Proteolytic activity, degradation, and dissolution of primary and permanent teeth.

BACKGROUND: Primary and permanent teeth composition may influence dissolution and degradation rates. AIM: To compare the dissolution and degradation of primary and permanent teeth. DESIGN: Enamel and dentin powders were obtained from primary molars and premolars and incubated within different pH buffers. Calcium and inorganic phosphate release was quantified in the buffers by atomic absorption and light spectrophotometry. A colorimetric assay was used to assess the MMP activity of primary dentin (PrD) and permanent dentin (PeD). Collagen degradation was assessed by dry mass loss, change in elastic modulus (E), and ICTP and CTX release. Data were submitted to ANOVA and Tukey's tests (α = 0.05). RESULTS: Similar dissolution was found between PrD and PeD after 256 hours. At pH 4.5, enamel released more minerals than dentin whereas at pH 5.5 the inverse result was observed. MMP activity was similar for both substrates. PrD showed higher dry mass loss after 1 week. In general, greater reduction in E was recorded for PrD. Higher quantities of ICTP and CTX were released from PrD after 1 week. CONCLUSIONS: Primary and permanent teeth presented similar demineralization rates. Collagen degradation, however, was faster and more substantial for PrD.

Collagen intrafibrillar mineralization as a result of the balance between osmotic equilibrium and electroneutrality.

Mineralization of fibrillar collagen with biomimetic process-directing agents has enabled scientists to gain insight into the potential mechanisms involved in intrafibrillar mineralization. Here, by using polycation- and polyanion-directed intrafibrillar mineralization, we challenge the popular paradigm that electrostatic attraction is solely responsible for polyelectrolyte-directed intrafibrillar mineralization. As there is no difference when a polycationic or a polyanionic electrolyte is used to direct collagen mineralization, we argue that additional types of long-range non-electrostatic interaction are responsible for intrafibrillar mineralization. Molecular dynamics simulations of collagen structures in the presence of extrafibrillar polyelectrolytes show that the outward movement of ions and intrafibrillar water through the collagen surface occurs irrespective of the charges of polyelectrolytes, resulting in the experimentally verifiable contraction of the collagen structures. The need to balance electroneutrality and osmotic equilibrium simultaneously to establish Gibbs-Donnan equilibrium in a polyelectrolyte-directed mineralization system establishes a new model for collagen intrafibrillar mineralization that supplements existing collagen mineralization mechanisms.

Zein based magnesium oxide nanoparticles: Assessment of antimicrobial activity for dental implications.

MgO nanoparticles have been recently discovered as an antibacterial, however, they limited by property degradation due to agglomeration. The addition of a coating agent, such as a zein polymer, is effective in preventing agglomeration without affecting nanosized properties. The aim of this study was to assess the antimicrobial property of MgO nanoparticles when coated with a zein polymer against several oral bacteria and fungi. This was done by utilizing various assessment techniques. The ultimate aim is to use these nanoparticles in dental preparations. The antimicrobial activity of zein-coated MgO nanoparticles at different concentrations of 0.5, 1 and 2% were tested against four different microorganisms: Staphylococcus aureus, Streptococcus mutans and Enterococcus faecalis (gram positive bacteria), and Candida albicans (as oral fungus). Two different techniques were utilized: the Kirby-Bauer test, and a modified direct contact test. The results indicated that the antibacterial effect of 1% or 2% zein-coated MgO nanowires were statistically significant (p<0.05) against the four organisms studied: S. mutans, S. aureus, E. faecalis and C. albicans. Zein-coated MgO nanoparticles are a new human friendly and potent antimicrobial agent that can be incorporated in the formulation of a variety of new dental materials and products that should provide improvements in dental care and oral health.

Effect of pH on dentin protease inactivation by carbodiimide.

A water-soluble crosslinking agent, 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC), has been used as a pretreatment of acid-etched dentin to inactivate matrix-bound endogenous dentin proteases. The aim of this study was to evaluate the effect of pH on the inactivation capacity of EDC. Demineralized dentin beams (1 × 2 × 6 mm) were divided into six groups (n = 8 per group). Then, EDC (0.3 M) was solubilized in distilled water with pH of 2, 4, 6, 7, 9, or 11. Control EDC was solubilized in 0.1 M 2-(N-morpholino) ethanesulfonic acid (MES) buffer and its pH was adjusted to 6. The dentin beams were pretreated for 1 min with EDC at each pH or with EDC in MES buffer at pH 6.0 and then incubated in 1 ml of simulated body fluid (pH 7.2) for 1, 3, 7, or 14 d. Untreated beams served as controls. At each study time-point, the dry mass of dentin beams was assessed and the incubation media were analyzed for carboxyterminal telopeptide of type-I collagen (ICTP) and C-terminal telopeptide of type I collagen (CTX) using specific ELISAs. Data were subjected to repeat-measures anova. The results of the study indicated that specimens pretreated with EDC in MES buffer showed the lowest collagen degradation in terms of mass loss and release of telopeptides, while specimens pretreated in alkaline media showed the highest collagen degradation. This study indicates that the pH of the EDC solution plays an important role in the stability of dentin protease inactivation.

Effect of a one-step self-etch adhesive on endogenous dentin matrix metalloproteinases.

Degradation of the hybrid layer created in dentin by dentin adhesives is caused by enzyme activities present within the dentin matrix that destroy unprotected collagen fibrils. The aim of the present study was to evaluate the effect of a one-step self-etch adhesive system on dentinal matrix metalloproteinases 2 and 4 (MMP-2 and MMP-9, respectively) using in situ zymography and an enzymatic activity assay. The null hypothesis tested was that there are no differences in the activities of dentinal MMPs before and after treatment with a one-step adhesive system. The MMP-2 and MMP-9 activities in dentin treated with the one-step adhesive, Adper Easy Bond, were quantified using an enzymatic activity assay system. The MMP activities within the hybrid layer created by the one-step adhesive tested were also evaluated using in situ zymography. The enzymatic assay revealed an increase in MMP-2 and MMP-9 activities after treatment with adhesive. In situ zymography indicated that gelatinolytic activity is present within the hybrid layer created with the one-step self-etch adhesive. The host-derived gelatinases were localized within the hybrid layer and remained active after the bonding procedure. It is concluded that the one-step self-etch adhesive investigated activates endogenous MMP-2 and MMP-9 with the dentin matrix, which may cause collagen degradation over time.

Zinc Inhibits Collagenolysis by Cathepsin K and Matrix Metalloproteinases in Demineralized Dentin Matrix.

The enzymatic degradation of dentin organic matrix occurs via both the action of matrix metalloproteinases (MMPs) and cysteine cathepsins (CCs). Zinc can prevent collagen hydrolysis by MMPs. However, its effect on the activity of dentin-bound CCs is not known. The aim of this study was to investigate the effect of zinc on matrix-bound cathepsin K and MMP activity in dentin. Completely demineralized dentin beams were divided into test groups (n = 9) and incubated at 37°C in an incubation media (1 mL) containing ZnCl2 of 0.02 (physiological level, control), 0.2, 0.5, 1, 5, 10, 20, 30, or 40 mM. The dry mass changes of the beams were determined, and incubation media were analyzed for cathepsin K- and MMP-specific collagen degradation end products - CTX (C-terminal cross-linked telopeptide of type I collagen) and ICTP (cross-linked carboxy-terminal telopeptide of type I collagen) - at 1, 3, and 7 days of incubation. The mass loss of the beams decreased when the zinc level in the incubation media was ≥5 mM (p < 0.05). The release of liberated collagen degradation telopeptides decreased in accordance with the decrease in the mass loss rates of the beams. Cathepsin K-induced dentin collagen degradation can be strongly inhibited by zinc. Zinc levels of ≥5 mM can be considered as a reliable threshold for the stabilization of dentin matrices.

Role of Chlorhexidine on Long-term Bond Strength of Self-adhesive Composite Cements to Intraradicular Dentin.

PURPOSE: To examine the effect of CHX pre-treatment on long-term bond strength of fiber posts luted with self-adhesive resin cements. MATERIALS AND METHODS: Seventy-two single-rooted teeth were selected for root canal treatment and post space preparation. The tested self-adhesive cement/post combinations were (N = 36): 1. RelyX Fiber-Posts luted with RelyX Unicem; 2. Rebilda Posts luted with Bifix SE Cement. For both self-adhesive cements, half of the specimens (experimental groups) were luted after the application of a solution of 2% CHX, while no CHX application was performed for the remaining specimens (control groups). Luted specimens were cut and used for push-out bond strength evaluation immediately, and after storage in artificial saliva for 6 months or 1 year. Additional specimens were processed for quantitative interfacial nanoleakage analysis. RESULTS: ANOVA showed that the variable times of storage had a significant influence on the results (p < 0.05), while no influence of the luting procedure (cements with or without CHX) on the final outcome (p > 0.05) was found. Tukey's pairwise post-hoc test showed that the radicular bond strength decreased with time of storage. In particular, a significant difference was found between T0 and T1y, but not between T0 and T6m. In contrast, in terms of pretreatment, no significant reduction in push-out bond strength was observed, irrespective of the aging time. CONCLUSION: CHX pretreatment did not prevent bond strength degradation of fiber posts luted with self-adhesive cements.

Dentin Bonding Durability of Two-step Self-etch Adhesives with Improved of Degree of Conversion of Adhesive Resins.

PURPOSE: To evaluate (1) the initial and long-term microtensile bond strengths of two-step self-etch adhesives with different degrees of conversion (DC); (2) the elastic modulus of the respective adhesive resins; (3) the water sorption of the respective adhesive resins. MATERIALS AND METHODS: Two two-step self-etch adhesives, Clearfil SE Bond (CSE) and Clearfil SE Bond 2 (CSE2) were used in this study. The DC was determined using ATR/FT-IR with a time-based spectrum analysis. Midcoronal flat dentin surfaces of 24 human molars were prepared with 600-grit SiC paper for microtensile bond strength (µTBS) testing. CSE and CSE2 were applied to the dentin surfaces according to the manufacturer's instructions, followed by composite buildups. The µTBS was measured after water storage for 24 h, 6 months, and 1 year. The elastic modulus (before and after 1 month of water immersion) was determined by the three-point flexural bending test and water sorption values by the water sorption test. RESULTS: CSE2 showed significantly higher DC than CSE. The µTBS of CSE2 was significantly higher than that of CSE in all water storage periods. One-year water storage decreased the µTBS of CSE; however, it did not decrease that of CSE2. Regarding the polymerized adhesive resins, the elastic modulus of CSE2 was significantly higher than that of CSE before and after water immersion (p < 0.001), and the water sorption of CSE was higher than that of CSE2. CONCLUSIONS: The higher DC of adhesive resins of two-step self-etch adhesives resists water aging and improves the initial bond strengths and durability of the resin-dentin bond.

Antibacterial properties of copper iodide-doped glass ionomer-based materials and effect of copper iodide nanoparticles on collagen degradation.

OBJECTIVES: This study investigated the antibacterial properties and micro-hardness of polyacrylic acid (PAA)-coated copper iodide (CuI) nanoparticles incorporated into glass ionomer-based materials, and the effect of PAA-CuI on collagen degradation. MATERIALS AND METHODS: PAA-CuI nanoparticles were incorporated into glass ionomer (GI), Ionofil Molar AC, and resin-modified glass ionomer (RMGI), Vitrebond, at 0.263 wt%. The antibacterial properties against Streptococcus mutans (n = 6/group) and surface micro-hardness (n = 5/group) were evaluated. Twenty dentin beams were completely demineralized in 10 wt% phosphoric acid and equally divided in two groups (n = 10/group) for incubation in simulated body fluid (SBF) or SBF containing 1 mg/ml PAA-CuI. The amount of dry mass loss and hydroxyproline (HYP) released were quantified. Kruskal-Wallis, Student's t test, two-way ANOVA, and Mann-Whitney were used to analyze the antibacterial, micro-hardness, dry mass, and HYP release data, respectively (p < 0.05). RESULTS: Addition of PAA-CuI nanoparticles into the glass ionomer matrix yielded significant reduction (99.999 %) in the concentration of bacteria relative to the control groups. While micro-hardness values of PAA-CuI-doped GI were no different from its control, PAA-CuI-doped RMGI demonstrated significantly higher values than its control. A significant decrease in dry mass weight was shown only for the control beams (10.53 %, p = 0.04). Significantly less HYP was released from beams incubated in PAA-CuI relative to the control beams (p < 0.001). CONCLUSIONS: PAA-CuI nanoparticles are an effective additive to glass ionomer-based materials as they greatly enhance their antibacterial properties and reduce collagen degradation without an adverse effect on their mechanical properties. CLINICAL RELEVANCE: The use of copper-doped glass ionomer-based materials under composite restorations may contribute to an increased longevity of adhesive restorations, because of their enhanced antibacterial properties and reduced collagen degradation.

NaF Inhibits Matrix-Bound Cathepsin-Mediated Dentin Matrix Degradation.

Matrix metalloproteinases (MMPs) and cysteine cathepsins (CCs) degrade the collagen fibrils of demineralized dentin. Sodium fluoride (NaF) has previously been shown to inhibit recombinant MMP-2 and MMP-9. This study aimed to evaluate the efficacy of NaF on the inhibition of dentin-bound MMPs and CCs. Dentin beams were completely demineralized in 10% phosphoric acid. The baseline total MMP activity and dry masses were measured. Beams were assigned to test groups based on similar MMP activity and dry mass (n = 10/group), and incubated in artificial saliva (control) or artificial saliva with NaF containing 6-238 mM fluoride for 1, 7 and 21 days. The dry mass loss and MMP activities were reassessed at each time point. The proteolytic activity was screened by gelatin zymography. ICTP and CTX released to the incubation medium were analyzed as indices of MMP and cathepsin K activity, respectively. The beams were examined under scanning electron microscopy. All NaF doses reduced the dry mass loss after 21 days (p < 0.05). NaF inhibition of the total MMP activity ranged between 5 and 80%. In gelatin zymography, the bands of MMP-2 and MMP-9 became less prominent with increasing NaF levels. NaF did not decrease the released ICTP (p > 0.05). Less CTX release was detected with F ≥179 mM (p < 0.05). CaF2-like minerals were observed on the beams. High levels of NaF may slow the degradation of the dentin matrix due to the inhibition of cathepsin K. Fluoride does not seem effective in the direct inhibition of proteolysis by dentin matrix-bound MMPs.

Uninfiltrated Collagen in Hybrid Layers produced after Reduced Acid-etching Time on Primary and Permanent Dentin.

AIM: This study evaluated the influence of acid-etching time on collagen exposure in adhesive interfaces established on primary and permanent dentin. MATERIALS AND METHODS: Flat dentin surfaces were produced on sound primary molars and premolars (n = 8). The surfaces were divided into mesial and distal halves, and each half was etched with phosphoric acid for 5 or 15 seconds. The teeth were randomly allocated into two groups according to the adhesive system applied: Prime & Bond NT or Prime & Bond 2.1. After the adhesive application, the specimens were processed for Goldner's trichrome staining. The thickness of the uninfiltrated collagen zone (UCZ) in the hybrid layer was measured under optical microscopy. Data were analyzed by analysis of variance and Tukey tests (α = 0.05). RESULTS: The thickness of UCZ was adhesive dependent. Within the same substrate, the specimens treated with Prime & Bond 2.1 presented thicker UCZ when etched for 15 seconds. Collagen exposure was significantly higher for the primary teeth etched for 5 seconds and treated with Prime & Bond 2.1. CONCLUSION: The thickness of UCZ in hybrid layers is directly affected by acid-etching time and by the adhesive system applied. Primary dentin seems to be more susceptible to collagen exposure than is permanent dentin. CLINICAL SIGNIFICANCE: Both acid-etching time and adhesive system can influence the amount of exposed collagen interfering on resin-dentin bond quality, especially on primary dentin.

Aging of adhesive interfaces treated with benzalkonium chloride and benzalkonium methacrylate.

Inhibition of endogenous dentin matrix metalloproteinases (MMPs) within incompletely infiltrated hybrid layers can contribute to the preservation of resin-dentin bonds. This study evaluated the bond stability of interfaces treated with benzalkonium chloride (BAC) and benzalkonium methacrylate (MBAC), and the inhibitory properties of these compounds on dentin MMP activity. Single-component adhesive ALL-BOND UNIVERSAL, modified with BAC or MBAC at concentrations of 0, 0.5, 1.0, and 2.0%, was used for microtensile bond strength (µTBS) evaluation after 24 h, 6 months, and 1 yr. Beams produced from human dentin were treated with 37% phosphoric acid, dipped in 0.5% BAC, 1.0% BAC, or water (control) for 60 s, and then incubated in SensoLyte generic MMP substrate to determine MMP activity. A significant decrease in the µTBS after 6 months and 1 yr was observed for the control group only. No significant differences among groups were shown at 24 h. After 6 months and 1 yr, the control group demonstrated significantly lower µTBS than all treatment groups. When applied for 60 s, 0.5% BAC inhibited total MMP activity by 31%, and 1.0% BAC inhibited total MMP activity by 54%. Both BAC and MBAC contributed to the preservation of resin-dentin bonds, probably because of their inhibitory properties of endogenous dentin proteinases.

Preservation of resin-dentin interfaces treated with benzalkonium chloride adhesive blends.

Reducing collagen degradation within hybrid layers may contribute to the preservation of adhesive interfaces. This study evaluated the stability of resin-dentin interfaces treated with benzalkonium chloride (BAC)-modified adhesive blends and assessed collagen degradation in dentin matrices treated with BAC. The etch-and-rinse adhesive, Adper Single Bond Plus, modified with 0.5% and 1.0% BAC, was evaluated for microtensile bond strength (µTBS) and nanoleakage (NL) after 24 h and 1 yr. Thirty completely demineralized dentin beams from human molars were dipped for 60 s in deionized water (DW; control), or in 0.5% or 1.0% BAC, and then incubated in simulated body fluid (SBF). Collagen degradation was assessed by quantification of the dry mass loss and the amount of hydroxyproline (HYP) released from hydrolyzed specimens after 1 or 4 wk. Although all groups demonstrated a significant increase in NL after 1 yr, adhesive modified with 0.5% BAC showed stable bond strength after 1 yr (9% decrease) relative to the control (44% decrease). Significantly less HYP release and dry mass loss were observed for both 0.5% and 1.0% BAC relative to the control. This in vitro study demonstrates that BAC contributes to the preservation of resin-dentin bonds for up to 1 yr by reducing collagen degradation.

Adhesive sealing of dentin surfaces in vitro: A review.

PURPOSE: This review describes the evolution of the use of dental adhesives to form a tight seal of freshly prepared dentin to protect the pulp from bacterial products, during the time between crown preparation and final cementation of full crowns. The evolution of these "immediate dentin sealants" follows the evolution of dental adhesives, in general. That is, they began with multiple-step, etch-and-rinse adhesives, and then switched to the use of simplified adhesives. METHODS: Literature was reviewed for evidence that bacteria or bacterial products diffusing across dentin can irritate pulpal tissues before and after smear layer removal. Smear layers can be solubilized by plaque organisms within 7-10 days if they are directly exposed to oral fluids. It is likely that smear layers covered by temporary restorations may last more than 1 month. As long as smear layers remain in place, they can partially seal dentin. Thus, many in vitro studies evaluating the sealing ability of adhesive resins use smear layer-covered dentin as a reference condition. Surprisingly, many adhesives do not seal dentin as well as do smear layers. RESULTS: Both in vitro and in vivo studies show that resin- covered dentin allows dentin fluid to cross polymerized resins. The use of simplified single bottle adhesives to seal dentin was a step backwards. Currently, most authorities use either 3-step adhesives such as Scotchbond Multi-Purpose or OptiBond FL or two-step self-etching primer adhesives, such as Clearfil SE, Unifil Bond or AdheSE.

Adopting the principles of collagen biomineralization for intrafibrillar infiltration of yttria-stabilized zirconia into three-dimensional collagen scaffolds.

In this paper, we report a process for generating collagen-yttria-stabilized amorphous zirconia hybrid scaffolds by introducing acetylacetone-inhibited zirconia precursor nanodroplets into a poly(allylamine)-coated collagen matrix. This polyelectrolyte coating triggers intrafibrillar condensation of the precursors into amorphous zirconia, which is subsequently transformed into tetragonal yttria-stabilized zirconia after calcination. Our findings represent a new paradigm in the synthesis of non-naturally occurring collagen-based hybrid scaffolds under alcoholic mineralizing conditions.

Ethanol-wet bonding and chlorhexidine improve resin-dentin bond durability: quantitative analysis using raman spectroscopy.

PURPOSE: To directly test the effectiveness of ethanol-wet bonding (EW) in improving monomer infiltration into demineralized dentin through quantitative measurement of bis-GMA and TEG-DMA molar concentrations within hybrid layers, and to comprehensively evaluate the effect of EW and chlorhexidine on durability of resin-dentin bonds compared to conventional water-wet bonding (WW). MATERIALS AND METHODS: A three-step etch-and-rinse adhesive (70% bis-GMA/28.75%TEG-DMA) was applied to coronal dentin using a clinically relevant ethanol-wet bonding protocol (EW) or the conventional water-wet bonding (WW) technique. Bis-GMA and TEG-DMA molar concentrations at various positions across the resin/dentin interfaces formed by EW and WW were measured using micro-Raman spectroscopy. The experiment was repeated at the same positions after 7-month storage in phosphate buffer solution containing 0.1% sodium azide. The µTBS and hybrid layer morphology (TEM) of bonding groups with and without chlorhexidine application were compared immediately and after 1-year storage in terms of nanoleakage, collagen fibril diameter, collagen interfibrillar width, and hybrid layer thickness. RESULTS: Specimens bonded with EW showed significantly higher monomer molar concentrations and µTBS throughout the hybrid layer immediately and after storage, providing direct evidence of superior infiltration of hydrophobic monomers in EW compared to WW. Microscopically, EW maintained interfibrillar width and hybrid layer thickness for resin infiltration and retention. The application of chlorhexidine further preserved collagen integrity and limited the degree of nanoleakage in EW after 1-year storage. CONCLUSION: EW enhances infiltration of hydrophobic monomers into demineralized dentin. The results suggest that a more durable resin-dentin bond may be achieved with combined usage of a clinically relevant EW and chlorhexidine.

Mechanisms regulating the degradation of dentin matrices by endogenous dentin proteases and their role in dental adhesion. A review.

PURPOSE: This systematic review provides an overview of the different mechanisms proposed to regulate the degradation of dentin matrices by host-derived dentin proteases, particularly as it relates to their role in dental adhesion. Significant developments have taken place over the last few years that have contributed to a better understanding of all the factors affecting the durability of adhesive resin restorations. The complexity of dentin-resin interfaces mandates a thorough understanding of all the mechanical, physical and biochemical aspects that play a role in the formation of hybrid layers. The ionic and hydrophilic nature of current dental adhesives yields permeable, unstable hybrid layers susceptible to water sorption, hydrolytic degradation and resin leaching. The hydrolytic activity of host-derived proteases also contributes to the degradation of the resin-dentin bonds. Preservation of the collagen matrix is critical to the improvement of resin-dentin bond durability. Approaches to regulate collagenolytic activity of dentin proteases have been the subject of extensive research in the last few years. A shift has occurred from the use of proteases inhibitors to the use of collagen cross-linking agents. Data provided by 51 studies published in peer-reviewed journals between January 1999 and December 2013 were compiled in this systematic review. RESULTS: Appraisal of the data provided by the studies included in the present review yielded a summary of the mechanisms which have already proven to be clinically successful and those which need further investigation before new clinical protocols can be adopted.

Intrafibrillar silicification of collagen scaffolds for sustained release of stem cell homing chemokine in hard tissue regeneration.

Traditional bone regeneration strategies relied on supplementation of biomaterials constructs with stem or progenitor cells or growth factors. By contrast, cell homing strategies employ chemokines to mobilize stem or progenitor cells from host bone marrow and tissue niches to injured sites. Although silica-based biomaterials exhibit osteogenic and angiogenic potentials, they lack cell homing capability. Stromal cell-derived factor-1 (SDF-1) plays a pivotal role in mobilization and homing of stem cells to injured tissues. In this work, we demonstrated that 3-dimensional collagen scaffolds infiltrated with intrafibrillar silica are biodegradable and highly biocompatible. They exhibit improved compressive stress-strain responses and toughness over nonsilicified collagen scaffolds. They are osteoconductive and up-regulate expressions of osteogenesis- and angiogenesis-related genes more significantly than nonsilicified collagen scaffolds. In addition, these scaffolds reversibly bind SDF-1α for sustained release of this chemokine, which exhibits in vitro cell homing characteristics. When implanted subcutaneously in an in vivo mouse model, SDF-1α-loaded silicified collagen scaffolds stimulate the formation of ectopic bone and blood capillaries within the scaffold and abrogate the need for cell seeding or supplementation of osteogenic and angiogenic growth factors. Intrafibrillar-silicified collagen scaffolds with sustained SDF-1α release represent a less costly and complex alternative to contemporary cell seeding approaches and provide new therapeutic options for in situ hard tissue regeneration.

Biomimetic silicification of demineralized hierarchical collagenous tissues.

Unlike man-made composite materials, natural biominerals containing composites usually demonstrate different levels of sophisticated hierarchical structures which are responsible for their mechanical properties and other metabolic functions. However, the complex spatial organizations of the organic-inorganic phases are far beyond what they achieved by contemporary engineering techniques. Here, we demonstrate that carbonated apatite present in collagen matrices derived from fish scale and bovine bone may be replaced by amorphous silica, using an approach that simulates what is utilized by phylogenetically ancient glass sponges. The structural hierarchy of these collagen-based biomaterials is replicated by the infiltration and condensation of fluidic polymer-stabilized silicic acid precursors within the intrafibrillar milieu of type I collagen fibrils. This facile biomimetic silicification strategy may be used for fabricating silica-based, three-dimensional functional materials with specific morphological and hierarchical requirements.

Influence of MDPB-containing primer on Streptococcus mutans biofilm formation in simulated Class I restorations.

PURPOSE: To evaluate the activity of a methacryloyloxydodecylpyridinium bromide (MDPB)-containing self-etching primer (Clearfil Protect Bond) against Streptococcus mutans and its ability to reduce biofilm formation on standardized experimental Class I restorations in vitro. MATERIALS AND METHODS: Forty experimental Class I round restorations were prepared on enamel-dentin slabs using different adhesive strategies: group 1 = MDPB-containing adhesive system (Clearfil Protect Bond); group 2 = MDPB-free self-etching adhesive system (Clearfil SE Bond); group 3: MDPB-containing self-etching primer in combination with a fluoride-free bonding agent; group 4: MDPB-free self-etching primer in combination with a fluoride-containing bonding agent; group 5: a three-step etch-and-rinse adhesive system (Adper Scotchbond Multi Purpose). A Streptococcus mutans biofilm was grown for 48 h on the restoration surfaces and subsequently evaluated using scanning electron microscopy on three different areas: enamel, composite, and interface surfaces. Statistical analysis was performed by multiple ANOVA after data transformation. RESULTS: Specimens in groups 2, 4 and 5 showed greater biofilm formation than those in groups 1 and 3 (p < 0.001) on all investigated substrates (enamel, composite, and interface areas). CONCLUSIONS: Specimens prepared with an MDPB-containing primer exhibited significant decreases in biofilm formation on Class I restorations in vitro. Further in vitro and in vivo studies are required to clarify the role of quaternary ammonium compounds in reducing bacterial biofilm formation on restoration surfaces.