Identification of stemness and differentially expressed genes in human cementum-derived cells.

PURPOSE: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. METHODS: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. RESULTS: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. CONCLUSIONS: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.

MicroRNAs as biomarkers for dental diseases.

MicroRNAs (miRNAs) are short, noncoding RNAs that act as key regulators of diverse biological processes by mediating translational repression or mRNA degradation of target genes. Recent studies discovered miRNAs in saliva, and these miRNAs are promising candidates for use as biomarkers of dental diseases. In this review, the results of miRNA studies in the dental field are presented, and a brief overview of the current progress, limitations, and perspectives regarding miRNA biomarkers for dental diseases is given.

Clinical Outcomes of Reamer- vs Osteotome-Mediated Sinus Floor Elevation with Simultaneous Implant Placement: A 2-Year Retrospective Study.

PURPOSE: This retrospective study evaluated and compared the 2-year outcomes for sinus floor elevation performed via either an osteotome-mediated sinus floor elevation (OSFE) technique or a reamer-mediated sinus floor elevation (RSFE) technique. Implant survival, as well as surgical and postoperative complications, were used as measures of comparison. MATERIALS AND METHODS: Patients were analyzed according to defined inclusion criteria. Orthopantography was used to assess preoperative; immediate postoperative; and 6-, 12-, and 24-month postoperative bone level changes. Implant survival and the incidence of complications, including sinus membrane perforation, were evaluated using appropriate statistical tests. RESULTS: From 2008 to 2010, 126 implants were placed simultaneously with sinus floor elevation in 85 patients (n = 43 women and 42 men; mean age ± standard deviation [SD] = 58.1 ± 10.2 years). The OSFE procedure (control) was used to place 65 implants in 45 patients, and the RSFE procedure (experimental) was used to place 61 implants in 40 patients. The mean maxillary residual bone height was 7.1 ± 1.6 mm. Endosinus bone gains were 5.7 ± 1.5 mm and 5.6 ± 2.3 mm for the experimental and control groups (P = .164), respectively, and the 2-year survival rates were 98.4% and 98.5%, respectively. Although no significant differences were observed between the two groups, three (6.7%) membrane perforations occurred in the OSFE group, and none occurred in the RSFE group. Other postoperative complications, including nasal bleeding, postoperative headache, and dizziness, were documented in 7 (15.6%) of 45 OSFE cases and 3 (7.5%) of 40 RSFE cases. CONCLUSION: The results presented herein indicate that comparable survival rates were achieved for implants placed in conjunction with a reamer- vs osteotome-mediated technique. Therefore, RSFE is a reliable and predictable procedure for implant placement in the posterior maxilla, with a low incidence of complications.

Complication incidence of two implant systems up to six years: a comparison between internal and external connection implants.

PURPOSE: This study was conducted to compare the cumulative survival rates (CSRs) and the incidence of postloading complications (PLCs) between a bone-level internal connection system (ICS-BL) and an external connection system (ECS). METHODS: The medical records of patients treated with either a ICS-BL or ECS between 2007 and 2010 at Asan Medical Center were reviewed. PLCs were divided into two categories: biological and technical. Biological complications included >4 mm of probing pocket depth, thread exposure in radiographs, and soft tissue complications, whereas technical complications included chipping of the veneering material, fracture of the implant, fracture of the crown, loosening or fracture of the abutment or screw, loss of retention, and loss of access hole filling material. CSRs were determined by a life-table analysis and compared using the log-rank chi-square test. The incidence of PLC was compared with the Pearson chi-squared test. RESULTS: A total of 2,651 implants in 1,074 patients (1,167 ICS-BLs in 551 patients and 1,484 ECSs in 523 patients) were analyzed. The average observation periods were 3.4 years for the ICS-BLs and 3.1 years for the ECSs. The six-year CSR of all implants was 96.1% (94.9% for the ICS-BLs and 97.1% for the ECSs, P=0.619). Soft tissue complications were more frequent with the ECSs (P=0.005) and loosening or fracture of the abutment or screw occurred more frequently with the ICS-BLs (P<0.001). CONCLUSIONS: Within the limitations of this study, the ICS-BL was more prone to technical complications while the ECS was more vulnerable to biological complications.

Rabbit maxillary sinus augmentation model with simultaneous implant placement: differential responses to the graft materials.

PURPOSE: This study was performed to establish an experimental rabbit model for single-stage maxillary sinus augmentation with simultaneous implant placement. METHODS: Twelve mature New Zealand white rabbits were used for the experiments. The rabbit maxillary sinuses were divided into 3 groups according to sinus augmentation materials: blood clot (BC), autogenous bone (AB), and bovine-derived hydroxyapatite (BHA). Small titanium implants were simultaneously placed in the animals during the sinus augmentation procedure. The rabbits were sacrificed 4 and 8 weeks after surgery and were observed histologically. Histomorphometric analyses using image analysis software were also performed to evaluate the parameters related to bone regeneration and implant-bone integration. RESULTS: The BC group showed an evident collapse of the sinus membrane and limited new bone formation around the original sinus floor at 4 and 8 weeks. In the AB group, the sinus membrane was well retained above the implant apex, and new bone formation was significant at both examination periods. The BHA group also showed retention of the elevated sinus membrane above the screw apex and evident new bone formation at both points in time. The total area of the mineral component (TMA) in the area of interest and the bone-to-implant contact did not show any significant differences among all the groups. In the AB group, the TMA had significantly decreased from 4 to 8 weeks. CONCLUSIONS: Within the limits of this study, the rabbit sinus model showed satisfactory results in the comparison of different grafting conditions in single-stage sinus floor elevation with simultaneous implant placement. We found that the rabbit model was useful for maxillary sinus augmentation with simultaneous implant placement.

Gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow.

PURPOSE: The aim of this study is to compare the gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow for characterization of dental stem cells. METHODS: We employed GeneChip analysis to the expression levels of approximately 32,321 kinds of transcripts in 5 samples of bone-marrow-derived mesenchymal stem cells (BMSCs) (n=1), periodontal ligament stem cells (PDLSCs) (n=2), and dental pulp stem cells (DPSCs) (n=2). Each cell was sorted by a FACS Vantage Sorter using immunocytochemical staining of the early mesenchymal stem cell surface marker STRO-1 before the microarray analysis. RESULTS: We identified 379 up-regulated and 133 down-regulated transcripts in BMSCs, 68 up-regulated and 64 down-regulated transcripts in PDLSCs, and 218 up-regulated and 231 down-regulated transcripts in DPSCs. In addition, anatomical structure development and anatomical structure morphogenesis gene ontology (GO) terms were over-represented in all three different mesenchymal stem cells and GO terms related to blood vessels, and neurons were over-represented only in DPSCs. CONCLUSIONS: This study demonstrated the genome-wide gene expression patterns of STRO-1(+) mesenchymal stem cells derived from dental tissues and bone marrow. The differences among the expression profiles of BMSCs, PDLSCs, and DPSCs were shown, and 999 candidate genes were found to be definitely up- or down-regulated. In addition, GOstat analyses of regulated gene products provided over-represented GO classes. These data provide a first step for discovering molecules key to the characteristics of dental stem cells.

A new resin-bonded retrograde filling material.

OBJECTIVE: This study determined the physical properties and cytotoxicity of a novel root-end filling material (NRC). STUDY DESIGN: NRC is a powder and liquid system. The liquid is composed of hydroxyethylmethacrylate, benzoyl peroxide, toluidine, and toluenesulfinate. And the powder is made of calcium oxide, calcium silicate, and triphenylbismuth carbonate. The setting time, compressive strength, and pH change of NRC and gray and white mineral trioxide aggregate (MTA) were determined according to ISO standardization. MC3T3-E1 cells were cultured on NRC and white MTA for determining MTT scores. The absorbance of formazan was measured at 570 nm with a spectrophotometer. The MTT assay was performed in triplicate and repeated in 2 cultures. One-way analysis of variance was used to determine statistical differences in physical properties and MTT assay (P < .05). RESULTS: Mean setting time of materials tested were: NRC 12.5 +/- 0.3 minutes, gray MTA 345.5 +/- 96.2 minutes, and white MTA 318.0 +/- 56.0 minutes. After 24 hours, the mean compressive strengths were: NRC, 21.6 +/- 5.5 MPa, gray MTA: 7.7 +/- 3.3 MPa, and white MTA, 18.9 +/- 3.2 MPa. The pH of the test materials were: NRC 12.0, gray MTA 12.2, and white MTA 11.9. There were no statistically significant differences in compressive strength and pH between white MTA and NRC. The compressive strength of gray MTA was significantly lower than white MTA and NRC (P < .05). The setting time of NRC was significantly lower than white and gray MTA. In MTT assay, both NRC and white MTA were not cytotoxic to MC3T3-E1 cells. CONCLUSIONS: It was concluded that the setting time, compressive strength, pH, and initial biocompatibility results of NRC are favorable for a root-end filling material.

Effect of calcium hydroxide-treated Prevotella nigrescens on the gene expression of matrix metalloproteinase and its inhibitor in MG63 cells.

The purpose of this study was to monitor the expression of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) produced by an osteoblastic cell line MG63 stimulated with Prevotella nigrescens lipopolysaccharides (LPS), and to compare the level of secretion before and after the P. nigrescens LPS was treated with calcium hydroxide [Ca(OH)2]. The underlying hypothesis is that the balance between MMP and TIMP secretion is the key to an understanding of the host degradative pathways involved in the pathogenesis of bacterial derived pulpal and periapical diseases. Confluent monolayers of MG63 human osteosarcoma cells were exposed to varying concentrations of P. nigrescens or Escherichia coli LPS. Alternately, confluent cultures were exposed to 10 microg/ml of bacterial LPS pretreated with Ca(OH)2 (12.5 mg/ml) for 72 hours. At the end of the experimental period, total RNA was extracted and real-time quantitative polymerase chain reaction (PCR) was performed for MMP-1, TIMP-1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The results showed that the expression of MMP-1 mRNA was low and invariant for the experimental period in the negative controls. However, exposure to P. nigrescens LPS increased expression after 48 hours. Expression of TIMP-1 mRNA was highly increased at 24 and 48 hours with lower concentrations of LPS in contrast to a suppression with a concentration of 10 microg/ml. Treatment of P. nigrescens LPS with Ca(OH)2 resulted in a down-regulation of MMP-1, whereas pretreated E. coli LPS demonstrated no stimulatory activity for MMP-1 gene expression. Both types of LPS when pretreated with Ca(OH)2 induced slightly up-regulated expression of TIMP-1.