viable alternative therapeutic method for treating bone diseases. Stem cells are essential to bone tissue regeneration; although, their availability is limited. One possible method of increasing the number of stem cells and promote osteogenic differentiation is the application of red flesh dragon fruit extract supplement. The present study was performed to identify and analyze proliferation and osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMMSCs) after exposure to red flesh dragon fruit extract. MATERIALS AND METHODS: This in vitro study was posttest only control group design. Red flesh dragon fruit extract was produced by means of water extraction method and subsequent dilution with different amounts of water to produce a range of concentrations. BMMSCs were obtained from the femurs of three White New Zealand rabbits. BMMSCs were then treated with 50, 100, 200, 300, and 400 µg/ml red flesh dragon fruit extract concentrations. The in vitro proliferation assay was determined by means of an 3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Osteogenic differentiation was determined by means of the degree of nodule mineralization. There were two groups as follows: group I with the addition of 50 µg/ml of red flesh dragon fruit extract and Group II without the addition of red flesh dragon fruit. Data were analyzed using analysis of variance and the Student's t-test (P = 0.05). RESULTS: 50, 100, 200, 300, and 400 µg/ml of red flesh dragon fruit extract demonstrated the capacity to significantly increase the proliferation of BMMSCs (P ≤ 0.05). Red flesh dragon fruit extract could significantly increase osteogenic differentiation (P ≤ 0.05). CONCLUSION: Red flesh dragon fruit extract enhances proliferation and osteogenic differentiation of BMMSCs.

Exfoliated Human Deciduous Tooth Stem Cells Incorporating Carbonate Apatite Scaffold Enhance BMP-2, BMP-7 and Attenuate MMP-8 Expression During Initial Alveolar Bone Remodeling in Wistar Rats (Rattus norvegicus).

BACKGROUND: Post-tooth extraction socket preservation is necessary due to alveolar bone resorptive patterns through regenerative dentistry approaches that involve the use of stem cells, scaffold and growth factor. Stem cells derived from human exfoliated deciduous teeth (SHED) are known to potentially possess the osteogenic ability. Meanwhile, carbonate apatite scaffold (CAS) can act as a biocompatible scaffold capable of supporting mesenchymal stem cells (MSCs) to proliferate and differentiate optimally. The aim of this study is to investigate the expression of bone morphogenic protein-2 and 7 (BMP2, BMP7) and Matrix Metalloproteinase-8 (MMP-8) after the transplantation of SHED-incorporated CAS during in vivo bone remodeling. MATERIAL AND METHODS: A total of 14 healthy, male, Wistar rats, whose mandible anterior teeth were extracted by means of sterile needle holder clamps, constituted the subjects of this study of alveolar bone defects. Two research groups were created: a control group (CAS) as group I and an experimental group (CAS + SHED) as group II. SHED with a density of 106 cells were incorporated into CAS before being transplanted into the experimental group. After 7 days, all the animals were sacrificed and their mandible anterior region extracted. The BMP2, BMP7 and MMP-8 expression were subsequently analyzed by means of immunostaining. An unpaired t-test was conducted to analyze the treatment and control group (p<0.01) data. RESULTS: The expression of BMP-2 and BMP-7 was higher in group II compared to group I. Meanwhile, the level of MMP-8 was lower in group II than group I. There was greater significant increased expression of BMP-2 and BMP-7 expression in Group II compared to Group I. There was significant decreased expression of MMP-8 between group II than group I (p<0.01). CONCLUSION: SHED-incorporated CAS can enhance BMP-2 and BMP-7 expression while attenuating MMP-8 expression during in vivo alveolar bone remodeling.

Concanavalin A Enhanced Proliferation and Osteogenic Differentiation of Dental Pulp Stem Cells.

OBJECTIVE: Dental pulp stem cells (DPSCs) can be used as a component in the formation of regenerative dentine during direct pulp capping therapy. Concanavalin A (ConA) is a type of lectin with a molecular weight of 26 kDa derived from the Canavalia ensiformis plant. Lectins possess strong proliferation and differentiation abilities in various animal cells including lymphocytes, osteoblasts, and chondrocytes. The aim of study was to determine the effect of ConA on the proliferation and osteogenic differentiation of DPSCs in vitro. MATERIALS AND METHODS: In this in vitro study, DPSCs were isolated from third molars before ConA induction was performed at concentrations of 5 and 10 µg/mL. The proliferation assay was determined by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Osteogenic differentiation was determined by means of mineralization. STATISTICAL ANALYSIS: Data were analyzed using analysis of variance and a Student's t-test. The p-value was set at 0.05. RESULTS: The addition of 5 and 10 µg/mL of ConA to DPSCs can significantly increase the proliferation and osteogenic differentiation of DPSCs (p ≤0.05). CONCLUSION: ConA can increase the proliferation and osteogenic differentiation of DPSCs.

Combined Hydroxyapatite Scaffold and Stem Cell from Human Exfoliated Deciduous Teeth Modulating Alveolar Bone Regeneration via Regulating Receptor Activator of Nuclear Factor-Κb and Osteoprotegerin System.

BACKGROUND: Tissue engineering using Stem cell from Human Exfoliated Deciduous Teeth (SHED) and a natural biomaterials biomaterial scaffold has become a promising therapy for the alveolar bone defect. The aim of this study was to analyze the Osteoprotegerin (OPG) and Receptor Activator of NF-Κb ligand (RANKL) expression after the application of Hydroxyapatite scaffold and SHED. METHODS: A laboratory experimental research with a post test-only control group. 14 male Wistar rats weighing from 260 to 280 g were used as the animal study. The animals were randomly assigned to an experimental group Hydroxyapatite scaffold (group I) and Hydroxyapatite scaffold combined with SHED (group II). The alveolar bone defect in the animal study model was affected by extracting anterior mandible teeth. Immunostaining was performed after 8 weeks in order to facilitate the examination of OPG and RANKL expression. Data were analyzed by independent t test. The correlation between OPG and RANKL expression were analyzed using Pearson's correlation test (P<0.05). Statistical analysis was performed using R statistical software version 3.4.0. RESULTS: The independent t test showed that the differences were statistically significant. OPG expression in Group I (6.0±1.00) was lower than in Group II (11.6±1.14) (P=0.0004). The independent t test showed that the differences were statistically significant. RANKL expression for Group I (12.67±2.08) and Group II (4.80±1.304) showed a statistically significant difference (P=0.0005). CONCLUSION: Hydroxyapatite scaffold and SHED increase Osteoprotegerin and decrease Receptor Activator of NF-κB Ligand expression with high potential as an effective agent in alveolar bone defect regeneration.

Extrusion of Irrigant in Open Apex Teeth with Periapical Lesions Following Laser-Activated Irrigation and Passive Ultrasonic Irrigation.

INTRODUCTION: Sodium hypochlorite (NaOCl) irrigation is critical for the success of endodontic treatment and several agitation techniques have been developed to improve the efficacy of this irrigation. Using a combination of contrast medium and radiographic examination, this study evaluated NaOCl extrusion during agitation of irrigant. Development of pressure, which may result in apical extrusion of the irrigant, has been described during laser-activated irrigation (LAI) and passive ultrasonic irrigation (PUI). METHODS AND MATERIALS: We examined 40 single root canals categorized as having open apices with apical lesions in 40 patients. For the final irrigation, the teeth were irrigated with a mixture of radiopaque contrast medium and 2.5% NaOCl in solution. The solution was activated for 60 sec in both groups [the Er, Cr: YSGG laser group (n=20) and the ultrasonic group (n=20)]. The teeth were imaged subsequently using radiography for the evaluation of contrast extrusion. RESULTS: Radiopaque contrast medium was absent from the periapical tissues in all cases. CONCLUSION: Use of LAI or PUI appears to be safe as used currently in endodontic treatment.

Characterization of human dental pulp cells grown in chemically defined serum-free medium.

Dental pulp cells (DPCs) are promising candidates for use as transplantable cells in regenerative medicine. However, ex vivo expansion of these cells typically requires culture media containing fetal bovine serum, which may cause infection and immunological reaction following transplantation. In addition, the proliferation and differentiation of DPCs markedly depend upon serum batches. Therefore, the present study examined whether DPCs could be expanded under serum-free conditions. DPCs obtained from four donors were identified to proliferate actively in the serum-free medium, STK2, when compared with those cells in control medium (Dulbecco's modified Eagle's medium containing 10% serum). The high proliferative potential with STK2 was maintained through multiple successive culture passages. DNA microarray analyses demonstrated that the gene expression profile of DPCs grown in STK2 was similar to that of cells grown in the control medium; however, a number of genes related to cell proliferation, including placental growth factor and inhibin-ßE, were upregulated in the STK2 cultures. Following induction of osteogenesis, DPCs grown in STK2 induced alkaline phosphatase activity and calcification at higher levels compared with the control medium cultures, indicating maintenance of differentiation potential in STK2. This serum-free culture system with DPCs may have applications in further experimental studies and as a clinical strategy in regenerative medicine.

DEC2 is a negative regulator for the proliferation and differentiation of chondrocyte lineage-committed mesenchymal stem cells.

Differentiated embryo chondrocyte 2 (DEC2) is a basic helix-loop-helix-Orange transcription factor that regulates cell differentiation in various mammalian tissues. DEC2 has been shown to suppress the differentiation of mesenchymal stem cells (MSCs) into myocytes and adipocytes. In the present study, we examined the role of DEC2 in the chondrogenic differentiation of human MSCs. The overexpression of DEC2 exerted minimal effects on the proliferation of MSCs in monolayer cultures with the growth medium under undifferentiating conditions, whereas it suppressed increases in DNA content, glycosaminoglycan content, and the expression of several chondrocyte-related genes, including aggrecan and type X collagen alpha 1, in MSC pellets in centrifuge tubes under chondrogenic conditions. In the pellets exposed to chondrogenesis induction medium, DEC2 overexpression downregulated the mRNA expression of fibroblast growth factor 18, which is involved in the proliferation and differentiation of chondrocytes, and upregulated the expression of p16INK4, which is a cell cycle inhibitor. These findings suggest that DEC2 is a negative regulator of the proliferation and differentiation of chondrocyte lineage-committed mesenchymal cells.

Characteristic expression of MSX1, MSX2, TBX2 and ENTPD1 in dental pulp cells.

Dental pulp cells (DPCs) are a promising source of transplantable cells in regenerative medicine. However, DPCs have not been fully characterized at the molecular level. The aim of the present study was to distinguish DPCs from various source-derived mesenchymal stem cells (MSCs), fibroblasts (FBs) and other cells by the expression of several DPC-characteristic genes. DPCs were isolated from human pulp tissues by the explant method or the enzyme digestion method, and maintained with media containing 10% serum or 7.5% platelet-rich plasma. RNA was isolated from the cells and from dental pulp tissue specimens. The mRNA levels were determined by DNA microarray and quantitative polymerase chain reaction analyses. The msh homeobox 1, msh homeobox 2, T-box 2 and ectonucleoside triphosphate diphosphohydrolase 1 mRNA levels in DPCs were higher than that of the levels identified in the following cell types: MSCs derived from bone marrow, synovium and adipose tissue; and in cells such as FBs, osteoblasts, adipocytes and chondrocytes. The enhanced expression in DPCs was consistently observed irrespective of donor age, tooth type and culture medium. In addition, these genes were expressed at high levels in dental pulp tissue in vivo. In conclusion, this gene set may be useful in the identification and characterization of DPCs in basic studies and pulp cell-based regeneration therapy.

Visualization of removal of trapped air from the apical region of the straight root canal models generating 2-phase intermittent counter flow during ultrasonically activated irrigation.

INTRODUCTION: The purpose of this in vitro study was to obtain a better understanding of the mechanism of irrigant traveling apically and generating 2-phase intermittent counter flow in straight root canal models during activation of the irrigant by ultrasonic means in an endodontic procedure. METHODS: A high-speed imaging system, with high temporal and spatial resolution (FastCam SA5; Photron, Tokyo, Japan) at a frame rate of 100,000 frames per second using a macro lens (60 mm, f/2.8; Nikon, Tokyo, Japan), was used to visualize, in glass models of root canals, an ultrasonically induced acoustic pressure wave in an EDTA solution environment. A 25-mm stainless steel noncutting file #20 driven by an ultrasonic device (P5 Newtron; Satelec Acteon, Mérignac, France) at power settings of 5 and 7 produced disturbances at the solution-air interface. RESULTS: We found that apically directed travel of the irrigant was caused by disruption of the surface tension at the solution-air interface. This disruption caused by ultrasonic activation energy displaced air in the form of bubbles from the apical region toward the solution. CONCLUSIONS: The apical movement of the solution may be attributed to ultrasonically induced wave generation at the solution-air interface, resulting in the removal of trapped air from the root canal and allowing the solution to travel apically in the opposite directions (via a 2-phase intermittent counter flow).

Prevalence of a second canal in the mesiobuccal root of permanent maxillary first molars from an Indonesian population.

A sample of 308 extracted human permanent maxillary first molars from an Indonesian population was randomly selected. A rhomboid access cavity was made in all teeth in anticipation of identifying a second mesiobuccal canal (MB2). Ultrasonic tips were used to open the subpulpal groove to locate the second canal in the mesiobuccal root. Dentin was carefully removed from the trifurcation area at the expense of the mesial wall. If a second canal was located, a 0.8 C+ file was inserted into it until the file reached the apex. The prevalence of a second canal in the mesiobuccal root of the permanent maxillary first molar was 68.5% (95% CI: 63.1%-73.4%) in this population. Sections of the mesial root showed that the MB2 was a separate canal in 52.6% of the sample and a joined canal in 47.4%. The mean (SD) distance between MB1 and MB2 was 1.55 (0.66) mm. The data obtained from this study provide theoretical and experimental evidence to aid in the clinical management of the MB2 canal and may increase the success rate for root canal treatment of the maxillary first molar.

Efficacy of smear layer removal at the root tip by using ethylenediaminetetraacetic acid and erbium, chromium: yttrium, scandium, gallium garnet laser.

INTRODUCTION: The purpose of this study was to compare the efficacy of laser-driven irrigation in removing the smear layer and debriding the apical region of the root canal (the root tip) with that of ultrasonic irrigation. METHODS: Forty extracted human teeth with straight single roots were randomized into 4 groups (n = 10). The specimens were shaped by using hand instruments up to a size 30/.02 file (Control, Laser 1, and Laser 2 groups) or a size 20/.02 file (Laser 3 group). During instrumentation, each canal was irrigated with 3% NaOCl and 17% ethylenediaminetetraacetic acid alternately between the use of successive files. The 4 groups of 10 teeth were processed as follows. In the Control group, teeth were irrigated with 17% ethylenediaminetetraacetic acid, and the irrigant was activated with an ultrasonic device for 60 seconds. In the Laser 1 and Laser 3 groups, the irrigant was activated with the laser for 60 seconds. In the Laser 2 group, the irrigant was activated with the laser for 30 seconds. RESULTS: There were significant differences between the smear layer and debris scores for the Laser 1 group and those for the Control (P < .001), Laser 2 (P = .002), and Laser 3 groups (P = .012 and P = .013, respectively). Completely clean root canals were found in the Laser 1 group. CONCLUSIONS: Use of a laser with a plain fiber tip, which produces cavitation in the irrigant, has potential as an improved alternative method for removing of the smear layer from the apical region of a straight root canal.

Transcriptional repression by the basic helix-loop-helix protein Dec2: multiple mechanisms through E-box elements.

Dec2, a member of the basic helix-loop-helix (bHLH) superfamily, has been shown to function as a transcriptional repressor and is implicated in cell proliferation and differentiation. In addition, Dec2 transcripts exhibit a striking circadian oscillation in the suprachiasmatic nucleus. To identify the molecular mechanisms by which Dec2 regulates gene expression, we carried out structure-function analyses. Gel retardation and luciferase assays showed that Dec2, as well as its related protein Dec1, preferentially binds to class B E-box elements (CACGTG) as a homodimer and represses the transcription of target genes in a histone deacetylase (HDAC)-dependent manner. Functional studies with the GAL4-DNA binding domain fusion proteins identified the domain responsible for the repression activity of Dec2 in its C-terminal region, which is also necessary to recruit HDAC1. In addition, the basic and HLH domains of Dec2 were required for DNA binding and homodimerization, respectively. In contrast, Dec proteins repressed a MyoD-activated promoter activity of muscle creatine kinase gene through class A E-box in an HDAC1-independent manner. Dec2 formed a heterodimer with MyoD through the basic and HLH domains. Consistent with this, both the basic and HLH domains were required for the ability of Dec2 to inhibit the transcriptional activity of MyoD. These findings indicate that Dec2 employs multiple mechanisms, including DNA-binding and protein-protein interactions, to achieve E-box-dependent transcriptional repressions.

Alveolar bone marrow as a cell source for regenerative medicine: differences between alveolar and iliac bone marrow stromal cells.

UNLABELLED: We isolated and expanded BMSCs from human alveolar/jaw bone at a high success rate (70%). These cells had potent osteogenic potential in vitro and in vivo, although their chondrogenic and adipogenic potential was less than that of iliac cells. INTRODUCTION: Human bone marrow stromal cells (BMSCs) have osteogenic, chondrogenic, and adipogenic potential, but marrow aspiration from iliac crest is an invasive procedure. Alveolar BMSCs may be more useful for regenerative medicine, because the marrow can be aspirated from alveolar bone with minimal pain. MATERIALS AND METHODS: In this study, alveolar bone marrow samples were obtained from 41 patients, 6-66 years of age, during the course of oral surgery. BMSCs were seeded and maintained in culture with 10% FBS and basic fibroblast growth factor. In addition, BMSCs were induced to differentiate into osteoblasts, chondrocytes, or adipocytes in appropriate medium. RESULTS AND CONCLUSION: From a small volume (0.1-3 ml) of aspirates, alveolar BMSCs expanded at a success ratio of 29/41 (70%). The success rate decreased with increasing donor age, perhaps because of age-dependent decreases in the number and proliferative capacity of BMSCs. The expanded BMSCs differentiated into osteoblasts under osteogenic conditions in 21-28 days: the mRNA levels of osteocalcin, osteopontin, and bone sialoprotein, along with the calcium level, in alveolar BMSC cultures were similar to those in iliac cultures. However, unlike iliac BMSC, alveolar BMSC showed poor chondrogenic or adipogenic potential, and similar differences were observed between canine alveolar and iliac BMSCs. Subsequently, human alveolar BMSCs attached to beta-tricalcium phosphate were transplanted into immunodeficient mice. In transplants, new bone formed with osteoblasts and osteocytes that expressed human vimentin, human osteocalcin, and human GAPDH. These findings suggest that BMSCs have distinctive features depending on their in vivo location and that alveolar BMSCs will be useful in cell therapy for bone diseases.

Anti-membrane-bound transferrin-like protein antibodies induce cell-shape change and chondrocyte differentiation in the presence or absence of concanavalin A.

Membrane-bound transferrin-like protein (MTf), a glycosylphosphatidylinositol-anchored protein, is expressed at high levels in many tumors and in several fetal and adult tissues including cartilage and the intestine, as well as in the amyloid plaques of Alzheimer's disease, although its role remains unknown. MTf is one of the major concanavalin A-binding proteins of the cell surface. In this study, we examined the effects of anti-MTf antibodies and concanavalin A on cell shape and gene expression, using cultures of chondrocytes and MTf-overexpressing ATDC5 and C3H10T1/2 cells. In cultures expressing MTf at high levels, concanavalin A induced cell-shape changes from fibroblastic to spherical cells, whereas no cell-shape changes were observed with wild-type ATDC5 or C3H10T1/2 cells expressing MTf at very low levels. The cell-shape changes were associated with enhanced proteoglycan synthesis and expression of cartilage-characteristic genes, including aggrecan and type II collagen. Some anti-MTf antibodies mimicked this action of concanavalin A, whereas other antibodies blocked the lectin action. The findings suggest that the crosslinking of MTf changes the cell shape and induces chondrogenic differentiation. MTf represents the first identification of a plant lectin receptor involved in cell-shape changes and the differentiation of animal cells.

Effects of overexpression of membrane-bound transferrin-like protein (MTf) on chondrogenic differentiation in Vitro.

Membrane-bound transferrin-like protein (MTf) is expressed in parallel with the expression of cartilage-characteristic genes during differentiation of chondrocytes, and the MTf level is much higher in cartilage than in other tissues. To investigate the role of MTf in cartilage, we examined the effects of growth factors on MTf expression in mouse prechondrogenic ATDC5 cells and the effect of MTf overexpression on differentiation of ATDC5 and mouse pluripotent mesenchymal C3H10T1/2 cells. In ATDC5 cultures, bone morphogenetic protein-2 and transforming growth factor-beta as well as insulin induced MTf mRNA expression when these peptides induced chondrogenic differentiation. Forced expression of rabbit MTf in ATDC5 cells induced aggrecan, type II collagen, matrilin-1, type X collagen mRNAs, and cell-shape changes from fibroblastic cells to spherical chondrocytes. Accordingly, the synthesis and accumulation of proteoglycans were higher in MTf-expressing cultures than in control cultures. These effects of MTf overexpression correlated with the MTf protein level on the cell surface and decreased in the presence of anti-MTf antibody. However, the aggrecan mRNA level in the ATDC5 cells overexpressing MTf was lower than that in wild type ATDC5 cells exposed to 10 microg/ml insulin. MTf overexpression in C3H10T1/2 cells also induced aggrecan and/or type II collagen mRNA but not the spherical phenotype. These findings suggest that the expression of MTf on the cell surface facilitates the differentiation of prechondrogenic cells, although MTf overexpression alone seems to be insufficient to commit pluripotent mesenchymal cells to the chondrocyte lineage.

Basic helix-loop-helix protein DEC1 promotes chondrocyte differentiation at the early and terminal stages.

The mRNA level of basic helix-loop-helix transcription factor DEC1 (BHLHB2)/Stra13/Sharp2 was up-regulated during chondrocyte differentiation in cultures of ATDC5 cells and growth plate chondrocytes, and in growth plate cartilage in vivo. Forced expression of DEC1 in ATDC5 cells induced chondrogenic differentiation, and insulin increased this effect of DEC1 overexpression. Parathyroid hormone (PTH) and PTH-related peptide (PTHrP) suppressed DEC1 expression and the differentiation of ATDC5 cells, but DEC1 overexpression antagonized this inhibitory action of PTH/PTHrP. Transforming growth factor-beta or bone morphogenetic protein-2, as well as insulin, induced DEC1 expression in ATDC5 cultures where it induced chondrogenic differentiation. In pellet cultures of bone marrow mesenchymal stem cells exposed to transforming growth factor-beta and insulin, DEC1 was induced at the earliest stage of chondrocyte differentiation and also at the hypertrophic stage. Overexpression of DEC1 in the mesenchymal cells induced the mRNA expressions of type II collagen, Indian hedgehog, and Runx2, as well as cartilage matrix accumulation; overexpression of DEC1 in growth plate chondrocytes at the prehypertrophic stage increased the mRNA levels of Indian hedgehog, Runx2, and type X collagen, and also increased alkaline phosphatase activity and mineralization. To our knowledge, DEC1 is the first transcription factor that can promote both chondrogenic differentiation and terminal differentiation.