Adsorption of Polystyrene from Theta Condition on Cellulose and Silica Studied by Quartz Crystal Microbalance.

Adsorption of hydrophobic polymers from a nonpolar solvent medium is an underutilized tool for modification of surfaces, especially of soft matter. Adsorption of polystyrene (PS) from a theta solvent (50/50 vol % toluene/heptane) on ultrathin model films of cellulose was studied with a quartz crystal microbalance with dissipation monitoring (QCM-D), using three different PS grades with monodisperse molecular weights (Mws). Comparison of cellulose to silica as an adsorbent was presented. Adsorption on both surfaces was mainly irreversible under the studied conditions. Characteristically to polymer monolayer formation, the mass of the adsorbing polymer increased with its Mw. The initial step of the layer formation was similar on both surfaces, but silica showed a stronger tendency for the formation of a loosely bound overlayer upon molecular rearrangements as the adsorption process proceeded. Despite the slightly less extended layers formed on cellulose at increasing Mw values, the overall thickness of the adsorbing wet layers on both surfaces was of the similar order of magnitude as the radius of gyration of the adsorbate molecule. Decent degree of hydrophobization of cellulose could be reached with all studied PS grades when the time allowed for adsorption was sufficient. QCM-D, a method conventionally utilized for studying aqueous systems, turned out to be a suitable tool for studying the adsorption process of hydrophobic polymers on soft polymeric matter such as cellulose taking place in a nonpolar solvent environment.

Assessment of the Alga Cladophora glomerata as a Source for Cellulose Nanocrystals.

Nanocellulose is isolated from cellulosic fibers and exhibits many properties that macroscale cellulose lacks. Cellulose nanocrystals (CNCs) are a subcategory of nanocellulose made of stiff, rodlike, and highly crystalline nanoparticles. Algae of the order Cladophorales are the source of the longest cellulosic nanocrystals, but manufacturing these CNCs is not well-studied. So far, most publications have focused on the applications of this material, with the basic manufacturing parameters and material properties receiving little attention. In this article, we investigate the entirety of the current manufacturing process from raw algal biomass (Cladophora glomerata) to the isolation of algal cellulose nanocrystals. Yields and cellulose purities are investigated for algal cellulose and the relevant process intermediates. Furthermore, the effect of sulfuric acid hydrolysis, which is used to convert cellulose into CNCs and ultimately determines the material properties and some of the sustainability aspects, is examined and compared to literature results on wood cellulose nanocrystals. Long (>4 µm) CNCs form a small fraction of the overall number of CNCs but are still present in measurable amounts. The results define essential material properties for algal CNCs, simplifying their future use in functional cellulosic materials.

Understanding Nanocellulose-Water Interactions: Turning a Detriment into an Asset.

Modern technology has enabled the isolation of nanocellulose from plant-based fibers, and the current trend focuses on utilizing nanocellulose in a broad range of sustainable materials applications. Water is generally seen as a detrimental component when in contact with nanocellulose-based materials, just like it is harmful for traditional cellulosic materials such as paper or cardboard. However, water is an integral component in plants, and many applications of nanocellulose already accept the presence of water or make use of it. This review gives a comprehensive account of nanocellulose-water interactions and their repercussions in all key areas of contemporary research: fundamental physical chemistry, chemical modification of nanocellulose, materials applications, and analytical methods to map the water interactions and the effect of water on a nanocellulose matrix.

Chitin-Active Lytic Polysaccharide Monooxygenases Are Rare in Cellulomonas Species.

Cellulomonas flavigena is a saprotrophic bacterium that encodes, within its genome, four predicted lytic polysaccharide monooxygenases (LPMOs) from Auxiliary Activity family 10 (AA10). We showed previously that three of these cleave the plant polysaccharide cellulose by oxidation at carbon-1 (J. Li, L. Solhi, E.D. Goddard-Borger, Y. Mattieu et al., Biotechnol Biofuels 14:29, 2021, https://doi.org/10.1186/s13068-020-01860-3). Here, we present the biochemical characterization of the fourth C. flavigena AA10 member (CflaLPMO10D) as a chitin-active LPMO. Both the full-length CflaLPMO10D-Carbohydrate-Binding Module family 2 (CBM2) and catalytic module-only proteins were produced in Escherichia coli using the native general secretory (Sec) signal peptide. To quantify chitinolytic activity, we developed a high-performance anion-exchange chromatography-pulsed amperometric detection (HPAEC-PAD) method as an alternative to the established hydrophilic interaction liquid ion chromatography coupled with UV detection (HILIC-UV) method for separation and detection of released oxidized chito-oligosaccharides. Using this method, we demonstrated that CflaLPMO10D is strictly active on the ß-allomorph of chitin, with optimal activity at pH 5 to 6 and a preference for ascorbic acid as the reducing agent. We also demonstrated the importance of the CBM2 member for both mediating enzyme localization to substrates and prolonging LPMO activity. Together with previous work, the present study defines the distinct substrate specificities of the suite of C. flavigena AA10 members. Notably, a cross-genome survey of AA10 members indicated that chitinolytic LPMOs are, in fact, rare among Cellulomonas bacteria. IMPORTANCE Species from the genus Cellulomonas have a long history of study due to their roles in biomass recycling in nature and corresponding potential as sources of enzymes for biotechnological applications. Although Cellulomonas species are more commonly associated with the cleavage and utilization of plant cell wall polysaccharides, here, we show that C. flavigena produces a unique lytic polysaccharide monooxygenase with activity on ß-chitin, which is found, for example, in arthropods. The limited distribution of orthologous chitinolytic LPMOs suggests adaptation of individual cellulomonads to specific nutrient niches present in soil ecosystems. This research provides new insight into the biochemical specificity of LPMOs in Cellulomonas species and related bacteria, and it raises new questions about the physiological function of these enzymes.

Controlled sulfation of mixed-linkage glucan by Response Surface Methodology for the development of biologically applicable polysaccharides.

Endogenous and exogenous sulfated polysaccharides exhibit potent biological activities, including inhibiting blood coagulation and protein interactions. Controlled chemical sulfation of alternative polysaccharides holds promise to overcome limited availability and heterogeneity of naturally sulfated polysaccharides. Here, we established reaction parameters for the controlled sulfation of the abundant cereal polysaccharide, mixed-linkage ß(1,3)/ß(1,4)-glucan (MLG), using Box-Behnken Design of Experiments (BBD) and Response Surface Methodology (RSM). The optimization of the degree-of-substitution (DS) was externally validated through the production of sulfated MLGs (S-MLGs) with observed DS and Mw values deviating less than 20% and 30% from the targeted values, respectively. Simultaneous optimization of DS and Mw resulted in the same range of deviation from the targeted value. S-MLGs with DS > 1 demonstrated a modest anticoagulation effect versus heparin, and a greater P-selectin affinity than fucoidan. As such, this work provides a route to medically important polymers from an economical agricultural polysaccharide.

On the properties of nanosilicate-based filled dental adhesives: Synthesis, characterization, and optimized formulation.

OBJECTIVE: In this study, we incorporated hybrid nanoparticles (poly (acrylic acid)-grafted nanoclay/nanosilica, respectively, with platelet and spherical morphologies, abbreviated as PAA-g-NC-Sil) in different concentrations (0, 0.2, 0.5, 1, 2 and 5 wt%) to an experimental dentin bonding system and investigated the physical properties of the filled adhesive and its shear bond strength (µ-SBS) to dentin. We subsequently compared the properties of the adhesives containing PAA-g-NC-Sil with previously studied adhesives containing poly (methacrylic acid)-g-nanoclay (PMA-g-NC) (Solhi et al., 2012a), poly (acrylic acid)-g-nanoclay (PAA-g-NC) (Solhi et al., 2012b), and the hybrid poly (methacrylic acid)-grafted-nanoclay-nanosilica (PMA-g-NC-Sil) (Solhi et al., 2020). MATERIALS AND METHODS: In a set of previous publications and the present paper, we grafted poly (acrylic acid) (PAA) or poly (methacrylic acid) (PMA) onto the surface of pristine Na-MMT nanoclay (Cloisite® Na+) through free radical polymerization of monomer in an aqueous media in the presence or absence of nanosilica particles. We characterized the resulting modified nanoparticles (PMA-g-NC, PAA-g-NC, PMA-g-NC-Sil and PAA-g-NC-Sil) using GPC, FTIR, TGA, and XRD. We then incorporated the modified particles as functionalized fillers to experimental dentin adhesives in different concentrations and studied the stability of modified fillers dispersion by separation analysis. We also studied the properties of the photo-cured adhesive matrices using FTIR, TEM, SEM, EDXA, and XRD. We examined the shear bond strength of the adhesives (containing different contents of each modified filler, separately) to human premolar teeth. The results were analysed and compared statistically. RESULTS: The results confirmed that the polymers have been grafted onto the surface of nanoclay. An exfoliated structure for the nanoclay platelets in the photo-cured adhesive containing PAA-g-NC-Sil was observed. Addition of 0.5 wt% of PAA-g-NC-Sil to the experimental adhesive increased the shear bond strength and the dispersion stability in comparison to unfilled adhesive. The same trend was also observed for adhesives containing PMA-g-NC, PAA-g-NC, and PMA-g-NC-Sil. The adhesive containing PAA-g-NC-Sil showed the best dispersion stability and subsequently the highest shear bond strength in the optimal concentration among adhesives containing the four available fillers (PMA-g-NC, PAA-g-NC, PMA-g-NC-Sil and PAA-g-NC-Sil). SIGNIFICANCE: Addition of poly (acrylic acid) modified nanoparticles to the experimental dentin adhesives resulted in higher shear bond strength due to the potential interactions between the carboxylic acid functional groups on the surface of the modified particles and the dentin structure. Between the poly (acrylic acid) and poly (methacrylic acid), the former acid with higher PKa performed better. Addition of the spherical nanosilica particles to the adhesives containing platelet nanoclay helped to better exfoliate the platelets resulting in improved µ-SBS and dispersion stability.

Four cellulose-active lytic polysaccharide monooxygenases from Cellulomonas species.

BACKGROUND: The discovery of lytic polysaccharide monooxygenases (LPMOs) has fundamentally changed our understanding of microbial lignocellulose degradation. Cellulomonas bacteria have a rich history of study due to their ability to degrade recalcitrant cellulose, yet little is known about the predicted LPMOs that they encode from Auxiliary Activity Family 10 (AA10). RESULTS: Here, we present the comprehensive biochemical characterization of three AA10 LPMOs from Cellulomonas flavigena (CflaLPMO10A, CflaLPMO10B, and CflaLPMO10C) and one LPMO from Cellulomonas fimi (CfiLPMO10). We demonstrate that these four enzymes oxidize insoluble cellulose with C1 regioselectivity and show a preference for substrates with high surface area. In addition, CflaLPMO10B, CflaLPMO10C, and CfiLPMO10 exhibit limited capacity to perform mixed C1/C4 regioselective oxidative cleavage. Thermostability analysis indicates that these LPMOs can refold spontaneously following denaturation dependent on the presence of copper coordination. Scanning and transmission electron microscopy revealed substrate-specific surface and structural morphological changes following LPMO action on Avicel and phosphoric acid-swollen cellulose (PASC). Further, we demonstrate that the LPMOs encoded by Cellulomonas flavigena exhibit synergy in cellulose degradation, which is due in part to decreased autoinactivation. CONCLUSIONS: Together, these results advance understanding of the cellulose utilization machinery of historically important Cellulomonas species beyond hydrolytic enzymes to include lytic cleavage. This work also contributes to the broader mapping of enzyme activity in Auxiliary Activity Family 10 and provides new biocatalysts for potential applications in biomass modification.

Dual modified nanosilica particles as reinforcing fillers for dental adhesives: Synthesis, characterization, and properties.

A facile procedure has been devised to develop a novel dentin bonding system containing poly (acrylic acid)-grafted-silanized fumed silica particles as reinforcing filler, with high stability of nanoparticle dispersion and enhanced bond strength and mechanical properties. In the first step, the silanization of fumed silica nanoparticles was performed in the following conditions: (i) ethanol-water solution with a pH of 5 and (ii) cyclohexane with a pH of 9 using trimethoxysilylpropyl methacrylate (γ-MPS) as a reactive silane coupling agent. FTIR and TGA analyses confirmed the presence of silane in the resultant structure and enhanced dispersion stability of modified particles was proved by a separation analyzer and also zeta potential analyses. In the second step, free radical polymerization of acrylic acid monomers in the presence of silanized nanoparticles was carried out and poly (acrylic acid) -grafted- silanized fumed silica were acquired. The flexural strength and fracture toughness of the adhesive containing 0.2 wt.% of the dual modified filler reached maximum of 70.4 MPa and 1.34 MPa m1/2, respectively, showing average improvements of 74% and 179%, respectively, in comparison with the adhesive without filler. Flexural modulus values did not significantly change with increasing the filler content except the adhesive containing 5 wt.% having the lowest flexural modulus. The highest microtensile bond strength was also observed at 0.2 wt.% filler content showing the average improvements of 197% as compared with the neat adhesive. Energy dispersive X-ray (EDX) mapping confirmed a homogenous and uniform distribution of the fillers in the adhesive matrix containing 0.2 wt.% and 0.5 wt.% of filler while incorporation of 5 wt.% led to large particle aggregates. SEM images of the fracture surface of the adhesive with different filler contents subjected to fracture toughness test showed rougher surface and longer crack path by increasing filler concentration. The adhesive containing 0.2 wt.% of filler perfectly penetrated into the dentin tubules proved by the SEM micrographs in microtensile bond strength test.

Poly (methacrylic acid) modified spherical and platelet hybrid nanoparticles as reinforcing fillers for dentin bonding systems: Synthesis and properties.

OBJECTIVE: In this study the mechanical and adhesion properties of an experimental methacrylate based dentin bonding system containing a combination of spherical and layered platelet nanoparticles were investigated. The nanoparticles were first modified through surface graft polymerization of methacrylic acid in order to make the particles surface compatible with the bonding matrix resin. MATERIALS AND METHODS: Graft free radical polymerization in aqueous media was performed to attach Poly (methacrylic acid) (PMA) chains onto the surface of Na-MMT nanoclay (Cloisite® Na+) and silica nanoparticles (Aerosil® 200). The hybrid PMA grafted nanoparticles (PMA-g-NC-Sil) were characterized using GPC, FTIR, TGA, and X-ray diffraction (XRD). Dentin adhesives containing different amounts of the hybrid modified nanoparticles were photopolymerized and their characteristics were studied using FTIR, TEM, SEM, EDXA, and XRD techniques. The adhesives containing different amounts of PMA-g-NC-Sil were applied to the conditioned human premolar dentin to bond a dental composite to the teeth. The bond strength was then measured by microshear bond strength testing method. The results were analyzed and compared statistically. The stability of PMA-g-NC-Sil dispersion in the dentin adhesive was investigated using separation analysis (LUMi Reader) techniques. RESULTS: The grafting of PMA chains onto the surface of nanoclay was confirmed by FTIR and TGA analytical techniques. The intercalated-exfoliated structure for the nanoclay platelets in the photo-cured adhesive was observed using XRD and TEM. The surface modification of the nanoparticles significantly increased the dispersion stability of the fillers in the adhesive solution. The microshear test results indicated that the incorporation of the PMA-g-NC-Sil nanoparticles significantly enhanced the bond strength to dentin with the highest shear bond strength observed at 0.5 wt%. SIGNIFICANCE: The incorporation of the PMA modified hybrid nanofillers into the dentin adhesive resulted in a dentin bonding agent with enhanced shear bond strength through reinforcing the adhesive matrix and potential interactions between their carboxylic acid groups and the tooth structure. The dispersion stability of the nanoparticles was also dramatically improved by the surface modification of the nanoparticles.

Structure-properties relationships in dental adhesives: Effect of initiator, matrix monomer structure, and nano-filler incorporation.

OBJECTIVES: This is a confirmatory study to evaluate the effect of photoinitiator type and concentration, matrix monomer chemical structure, and nanoparticle incorporation on the physical and mechanical properties of an experimental dentin bonding agent. MATERIALS AND METHODS: Different concentrations of camphorquinone-amine (CQ-A) system, butanedione (BD), and phenylpropanedione (PPD), as photoinitiator, BTDMA, as a comonomer containing carboxylic acid groups, and silica nanoparticles as reinforcing inorganic filler were incorporated into a methacrylate base experimental dental adhesive. The effect of these ingredients, as independent variables, on the shrinkage kinetics, flexural strength and modulus, and microshear bond strength of the adhesives were then investigated. The results were analyzed using one-way ANOVA and Tukey's post-hoc test at the significance level of 0.05. RESULTS: The results indicate that the efficiency of CQ-A initiator system is diminished in the presence of the acidic monomer BTDMA while the photopolymerization is efficiently progressed with BD as initiator. PPD shows the lowest efficiency in the photopolymerization of the adhesives. BTDMA as a monomer with the capability of interaction with tooth structure provides adhesive with improved microshear bond strength to dentin. Incorporation of silica nanoparticles at low concentrations enhances the flexural and microshear strength of the dentin bonding agent. SIGNIFICANCE: Understanding the structure-property relationship in dental adhesives may help the material selection in clinical dentistry. The study elucidates the relationship between monomer structure, initiator type, and nanofiller and physical and mechanical properties in dental adhesives.

Physical characterization of unfilled and nanofilled dental resins: Static versus dynamic mechanical properties.

OBJECTIVE: The aim of this study was to evaluate the mechanical properties of dental resins and dental nanocomposites by means of 3-point bending, 4-point bending and piston-on-three ball biaxial static tests and also to investigate their dynamic mechanical properties. The obtained results from the static 3-point bending test also were compared with the corresponding mode in DMTA. METHODS: The hydrophilic surface of the inorganic OX-50 particles was treated with γ-MPS and the resulting silanized OX-50 was characterized using FTIR and TGA. The test specimens with a specified shape and dimensions for each type of flexural static and dynamic tests were prepared for Bis-GMA/TEGDMA (70/30wt%/wt%) and the corresponding nanocomposite containing 50wt% of silanized OX-50. The specimens were stored in distilled water at 37°C for 24h prior to the mechanical tests and then they were subjected to different types of static mode and also to dynamic mode of flexural test. The effect of test speed, type of the test and the presence of silanized nano-particles were investigated for specimens under static flexural tests. The results were then statistically analyzed and compared using one-way ANOVA and the Tukey's post hoc test (significance level=0.05). Fractured surfaces were observed by means of scanning electron microscopy. Finite element analyses were also performed to compare the static tests. On the other hand, the effect of frequency, temperature and the presence of silanized nano particles on the viscoelastic properties were investigated for dynamic mechanical tests. RESULTS: The grafting of γ-MPS onto the OX-50 nano particles was confirmed. The results of 3-point bending and 4-point bending uniaxial and biaxial flexural tests of resin and composite showed the highest strength values for biaxial test specimens and the lowest strength value for 4-point bending specimens in accordance with finite element analysis results. Also, an increasing trend was observed for flexural strength of all resin samples with increasing the test speed up a critical speed which decreased beyond it. The nanocomposite specimens showed higher modulus and lower tanδ than the unfilled resin. The storage modulus measured in the dynamic tests approached the static elastic modulus values with decreasing the frequency. SIGNIFICANCE: Different static and dynamic test methods are used to evaluate the mechanical properties of dental resins and composites. This study provides an insight into the tests and presents more details on the effect of test conditions.

Synthesis and in vitro characterization of a preactivated thiomer via polymerization reaction.

The objective of this study was to synthesize 6-(2-acryloylamino-ethyldisulfanyl)-nicotinic acid (ACENA) for subsequent copolymerization with acrylic acid (AA) as a new method for synthesis of preactivated thiomers. Copolymerization reactions of ACENA and AA with different molar ratios were performed and the molecular weight (M(w)) values of the resulting copolymers were calculated and reported from 3046 to 3271 Da. The disulfide bond content values in the polymer chain were determined from 400 to 544 µmol disulfide bond per gram polymer. The transport enhancement ratio for 0.5% (m/v) solution of poly(acrylic acid) (PAA) was 1.1 using sodium fluorescein (Na-Flu) as model drug, in Ussing-type chambers, whereas it was over 1.9 for 0.5% (m/v) solution of ACENA and AA copolymers. Resazurin cell-viability test showed no significant toxicity for the polymers. Copolymerization of AA and disulfide-bond-containing monomers can open new horizons for the preparation of preactivated thiomers taking the better controllability and the huge variety of available monomers and combinations thereof into account.

A novel dentin bonding system containing poly(methacrylic acid) grafted nanoclay: synthesis, characterization and properties.

OBJECTIVES: Developing a novel dentin bonding system containing poly(methacrylic acid)-grafted-nanoclay (PMAA-g-nanoclay) as reinforcing filler, with high stability of nanoparticle dispersion and improved bond strength and mechanical properties were the main objectives of this study. MATERIALS AND METHODS: Poly(methacrylic acid) (PMAA) was grafted onto the pristine sodium montmorrillonite (Na-MMT) nanoclay surface and characterized using FTIR, TGA, and X-ray diffraction (XRD). The PMAA-g-nanoclay was incorporated into an experimental dentin bonding system as filler in different concentrations and stability of nanoclay dispersion in the dilute adhesive, morphology of nanoclay layers in the photocured adhesive matrix, shear bond strength to caries-free extracted human premolar teeth, and mode of failure were studied. The mechanical properties including diametral tensile strength (DTS), flexural strength (FS), and flexural modulus (FM) were also investigated. The measured FM was also compared to theoretical prediction models. RESULTS: The grafting of PMAA onto the nanoclay surface was confirmed and the results revealed a partially exfoliated structure for PMAA-g-nanoclay. The dispersion stability of the modified nanoparticles in the dilute adhesive increased more than 45 times in comparison with the pristine nanoclay. The incorporation of 0.5wt.% PMAA-g-nanoclay to the adhesive resulted in a significant increase in microshear bond strength, DTS, and FS. Higher PMAA-g-nanoclay contents resulted in increased flexural modulus. The experimental flexural modulus was in good agreement with the Halpin-Tsai theoretical model. SIGNIFICANCE: Incorporation of PMAA-g-nanoclay particles as novel functional fillers into dental adhesive could result in the development of bonding systems with improved physical, mechanical, and adhesion properties.

Poly(acrylic acid) grafted montmorillonite as novel fillers for dental adhesives: synthesis, characterization and properties of the adhesive.

OBJECTIVE: This work investigates the graft polymerization of acrylic acid onto nanoclay platelets to be utilized as reinforcing fillers in an experimental dental adhesive. Physical and mechanical properties of the adhesive and its shear bond strength to dentin are studied. The effect of the modification on the stability of the nanoparticle dispersion in the dilute adhesive is also investigated. MATERIALS AND METHODS: Poly(acrylic acid) (PAA) was grafted onto the pristine Na-MMT nanoclay (Cloisite(®) Na(+)) through the free radical polymerization of acylic acid in an aqueous media. The resulting PAA-g-nanoclay was characterized using FTIR, TGA and X-ray diffraction (XRD). The modified nanoclays were added to an experimental dental adhesive in different concentrations and the morphology of the nanoclay layers in the photocured adhesive matrix was studied using TEM and XRD. Shear bond strength of the adhesives containing different filler contents was tested on the human premolar teeth. The stability of nanoclay dispersion in the dilute adhesive was also studied using a separation analyzer. The results were then statistically analyzed and compared. RESULTS: The results confirmed the grafting reaction and revealed a partially exfoliated structure for the PAA-g-nanoclay. Incorporation of 0.2 wt.% of the modified nanoclay into the experimental adhesive provided higher shear bond strength. The dispersion stability of the modified nanoparticles in the dilute adhesive was also enhanced more than 25 times. SIGNIFICANCE: Incorporation of the modified particles as reinforcing fillers into the adhesive resulted in higher mechanical properties. The nanofiller containing bonding agent also showed higher shear bond strength due to the probable interaction of the carboxylic acid functional groups on the surface of the modified particles with hydroxyapatite of dentin.

PMMA-grafted nanoclay as novel filler for dental adhesives.

OBJECTIVE: The aim of this study was to investigate the benefits of incorporation of poly(methyl methacrylate)-grafted-nanoclay on the bond strength of an experimental one-bottle dentin bonding system. The effect of the modification on the stability of the nanoparticle dispersion in the dilute adhesive was also studied. MATERIALS AND METHODS: Poly(methyl methacrylate) was grafted onto the pristine Na-MMT nanoclay (Cloisite Na(+)) through the free radical polymerization of methyl methacrylate in an aqueous media in the presence of ammonium persulfate as initiator. A reactive surfactant (AMPS) was also used in the reaction recipe to provide active sites on the surface of the nanoclay. The grafting polymerization reaction was carried out at 70 degrees C. The PMMA-g-nanoclay was then coagulated in methanol and filtered. The resulting PMMA-g-nanoclay was characterized using FTIR, TGA, X-ray diffraction (XRD) and particle size distribution analysis. The modified nanoclay was added to an experimental dentin bonding system as filler and the morphology of the nanoclay layers in the adhesive matrix was studied using TEM and XRD. Shear bond strength of the adhesives containing different filler contents was tested on the caries-free extracted human premolar teeth. The mode of failure was studied by scanning electron microscopy. The stability of the nanoclay dispersion in the dilute adhesive was also studied using a separation analyzer. The results were then statistically analyzed and compared. RESULTS: The grafting of poly(methylmethacrylate) onto the nanoclay was confirmed and the results revealed a partially exfoliated structure for the PMMA-g-nanoclay. Incorporation of the modified nanoclay provided a dentin bonding system with higher shear bond strength. The dispersion stability of the modified nanoparticles in the dilute adhesive was also increased more than 40 times in comparison with the pristine nanoclay. SIGNIFICANCE: The grafting modification provided nanoclay particles with higher dispersion stability than pristine Na-MMT nanoclay in a dilute dentin bonding system. Incorporation of the modified nanoclay into the bonding system provided higher shear bond strength. The finding would be beneficial in producing nano-filler containing adhesive systems.