Pan PPAR agonist stimulation of induced MSCs produces extracellular vesicles with enhanced renoprotective effect for acute kidney injury.

BACKGROUND: Acute kidney injury (AKI) has a complex pathophysiology and imposes serious health concerns worldwide. Extracellular vesicles (EVs) derived from induced mesenchymal stem cells (iMSCs) have been recognized as novel cell-free therapeutics for various inflammatory and degenerative disorders. In this study, we investigated whether iMSCs stimulated with a pan-peroxisome proliferator-activated receptor (PPAR) agonist could enhance the therapeutic efficacy of EVs against AKI. METHODS: Human iMSCs were primed with or without lanifibranor, a PPAR agonist for 24 h, and EVs were collected after an additional 24 h. The basic characteristics of EVs were evaluated using cryo-transmission electron microscopy imaging, immunoblot detection of EV markers, nanoparticle tracking analysis, and localization in AKI kidneys. In vitro, the potential of the EVs to promote the growth and survival of HK-2 cells undergoing cisplatin-induced apoptosis and anti-inflammatory effects in M1-polarized THP-1 was compared. Subsequently, AKI was induced in BALB/c mice using cisplatin. After 8 and 24 h of cisplatin treatment, iMSC-EVs or pan-PPAR-iMSC-EVs were injected intravascularly. At 96 h after cisplatin administration, the renoprotective effects of iMSC-EVs or pan-PPAR-iMSC-EVs in inhibiting inflammation and apoptosis were compared using serum biochemistry, histology, immunohistochemistry, and gene expression analysis by qPCR. RESULTS: Both EV types expressed EV markers and had typical EV morphology, and their localization in the renal tissue was confirmed. The proliferation and survival of HK-2 cells were higher in pan-PPAR-iMSC-EVs than those in iMSC-EVs. In M1-polarized THP-1 cells, the reduction in the mRNA expression of inflammatory cytokines was more significant in pan-PPAR-iMSC-EVs than that in iMSC-EVs. In the mouse model of cisplatin-induced AKI, pan-PPAR-iMSC-EVs markedly enhanced renoprotective effects compared to iMSC-EVs. Specifically, pan-PPAR-iMSC-EVs reduced tissue inflammation, immune cell infiltration, and apoptosis. Pan-PPAR-iMSC-EVs also increased renal capillary density. CONCLUSION: Priming iMSCs with a PPAR agonist significantly improved the therapeutic potential of EVs by reducing inflammation and apoptosis. The reported strategy may contribute to the development of a novel cell-free option for AKI treatment. TRIAL REGISTRATION: Not applicable.

Extracellular vesicles from induced pluripotent stem cell-derived mesenchymal stem cells enhance the recovery of acute kidney injury.

BACKGROUND AIMS: To investigate whether the extracellular vesicles (EVs) from mesenchymal stem cell-like cells derived from induced pluripotent stem cells (iMSC-EVs) can inhibit the progression of acute kidney injury (AKI). METHODS: The characteristics of iMSC-EVs were confirmed by immunoblotting, cryo-transmission electron microscopy, nanoparticle tracking analysis, and their localization in kidneys. Using human renal epithelial cells, the potential of iMSC-EVs to stimulate the growth and survival of HK-2 cells undergoing cisplatin-induced cell death was investigated. The anti-inflammatory effects of iMSC-EVs was examined in M1-polarized THP-1 macrophages. Subsequently, the therapeutic potential of iMSC-EVs was assessed in cisplatin-induced acute kidney injury in BALB/c mice. The anti-apoptotic and anti-inflammatory effect of iMSC-EVs was evaluated using serum biochemistry, histology, immunohistochemistry, and gene expression analysis. RESULTS: iMSC-EVs promoted the growth of renal epithelial cell (HK-2) and enhanced the survival of HK-2 undergoing cisplatin-induced cell death. In cisplatin-induced mice with AKI, iMSC-EVs alleviated AKI, as shown by reduced blood nitrogen urea/creatinine and increased body weight. Also, iMSC-EVs enhanced renal tissue integrity and the number of proliferating cell nuclear antigen-positive tubules. iMSC-EVs decreased the infiltration of immune cells, reduced the expression of inflammatory genes in M1-induced THP-1 cells and enhanced capillary density in the kidney of AKI mice. Real-time quantitative polymerase chain reaction analysis showed that the expression of inflammatory genes in the kidney of AKI mice was reduced compared with that received vehicle. Immunoblotting revealed that iMSC-EVs led to a decreased protein expression of key inflammatory genes. Also, iMSC-EVs reversed the activation of ERK1/2 signaling induced by AKI. Finally, iMSC-EVs inhibited the apoptosis of HK-2 cells induced by cisplatin as well as that of renal tissue of AKI mice. CONCLUSIONS: Our data suggest that iMSC-EVs have potential to become a novel, cell-free therapeutic for cisplatin-induced AKI.

Nanophotosensitizers Composed of Phenyl Boronic Acid Pinacol Ester-Conjugated Chitosan Oligosaccharide via Thioketal Linker for Reactive Oxygen Species-Sensitive Delivery of Chlorin e6 against Oral Cancer Cells.

Chlorin E6 (Ce6)-incorporated nanophotosensitizers were fabricated for application in photodynamic therapy (PDT) of oral cancer cells. For this purpose, chitosan oligosaccharide (COS) was conjugated with hydrophobic and reactive oxygen species (ROS)-sensitive moieties, such as phenyl boronic acid pinacol ester (PBAP) via a thioketal linker (COSthPBAP). ThdCOOH was conjugated with PBAP to produce ThdCOOH-PBAP conjugates and then attached to amine groups of COS to produce a COSthPBAP copolymer. Ce6-incorporated nanophotosensitizers using the COSthPBAP copolymer were fabricated through the nanoprecipitation and dialysis methods. The Ce6-incorporated COSthPBAP nanophotosensitizers had a small diameter of less than 200 nm with a mono-modal distribution pattern. However, it became a multimodal and/or irregular distribution pattern when H2O2 was added. In a morphological observation using TEM, the nanophotosensitizers were disintegrated by the addition of H2O2, indicating that the COSthPBAP nanophotosensitizers had ROS sensitivity. In addition, the Ce6 release rate from the COSthPBAP nanophotosensitizers accelerated in the presence of H2O2. The SO generation was also higher in the nanophotosensitizers than in the free Ce6. Furthermore, the COSthPBAP nanophotosensitizers showed a higher intracellular Ce6 uptake ratio and ROS generation in all types of oral cancer cells. They efficiently inhibited the viability of oral cancer cells under light irradiation, but they did not significantly affect the viability of either normal cells or cancer cells in the absence of light irradiation. The COSthPBAP nanophotosensitizers showed a tumor-specific delivery capacity and fluorescence imaging of KB tumors in an in vivo animal tumor imaging study. We suggest that COSthPBAP nanophotosensitizers are promising candidates for the imaging and treatment of oral cancers.

A large invasive chondroblastoma on the temporomandibular joint and external auditory canal: a case report and literature review.

BACKGROUND: Chondroblastomas, which account for approximately 1% of all bone tumors, typically occur in long bones, such as the femur, humerus, and tibia. However, in extremely rare cases, they may also occur in the craniofacial region where the tumor is often found in the squamous portion of the temporomandibular joint (TMJ) and in the temporal bone. CASE PRESENTATION: This case report describes a large chondroblastoma (diameter, approximately 37 mm) that occurred in the TMJ. The tumor was sufficiently aggressive to destroy the TMJ, mandibular condyle neck, external auditory canal (EAC), mandibular fossa of the temporal bone, and facial nerve. The tumor was completely excised using a pre-auricular approach. The EAC and surgical defect were successfully reconstructed using a temporoparietal fascia flap (TPFF) and an inguinal free fat graft. There was no local tumor recurrence at the 18-month follow-up visits. However, the patient developed sensory neural hearing loss, and his eyebrow paralysis worsened, eventually requiring plastic surgery. CONCLUSION: Large, invasive chondroblastomas of the TMJ can be completely removed through a pre-auricular approach, and the resulting surgical defect can be reconstructed using TPFF and free fat grafts. However, preoperative evaluation of the facial nerve and auditory function is necessary. Therefore, a multidisciplinary approach is essential.

The effect of contextual factors on unintentional injury hospitalization: from the Korea National Hospital Discharge Survey.

BACKGROUND: It has been suggested that health risks are affected by geographical area, but there are few studies on contextual effects using multilevel analysis, especially regarding unintentional injury. This study investigated trends in unintentional injury hospitalization rates over the past decade in Korea, and also examined community-level risk factors while controlling for individual-level factors. METHODS: Using data from the 2004 to 2013 Korea National Hospital Discharge Survey (KNHDS), trends in age-adjusted injury hospitalization rate were conducted using the Joinpoint Regression Program. Based on the 2013 KNHDS, we collected community-level factors by linking various data sources and selected dominant factors related to injury hospitalization through a stepwise method. Multilevel analysis was performed to assess the community-level factors while controlling for individual-level factors. RESULTS: In 2004, the age-adjusted unintentional injury hospitalization rate was 1570.1 per 100,000 population and increased to 1887.1 per 100,000 population in 2013. The average annual percent change in rate of hospitalizations due to unintentional injury was 2.31% (95% confidence interval: 1.8-2.9). It was somewhat higher for females than for males (3.25% vs. 1.64%, respectively). Both community- and individual-level factors were found to significantly influence unintentional injury hospitalization risk. As community-level risk factors, finance utilization capacity of the local government and neighborhood socioeconomic status, were independently associated with unintentional injury hospitalization after controlling for individual-level factors, and accounted for 19.9% of community-level variation in unintentional injury hospitalization. CONCLUSION: Regional differences must be considered when creating policies and interventions. Further studies are required to evaluate specific factors related to injury mechanism.

Healing outcomes of root canal treatment for C-shaped mandibular second molars: a retrospective analysis.

OBJECTIVES: This study aimed to evaluate the healing rate of non-surgical endodontic treatment between C-shaped and non-C-shaped mandibular second molars. MATERIALS AND METHODS: Clinical records and radiological images of patients who had undergone endodontic treatment on mandibular second molars between 2007 and 2014 were screened. The periapical index scoring system was applied to compare healing outcomes. Information about preoperative and postoperative factors as well as the demographic data of the patients was acquired and evaluated using chi-square and multinomial logistic regression tests. RESULTS: The total healing rate was 68.4%. Healing rates for the mandibular second molar were 70.9% in C-shaped canals (n = 79) and 66.6% in non-C-shaped ones (n = 117). The difference was not statistically significant. CONCLUSIONS: The presence of a C-shaped canal in the mandibular second molar did not have a significantly negative effect on healing after treatment. Instead, proper pulpal diagnosis and final restoration were indicated as having significantly greater influence on the healing outcomes of C-shaped and non-C-shaped canals, respectively.

Finite element analysis on stress distribution of maxillary implant-retained overdentures depending on the Bar attachment design and palatal coverage.

PURPOSE: The purpose of this study was to determine the effect of anchorage systems and palatal coverage of denture base on load transfer in maxillary implant-retained overdenture. MATERIALS AND METHODS: Maxillary implant-retained overdentures with 4 implants placed in the anterior region of edentulous maxilla were converted into a 3-D numerical model, and stress distribution patterns in implant supporting bone in the case of unilateral vertical loading on maxillary right first molar were compared with each other depending on various types of anchorage system and palatal coverage extent of denture base using three-dimensional finite element analysis. RESULTS: In all experimental models, the highest stress was concentrated on the most distal implant and implant supporting bone on loaded side. The stress at the most distal implant-supporting bone was concentrated on the cortical bone. In all anchorage system without palatal coverage of denture base, higher stresses were concentrated on the most distal implant and implant supporting bone on loaded side. CONCLUSION: It could be suggested that when making maxillary implant retained overdenture, using Hader bar instead of milled bar and full palatal coverage rather than partial palatal coverage are more beneficial in distributing the stress that is applied on implant supporting bone.

The effect of saliva decontamination procedures on dentin bond strength after universal adhesive curing.

OBJECTIVES: The purpose of this study was to investigate the effectiveness of multiple decontamination procedures for salivary contamination after curing of a universal adhesive on dentin bond strength according to its etch modes. MATERIALS AND METHODS: Forty-two extracted bovine incisors were trimmed by exposing the labial dentin surfaces and embedded in cylindrical molds. A universal adhesive (All-Bond Universal, Bisco) was used. The teeth were randomly divided into groups according to etch mode and decontamination procedure. The adhesive was applied according to the manufacturer's instructions for a given etch mode. With the exception of the control groups, the cured adhesive was contaminated with saliva for 20 sec. In the self-etch group, the teeth were divided into three groups: control, decontamination with rinsing and drying, and decontamination with rinsing, drying, and adhesive. In the etch-and-rinse group, the teeth were divided into four groups: control, decontamination with rinsing and drying, decontamination with rinsing, drying, and adhesive, and decontamination with rinsing, drying, re-etching, and reapplication of adhesive. A composite resin (Filtek Z350XT, 3M ESPE) was used for filling and was cured on the treated surfaces. Shear bond strength was measured, and failure modes were evaluated. The data were subjected to one-way analysis of variation and Tukey's HSD test. RESULTS: The etch-and-rinse subgroup that was decontaminated by rinse, drying, re-etching, and reapplication of adhesive showed a significantly higher bond strength. CONCLUSIONS: When salivary contamination occurs after curing of the universal adhesive, additional etching improves the bond strength to dentin.

Odontogenic effect of a fast-setting pozzolan-based pulp capping material.

INTRODUCTION: Mineral 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. METHODS: Human 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. RESULTS: The 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. CONCLUSIONS: Our results indicate that ProRoot and Endocem have similar biocompatibility and odontogenic effects. Therefore, Endocem is as effective a pulp capping material as ProRoot.

Magnesium vs. machined surfaced titanium - osteoblast and osteoclast differentiation.

PURPOSE: This study focused on in vitro cell differentiation and surface characteristics in a magnesium coated titanium surface implanted on using a plasma ion source. MATERIALS AND METHODS: 40 commercially made pure titanium discs were prepared to produce Ti oxide machined surface (M) and Mg-incorporated Ti oxide machined surface (MM). Surface properties were analyzed using a scanning electron microscopy (SEM). On each surface, alkaline phosphatase (ALP) activity, alizarin red S staining for mineralization of MC3T3-E1 cells, and quantitative analysis of osteoblastic gene expression, were evaluated. Actin ring formation assay and gene expression analysis of TRAP and GAPDH performing RT-PCR were performed to characterize osteoclast differentiation on mouse bone marrow-derived macrophages (BMMs). RESULTS: MM showed similar surface morphology and surface roughness with M, but was slightly smoother after ion implantation at the micron scale. M was more hydrophobic than MM. No significant difference between surfaces on ALP activity at 7 and 14 days were observed. Real-time PCR analyses showed similar levels of mRNA expression of the osteoblast phenotype genes; osteopontin (OPN), osteocalcin (OCN), bone sialoprotein (BSP), and collagen 1 (Col 1) in cell grown on MM at 7, 14 and 21 days. Alizarin red S staining at 21 days showed no significant difference. BMMs differentiation increased in M and MM. Actin ring formation assay and gene expression analysis of TRAP showed osteoclast differentiation to be more active on MM. CONCLUSION: Both M and MM have a good effect on osteoblastic cell differentiation, but MM may speed the bone remodeling process by activating on osteoclast differentiation.

Effects of layering technique on the shade of resin overlays and the microhardness of dual cure resin cement.

The purpose of this study was to assess the color of layered resin overlays and to test the early microhardness of dual cure resin cement (DCRC) light cured through the layered resin overlays. Resin overlays of 1.5 mm thickness were fabricated with the A3 shade of Z350 (Group 1L), the A3B and A3E shades of Supreme XT (Group 2L), and the A3, E3, and T1 shades of Sinfony (Group 3L) using one, two, and three layers, respectively (n=7). Each layer of the resin overlays was set in equal thickness. The color of the resin overlays was measured with a colorimeter and compared with an A3 shade resin denture tooth. DCRC was light cured through the resin overlays, and the early microhardness of the DCRC was measured. The ΔE value between the denture tooth and the resin overlays and the Vickers hardness number (VHN) of the DCRC were analyzed with one-way ANOVA and Tukey's HSD test. The color differences were 8.9±0.5, 5.3±1.0, and 7.3±0.5 and the VHNs were 19.4±1.1, 21.1±0.9, and 29.3±0.6 for Groups 1L, 2L, and 3L, respectively. Therefore, to match the designated tooth color of resin inlays and to increase the early microhardness of DCRC, layered resin inlays are more appropriate than single-dentin-layer resin inlays. However, the translucent layer should be used cautiously because the color difference of resin inlays with a translucent layer was affected more than those without a translucent layer.

Healing after horizontal root fractures: 3 cases with 2-year follow-up.

Among dental traumas, horizontal root fractures are relatively uncommon injuries. Proper initial management and periodical evaluation is essential for the successful treatment of a root-fractured tooth. If pulpal necrosis develops, endodontic treatment is indicated, exclusively for the coronal fragment. Fragment diastases exert a great influence on healing at the fracture line and on pulpal necrosis. An adequately treated root-fractured tooth has a good prognosis. This case report describes the treatment and 2-yr follow up of 3 maxillary central incisors, first with horizontal root fracture, second with horizontal root fracture and avulsion, and third with horizontal root fracture and lateral luxation. All three cases were treated with mineral trioxide aggregate (ProRoot, Dentsply). During 2 yr of follow-up evaluation, the root-fractured teeth of the present patients were well retained in the arch, showing periodontal healing, even after endodontic treatment.

Effect of a multi-layer infection control barrier on the micro-hardness of a composite resin.

OBJECTIVE: The aim of this study was to evaluate the effect of multiple layers of an infection control barrier on the micro-hardness of a composite resin. MATERIAL AND METHODS: One, two, four, and eight layers of an infection control barrier were used to cover the light guides of a high-power light emitting diode (LeD) light curing unit (LCU) and a low-power halogen LCU. The composite specimens were photopolymerized with the LCUs and the barriers, and the micro-hardness of the upper and lower surfaces was measured (n=10). The hardness ratio was calculated by dividing the bottom surface hardness of the experimental groups by the irradiated surface hardness of the control groups. The data was analyzed by two-way ANOVA and Tukey's HSD test. RESULTS: The micro-hardness of the composite specimens photopolymerized with the LED LCU decreased significantly in the four- and eight-layer groups of the upper surface and in the two-, four-, and eight-layer groups of the lower surface. The hardness ratio of the composite specimens was <80% in the eight-layer group. The micro-hardness of the composite specimens photopolymerized with the halogen LCU decreased significantly in the eight-layer group of the upper surface and in the two-, four-, and eight-layer groups of the lower surface. However, the hardness ratios of all the composite specimens photopolymerized with barriers were <80%. CONCLUSIONS: The two-layer infection control barrier could be used on high-power LCUs without decreasing the surface hardness of the composite resin. However, when using an infection control barrier on the low-power LCUs, attention should be paid so as not to sacrifice the polymerization efficiency.

Effect of a multi-layer infection control barrier on the micro-hardness of a composite resin

OBJECTIVE: The aim of this study was to evaluate the effect of multiple layers of an infection control barrier on the micro-hardness of a composite resin. MATERIAL AND METHODS: One, two, four, and eight layers of an infection control barrier were used to cover the light guides of a high-power light emitting diode (LeD) light curing unit (LCU) and a low-power halogen LCU. The composite specimens were photopolymerized with the LCUs and the barriers, and the micro-hardness of the upper and lower surfaces was measured (n=10). The hardness ratio was calculated by dividing the bottom surface hardness of the experimental groups by the irradiated surface hardness of the control groups. The data was analyzed by two-way ANOVA and Tukey's HSD test. RESULTS: The micro-hardness of the composite specimens photopolymerized with the LED LCU decreased significantly in the four- and eight-layer groups of the upper surface and in the two-, four-, and eight-layer groups of the lower surface. The hardness ratio of the composite specimens was <80% in the eight-layer group. The micro-hardness of the composite specimens photopolymerized with the halogen LCU decreased significantly in the eight-layer group of the upper surface and in the two-, four-, and eight-layer groups of the lower surface. However, the hardness ratios of all the composite specimens photopolymerized with barriers were <80%. CONCLUSIONS: The two-layer infection control barrier could be used on high-power LCUs without decreasing the surface hardness of the composite resin. However, when using an infection control barrier on the low-power LCUs, attention should be paid so as not to sacrifice the polymerization efficiency.

Endodontic management of a C-shaped maxillary first molar with three independent buccal root canals by using cone-beam computed tomography.

The aim of this study was to present a method for endodontic management of a maxillary first molar with unusual C-shaped morphology of the buccal root verified by cone-beam computed tomography (CBCT) images. This rare anatomical variation was confirmed using CBCT, and nonsurgical endodontic treatment was performed by meticulous evaluation of the pulpal floor. Posttreatment image revealed 3 independent canals in the buccal root obturated efficiently to the accepted lengths in all 3 canals. Our study describes a unique C-shaped variation of the root canal system in a maxillary first molar, involving the 3 buccal canals. In addition, our study highlights the usefulness of CBCT imaging for accurate diagnosis and management of this unusual canal morphology.

Power density of various light curing units through resin inlays with modified layer thickness.

OBJECTIVES: The purpose of this study was to enhance curing light penetration through resin inlays by modifying the thicknesses of the dentin, enamel, and translucent layers. MATERIALS AND METHODS: To investigate the layer dominantly affecting the power density of light curing units, resin wafers of each layer with 0.5 mm thickness were prepared and power density through resin wafers was measured with a dental radiometer (Cure Rite, Kerr). The dentin layer, which had the dominant effect on power density reduction, was decreased in thickness from 0.5 to 0.1 mm while thickness of the enamel layer was kept unchanged at 0.5 mm and thickness of the translucent layer was increased from 0.5 to 0.9 mm and vice versa, in order to maintain the total thickness of 1.5 mm of the resin inlay. Power density of various light curing units through resin inlays was measured. RESULTS: Power density measured through 0.5 mm resin wafers decreased more significantly with the dentin layer than with the enamel and translucent layers (p < 0.05). Power density through 1.5 mm resin inlays increased when the dentin layer thickness was reduced and the enamel or translucent layer thickness was increased. The highest power density was recorded with dentin layer thickness of 0.1 mm and increased translucent layer thickness in all light curing units. CONCLUSIONS: To enhance the power density through resin inlays, reducing the dentin layer thickness and increasing the translucent layer thickness would be recommendable when fabricating resin inlays.

Simvastatin promotes odontoblastic differentiation and expression of angiogenic factors via heme oxygenase-1 in primary cultured human dental pulp cells.

INTRODUCTION: Although simvastatin has multiple demonstrable effects, its function in dentinogenesis remains unclear. In this study, we tested the hypothesis that the addition of simvastatin to human dental pulp cells (HDPCs) stimulates odontogenesis both by promoting odontoblastic differentiation and by favoring the release of angiogenic factors. In addition, the role of heme oxygenase-1 (HO-1) in these effects was investigated. METHODS: The expression of markers for odontoblastic differentiation and angiogenesis was analyzed by means of alkaline phosphatase (ALP) activity, alizarin red staining, and Western blotting. RESULTS: Simvastatin enhanced the differentiation of HDPCs by up-regulating mineralization nodules and odontogenic markers as well as angiogenic markers. These phenomena were then correlated with the induction of HO-1 protein levels. The inducing effect of simvastatin on odontoblastic differentiation and angiogenesis was nullified by an HO-1 inhibitor and a carbon monoxide (CO) scavenger. CONCLUSIONS: These results suggested that simvastatin exerts its odontoblastic differentiation and angiogenesis-inducing effects in HDPCs through a mechanism that involves the action of HO-1 and its product CO.