Influence of surface pre-reacted glass-ionomer (S-PRG) filler eluate on collagen morphology, remineralization, and ultimate tensile strength of demineralized dentin.

OBJECTIVE: To evaluate the effect of ions released from surface pre-reacted glass-ionomer (S-PRG) filler on collagen morphology, remineralization, and ultimate tensile strength (UTS) of demineralized dentin. MATERIALS AND METHODS: Bovine incisor root dentins were demineralized with EDTA and divided into three treatment groups: 1) water (control); 2) S-PRG filler eluate; 3) 125 ppm sodium fluoride (NaF). After a 3-min treatment, the specimens were stored in simulated body fluid (SBF) for 3 months. Collagen morphology and remineralization were assessed using transmission electron microscopy (TEM), scanning electron microscopy (SEM), and Fourier-transform infrared spectroscopy (FTIR). Additionally, ultimate tensile strength (UTS) was measured. RESULTS: TEM and SEM demonstrated that S-PRG induced more effective remineralization compared to NaF, while the control group exhibited faint mineral deposition with collagen degradation. S-PRG displayed the most homogenous mineral deposition in collagen fibrils, along with closure of interfibrillar spaces. Extensive mineral precipitation was observed within dentinal tubules in the S-PRG group. In addition, S-PRG filler eluate demonstrated significantly higher phosphate-to-amide ratio and UTS compared to NaF and control groups (p < 0.05). CONCLUSIONS: Ion released from S-PRG filler positively influenced collagen morphology, remineralization, and ultimate tensile strength of demineralized dentin. CLINICAL SIGNIFICANCE: S-PRG filler enhances remineralization and improve the biomechanics of demineralized dentin.

Addition of metal chlorides to a HOCl conditioner can enhance bond strength to smear layer deproteinized dentin.

OBJECTIVES: To evaluate the effect of smear layer deproteinization using hypochlorous acid (HOCl) with/without metal chlorides (SrCl2 and ZnCl2) on the microtensile bond strength (µTBS) of two simplified adhesives to dentin. METHODS: Human dentin surfaces with a standardized smear layer were pretreated using a 105 ppm HOCl solution with/without SrCl2 (0.05 M, 0.1 M, 0.2 M, 0.4 M) or ZnCl2 (0.05 M, 0.1 M, 0.2 M) for 5 s, 15 s, or 30 s. After the deproteinizing solution was washed out with water for 5 s, 15 s, or 30 s, pretreated surfaces were bonded with one-step self-etch adhesive Bond Force II or universal adhesive Clearfil Universal Bond Quick, and µTBS was measured after 24 h. Additionally, the deproteinizing effects of HOCl solutions with/without the metal chlorides were compared by measuring changes in the amide:phosphate ratio using attenuated total reflection Fourier transform infrared spectroscopy. Statistical analysis was performed using multifactor ANOVA, Tukey's post hoc tests and t-tests (p < 0.05). RESULTS: Pretreatment with pure HOCl for 15 s and 30 s significantly decreased the amide:phosphate ratio (p < 0.05), indicating effective deproteinization, but the µTBS of both adhesives increased significantly only if HOCl was washed out for 30 s (p < 0.05). Increasing the concentrations of metal chlorides enabled shortening of the wash-out time down to 5 s while maintaining the improved µTBS (p < 0.05). The deproteinizing effect of HOCl was not significantly altered by the addition of metal chlorides (p > 0.05). SIGNIFICANCE: The effectiveness of smear layer deproteinization using HOCl can be improved by the addition of metal chlorides, as their increasing concentration allowed to shorten the wash-out time from 30 s down to 5 s.

Application of Sulfinate Agent in Conjunction with HOCl Smear-Layer Deproteinization Improves Dentin Bonding Durability of One-step Self-etch Adhesives.

PURPOSE: To evaluate the effect of a sulfinate agent on the bonding durability of one-step self-etch adhesives (1-SEAs) to smear-layer-covered dentin deproteinized with hypochlorous acid (HOCl). MATERIALS AND METHODS: Human coronal dentin disks with a standardized smear layer were deproteinized with 100 ppm HOCl solution for 0 s (control), 15 s or 30 s. After rinsing with water for 30 s and air drying, half of the specimens were treated with a sulfinate agent (Scotchbond Universal Dual Cure Activator; SDA) prior to the application of a 1-SEA (Bond Force II [Tokuyama Dental] or Clearfil Universal Bond Quick [Kuraray Noritake]). Microtensile bond strength (µTBS) was measured after 24 h or 10,000 thermal cycles (TC). The data were analyzed by three-way ANOVA with Tukey's post-hoc tests and t-tests at the 0.05 significance level. RESULTS: The 24-h µTBS of both adhesives increased statistically significantly with the HOCl pretreatment for 15 s or 30 s (p < 0.05), but it was not statistically significantly affected by the application of SDA (p > 0.05). However, after TC, the groups treated with the combination of HOCl and SDA maintained their µTBS (p > 0.05), as opposed to untreated dentin and dentin treated with either HOCl or SDA, whose µTBS decreased significantly (p < 0.05). CONCLUSION: The application of the sulfinate agent did not statistically significantly affect the immediate bond strength of 1-SEAs, and it could not prevent a significant decrease in the bond strength to untreated dentin after thermocycling. However, the sulfinate agent significantly improved the bonding durability of 1-SEAs to HOCl smear-layer deproteinized dentin.

Smear layer deproteinization with NaOCl and HOCl: Do application/wash-out times affect dentin bonding of one-step self-etch adhesives?

This study investigated the influence of application/wash-out times of sodium hypochlorite (NaOCl) and hypochlorous acid (HOCl) on dentin bond strength of one-step self-etch adhesives (1-SEAs). Human coronal dentin discs with a standardized smear layer were pretreated with 6% NaOCl or 100 ppm HOCl for 5 s, 15 s, or 30 s, and washed out with water for 5 s, 15 s or 30 s with or without the application of Clearfil DC Activator (CDA). No pretreatment was used as a control. The discs were bonded with a 1-SEA (Bond Force II or Clearfil Universal Bond Quick) and microtensile bond strength (µTBS) was measured after 24 h. Pretreatment with NaOCl for 15 s and 30 s significantly decreased µTBS (p<0.05), irrespective of wash-out time. The application of CDA recovered µTBS but did not outperform the control group. Conversely, pretreatment with HOCl for 15 s and 30 s followed by 30 s wash-out time significantly increased µTBS of 1-SEAs (p<0.05), regardless of CDA application.

Grape Seed Extract Reduces Active Gelatinases Using an Etch-and-Rinse Mode Universal Adhesive.

PURPOSE: To examine the effect of grape seed extract (GSE) pretreatment on the reduction of active gelatinases when using universal adhesives in etch-and-rinse mode. MATERIALS AND METHODS: Fifty extracted non-carious human teeth were used in this study. Dentin powder was prepared for analysis of active MMP-2 by ELISA. Resin-dentin slices were prepared for in situ zymography in order to localize active gelatinases by quenched fluorescein-conjugated gelatin under a confocal microscope. Fluorescence intensity was analyzed quantitatively. Specimens were allocated into 6 groups: a non-treated control, phosphoric acid etched (PA), PA followed by Single Bond Universal (SBU, 3M Oral Care) or G-Premio Bond (GPB, GC), and PA followed by GSE for 1 min prior to SBU or GPB. The data were analyzed with one-way ANOVA and Bonferroni's test at a significance level of 0.05. RESULTS: Application of the universal adhesives SBU and GPB in etch-and-rinse mode reduced active MMP-2 in dentin matrices. However, the only further significant reduction was found in GPB pre-treated with GSE, as analyzed by ELISA. In situ zymography demonstrated the location of active gelatinases, and fluorescence intensity analysis confirmed significant reduction of active gelatinases in the hybrid layer of GPB pre-teated with GSE compared to GPB alone. CONCLUSION: The present study demonstrated the potential advantage of applying GSE to reduce active gelatinases especially at the hybrid layer, with greater benefit achieved for hydrophobic adhesives. Nevertheless, the mechanism of action, analysis of bond strength, and long-term efficacy require further study.

Compromised alveolar bone cells in a patient with dentinogenesis imperfecta caused by DSPP mutation.

OBJECTIVES: Dentin sialophosphoprotein (DSPP) plays an important role in the mineralization of both dentin and bones. The Dspp null mice developed periodontal diseases. Patients with DSPP mutations have dentinogenesis imperfecta (DGI), but very little is known about their bone characteristics. This study aims to characterize alveolar bone cells of a DGI patient with DSPP mutation. MATERIALS AND METHODS: Pathogenic variants were identified by whole exome and sanger sequencing. Cells isolated from the alveolar bones of a DSPP patient were investigated for their characteristics including cell morphology, attachment, spreading, proliferation, colony formation, mineralization, and osteogenic differentiation. RESULTS: We identified a Thai family with three members affected with autosomal dominant DGI harboring a heterozygous pathogenic missense mutation, c.50C > T, p.P17L, in exon 2 of the DSPP gene. The patients' phenotypes presented deteriorated opalescent teeth with periapical lesions, thickening of lamina dura, furcation involvement, alveolar bone loss, and bone exostoses. The alveolar bone cells isolated from DSPP patient exhibited compromised proliferation and colony formation. Scanning electron microscope revealed altered cellular morphology and spreading. The DSPP cells showed deviated mRNA levels of OCN, ALP, and COL1 but maintained in vitro mineralization ability compared to the control. CONCLUSIONS: We demonstrate that the DSPP p.P17L mutant alveolar bone cells had compromised cell spreading, proliferation, colony formation, and osteogenic induction, suggesting abnormal bone characteristics in the patient with DGI caused by DSPP mutation. CLINICAL RELEVANCE: DSPP mutation can induce the behavior alterations of alveolar bone cells.