The molecular weight of hyaluronic acid influences metabolic activity and osteogenic differentiation of periodontal ligament cells.

OBJECTIVE: While HA is present naturally in periodontal tissues, its molecular weight can vary widely in vivo. The objective of this study was to directly compare the biological reactions of periodontal ligament cells to four distinct molecular weights of hyaluronic acid (HA). MATERIALS AND METHODS: Immortalized human periodontal ligament cells (PDL-hTERT) were cultured for 21 days in culture medium alone (control) or enriched with osteogenic supplements (OS group). Other 4 experimental groups were cultured in OS medium with the addition of HA with different molecular weights (HMW, MMW, LMW, and ULMW). The cell morphology was examined daily. WST1 assays were performed to evaluate metabolic activity. Von Kossa staining and calcium deposition assay were used to analyze osteogenic differentiation and mineralization. RESULTS: Cell morphology remained unaltered in all groups. Cells stimulated with OS alone or with the addition of hyaluronan showed all the typical microscopic appearance of osteogenic differentiation. Metabolic activity increased in all groups over time. Hyaluronan stimulated greater metabolic activity than the control group, with LMW HA and MMW HA showing the most significant increase. All groups showed mineral deposits and calcium deposition after 21 days of stimulation. CONCLUSION: Our results suggest that hyaluronan can promote metabolic activity and mineralization of PDL-hTERT cells, with LMW HA being the most effective. CLINICAL RELEVANCE: These results shed light on how the various molecular weight fractions of HA promote tissue regeneration and repair, as well as help to identify an optimal molecular weight range for this application in periodontal tissues.

NAC ameliorates dental composite-induced DNA double-strand breaks and chromatin condensation.

Released (co)monomers from dental composite components can induce DNA damage of which DNA double-strand breaks (DSBs) threaten genome integrity. Here, we tested whether the administration of the antioxidant N-acetylcysteine (NAC) is able to reduce the dental composite-induced DSBs in primary human gingiva fibroblasts. The dental composites Bis-GMA (bisphenol-A-glycerolate dimethacrylate), GMA (glycidyl methacrylate), HEMA (2-hydroxyethyl methacrylate) and TEGDMA (triethyleneglycol dimethacrylate) were found to induce co-localizing microscopic nuclear foci numbers of the DSB markers γ-H2AX and 53BP1 per cell in the order: GMA>Bis-GMA>TEGDMA>HEMA. Supplementation of (co)monomer-containing culture medium with NAC led to a significant reduction of resin-induced DSBs as well as to an amelioration of dental monomer-induced nuclear chromatin condensation in gingival fibroblasts. Thus, antioxidant treatment can reduce radical-induced chromatin and DNA damage and open avenues to mitigate genotoxic effects of dental composite compounds.

Fluoride/vitamin D tablet supplementation in infants-effects on dental health after 10 years.

OBJECTIVE: We examined whether fluoride/vitamin D supplementation in the first year of life is associated with caries or molar incisor hypomineralization (MIH) at 10 years of age. METHODS: The study population consisted of 406 children for whom information on fluoride/vitamin D supplementation during the first year of life was available. Dental examination at the age of 10 included caries and MIH registration. The results of logistic regression models were adjusted for gender, age, BMI, parental education, and equivalent income. RESULTS: Children receiving supplementation during the entire first year of life had a significantly lower probability of having caries-related restorations in primary teeth in comparison to those who received supplementation for less than 6 months (fluoride supplementation: odds ratio (OR) for d3-4mfs 2.47 (1.32-4.63), for fs 2.70 (1.43-5.10); vitamin D supplementation: OR for d3-4mfs 2.08 (1.00-4.32), fs 2.50 (1.19-5.25)). The majority of logistic regression analyses indicated no association between supplementation and MIH. CONCLUSIONS: It was found a consistent significant caries-preventive effect in the primary dentition of children who received fluoride (256/372)/vitamin D supplementation (274/376) in all 12 months over the first year of life; no effects were observed for permanent dentition. The high parental interest in supplementation is linked to an imbalance of the study groups. Furthermore, tooth brushing frequency, use of fluoride toothpastes and/or other oral hygiene products were not recorded during the observation period which may also confound the results. CLINICAL RELEVANCE: Fluoride/vitamin D supplementation can be used in children for preventing caries in the primary dentition.

Comparison of ozone gas and sodium hypochlorite/chlorhexidine two-visit disinfection protocols in treating apical periodontitis: a randomized controlled clinical trial.

OBJECTIVES: In this single-blinded, prospective, randomized, controlled clinical trial, the effectiveness of an ozone gas or NaOCl/CHX disinfection protocol was compared within the root canal treatment of apical periodontitis. MATERIALS AND METHODS: Sixty permanent teeth were randomly allocated to one technique. The clinical/radiographic assessment criteria included clinical symptoms, periapical index (PAI), and size of the apical lesion. In both groups, the root canal was mechanically cleaned and irrigated with NaCl and EDTA. Ozone gas (32 g m-3) or NaOCl (3 %) was applied followed by a 1-week inter-appointment dressing (Ca(OH)2). As final disinfection, ozone gas (ozone group) or CHX 2 % (NaOCl group) was applied. Microbial samples were taken after preparing the access cavity, after chemo-mechanical treatment and after inter-appointment dressing by sterile paper points. Microbial identification was performed by mass spectroscopy (MALDI-TOF-MS) and 16S-rRNA gene sequencing. The treated teeth were blindly re-evaluated after 6/12 months. Success rates, the decrease in PAI, the size of apical lesions and bacterial reduction were compared between groups (Fischer's exact test, Mann-Whitney U test). RESULTS: There were no significant differences between the success rates (ozone group: 96.2/95.5 % after 6/12 months; NaOCl group: 95.5/95.2 % after 6/12 months). The differences in the decreases in PAI values and apical lesion sizes were also insignificant after 6 and 12 months. The bacterial reduction showed no significant differences between groups after chemo-mechanical treatment and after inter-appointment dressing. The most commonly found bacterial genera were Streptococcus spp., Parvimonas spp. and Prevotella spp. CONCLUSIONS: The here used ozone gas and NaOCl/CHX protocols showed no difference in bacterial reduction in the sampled areas of the root canals. CLINICAL RELEVANCE: Within the limitations of the study, ozone gas seems to be a possible alternative disinfection agent within the root canal treatment of apical periodontitis.

Full-length amelogenin influences the differentiation of human dental pulp stem cells.

BACKGROUND: Amelogenin is an extracellular matrix protein well known for its role in the organization and mineralization of enamel. Clinically, it is used for periodontal regeneration and, due to its finding also in predentin and intercellular spaces of dental pulp cells, it has recently been suggested for pulp capping procedures. The aim of this study was to analyse in vitro the effect of the recombinant human full-length amelogenin on the growth and differentiation of human dental pulp stem cells (hDPSCs). METHODS: Human DPSCs were treated with a supplement of amelogenin at a concentration of 10 ng/ml, 100 ng/ml and 1000 ng/ml. The groups were compared to the unstimulated control in terms of cell morphology and proliferation, mineralization and gene expression for ALP (alkaline phosphatase), DMP1 (dentin matrix protein-1) and DSPP (dentin sialophosphoprotein). RESULTS: Amelogenin affects hDPSCs differently than PDL (periodontal ligament) cells and other cell lines. The proliferation rate at two weeks is significantly reduced in presence of the highest concentration of amelogenin as compared to the unstimulated control. hDPSCs treated with low concentrations present a downregulation of DMP1 and DSPP, which is significant for DSPP (p = 0.011), but not for DMP1 (p = 0.395). CONCLUSIONS: These finding suggest that the role of full-length amelogenin is not restricted to participation in tooth structure. It influences the differentiation of hDPSC according to various concentrations and this might impair the clinical results of pulp capping.

Effect of ozone on oral cells compared with established antimicrobials.

Ozone has been proposed as an alternative antiseptic agent in dentistry based on reports of its antimicrobial effects in both gaseous and aqueous forms. This study investigated whether gaseous ozone (4 x 10(6) microg m(-3)) and aqueous ozone (1.25-20 microg ml(-1)) exert any cytotoxic effects on human oral epithelial (BHY) cells and gingival fibroblast (HGF-1) cells compared with established antiseptics [chlorhexidine digluconate (CHX) 2%, 0.2%; sodium hypochlorite (NaOCl) 5.25%, 2.25%; hydrogen peroxide (H(2)O(2)) 3%], over a time of 1 min, and compared with the antibiotic, metronidazole, over 24 h. Cell counts, metabolic activity, Sp-1 binding, actin levels, and apoptosis were evaluated. Ozone gas was found to have toxic effects on both cell types. Essentially no cytotoxic signs were observed for aqueous ozone. CHX (2%, 0.2%) was highly toxic to BHY cells, and slightly (2%) and non-toxic (0.2%) to HGF-1 cells. NaOCl and H(2)O(2) resulted in markedly reduced cell viability (BHY, HGF-1), whereas metronidazole displayed mild toxicity only to BHY cells. Taken together, aqueous ozone revealed the highest level of biocompatibility of the tested antiseptics.

Effect of ozone on non-cavitated fissure carious lesions in permanent molars. A controlled prospective clinical study.

PURPOSE: To investigate, with a randomized controlled clinical study, the effect of ozone on non-cavitated initial occlusal fissure caries compared with untreated contra-lateral control lesions (split mouth) considering the patient's current caries risk. METHODS: Forty-one patients with 57 pairs of lesions were enrolled in the study (mean age 7.7 +/- 2.2 years; upper jaw n=29, lower jaw n=28). Gaseous ozone (HealOzone) was applied once for 40 seconds to the randomly assigned test molar of each pair without the use of remineralizing solutions. Lesion progression or reversal was monitored by the laser fluorescence system DIAGNOdent for up to 3 months and the deterioration or improvement compared between the ozone-treated lesions and the untreated control lesions (in pairs). This was done for the whole study population and a subgroup of patients with high current caries risk (lesion pairs n=26). RESULTS: After 3 months, explorative data analysis revealed that the ozone-treated lesions showed significantly more caries reversal or reduced caries progression than the untreated control lesions within the group of patients at high current caries risk (Wilcoxon-Test, P= 0.035). There was no statistical significance examining the whole study population. From the data it can be concluded that ozone application significantly improved non-cavitated initial fissure caries in patients at high caries risk over a 3-month period.

The effectiveness of InGaAsP diode laser radiation to detect subgingival calculus as compared to an explorer.

BACKGROUND: The aim of the present study was to compare the ability of the diode laser to detect residual calculus with that of an explorer. METHODS: The root surface of 40 extracted human teeth, each partially covered with subgingival calculus, was instrumented with curets under simulated clinical conditions in a manikin. The samples were randomly assigned to two study groups. In group A, the root surface was treated with an explorer until it appeared free of mineralized deposits upon examination. The samples in group B were instrumented until the relative intensity of fluorescence as induced with diode laser radiation was below a threshold value of 5. The root surface of each sample was then examined for residual calculus using standardized digital images. The statistical analysis was performed with a non-paired t test at a level of significance of 5% (P < 0.05). RESULTS: The root surface of single-rooted teeth showed residual calculus on 0.19 +/- 0.37 x 10(7) microm2 in the laser group and on 0.11 +/- 0.26 x 10(7) microm2 in the explorer group (P = 0.19). For multirooted teeth, the mean calculus-covered area was 0.50 +/- 0.48 x 10(7) microm2 for the teeth evaluated with an explorer and 0.27 +/- 0.43 x 10(7) microm2 for the diode laser group (P = 0.02). CONCLUSION: The present findings indicate that the detection of subgingival calculus is significantly improved using 655 nm diode laser radiation compared to an explorer for molars but not for single-rooted teeth.