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  1. Daood U, Fawzy A
    J Mech Behav Biomed Mater, 2023 Apr;140:105737.
    PMID: 36827934 DOI: 10.1016/j.jmbbm.2023.105737
    The aim is to evaluate the development of an experimental multi-mode/Universal resin-based dentin adhesive modified with synthetic Mg2+ doped hydroxyapatite crystals (HAp) having self-remineralization and antibiofilm properties. HAp doped with Mg2+ was prepared by the precipitation method. Experimental adhesives were subjected to degree of conversion and X-ray diffraction test for size and crystal structure. Bond strength was tested, and electron microscopy (SEM/TEM) imaging of resin-dentin interface was done along with nanoleakage, nanoindentation, confocal and Raman analyses. S. mutans was analysed using CLSM images against modified adhesive specimens. Nucleating abilities within the resin-dentin specimens are determined by measuring Ca2+. Alkaline phosphatase, Runx2, and Ocn transcripts are amplified using quantitative polymerase chain reaction (q-PCR). A calcium assay is performed to quantify level of mineralisation. When compared to control adhesives, the 0.5% Hap/Mg2+ containing experimental dentin adhesive demonstrated improved interaction with dentin. The preservation of uniform intact hybrid layer with the absence of nanoleakage indicated dentin bond integrity with 0.5% HAP/Mg2+ modified adhesive. Self-remineralization and antibiofilm potentials are supported.
  2. Bhatia S, Nagendrababu V, Peters OA, Fawzy A, Daood U
    Sci Rep, 2021 04 27;11(1):9027.
    PMID: 33907252 DOI: 10.1038/s41598-021-88570-4
    To evaluate structural profiles and mechanical behaviour of WaveOne Gold (WOG), Twisted File Adaptive (TFA) and XP-endo shaper (XPS) instruments after root canal preparation. Standardized in vitro shaping was performed in presence of 5.25% sodium hypochlorite. File morphology was analyzed using scanning electron microscopy; X-ray diffraction analysis was performed before and after use along with Raman spectroscopy. Nanoindentation was carried out to characterize surface topography. Ni2+ release was measured at 1, 3, 5 and 7 days. X-ray photoelectron spectroscopy (XPS) analysis was done before and after use. After allocating scan line shifts like in WOG, mechanical deformation was shown using first order polynomials. XPS file system showed minimal grooves on surface. SEM of WOG instrument showed scraping surface defects. Hardness varied from 8.11 ± 0.99 GPa in TFA system to 6.7 ± 1.27 GPa and 4.06 ± 4.1 GPa in XPS and WOG. Ni2+ concentration from WOG was 171.2 μg/L. Raman peak at 540-545 cm-1 is attributed to Cr2O3. High resolution of Ti 2p spectrum show distinctive peaks with binding energies dominating in WOG, XPS and TFA file system. XRD exhibited NiTi phases with diffraction peaks. WOG files showed more surface deterioration and less passive layer formation as compared to TFA and XPS systems.
  3. Kok ESK, Lim XJ, Chew SX, Ong SF, See LY, Lim SH, et al.
    BMC Oral Health, 2021 03 12;21(1):116.
    PMID: 33711992 DOI: 10.1186/s12903-021-01470-x
    BACKGROUND: Compare antimicrobial efficacy of a quarternary ammonium silane (QAS)/k21 as an intracanal medicament against E. faecalis and C. albicans biofilms formed on root dentin.

    METHODOLOGY: Dentin blocks were sterilized and E. faecalis and C. albicans microbial colonies were counted for colony-forming-units against 2%k21, 2%CHX and Ca(OH)2 medicaments. Biofilm colonies after 7 days on dentin were analysed using confocal laser scanning microscopy with live/dead bacterial viability staining. TEM was done to study dentin collagen matrix. Dentin discs from 3rd day and 7th day well plate was used for Raman spectra and observed under fluorescent-microscope. Docking studies were carried out on MMP-2 S1 binding-domain with k21.

    RESULTS: There was reduction of E. faecalis/C. albicans when k21, chlorhexidine and calcium hydroxide were used with highest percentage in 2%k21 treated specimens. 2%k21 showed dense and regular collagen network with intact cross-banding and decreased Raman intensity for 2%k21 on 3rd day. NaOCl + k21 showed least adherence, whereas saline groups showed highest adherence of E. faecalis and C. albicans to root-canal dentin. Alizarin red staining of hDPSCs revealed calcium deposition in all groups with significant difference seen amongst 2%k21 groups. MMP-2 ligand binding was seen accurately indicating possible target sites for k21 intervention.

    CONCLUSION: 2%k21 can be considered as alternative intracanal medicament.

  4. Bourgi R, Daood U, Bijle MN, Fawzy A, Ghaleb M, Hardan L
    Polymers (Basel), 2021 Feb 26;13(5).
    PMID: 33652596 DOI: 10.3390/polym13050704
    Enzymatic biodegradation of demineralized collagen fibrils could lead to the reduction of resin-dentin bond strength. Therefore, methods that provide protection to collagen fibrils appear to be a pragmatic solution to improve bond strength. Thus, the study's aim was to investigate the effect of ribose (RB) on demineralized resin-dentin specimens in a modified universal adhesive. Dentin specimens were obtained, standardized and then bonded in vitro with a commercial multi-mode adhesive modified with 0, 0.5%, 1%, and 2% RB, restored with resin composite, and tested for micro-tensile bond strength (µTBS) after storage for 24 h in artificial saliva. Scanning electron microscopy (SEM) was performed to analyze resin-dentin interface. Contact angles were analyzed using a contact angle analyzer. Depth of penetration of adhesives and nanoleakage were assessed using micro-Raman spectroscopy and silver tracing. Molecular docking studies were carried out using Schrodinger small-molecule drug discovery suite 2019-4. Matrix metalloproteinases-2 (MMP-2) and cathepsin-K activities in RB-treated specimens were quantified using enzyme-linked immunosorbent assay (ELISA). The significance level was set at α = 0.05 for all statistical analyses. Incorporation of RB at 1% or 2% is of significant potential (p < 0.05) as it can be associated with improved wettability on dentin surfaces (0.5% had the lowest contact angle) as well as appreciable hybrid layer quality, and higher resin penetration. Improvement of the adhesive bond strength was shown when adding RB at 1% concentration to universal adhesive (p < 0.05). Modified adhesive increased the resistance of collagen degradation by inhibiting MMP-2 and cathepsin-K. A higher RB concentration was associated with improved results (p < 0.01). D-ribose showed favorable negative binding to collagen. In conclusion, universal adhesive using 1% or 2% RB helped in maintaining dentin collagen scaffold and proved to be successful in improving wettability, protease inhibition, and stability of demineralized dentin substrates. A more favorable substrate is created which, in turn, leads to a more stable dentin-adhesive bond. This could lead to more advantageous outcomes in a clinical scenario where a stable bond may result in longevity of the dental restoration.
  5. Daood U, Parolia A, Matinlinna J, Yiu C, Ahmed HMA, Fawzy A
    Dent Mater, 2020 12;36(12):e386-e402.
    PMID: 33010944 DOI: 10.1016/j.dental.2020.09.008
    OBJECTIVES: Evaluate a new modified quaternary ammonium silane irrigant solution for its antimicrobial, cytotoxic and mechanical properties of dentine substrate.

    METHODS: Root canal preparation was performed using stainless steel K-files™ and F4 size protaper with irrigation protocols of 6% NaOCl + 2% CHX; 3.5% QIS; 2% QIS and sterile saline. Biofilms were prepared using E. faecalis adjusted and allowed to grow for 3 days, treated with irrigants, and allowed to grow for 7 days. AFM was performed and surface free energy calculated. MC3T3 cells were infected with endo irrigant treated E. faecalis biofilms. Raman spectroscopy of biofilms were performed after bacterial re-growth on root dentine and exposed to different irrigation protocols and collagen fibers analysed collagen fibers using TEM. Antimicrobial potency against E. faecalis biofilms and cytoxicity against 3T3 NIH cells were also. Resin penetration and MitoTracker green were also evaluated for sealer penetration and mitochondrial viability. Data were analysed using One-way ANOVA, principal component analysis and post-hoc Fisher's least-significant difference.

    RESULTS: Elastic moduli were maintained amongst control (5.5 ± 0.9) and 3.5% QIS (4.4 ± 1.1) specimens with surface free energy higher in QIS specimens. MC3T3 cells showed reduced viability in 6%NaOCl+2%CHX specimens compared to QIS specimens. DNA/purine were expressed in increased intensities in control and 6% NaOCl + 2% CHX specimens with bands around 480-490 cm-1 reduced in QIS specimens. 3.5% QIS specimens showed intact collagen fibrillar network and predominantly dead bacterial cells in confocal microscopy. 3.5% QIS irrigant formed a thin crust-type surface layer with cytoplasmic extensions of 3T3NIH spread over root dentine. Experiments confirmed MitoTracker accumulation in 3.5% treated cells.

    SIGNIFICANCE: Novel QIS root canal irrigant achieved optimum antimicrobial protection inside the root canals facilitating a toxic effect against the Enterococcus faecalis biofilm. Root dentine substrates exhibited optimum mechanical properties and there was viability of fibroblastic mitochondria.

  6. Fu C, Deng S, Koneski I, Awad MM, Akram Z, Matinlinna J, et al.
    J Mech Behav Biomed Mater, 2020 12;112:104082.
    PMID: 32979607 DOI: 10.1016/j.jmbbm.2020.104082
    OBJECTIVE: To investigate the effect of blue light photoactivated riboflavin modified universal adhesives on dentin collagen biodegradation resistance, dentin apparent elastic modulus, and resin-dentin bond strength with interfacial morphology.

    METHODS: Dentin slabs were treated with 0.1% riboflavin-5-phosphate modified (powder added slowly while shaking and then sonicated to enhance the dispersion process) Universal Adhesive Scotch Bond and Zipbond™ along with control (non-modified) and experimental adhesives, photoactivated with blue light for 20s. Hydroxyproline (HYP) release was assessed after 1-week storage. Elastic-modulus testing was evaluated using universal testing machine at 24 h. Resin-dentin interfacial morphology was assessed with scanning electron-microscope, after 6-month storage. 0.1% rhodamine dye was added into each adhesive and analyzed using CLSM. Detection of free amino groups was carried out using ninhydrin and considered directly proportional to optical absorbance. Collagen molecular confirmation was determined using spectropolarimeter to evaluate and assess CD spectra. For molecular docking studies with riboflavin (PDB ID file), the binding pocket was selected with larger SiteScore and DScore using Schrodinger PB software. After curing, Raman shifts in Amide regions were obtained at 8 μm levels. Data were analyzed using Two-way analysis of variance (ANOVA, p ≤ 0.05) and Tukey-Kramer multiple comparison post hoc tests.

    RESULTS: At baseline, bond strength reduced significantly (p ≤ 0.05) in control specimens. However, at 6 months' storage, UVA Zipbond™ had significantly higher μTBS. Resin was able to diffuse through the porous demineralized dentin creating adequate hybrid layers in both 0.1%RF modified adhesives in CLSM images. In riboflavin groups, hybrid layer and resin tags were more pronounced. The circular dichroism spectrum showed negative peaks for riboflavin adhesive specimens. Best fitted poses adopted by riboflavin compound are docked with MMP-2 and -9 proteases. Amide bands and CH2 peaks followed the trend of being lowest for control UA Scotch bond adhesive specimens and increasing in Amides, proline, and CH2 intensities in 0.1%RF modified adhesive specimens. All 0.1%RF application groups showed statistically significant (p 

  7. Daood U, Fawzy AS
    Dent Mater, 2020 03;36(3):456-467.
    PMID: 32008748 DOI: 10.1016/j.dental.2020.01.005
    OBJECTIVE: The aim is to investigate the potential significance of combining minimally invasive high-intensity focused ultrasound (HIFU) with hydroxyapatite (HA) nanorods treatment for the remineralization of demineralized coronal dentine-matrix.

    METHODS: HA having nanorods structure were synthetized using ultrasonication with precipitation method. HA nanorods were characterized by TEM for average-size/shape. Following phosphoric acid demineralization, dentine specimens were treated with HA-nanorods with/without subsequent HIFU exposure for 5 s, 10 s and 20 s then stored in artificial saliva for 1-month. Dentine specimens were characterized using different SEM and Raman spectroscopic techniques. In addition, the biochemical stability and HA-nanorods were examined using ATR-FTIR to observe attachment of nanoparticles. Also, surface nanoindentation properties were evaluated using AFM in tapping-mode.

    RESULTS: HA-nanorods displayed well-defined, homogenous plate-like nanostructure. TEM revealed intact collagen-fibrils network structure with high density due to obliteration of interfibrillar spaces with clear evidence of remineralization in combined HA/HIFU treatment. With HA-nanorods treatment collagen-network structure was visible, consisting of fibrils interlaced into a compact pattern with evidence of minerals deposition. AFM investigation revealed clear mineral formation with the increase of HIFU exposure time. Bands associated with inorganic phase dominate well in HIFU exposed specimens with PO stretching within dentine mineral identified at 960 cm-1. Characteristic dentine structure for control and HIFU 20 s specimens is reflected as oscillatory mean Amide-I intensity with measurement giving a precise sinusoidal response of polarization angle β within dentinal tissue. Nanoindentation testing showed a gradual significant increase in elastic-modulus with the increase in HIFU exposure time after 1-month storage. FTIR spectrum of the HIFU exposed dentine displayed bands at 1650 cm-1, 1580 cm-1 and 1510 cm-1 that can be attributed to Amide-I, II and III.

    SIGNIFICANCE: The synergetic effect of HIFU exposure on remineralization potential of demineralized dentine-matrix following nano-hydroxyapatite treatment was revealed. This synergetic effect is dependent on HIFU exposure time.

  8. Daood U, Sauro S, Pichika MR, Omar H, Liang Lin S, Fawzy AS
    Dent Mater, 2020 01;36(1):145-156.
    PMID: 31818524 DOI: 10.1016/j.dental.2019.11.003
    OBJECTIVE: To modify a universal dentine adhesive with different concentrations of riboflavin and D-Alpha 1000 Succinate polyethylene (VE-TPGS) as a chemical enhancer and to assess the micro-tensile bond strength (24h/12 months), determine resin penetration, measurement of intermolecular interactions and cytotoxicity.

    MATERIALS AND METHODS: An experimental adhesive system based on bis-GMA, HEMA and hydrophobic monomer was doped with RF0.125 (RF - Riboflavin) or RF/VE-TPGS (0.25/0.50) and submitted to μTBS evaluation. Resin dentine slabs were prepared and examined using SEM and TEM. Adhesion force was analysed on ends of AFM cantilevers deflection. Quenched peptide assays were performed using fluorescence scanner and wavelengths set to 320nm and 405nm. Cytotoxicity was assessed using human peripheral blood mononuclear cell line. Molecular docking studies were carried out using Schrödinger small-molecule drug discovery suite 2018-2. Data from viable cell results was analyzed using one-way ANOVA. Bond strength values were analysed by two-way ANOVA. Nonparametric results were analyzed using a Kruskal-Wallis test at a 0.05 significance level.

    RESULTS: RF/VE-TPGS0.25 groups showed highest bond strength results after 24-h storage in artificial saliva (p<0.05). RF/VE-TPGS0.50 groups showed increased bond strength after 12-months of ageing. RF/VE-TPGS modified adhesives showed appreciable presence of a hybrid layer. Packing fraction indicated solid angle profiles describing well sized density and topology relations for the RF/VE-TPGS adhesives, in particular with the RF/VE-TPGS0.50 specimens. Qualitative analysis of the phenotype of macrophages was prominently CD163+ in the RF/VE-TPGS0.50. Both the compounds showed favourable negative binding energies as expressed in terms of 'XP GScore'.

    CONCLUSION: New formulations based on the incorporation of RF/VE-TPGS in universal adhesives may be of significant potential in facilitating penetration, distribution and uptake of riboflavin within the dentine surface.

  9. Daood U, Parolia A, Elkezza A, Yiu CK, Abbott P, Matinlinna JP, et al.
    Dent Mater, 2019 09;35(9):1264-1278.
    PMID: 31201019 DOI: 10.1016/j.dental.2019.05.020
    OBJECTIVE: To analyze effect of NaOCl+2% quaternary ammonium silane (QAS)-containing novel irrigant against bacteria impregnated inside the root canal system, and to evaluate its antimicrobial and mechanical potential of dentine substrate.

    METHODS: Root canal was prepared using stainless steel K-files™ and ProTaper™ and subjected to manual and ultrasonic irrigation using 6% NaOCl+2% CHX, 6% NaOCl+2% QAS and saline as control. For confocal-microscopy, Raman spectroscopy and SEM analysis before and after treatment, Enterococcus faecalis cultured for 7 days. Raman spectroscopy analysis was done across cut section of gutta percha/sealer-dentine to detect resin infiltration. Indentation of mechanical properties was evaluated using a Berkovich indenter. The contact angle of irrigants and surface free energy were evaluated. Mineralization nodules were detected through Alazarin red after 14 days.

    RESULTS: Control biofilms showed dense green colonies. Majority of E. faecalis bacteria were present in biofilm fluoresced red in NaOCl+2% QAS group. There was reduction of 484cm-1 Raman band and its intensity reached lowest with NaOCl+2% QAS. There was an increase in 1350-1420cm-1 intensity in the NaOCl+2% CHX groups. Gradual decrease in 1639cm-1 and 1609cm-1 Raman signal ratios were seen in the resin-depth region of 17μm>, 14.1μm> and 13.2μm for NaOCl+2% QAS, NaOCl+2% CHX and control groups respectively. All obturated groups showed an intact sealer/dentine interface with a few notable differences. 0.771 and 83.5% creep indentation distance for NaOCl+2% QAS ultrasonic groups were observed. Highest proportion of polar component was significantly found in the NaOCl+2% QAS groups which was significantly higher as compared to other groups. Mineralized nodules were increased in NaOCl+2% QAS.

    SIGNIFICANCE: Favorable antimicrobial and endodontic profile of the NaOCl+2% QAS solution might suggest clinical use for it for more predictable reduction of intracanal bacteria.

  10. Al-Nabulsi M, Daud A, Yiu C, Omar H, Sauro S, Fawzy A, et al.
    Materials (Basel), 2019 Aug 07;12(16).
    PMID: 31394743 DOI: 10.3390/ma12162504
    Objective: To evaluate the effect of a new application method of bulk-fill flowable composite resin material on bond-strength, nanoleakage, and mechanical properties of dentine bonding agents.

    MATERIALS AND METHODS: Sound extracted human molars were randomly divided into: manufacturer's instructions (MI), manual blend 2 mm (MB2), and manual blend 4 mm (MB4). Occlusal enamel was removed and flattened, dentin surfaces were bonded by Prime & Bond universal (Dentsply and Optibond FL, Kerr). For the MI group, adhesives were applied following the manufacturer's instructions then light-cured. For MB groups, SDR flow+ bulk-fill flowable composite resin was applied in 2- or 4-mm increment then manually rubbed by a micro brush for 15 s with uncured dentine bonding agents and the mixture was light-cured. Composite buildup was fabricated incrementally using Ceram.X One, Dentsply nanohybrid composite resin restorative material. After 24-h water storage, the teeth were sectioned to obtain beams of about 0.8 mm2 for 24-h and thermocycled micro-tensile bond strength at 0.5 mm/min crosshead speed. Degree of conversion was evaluated with micro-Raman spectroscopy. Contraction gaps at 24 h after polymerization were evaluated and atomic force microscopy (AFM) nano-indentation processes were undertaken for measuring the hardness across the interface. Depth of resin penetration was studied using a scanning electron microscope (SEM). Bond strength data was expressed using two-way ANOVA followed by Tukey's test. Nanoindentation hardness was separately analyzed using one-way ANOVA.

    RESULTS: Factors "storage F = 6.3" and "application F = 30.11" significantly affected the bond strength to dentine. For Optibond FL, no significant difference in nanoleakage was found in MI/MB4 groups between baseline and aged specimens; significant difference in nanoleakage score was observed in MB2 groups. Confocal microscopy analysis showed MB2 Optibond FL and Prime & Bond universal specimens diffusing within the dentine. Contraction gap was significantly reduced in MB2 specimens in both adhesive systems. Degree of conversion (DC) of the MB2 specimens were numerically more compared to MS1 in both adhesive systems.

    CONCLUSION: Present study suggests that the new co-blend technique might have a positive effect on bond strengths of etch-and-rinse adhesives to dentine.

  11. Fawzy AS, Daood U, Matinlinna JP
    Dent Mater, 2019 07;35(7):979-989.
    PMID: 31003759 DOI: 10.1016/j.dental.2019.04.001
    OBJECTIVE: This study introduced the potential and proof-of-concept of high intensity focused ultrasound (HIFU) technology for dentin-surface treatment for resin-dentin bonding without acid-aided demineralization. This new strategy could provide a way to enhance interface-integrity and bond-durability by changing the nature of dentin-substrate; bonded-interface structure and properties; and minimizing denuded-collagen exposure.

    METHODS: The interaction between HIFU waves and dentin-surface in terms of structural, mechanical and chemical variations were investigated by SEM, TEM, AFM, nano-indentation and Raman-analysis. The bonding between HIFU-treated dentin and two-step, etch-and-rinse, adhesive was preliminary explored by characterizing dentin-bound proteases activities, resin-dentin interfacial morphology and bond-durability with HIFU exposure at different time-points of 60, 90 and 120 s compared to conventional acid-etching technique.

    RESULTS: With the increase in HIFU exposure-time from 60-to-120 s, HIFU waves were able to remove the smear-layer, expose dentinal-tubules and creating textured/rough dentin surface. In addition, dentin surfaces showed a pattern of interlocking ribbon-like minerals-coated collagen-fibrils protruding from the underlaying amorphous dentin-background with HIFU exposure for 90 s and 120 s. This characteristic pattern of dentin-surface showing inorganic-minerals associated/aligned with collagen-fibrils, with 90-to-120 s HIFU-treatment, was confirmed by the Raman-analysis. HIFU-treated specimens showed higher nano-indentation properties and lower concentrations of active MMP-2 and Cathepsin-K compared to the acid-etched specimens. The resin-dentin bonded interface revealed the partial/complete absence of the characteristic hybrid-layer formed with conventional etch-and-rinse bonding strategy. Additionally, resin-infiltration and resin-tags formation were enhanced with the increase in HIFU exposure-time to 120 s. Although, all groups showed significant decrease in bond-strength after 12 months compared to 24 h storage in artificial saliva, groups exposed to HIFU for 90 s and 120 s showed significantly higher μTBS compared to the control acid-etched group.

    SIGNIFICANCE: The implementation of HIFU-technology for dental hard-tissues treatment could be of potential significance in adhesive/restorative dentistry owing to its ability of controlled, selective and localised combined tissue alteration/ablation effects.

  12. Daood U, Akram Z, Matinlinna JP, Fawzy AS
    Dent Mater, 2019 07;35(7):1017-1030.
    PMID: 31064669 DOI: 10.1016/j.dental.2019.04.005
    OBJECTIVE: The aim of this study was to investigate EDC-assisted collagen crosslinking effect with different concentrations of tiopronin-protected gold (TPAu) nanoparticles on demineralized dentine.

    METHODS: TPAu nanoparticles were fabricated from 0.31-g tetrachloroauric acid and 0.38-g of N-(2-mercaptopropionyl) glycine (2.4-mmol). Then co-dissolved using 35-mL of 6:1 methanol/acetic acid and mixed using NaBH4. EDC (0.3-M) was conjugated to TPAu nanoparticles at TPAU/EDC-0.25:1, and TPAU/EDC-0.5:1 treatment formulations ratios. Dentin specimens treated with 0.3-M EDC solution alone or left untreated were used as control. Nanoparticles formulations were characterized in term of particles morphology and size, Zeta potential, thermogravimetric analysis and small-angle X-ray scattering. Dentin substrates were characterized in term of TEM investigation, dentin proteases characterization, hydroxyproline liberation, elastic modulus measurement, Raman analysis and confocal microscopy viewing.

    RESULTS: TEM evaluation of tiopronin protected gold nanoparticles dispersion revealed nano-clusters formations in both groups. However, based on our TEM measurements, the particle-size was ranging from ˜20 to 50 nm with spherical core-shape which were almost similar for both TPAu/EDC ratios (0.5:1 and 0.25:1). Zeta potential measurements indicate negative nanoparticles surface charge. SAXS profiles for both formulations, suggest a typical profile for uni-lamellar nanoparticles. Superior dentin collagen cross-linking effect was found with the TPAu/EDC nanoparticles formulations compared to the control and EDC treated groups.

    SIGNIFICANCE: Cross-linking of dentin collagen using TPAu coupled with EDC through TPAu/EDC nanoparticles formulations is of potential significance in improving the biodegradation resistance, proteases inhibition, mechanical and structural stability of demineralized dentin substrates. In addition, the cross-linking effect is dependent on TPAu/EDC ratio, whereas higher cross-linking effect was found at TPAu/EDC ratio of 0.5:1.

  13. Daood U, Matinlinna JP, Fawzy AS
    Dent Mater, 2019 02;35(2):356-367.
    PMID: 30528297 DOI: 10.1016/j.dental.2018.11.031
    OBJECTIVE: Effect of d-alpha-tocopheryl poly(ethyleneglycol)-1000-succinate (VE-TPGS) with riboflavin-5'-phosphate solution on crosslinking of dentine collagen was investigated to analyze collagen's structural integrity.

    METHODS: VE-TPGS was added to RF-solution, at RF/VE-TPGS (w/w) ratios of 0.125/0.250 and 0.125/0.500. Demineralized dentine beams were used (10wt.% phosphoric acid), rinsed using deionized-water and analysed using ELISA (Human MMP2 ELISA; Human CTSK/Cathepsin-K for MMP2 and Cathepsin K analysis). AFM of dentine collagen-fibrils structure was done before and after dentine specimens' placement in mineralization solution and tested after 14days in artificial saliva/collagenase (AS/Co) solution. The specimens were tested after 24h in mineralization solution for surface/bulk elastic modulus. Nano-indentation was carried out for each specimen on intertubular-dentine with lateral spacing of 400nm. Reduced elastic-modulus and nano-hardness were calculated and collagen content was determined using hydroxyproline-assay. Micro-Raman were performed. TEM was carried out to study structural variations of dentine-collagen in artificial-saliva (collagenase). Data were presented as mean±standard deviation and analyzed by SPSS v.15, by analysis of variance.

    RESULTS: Synergetic effect of VE-TPGS was observed with RF through higher structural integrity of dentine collagen-fibrils shown by TEM/AFM. Superior surface/bulk mechanical stability was shown by nano-indentation/mechanical testing. Improvement in collagenase degradation resistance for hydroxyproline release was observed and lower endogenous-protease release of MMP-2/Cathepsin-K. Raman-analysis analysed chemical interactions between RF and collagen confirming structural-integrity of collagen fibrils after crosslinking. After 24h mineralization, AFM showed mineral depositions in close association with dentine-collagen fibrils with RF/VE-TPGS formulations.

    SIGNIFICANCE: Potential synergetic effect of RF/VE-TPGS was observed by reflection of higher structural integrity and conformational-stability of dentine-collagen fibrils.

  14. Daood U, Fawzy AS
    Arch Oral Biol, 2019 Feb;98:195-203.
    PMID: 30502562 DOI: 10.1016/j.archoralbio.2018.10.019
    OBJECTIVE: To investigate effects of HIFU on macrophage phenotype, surface micro-topography and nano-scale surface mechanical properties of dental cementum.

    MATERIALS AND METHODS: Root discs (2 mm thickness) were cut apical to CEJ and sectioned into quadrants. HIFU setup with bowl-shaped piezo ceramic transducer submerged in a water tank was used for exposure on each specimen for 15 s, 30 s or 60 s. The specimens of the control group were left without any HIFU exposure. HIFU was generated with a continuous sinusoidal wave of 120Vpp amplitude, 250 KHZ resonance-frequency and highest ultrasonic pressure of ∼10 bar at the focus. Specimens for SEM were viewed, and micro-topography characterization performed, using AFM and Ra parameter and surface area (SA) calculated by specialized SPM surface analysis software. For nano-indentation testing, experiments were carried out using AFM. Macrophage cell isolation and culturing was performed on cementum to receive the HIFU treatment at different time periods. Raman spectroscopy were scanned to create spectra perpendicular to the cementum substrate to analyze generation of standard spectra for Raman intensity ratio of hydroxyapatite normalized to the peaks ν1 960 cm-1. Data was expressed as means ± standard deviations and analyzed by one-way ANOVA in term of Ra, SA, H and Er. Different points for fluorescence intensity ratio were analyzed by Raman using Wilcoxon rank sum test.

    RESULTS: HIFU exposure at 60 s removed the smear layer and most of cementum appeared smoothened. AFM characterisation, showed a slight decrease in the irregularity of the surface as exposure time increased. Intact macrophages can be identified in control and all experimental HIFU groups. The level of fluorescence for the control and HIFU 15 and 30 s were low as compared to HIFU 60 s.

    CONCLUSION: If HIFU can be successfully implemented, it may be a possible alternative to current methods used in periodontal therapy to achieve smooth root surfaces.

  15. Daood U, Tsoi JKH, Neelakantan P, Matinlinna JP, Omar HAK, Al-Nabulsi M, et al.
    Dent Mater, 2018 08;34(8):1175-1187.
    PMID: 29779627 DOI: 10.1016/j.dental.2018.05.005
    OBJECTIVE: Collagen fibrils aid in anchoring resin composite restorations to the dentine substrate. The aim of the study was to investigate effect of non-enzymatic glycation on bond strength and durability of demineralized dentine specimens in a modified two-step etch-and-rinse dentine adhesive.

    METHODS: Dentine surfaces were etched with 37% phosphoric acid, bonded with respective in vitro ethanol and acetone adhesives modified with (m/m, 0, 1%, 2% and 3% ribose), restored with restorative composite-resin, and sectioned into resin-dentine slabs and beams to be stored for 24h or 12 months in artificial saliva. Bond-strength testing was performed with bond failure analysis. Pentosidine assay was performed on demineralized ribose modified dentine specimens with HPLC sensitive fluorescent detection. The structural variations of ribose-modified dentine were analysed using TEM and human dental pulpal cells were used for cell viability. Three-point bending test of ribose-modified dentine beams were performed and depth of penetration of adhesives evaluated with micro-Raman spectroscopy. The MMP-2 and cathepsin K activities in ribose-treated dentine powder were also quantified using ELISA. Bond strength data was expressed using two-way ANOVA followed by Tukey's test. Paired T tests were used to analyse the specimens for pentosidine crosslinks. The modulus of elasticity and dentinal MMP-2 and cathepsin K concentrations was separately analyzed using one-way ANOVA.

    RESULTS: The incorporation of RB in the experimental two-step etch-and-rinse adhesive at 1% improved the adhesive bond strength without adversely affecting the degree of polymerisation. The newly developed adhesive increases the resistance of dentine collagen to degradation by inhibiting endogenous matrix metalloproteinases and cysteine cathepsins. The application of RB to acid-etched dentine helps maintain the mechanical properties.

    SIGNIFICANCE: The incorporation of 1%RB can be considered as a potential candidate stabilizing resin dentine bond.

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