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1.  Effect of salivary pellicle on antibacterial activity of novel antibacterial dental adhesives using a dental plaque microcosm biofilm model 
Objectives
Antibacterial primer and adhesive are promising to inhibit biofilms and caries. Since restorations in vivo are exposed to saliva, one concern is the attenuation of antibacterial activity due to salivary pellicles. The objective of this study was to investigate the effects of salivary pellicles on bonding agents containing a new monomer dimethylaminododecyl methacrylate (DMADDM) or nanoparticles of silver (NAg) against biofilms for the first time.
Methods
DMADDM and NAg were synthesized and incorporated into Scotchbond Multi-Purpose adhesive and primer. Specimens were either coated or not coated with salivary pellicles. A microcosm biofilm model was used with mixed saliva from ten donors. Two types of culture medium were used: an artificial saliva medium (McBain), and Brain Heart Infusion (BHI) medium without salivary proteins. Metabolic activity, colony-forming units (CFU), and lactic acid production of plaque microcosm biofilms were measured (n = 6).
Results
Bonding agents containing DMADDM and NAg greatly inhibited biofilm activities, even with salivary pellicles. When using BHI, the pre-coating of salivary pellicles on resin surfaces significantly decreased the antibacterial effect (p < 0.05). When using artificial saliva medium, pre-coating of salivary pellicles on resin did not decrease the antibacterial effect. These results suggest that artificial saliva yielded medium-derived pellicles on resin surfaces, which provided attenuating effects on biofilms similar to salivary pellicles. Compared with the commercial control, the DMADDM-containing bonding agent reduced biofilm CFU by about two orders of magnitude.
Significance
Novel DMADDM- and NAg-containing bonding agents substantially reduced biofilm growth even with salivary pellicle coating on surfaces, indicating a promising usage in saliva-rich environment. DMADDM and NAg may be useful in a wide range of primers, adhesives and other restoratives to achieve antibacterial and anti-caries capabilities.
doi:10.1016/j.dental.2013.11.004
PMCID: PMC4023513  PMID: 24332270
Antibacterial dental adhesive; Quaternary ammonium; Silver nanoparticles; Dental plaque microcosm biofilm; Salivary pellicle
2.  Fatigue of the Resin-Dentin Interface: A New Approach for Evaluating the Durability of Dentin Bonds 
There are concerns regarding the longevity of resin composite restorations and the clinical relevance of in vitro bond strength testing to the durability of dentin bonds in vivo.
Objective
The objectives of this investigation were to: 1) develop a new method of experimental evaluation for quantifying the durability of dentin bonds, 2) apply this method to characterize the interfacial strength of a selected commercial system under both monotonic and cyclic loading, and 3) distinguish mechanisms contributing to the interface degradation and failure.
Methods
A new method for fatigue testing the resin-dentin interface was developed based on a four-point flexure arrangement that includes two identical bonded interfaces. Cyclic loading of specimens comprised of coronal dentin bonded to a commercial resin composite and controls of resin composite was performed to failure within a hydrated environment. Scanning electron microscopy and nanoscopic dynamic mechanical analysis were used to evaluate failure mechanisms.
Results
The fatigue strength of the resin-dentin interface was significantly lower (p≤0.0001) than that of the resin composite and reported for dentin over the entire finite life regime. Defined at 1×107 cycles, the apparent endurance limit of the resin-dentin interface was 13 MPa, in comparison to 48 MPa and 44 MPa for the resin composite and dentin, respectively. The ratio of fully reversed endurance limit to ultimate strength of the interface (0.26) was the lowest of the three materials.
Significance
The proposed approach for characterizing the fatigue strength of resin-dentin bonds may offer new insights concerning durability of the bonded interface.
doi:10.1016/j.dental.2013.01.008
PMCID: PMC3602384  PMID: 23434232
Adhesive Resin; Bonding; Durability; Fatigue; Hybrid Layer; Resin Composite
3.  Enhancement in dentin collagen’s biological stability after proanthocyanidins treatment in clinically relevant time periods 
Objective
To investigate whether proanthocyanidins (PA) is capable of improving dentin collagen’s biological stability through cross-linking within time periods that are clinically relevant.
Materials and methods
Demineralized dentin collagen slabs were treated with 3.75 wt% PA solution for 10 s, 1 min, 30 min, 60 min, 120 min, 360 min, and 720 min, respectively. The resultant cross-linked collagen samples were subject to digestion with 0.1% collagenase at 37 °C for 2 h, 6 h, 12 h, 24 h, 36 h, and 48 h. The percentage of weight loss after digestion was calculated to evaluate PA-treated collagen’s resistance toward enzymatic degradation. Fourier-Transformed Infrared (FTIR) spectroscopy was used to probe evidences of PA-collagen interactions after various periods of PA treatment.
Results
The collagenase digestion assay suggests that PA treatment as short as 10 s can enhance collagen’s resistance toward enzymatic challenge. The FTIR spectroscopy further verifies that PA is indeed incorporated into collagen regardless of treatment time, possibly via a mechanism involving the chemical interactions between PA and collagen.
Significance
This study confirmed that PA can effectively cross-link collagen and improve its biological stability in time periods as short as 10 s. The use of PA as a priming agent is therefore clinically feasible and is a promising approach to improving the durability of current dentin bonding systems.
doi:10.1016/j.dental.2013.01.013
PMCID: PMC3640802  PMID: 23434233
Dentin collagen; Cross-linking; Proanthocyanidins; Biodegradation; FTIR
4.  Novel dental adhesives containing nanoparticles of silver and amorphous calcium phosphate 
Objectives
Secondary caries is the main reason for restoration failure, and replacement of the failed restorations accounts for 50–70% of all restorations. Antibacterial adhesives could inhibit residual bacteria in tooth cavity and invading bacteria along the margins. Calcium (Ca) and phosphate (P) ion release could remineralize the lesions. The objectives of this study were to incorporate nanoparticles of silver (NAg) and nanoparticles of amorphous calcium phosphate (NACP) into adhesive for the first time, and to investigate the effects on dentin bond strength and plaque microcosm biofilms.
Methods
Scotchbond Multi-Purpose adhesive was used as control. NAg were added into primer and adhesive at 0.1% by mass. NACP were mixed into adhesive at 10%, 20%, 30% and 40%. Microcosm biofilms were grown on disks with primer covering the adhesive on a composite. Biofilm metabolic activity, colony-forming units (CFU) and lactic acid were measured.
Results
Human dentin shear bond strengths (n=10) ranged from 26 to 34 MPa; adding NAg and NACP into adhesive did not decrease the bond strength (p > 0.1). SEM examination revealed resin tags from well-filled dentinal tubules. Numerous NACP infiltrated into the dentinal tubules. While NACP had little antibacterial effect, NAg in bonding agents greatly reduced the biofilm viability and metabolic activity, compared to the control (p < 0.05). CFU for total microorganisms, total streptococci, and mutans streptococci on bonding agents with NAg were an order of magnitude less than those of the control. Lactic acid production by biofilms for groups containing NAg was 1/4 of that of the control.
Significance
Dental plaque microcosm biofilm viability and acid production were greatly reduced on bonding agents containing NAg and NACP, without compromising dentin bond strength. The novel method of incorporating dual agents (remineralizing agent NACP and antibacterial agent NAg) may have wide applicability to other dental bonding systems.
doi:10.1016/j.dental.2012.10.005
PMCID: PMC3552134  PMID: 23138046
Antibacterial adhesive; dentin bond strength; silver nanoparticles; calcium phosphate nanoparticles; human saliva microcosm biofilm; caries inhibition
5.  Novel calcium phosphate nanocomposite with caries-inhibition in a human in situ model 
Objectives
Secondary caries at the restoration margins remains the main reason for failure. Although calcium phosphate (CaP) composites are promising for caries inhibition, there has been no report of CaP composite to inhibit caries in situ. The objectives of this study were to investigate the caries-inhibition effect of nanocomposite containing nanoparticles of amorphous calcium phosphate (NACP) in a human in situ model for the first time, and to determine colony-forming units (CFU) and Ca and P ion concentrations of biofilms on the composite restorations.
Methods
NACP with a mean particle size of 116 nm were synthesized via a spray-drying technique. Two composites were fabricated: NACP nanocomposite, and control composite filled with glass particles. Twenty-five volunteers wore palatal devices containing bovine enamel slabs with cavities restored with NACP or control composite. After 14 days, the adherent biofilms were collected for analyses. Transverse microradiography determined the enamel mineral profiles at the margins, and the enamel mineral loss ! Z was measured.
Results
NACP nanocomposite released Ca and P ions and the release significantly increased at cariogenic low pH (p < 0.05). Biofilms on NACP nanocomposite contained higher Ca (p = 0.007) and P ions (p = 0.005) than those of control (n = 25). There was no significant difference in biofilm CFU between the two composites (p > 0.1). Microradiographs showed typical subsurface lesions in enamel next to control composite, but much less lesion around NACP nanocomposite. Enamel mineral loss ! Z (mean ± sd; n = 25) around NACP nanocomposite was 13.8 ± 9.3 μm, much less than 33.5 ± 19.0 μm of the control (p = 0.001).
Significance
Novel NACP nanocomposite substantially reduced caries formation in a human in situ model for the first time. Enamel mineral loss at the margins around NACP nanocomposite was less than half of the mineral loss around control composite. Therefore, the Ca and P ion-releasing NACP nanocomposite is promising for caries-inhibiting restorations.
doi:10.1016/j.dental.2012.10.010
PMCID: PMC3561736  PMID: 23140916
human in situ model; dental nanocomposite; calcium phosphate nanoparticles; dental biofilms; ion release; secondary caries inhibition
6.  Measurement of micro-strain across resin-dentin interfaces using microscopic moiré interferometry 
Little is known about the mechanical behavior of resin-dentine interfaces during loading. The presence of relatively compliant hybrid and adhesive layers between stiffer dentin and resin composite should deform more during compressive loading.
Objective
The objective of this study was to measure changes in microstrain across bonded dentine interfaces in real time using a recently developed microscope moiré interferometer.
Method
This system used a miniature moiré interferometer, using two CCD cameras for simultaneous recording of longitudinal and transverse deformation fields, a piezotransducer for fringe shifting and use of a microscope objective with magnification up to 600×. Small beams (1 × 2 × 6 mm) of moist resin-bonded dentine covered with a diffraction grating replica were placed in a miniature compression tester to allow controlled loading from 2-37 N while imaging the interference fringe patterns.
Results
Dentine beams bonded with Single Bond/Z100 under compressive loading of resin-dentine interfaces exhibited comparatively large strains across the adhesive-hybrid layer interface. When the wrapped phase maps were unwrapped to permit conversion of fringe order to displacements, the 2-D profiles of strain fields revealed nonuniform strains across the adhesive interface. In the adhesive/hybrid layer zone, the negative strain was larger (i.e. -6000 με) than that seen in the adjacent resin composite or underlying mineralized dentin. The changes were elastic because they disappeared when the load was removed.
Significance
Microscopic moiré interferometry can be very useful in revealing real-time strain of bonded interfaces under load.
doi:10.1016/j.dental.2007.10.005
PMCID: PMC3901046  PMID: 18045678
Composite; Compression; Dentin; Dentin bonding agent; Mechanical properties; Moiré interferometry
7.  Optimizing dentin bond durability: strategies to prevent hydrolytic degradation of the hybrid layer 
Objectives
Endogenous dentin collagenolytic enzymes, matrix metalloproteinases (MMPs) and cysteine cathepsins, are responsible for the time-related hydrolysis of collagen matrix of the hybrid layers. As the integrity of the collagen matrix is essential for the preservation of long-term dentin bond strength, inhibition or inactivation of endogenous dentin proteases is necessary for durable resin-bonded composite resin restorations.
Methods
Dentin contains collagenolytic enzymes, matrix metalloproteinases (MMPs) and cysteine cathepsins, which are responsible for the hydrolytic degradation of collagen matrix in the bonded interface. Several tentative approaches to prevent enzyme function either directly or indirectly have been proposed in the literature.
Results
Chlorhexidine, a general inhibitor of both MMPs and cysteine cathepsins, applied before primer/adhesive application is the most tested method. In general, these experiments have shown that enzyme inhibition is a promising scheme to improve hybrid layer preservation and bond strength durability. Other enzyme inhibitors, e.g. enzyme-inhibiting monomers and antimicrobial compounds, may be considered promising alternatives that would allow more simple clinical application than chlorhexidine. Cross-linking collagen and/or dentin organic matrix-bound enzymes could render hybrid layer organic matrix resistant to degradation, and complete removal of water from the hybrid layer with ethanol wet bonding or biomimetic remineralization should eliminate hydrolysis of both collagen and resin components.
Significance
Identification of the enzymes responsible for the hydrolysis of hybrid layer collagen and understanding their function has prompted several innovative approaches to retain the hybrid layer integrity and strong dentin bonding. The ultimate goal, prevention of collagen matrix degradation with techniques and commercially available materials that are simple and effective in clinical settings may be achievable in several ways, and will likely become reality in the near future.
doi:10.1016/j.dental.2013.07.016
PMCID: PMC3899917  PMID: 23953737
8.  Transdentinal cytotoxicity of experimental adhesive systems of different hydrophilicity applied to ethanol-saturated dentin 
The aim of this study was to evaluate the transdentinal cytotoxicity of experimental adhesive systems (EASs) with different hydrophilicity and dentin saturation solutions (ethanol and water) on odontoblast-like cells. One hundred 0.4-mm-thick dentin discs were mounted in in vitro pulp chambers and assigned to 10 groups. Odontoblast-like cells MDPC-23 were seeded onto the pulpal side of the discs, incubated for 48h. The EASs with increasing hydrophilicity (R2, R3, R4 and R5) were applied to the occlusal side of the discs after acid etching and saturation of demineralized dentin with water or ethanol. R0 (water and ethanol- no adhesive) served as controls. After 24h, cell metabolism was evaluated by SDH enzyme production (MTT assay; n=8 discs) and cell morphology was examined by SEM (n=2 discs). The type of cell death was identified by flow cytometry and the degree of monomer conversion (%DC) was determined by infrared spectroscopy (FTIR) after two photoactivation times (10 s or 20 s). Data were analyzed statistically by the Kruskal-Wallis and Mann-Whitney tests (α=0.05). Dentin saturation with ethanol resulted in higher necrotic cell death ratios for R3, R4 and R5 compared with water saturation, although R3 and R4 induced higher SDH production. Photoactivation for 20 s significantly improved the %DC of all EASs compared with 10 s. A significant positive correlation was observed between the degree of hydrophilicity and %DC, for both photoactivation times. In conclusion, except for R2, dentin saturation with ethanol increased the cytotoxicity of EASs, as expressed by the induction of necrotic cell death.
doi:10.1016/j.dental.2013.07.006
PMCID: PMC3896916  PMID: 23906501
adhesive systems; cytotoxicity; ethanol; dentin; odontoblast-like MDPC-23 cells
9.  Chlorhexidine Release and Water Sorption Characteristics of Chlorhexidine-Incorporated Hydrophobic/Hydrophilic Resins 
doi:10.1016/j.dental.2008.03.011
PMCID: PMC3886642  PMID: 18439668
Chlohexidine; Drug Release; Water Sorption; Solubility; Hydrophobic resin; Hydrophilic resin; Hoy's solubility parameters
10.  Use of a specific MMP inhibitor (Galardin) for preservation of hybrid layer 
Objective
Dentinal MMPs have been claimed to contribute to the auto-degradation of collagen fibrils within incompletely resin-infiltrated hybrid layers and their inhibition may, therefore, slow the degradation of hybrid layer. This study aimed to determine the contribution of a synthetic MMPs inhibitor (Galardin) to the proteolytic activity of dentinal MMPs and to the morphological and mechanical features of hybrid layers after aging.
Methods
Dentin powder obtained from human molars was treated with Galardin or chlorhexidine digluconate and zymographically analyzed. Microtensile bond strength was also evaluated in extracted human teeth. Exposed dentin was etched with 35% phosphoric acid and specimens were assigned to (1) pre-treatment with Galardin as additional primer for 30s; (2) no pre-treatment. A two-step etch-and-rinse adhesive (Adper Scotchbond 1XT, 3M ESPE) was then applied in accordance with manufacturer's instructions and resin composite build-ups were created. Specimens were immediately tested for their microtensile bond strength or stored in artificial saliva for 12 months prior to being tested. Data were evaluated by two-way ANOVA and Tukey's tests (〈=0.05). Additional specimens were prepared for interfacial nanoleakage analysis under light microscopy and TEM, quantified by two independent observers and statistically analyzed (|2 test, 〈=0.05).
Results
The inhibitory effect of Galardin on dentinal MMPs was confirmed by zymographic analysis, as complete inhibition of both MMP-2 and -9 was observed. The use of Galardin had no effect on immediate bond strength, while it significantly decreased bond degradation after 1 year (p<0.05). Interfacial nanoleakage expression after aging revealed reduced silver deposits in galardin-treated specimens compared to controls (p<0.05).
Conclusions
This study confirmed that the proteolytic activity of dentinal MMPs was inhibited by the use of Galardin in a therapeutic primer. Galardin also partially preserved the mechanical integrity of the hybrid layer created by a two-step etch-and-rinse adhesive after artificial aging.
doi:10.1016/j.dental.2010.02.007
PMCID: PMC3881003  PMID: 20299089
Galardin; dental bonding systems; hybrid layer; aging; dentin
11.  Electrical properties of resin monomers used in restorative dentistry 
Objectives
The application of an electric field has been shown to positively influence the impregnation of the resin monomers currently used in dentin bonding systems during hybrid layer formation. This study presents an experimental characterization of the electrical properties of these monomers with the aim of both correlating them to their chemical structures and seeking an insight into the mechanisms of the monomer migration under an applied electric field.
Methods
Some common monomers examined were TEGDMA (triethyleneglycol dimethacrylate), HEMA (2-hydroxyethyl methacrylate), UDMA (urethane dimethacrylate), 2-MP (bis[2-(methacryloyloxy)ethyl] phosphate, TCDM di(hydroxyethyl methacrylate) ester of 5-(2,5-dioxotetrahydrofurfuryl)-3-methyl-3-cyclohexenyl-1,2-dicarboxylic anhydride) and Bis-GMA [2,2-bis(4-2-hydroxy-3-methacryloyloxypropoxyphenyl)propane]. A customized cell produced for the measurement of the electrical properties of monomers was manufactured and electrical conductivity and permittivity of resin monomers were measured.
Results
The permittivity of the tested monomers is largely affected by electrical frequency. The large values of permittivity and dielectric losses observed as frequency decreased, indicate a dominant effect of ionic polarization, particularly evident in materials showing the highest conductivity. Permittivity and conductivity of the tested monomers showed a similar behavior, i.e. materials with the lowest permittivity also show small values of conductivity and vice versa.
Significance
The results of the present study revealed a good correlation between electrical properties and Hoy solubility parameters and, in particular, the higher the polar contribution (polar forces plus hydrogen bonding) the higher the permittivity and conductivity. The most relevant outcome of this study is that the electrophoretic mechanism prevails on the electroendoosmotic effect in determining the monomer migration under the application of electric fields.
doi:10.1016/j.dental.2012.05.009
PMCID: PMC3881004  PMID: 22770675
composite resins; dentin bonding systems; electrical conductivity; electrical permittivity
12.  Effects of solubilizing surfactants and loading of antiviral, antimicrobial, and antifungal drugs on their release rates from ethylene vinyl acetate copolymer 
Objectives
This study investigates the effects of surfactants and drug loading on the drug release rate from ethylene vinyl acetate (EVA) copolymer. The release rate of nystatin from EVA was studied with addition of non-ionic surfactants Tween 60 and Cremophor RH 40. In addition, the effect of increasing drug load on the release rates of nystatin, chlorhexidine diacetate and acyclovir is also presented.
Method
Polymer casting solutions were prepared by stirring EVA copolymer and nystatin (2.5 wt %) in dichloromethane. Nystatin and surfactants were added in ratios of (1:1), (1:2) and (1:3). Drug loading was studied with 2.5, 5.0, 7.5, and 10.0% wt. proportions of nystatin, chlorhexidine diacetate and acyclovir incorporated into a separate polymer. Three drug loaded polymer square films (3cm × 3cm × 0.08 cm) were cut from dry films to follow the kinetics of drug release at 37°C. 10 ml of either distilled water or PBS was used as the extracting medium that was replaced daily. PBS was used for nystatin release with addition of surfactants and water was used for the study on drug loading and surfactant release. The rate of drug release was measured by UV-spectrophotometer. The amount of surfactant released was determined by HPLC.
Results
The release of nystatin was low in PBS and its release rate increased with the addition of surfactants. Also, increasing surfactant concentrations resulted in increased drug release rates. The release rates of chlorhexidine diacetate (p<0.0001), acyclovir (p<0.0003) and nystatin (p<0.0017) linearly increased with increasing drug loads. The amount of surfactants released was above the CMC.
Significance
This study demonstrates that the three therapeutic agents show a sustained rate of drug release from EVA copolymer over extended periods of time. Nystatin release in PBS is low owing to its poor solubility. Its release rate is enhanced by addition of surfactants and increasing the drug load as well.
doi:10.1016/j.dental.2006.08.005
PMCID: PMC3881552  PMID: 17049593
Drug delivery; EVA matrix; nystatin; surfactants; chlorhexidine diacetate; acyclovir; drug loading
13.  Resin-based composite performance: Are there some things we can’t predict? 
Objective
The objective of this manuscript is to address the following questions: Why do direct dental composite restorative materials fail clinically? What tests may be appropriate for predicting clinical performance? Does in vitro testing correlate with clinical performance?
Materials and methods
The literature relating to the clinical and laboratory performance of dental composite restorative materials was reviewed. The main reasons for failure and replacement of dental composite restorations provided the guidance for identifying specific material’s properties that were likely to have the greatest impact on clinical outcomes.
Results
There are few examples of studies showing correlation between laboratory tests of physical or mechanical properties and clinical performance of dental composites. Evidence does exist to relate clinical wear to flexure strength, fracture toughness and degree of conversion of the polymer matrix. There is evidence relating marginal breakdown to fracture toughness. Despite the fact that little confirmatory evidence exists, there is the expectation that clinical fracture and wear relates to resistance to fatigue. Only minimal evidence exists to correlate marginal quality and bond strength in the laboratory with clinical performance of bonded dental composites.
Conclusions
The use of clinical trials to evaluate new dental composite formulations for their performance is expensive and time consuming, and it would be ideal to be able to predict clinical outcomes based on a single or multiple laboratory tests. However, though certain correlations exist, the overall clinical success of dental composites is multi-factorial and therefore is unlikely to be predicted accurately by even a battery of in vitro test methods.
doi:10.1016/j.dental.2012.06.013
PMCID: PMC3495244  PMID: 22809582
dental materials; testing; physical properties; clinical performance; dental composites; direct restoratives
14.  A Hydrogel Scaffold That Maintains Viability and Supports Differentiation of Dental Pulp Stem Cells 
Objectives
The clinical translation of stem cell-based Regenerative Endodontics demands further development of suitable injectable scaffolds. Puramatrix™ is a defined, self-assembling peptide hydrogel which instantaneously polymerizes under normal physiological conditions. Here, we assessed the compatibility of Puramatrix™ with dental pulp stem cell (DPSC) growth and differentiation.
Methods
DPSC cells were grown in 0.05 to 0.25% Puramatrix™. Cell viability was measured colorimetrically using the WST-1 assay. Cell morphology was observed in 3-D modeling using confocal microscopy. In addition, we used the human tooth slice model with Puramatrix™ to verify DPSC differentiation into odontoblast-like cells, as measured by expression of DSPP and DMP-1.
Results
DPSC survived and proliferated in Puramatrix™ for at least three weeks in culture. Confocal microscopy revealed that cells seeded in Puramatrix™ presented morphological features of healthy cells, and some cells exhibited cytoplasmic elongations. Notably, after 21 days in tooth slices containing Puramatrix™, DPSC cells expressed DMP-1 and DSPP, putative markers of odontoblastic differentiation.
Significance
Collectively, these data suggest that self-assembling peptide hydrogels might be useful injectable scaffolds for stem cell-based Regenerative Endodontics.
doi:10.1016/j.dental.2012.08.002
PMCID: PMC3515741  PMID: 22901827
Tissue engineering; Hydrogel; Dental pulp; Regenerative Endodontics; Odontoblast; Stem cells
15.  Translating research and into everyday clinical practice: Lessons learned from a USA national dental practice-based research network 
Clinical studies are of paramount importance for testing and translation of the research findings to the community. Despite the existence of clinical studies, a significant delay exists between the generation of new knowledge and its application into the medical/dental community and their patients. One example is the repair of defective dental restorations. About 75% of practitioners in general dental practices do not consider the repair of dental restorations as a viable alternative to the replacement of defective restorations. Engaging and partnering with health practitioners in the field on studies addressing everyday clinical research questions may offer a solution to speed up the translation of the research findings. Practice-based research (PBR) offers a unique opportunity for practitioners to be involved in the research process, formulating clinical research questions. Additionally, PBR generates evidence-based knowledge with a broader spectrum that can be more readily generalized to the public. With PBR, clinicians are involved in the entire research process from its inception to its dissemination. Early practitioner interaction in the research process may result in ideas being more readily incorporated into practice. This paper discusses PBR as a mean to speed up the translation of research findings to clinical practice. It also reviews repair versus replacement of defective restorations as one example of the delay in the application of research findings to clinical practice.
doi:10.1016/j.dental.2012.07.159
PMCID: PMC3524332  PMID: 22889478
16.  Perspectives on the Role of Nanotechnology in Bone Tissue Engineering 
Objective
This review surveys new developments in bone tissue engineering, specifically focusing on the promising role of nanotechnology and describes future avenues of research.
Methods
The review first reinforces the need to fabricate scaffolds with multi-dimensional hierarchies for improved mechanical integrity. Next, new advances to promote bioactivity by manipulating the nano-level internal surfaces of scaffolds are examined followed by an evaluation of techniques to using scaffolds as a vehicle for local drug delivery to promote bone regeneration/integration and methods of seeding cells into the scaffold.
Results
Through a review of the state of the field, critical questions are posed to guide future research towards producing materials and therapies to bring state-of-the-art technology to clinical settings.
Significance
The development of scaffolds for bone regeneration requires a material able to promote rapid bone formation while possessing sufficient strength to prevent fracture under physiological loads. Success in simultaneously achieving mechanical integrity and sufficient bioactivity with a single material has been limited. However, the use of new tools to manipulate and characterize matter down to the nano-scale may enable a new generation of bone scaffolds that will surpass the performance of autologous bone implants.
doi:10.1016/j.dental.2012.08.001
PMCID: PMC3638810  PMID: 22901861
bone; nanotechnology; bone scaffolds; composites; drug delivery; cell seeding
17.  Optimizing dentin bond durability: control of collagen degradation by matrix metalloproteinases and cysteine cathepsins 
Objectives
Contemporary adhesives lose their bond strength to dentin regardless of the bonding system used. This loss relates to the hydrolysis of collagen matrix of the hybrid layers. The preservation of the collagen matrix integrity is a key issue in the attempts to improve the dentin bonding durability.
Methods
Dentin contains collagenolytic enzymes, matrix metalloproteinases (MMPs) and cysteine cathepsins, which are responsible for the hydrolytic degradation of collagen matrix in the bonded interface.
Results
The identities, roles and function of collagenolytic enzymes in mineralized dentin has been gathered only within last 15 years, but they have already been demonstrated to have an important role in dental hard tissue pathologies, including the degradation of the hybrid layer. Identifying responsible enzymes facilitates the development of new, more efficient methods to improve the stability of dentin-adhesive bond and durability of bond strength.
Significance
Understanding the nature and role of proteolytic degradation of dentin-adhesive interfaces has improved immensely and has practically grown to a scientific field of its own within only 10 years, holding excellent promise that stable resin-dentin bonds will be routinely available in a daily clinical setting already in a near future.
doi:10.1016/j.dental.2012.08.004
PMCID: PMC3684081  PMID: 22901826
18.  Present and future of glass-ionomers and calcium-silicate cements as bioactive materials in dentistry: Biophotonics-based interfacial analyses in health and disease 
Dental Materials  2014;30(1):50-61.
Objective
Since their introduction, calcium silicate cements have primarily found use as endodontic sealers, due to long setting times. While similar in chemistry, recent variations such as constituent proportions, purities and manufacturing processes mandate a critical understanding of service behavior differences of the new coronal restorative material variants. Of particular relevance to minimally invasive philosophies is the potential for ion supply, from initial hydration to mature set in dental cements. They may be capable of supporting repair and remineralization of dentin left after decay and cavity preparation, following the concepts of ion exchange from glass ionomers.
Methods
This paper reviews the underlying chemistry and interactions of glass ionomer and calcium silicate cements, with dental tissues, concentrating on dentin–restoration interface reactions. We additionally demonstrate a new optical technique, based around high resolution deep tissue, two-photon fluorescence and lifetime imaging, which allows monitoring of undisturbed cement–dentin interface samples behavior over time.
Results
The local bioactivity of the calcium-silicate based materials has been shown to produce mineralization within the subjacent dentin substrate, extending deep within the tissues. This suggests that the local ion-rich alkaline environment may be more favorable to mineral repair and re-construction, compared with the acidic environs of comparable glass ionomer based materials.
Significance
The advantages of this potential re-mineralization phenomenon for minimally invasive management of carious dentin are self-evident. There is a clear need to improve the bioactivity of restorative dental materials and these calcium silicate cement systems offer exciting possibilities in realizing this goal.
doi:10.1016/j.dental.2013.08.202
PMCID: PMC3885799  PMID: 24113131
Bioactivity; Calcium silicate; Glass ionomer; Cements; Caries; Remineralization; Biophotonic imaging
19.  Durability of bonds and clinical success of adhesive restorations 
Resin-dentin bond strength durability testing has been extensively used to evaluate the effectiveness of adhesive systems and the applicability of new strategies to improve that property. Clinical effectiveness is determined by the survival rates of restorations placed in non-carious cervical lesions (NCCL). While there is evidence that the bond strength data generated in laboratory studies somehow correlates with the clinical outcome of NCCL restorations, it is questionable whether the knowledge of bonding mechanisms obtained from laboratory testing can be used to justify clinical performance of resin-dentin bonds. There are significant morphological and structural differences between the bonding substrate used in in vitro testing versus the substrate encountered in NCCL. These differences qualify NCCL as a hostile substrate for bonding, yielding bond strengths that are usually lower than those obtained in normal dentin. However, clinical survival time of NCCL restorations often surpass the durability of normal dentin tested in the laboratory. Likewise, clinical reports on the long-term survival rates of posterior composite restorations defy the relatively rapid rate of degradation of adhesive interfaces reported in laboratory studies. This article critically analyzes how the effectiveness of adhesive systems is currently measured, to identify gaps in knowledge where new research could be encouraged. The morphological and chemical analysis of bonded interfaces of resin composite restorations in teeth that had been in clinical service for many years, but were extracted for periodontal reasons, could be a useful tool to observe the ultrastructural characteristics of restorations that are regarded as clinically acceptable. This could help determine how much degradation is acceptable for clinical success.
doi:10.1016/j.dental.2011.09.011
PMCID: PMC3863938  PMID: 22192252
Dentin; Adhesives; Durability; Clinical Outcome
20.  State of the art etch-and-rinse adhesives 
Etch-and-rinse adhesive systems are the oldest of the multi-generation evolution of resin bonding systems. In the 3-step version, they involve acid-etching, priming and application of a separate adhesive. Each step can accomplish multiple goals. This review explores the therapeutic opportunities of each separate step. Acid-etching, using 32-37% phosphoric acid (pH 0.1-0.4) not only simultaneously etches enamel and dentin, but the low pH kills many residual bacteria. Some etchants include anti-microbial compounds such as benzalkonium chloride that also inhibits matrix metalloproteinases (MMPs) in dentin. Primers are usually water and HEMA-rich solutions that ensure complete expansion of the collagen fibril meshwork and wet the collagen with hydrophilic monomers. However, water alone can re-expand dried dentin and can also serve as a vehicle for protease inhibitors or protein cross-linking agents that may increase the durability of resin-dentin bonds. In the future, ethanol or other water-free solvents may serve as dehydrating primers that may also contain antibacterial quaternary ammonium methacrylates to inhibit dentin MMPs and increase the durability of resin-dentin bonds. The complete evaporation of solvents is nearly impossible. Manufacturers may need to optimize solvent concentrations. Solvent-free adhesives can seal resin-dentin interfaces with hydrophobic resins that may also contain fluoride and antimicrobial compounds. Etch-and-rinse adhesives produce higher resin-dentin bonds that are more durable than most 1 and 2-step adhesives. Incorporation of protease inhibitors in etchants and/or cross-linking agents in primers may increase the durability of resin-dentin bonds. The therapeutic potential of etch-and-rinse adhesives has yet to be fully exploited.
doi:10.1016/j.dental.2010.10.016
PMCID: PMC3857593  PMID: 21112620
Acid-etchants; Primers; Adhesives; Durability; MMPs
21.  Effect of a non-thermal, atmospheric-pressure, plasma brush on conversion of model self-etch adhesive formulations compared to conventional photo-polymerization 
Objective
To determine the effectiveness and efficiency of non-thermal, atmospheric plasmas for inducing polymerization of model dental self-etch adhesives.
Methods
The monomer mixtures used were bis-[2-(methacryloyloxy)ethyl] phosphate (2MP) and 2-hydroxyethyl methacrylate (HEMA), with mass ratios of 70/30, 50/50 and 30/70. Water was added to the above formulations: 10–30 wt%. These monomer/water mixtures were treated steadily for 40 s under a non-thermal atmospheric plasma brush working at temperatures from 32° to 35°C. For comparison, photo-initiators were added to the above formulations for photo-polymerization studies, which were light-cured for 40 s. The degree of conversion (DC) of both the plasma- and light-cured samples was measured using FTIR spectroscopy with an attenuated total reflectance attachment.
Results
The non-thermal plasma brush was effective in inducing polymerization of the model self-etch adhesives. The presence of water did not negatively affect the DC of plasma-cured samples. Indeed, DC values slightly increased, with increasing water content in adhesives: from 58.3% to 68.7% when the water content increased from 10% to 30% in the adhesives with a 50/50 (2MP/HEMA) mass ratio. Conversion values of the plasma-cured groups were higher than those of light-cured samples with the same mass ratio and water content. Spectral differences between the plasma- and light-cured groups indicate subtle structural distinctions in the resultant polymer networks.
Significance
This research if the first to demonstrate that the non-thermal plasma brush induces polymerization of model adhesives under clinical settings by direct/indirect energy transfer. This device shows promise for polymerization of dental composite restorations having enhanced properties and performance.
doi:10.1016/j.dental.2012.09.005
PMCID: PMC3482274  PMID: 23018084
non-thermal plasmas; plasma-induced polymerization; self-etch adhesives; FTIR
22.  Resin–dentin bonds to EDTA-treated vs. acid-etched dentin using ethanol wet-bonding 
Objective
To compare resin–dentin bond strengths and the micropermeability of hydrophobic vs. hydrophilic resins bonded to acid-etched or EDTA-treated dentin, using the ethanol wet-bonding technique.
Methods
Flat dentin surfaces from extracted human third molars were conditioned before bonding with: 37% H3PO4 (15 s) or 0.1 M EDTA (60 s). Five experimental resin blends of different hydrophilicities and one commercial adhesive (SBMP: Scotchbond Multi-Purpose) were applied to ethanol wet-dentin (1 min) and light-cured (20 s). The solvated resins were used as primers (50% ethanol/50% comonomers) and their respective neat resins were used as the adhesive. The resin-bonded teeth were stored in distilled water (24 h) and sectioned in beams for microtensile bond strength testing. Modes of failure were examined by stereoscopic light microscopy and SEM. Confocal tandem scanning microscopy (TSM) interfacial characterization and micropermeability were also performed after filling the pulp chamber with 1 wt% aqueous rhodamine-B.
Results
The most hydrophobic resin 1 gave the lowest bond strength values to acid-etched dentin and all beams failed prematurely when the resin was applied to EDTA-treated dentin. Resins 2 and 3 gave intermediate bond strengths to both conditioned substrates. Resin 4, an acidic hydrophilic resin, gave the highest bond strengths to both EDTA-treated and acid-etched dentin. Resin 5 was the only hydrophilic resin showing poor resin infiltration when applied on acid-etched dentin.
Significance
The ethanol wet-bonding technique may improve the infiltration of most of the adhesives used in this study into dentin, especially when applied to EDTA-treated dentin. The chemical composition of the resin blends was a determining factor influencing the ability of adhesives to bond to EDTA-treated or 37% H3PO4 acid-etched dentin, when using the ethanol wet-bonding technique in a clinically relevant time period.
doi:10.1016/j.dental.2009.12.008
PMCID: PMC3824620  PMID: 20074787
Microtensile bond strength; Micropermeability Confocal microscopy; Ethanol-saturated dentin; Hydrophobic hybrid layer
23.  Chlorhexidine stabilizes the adhesive interface: a 2 year in vitro study 
Objectives
This study evaluated the role of endogenous dentin MMPs in auto-degradation of collagen fibrils within adhesive-bonded interfaces. The null hypotheses tested were that adhesive blends or chlorhexidine digluconate (CHX) application does not modify dentin MMPs activity and that CHX used as therapeutic primer does not improve the stability of adhesive interfaces over time.
Methods
Zymograms of protein extracts from human dentin powder incubated with Adper Scotchbond 1XT (SB1XT) on untreated or 0.2–2% CHX treated dentin were obtained to assay dentin MMPs activity. Microtensile bond strength and interfacial nanoleakage expression of SB1XT bonded interfaces (with or without CHX pre-treatment for 30s on the etched surface) were analyzed immediately and after 2 yr of storage in artificial saliva at 37°C.
Results
Zymograms showed that application of SB1XT to human dentin powder increases MMP-2 activity, while CHX pre-treatment inhibited all dentin gelatinolytic activity, irrespective from the tested concentration. CHX significantly lowered the loss of bond strength and nanoleakage seen in acid-etched resin-bonded dentin artificially aged for 2 yr.
Significance
The study demonstrates the active role of SB1XT in dentin MMP-2 activation and the efficacy of CHX inhibition of MMPs even if used at low concentration (0.2%).
doi:10.1016/j.dental.2009.11.153
PMCID: PMC3819099  PMID: 20045177
chlorhexidine; dental bonding systems; hybrid layer; aging; dentin
24.  Influence of different initiators on the degree of conversion of experimental adhesive blends in relation to their hydrophilicity and solvent content 
Objectives
The aim of this study was to compare the curing reaction of five experimental adhesive blends containing different photo-initiating systems. The hypothesis tested was that degree of conversion (DC) of resin blends is affected by resin type, solvent content and photo-initiating system.
Methods
The experimental methacrylate resin blends were ranked from hydrophobic (R2) to hydrophilic (R3 and R4) and tested as neat, or solvated with 10% or 20% ethanol, or 10% ethanol and 10% water. Three different photo-initiators were used: IS-1 = 0.25% CQ (camphorquinone) + 1% EDMAB (ethyl 4-dimethylaminobenzoate); IS-2 = 1.25% TPO (diphenyl(2,4,6-trimethylbenzoyl)-phosphine oxide); IS-3 = 0.25% CQ + 0.50% EDMAB + 0.50% TPO. DC of resin blends was measured with a differential scanning calorimeter. Data were analyzed with a three-way ANOVA.
Results
Neat resin type influenced DC, as R4 showed the highest values compared to R2 and R3 (p<0.05). Solvent had a significant effect on DC (p<0.05): dilution of resin blends with 10% or 20% ethanol or 10% ethanol + 10% water increased the DC of all resins, except for R4. Initiators influenced the polymerization since neat resins and mixtures solvated with 10 or 20% ethanol showed their highest DC values when polymerized with IS-1 or IS-3 (p>0.05), while IS-2 or IS-3 increased the DC values of resins diluted with 10% ethanol and 10% water (p<0.05).
Conclusions
Water-compatible photo-initiators such as TPO should be included in the hydrophilic solvated adhesive formulation to ensure an appropriate DC of the adhesive layer.
doi:10.1016/j.dental.2009.11.078
PMCID: PMC3817491  PMID: 20018363
dentin-bonding agents; degree of conversion; initiators; hydrophilicity; solvent
25.  Effects of residual ethanol on the rate and degree of conversion of five experimental resins 
Objectives
This study examined the extent of ethanol retention in five comonomer blends of experimental methacrylate-based dental adhesives, containing (10, 20, or 30 wt%) ethanol, after solvent evaporation, as well as observing the effect of residual ethanol and exposure duration on degree of conversion (DC). The null hypothesis that was tested was that residual, unevaporated ethanol has no effect on the rate or extent of DC of polymerized adhesive resins.
Methods
A known mass of each mixture was placed in glass wells and evaporated for 60 sec. The mass of the mixtures before and after evaporation was measured, allowing calculation of the gravimetric ethanol loss/retention.
Results
The concentration of retained ethanol increased significantly with ethanol concentration (p<0.01): 1.1–1.9 moles/L for 10% ethanol/90% comonomers, 2.2–3.5 moles/L for 20% ethanol, and 2.6–3.7 moles/L for 30% ethanol/70% comonomers. As ethanol is evaporated from solvated comonomer mixtures, the molar concentration of comonomers increases, reducing the vapor pressure of the remaining ethanol. Thus, the fractional loss of ethanol solvent decreases as the comonomer concentration increases.
The DC of 10, 20, and 30 wt% ethanol blends increased with ethanol concentration in 4 of the 5 experimental resins (p<0.05), increasing by 30 to 45% when 10 or 20 wt% ethanol was added to neat resins, regardless of exposure duration. Depending on the resin system, inclusion of 30% ethanol lowered DC at 20 s but increased DC after 40–60 sec of light exposure.
Conclusion
Since 10 and 20 wt% ethanol-resin blends increased the DC of solvated resins by 30–45% over neat resins, the test null hypothesis is rejected. Even with prolonged evaporation, 4–9% residual ethanol concentration can remain in 90/10 (wt/wt) comonomer/ethanol mixtures. This is thought to be because comonomers lower the vapor pressure of ethanol. This amount of residual ethanol facilitates DC but lowers the rate of polymerization.
doi:10.1016/j.dental.2008.11.005
PMCID: PMC3817492  PMID: 19111335
ethanol; DC; methacrylate resins; vapor pressure

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