An osteoinductive surface by adhesive bone morphogenetic protein-2 prepared using the bioorthogonal approach for tight binding of titanium with bone.

Inorganic biomaterials are used in various orthopedic and dental implants. Nevertheless, they cause clinical issues such as loosening of implants and patient morbidity. Therefore, inspired by mussel adhesive proteins, we aimed to design an adhesive and dimer-forming highly active bone morphogenetic protein-2 (BMP-2) using bioorthogonal chemistry, in which recombinant DNA technology was combined with enzymatic modifications, to achieve long-term osseointegration with titanium. The prepared BMP-2 exhibited substantially higher binding activity than wild-type BMP-2, while the adhered BMP-2 was more active than soluble BMP-2. Therefore, the adhesive BMP-2 was immobilized onto titanium wires and screws and implanted into rat bones, and long-term osteogenesis was evaluated. Adhesive BMP-2 promoted the mechanical binding of titanium to bones, enabling efficient bone regeneration and effective stabilization of implants. Thus, such adhesive biosignaling proteins can be used in regenerative medicine.

Bundling of collagen fibrils influences osteocyte network formation during bone modeling.

Osteocytes form a cellular network by gap junctions between their cell processes. This network is important since intercellular communication via the network is essential for bone metabolism. However, the factors that influence the formation of this osteocyte network remain unknown. As the early stage of osteocyte network formation occurs on the bone surface, we observed a newly formed trabecular bone surface by orthogonal focused ion beam-scanning electron microscopy. The embedding late osteoblast processes tended to avoid bundled collagen fibrils and elongate into sparse collagen fibrils. Then, we examined whether the inhibition of bundling of collagen fibrils using a potent lysyl oxidase inhibitor, ß-aminopropionitrile (BAPN) changed the cellular network of the chick calvaria. The osteocyte shape of the control group was spindle-shape, while that of the BAPN group was sphere-shaped. In addition, the osteocyte processes of the control group were elongated vertically to the long axis of the cell body, whereas the osteocyte processes of the BAPN group were elongated radially. Therefore, it was suggested that the bundling of collagen fibrils influences normal osteocyte network formation during bone modeling.

Fabrication of a Fish-Bone-Inspired Inorganic-Organic Composite Membrane.

Biological materials have properties like great strength and flexibility that are not present in synthetic materials. Using the ribs of crucian carp as a reference, we investigated the mechanisms behind the high mechanical properties of this rib bone, and found highly oriented layers of calcium phosphate (CaP) and collagen fibers. To fabricate a fish-rib-bone-mimicking membrane with similar structure and mechanical properties, this study involves (1) the rapid synthesis of plate-like CaP crystals, (2) the layering of CaP-gelatin hydrogels by gradual drying, and (3) controlling the shape of composite membranes using porous gypsum molds. Finally, as a result of optimizing the compositional ratio of CaP filler and gelatin hydrogel, a CaP filler content of 40% provided the optimal mechanical properties of toughness and stiffness similar to fish bone. Due to the rigidity, flexibility, and ease of shape control of the composite membrane materials, this membrane could be applied as a guided bone regeneration (GBR) membrane.

Long-Term Antibacterial Efficacy of Cetylpyridinium Chloride-Montmorillonite Containing PMMA Resin Cement.

Despite being able to adhesively restore teeth, adhesives and cement do not possess any anticariogenic protection potential, by which caries recurrence may still occur and reduce the clinical lifetime of adhesive restorations. Several antibacterial agents have been incorporated into dental adhesives and cement to render them anticariogenic. Due to an additional therapeutic effect, such materials are classified as 'dental combination products' with more strict market regulations. We incorporated cetylpyridinium chloride (CPC), often used for oral hygiene applications, into montmorillonite (CPC-Mont), the latter to serve as a carrier for controlled CPC release. CPC-Mont incorporated into tissue conditioner has been approved by the Pharmaceuticals and Medical Devices Agency (PmontMDA) in Japan. To produce a clinically effective dental cement with the antibacterial potential to prevent secondary caries, we incorporated CPC-Mont into PMMA resin cement. We measured the flexural strength, shear bond strength onto dentin, CPC release, and the biofilm-inhibition potential of the experimental CPC-Mont-containing PMMA cement. An 8 and 10 wt% CPC-Mont concentration revealed the antibacterial potential without reducing the mechanical properties of the PMMA cement.

Important roles of odontoblast membrane phospholipids in early dentin mineralization.

The objective of this study was to first identify the timing and location of early mineralization of mouse first molar, and subsequently, to characterize the nucleation site for mineral formation in dentin from a materials science viewpoint and evaluate the effect of environmental cues (pH) affecting early dentin formation. Early dentin mineralization in mouse first molars began in the buccal central cusp on post-natal day 0 (P0), and was first hypothesized to involve collagen fibers. However, elemental mapping indicated the co-localization of phospholipids with collagen fibers in the early mineralization area. Co-localization of phosphatidylserine and annexin V, a functional protein that binds to plasma membrane phospholipids, indicated that phospholipids in the pre-dentin matrix were derived from the plasma membrane. A 3-dimensional in vitro biomimetic mineralization assay confirmed that phospholipids from the plasma membrane are critical factors initiating mineralization. Additionally, the direct measurement of the tooth germ pH, indicated it to be alkaline. The alkaline environment markedly enhanced the mineralization of cell membrane phospholipids. These results indicate that cell membrane phospholipids are nucleation sites for mineral formation, and could be important materials for bottom-up approaches aiming for rapid and more complex fabrication of dentin-like structures.

Osteoblast Response of Additively Manufactured Zirconia and Alumina-Toughened Zirconia.

Zirconia ceramics have been widely used in dentistry. Herein, we assess the surface morphology, surface texture, and osteoblast response of additively manufactured zirconia and alumina-toughened zirconia (ATZ) in comparison with titanium. The surface roughness, contact angle, and surface microstructure of titanium sandblasted with large-grit alumina and subsequently acid-etched using 18% HCl and 49% H2SO4 (SLA-titanium), uniaxially pressed zirconia (UP zirconia), additively manufactured zirconia (AM zirconia), and additively manufactured ATZ (AM ATZ) were investigated. Moreover, the cell viability, alkaline phosphatase (ALP) activity, and gene expression of type I collagen on these materials were evaluated. The data were statistically analyzed using one-way ANOVA with Tukey's post hoc test. SLA-titanium showed the highest surface roughness and contact angle. The other three materials showed comparable surface roughness and contact angles. Micro- and nanoroughness were observed on the surface of SLA-titanium. UP zirconia and AM zirconia had similar surface morphologies. The cell viability, ALP activity, and gene expression of type I collagen on AM zirconia were comparable to or better than those on SLA-titanium. Our results indicate that AM zirconia is a promising material for zirconia dental implants.

Does Multifunctional Acrylate's Addition to Methacrylate Improve Its Flexural Properties and Bond Ability to CAD/CAM PMMA Block?

This study investigated the effects of a multifunctional acrylate copolymer-Trimethylolpropane Triacrylate (TMPTA) and Di-pentaerythritol Polyacrylate (A-DPH)-on the mechanical properties of chemically polymerized acrylic resin and its bond strength to a CAD/CAM polymethyl methacrylate (PMMA) disk. The methyl methacrylate (MMA) samples were doped with one of the following comonomers: TMPTA, A-DPH, or Trimethylolpropane Trimethacrylate (TMPTMA). The doping ratio ranged from 10 wt% to 50 wt% in 10 wt% increments. The flexural strength (FS) and modulus (FM) of PMMA with and without comonomer doping, as well as the shear bond strength (SBS) between the comonomer-doped PMMA and CAD/CAM PMMA disk, were evaluated. The highest FS (93.2 ± 4.2 MPa) was obtained when doped with 20 wt% of TMPTA. For TMPTMA, the FS decreased with the increase in the doping ratio. For SBS, TMPTA showed almost constant values (ranging from 7.0 to 8.2 MPa) regardless of the doping amount, and A-DPH peaked at 10 wt% doping (8.7 ± 2.2 MPa). TMPTMA showed two peaks at 10 wt% (7.2 ± 2.6 MPa) and 40 wt% (6.5 ± 2.3 MPa). Regarding the failure mode, TMPTMA showed mostly adhesive failure between the CAD/CAM PMMA disk and acrylic resin while TMPTA and A-DPH showed an increased rate of cohesive or mixed failures. Acrylate's addition as a comonomer to PMMA provided improved mechanical properties and bond strength to the CAD/CAM PMMA disk.

Distinct Morphologies of Bone Apatite Clusters in Endochondral and Intramembranous Ossification.

Bone apatite crystals grow in clusters, but the microstructure of these clusters is unknown. This study compares the structural and compositional differences between bone apatite clusters formed in intramembranous (IO) and endochondral ossification (EO). Calvaria (IO) and femurs (EO) are isolated from mice at embryonic days (E) 14.5 to 15.5 and post-natal days (P) 6 to 7, respectively. Results show that the initially formed bone apatite clusters in EO (≅1.2 µm2 ) are >10 times larger than those in IO (≅0.1 µm2 ), without significant changes in ion composition. In IO (E14.5 calvarium), early minerals are formed inside matrix vesicles (MVs). In contrast, in EO (P6 femur epiphysis), no MVs are observed, and chondrocyte-derived plasma membrane nanofragments (PMNFs) are the nucleation site for mineralization. Apatite cluster size difference is linked with the different nucleation sites. Moreover, an alkaline pH and slow P supply into a Ca-rich microenvironment are suggested to facilitate apatite cluster growth, as demonstrated in a biomimetic mineralization system. Together, the results reveal for the first time the distinct and exquisite microstructures of bone apatite clusters in IO and EO, and provide insightful inspirations for the design of more efficient materials for bone tissue engineering and repair.

Preliminary Study on the Optimization of Femtosecond Laser Treatment on the Surface Morphology of Lithium Disilicate Glass-Ceramics and Highly Translucent Zirconia Ceramics.

All-ceramic restorations have become increasingly popular in dentistry. Toward ensuring that these restorations adhere to the tooth structure, this study determines the optimal femtosecond laser (FL) treatment parameters for lithium disilicate glass-ceramics and highly translucent zirconia ceramics with respect to surface morphology. For both the ceramics, the following surface conditions were investigated: (1) as-sintered; (2) Al2O3 sandblasted; (3) FL treatment (dot pattern with line distances of 14, 20, and 40 µm); (4) FL treatment (crossed-line pattern with a line distance of 20 and 40 µm). Surface roughness parameters were estimated using a 3D confocal laser microscope; microstructures were analyzed using a scanning electron microscope. Peak fluence (Fpeak) values of 4 and 8 J/cm2 and irradiation numbers (N) of 20 and 10 shots were selected to create dot patterns in highly translucent zirconia and lithium disilicate glass-ceramics, respectively. Furthermore, Fpeak = 8 J/cm2 and N = 20 shots were chosen to obtain crossed-line patterns in both ceramics. Our results show that lithium disilicate glass-ceramics and highly translucent zirconia exhibit a similar surface morphology under each of the surface treatment conditions. Therefore, FL irradiation of dot or crossed-line patterns (at a distance of 20 and 40 µm) are potential candidates for future investigations.

Eco-Benign Orange-Hued Pigment Derived from Aluminum-Enriched Biogenous Iron Oxide Sheaths.

Inorganic pigments have been widely used due to their low cost of production, strong hiding power, and chemical resistance; nevertheless, they have limited hue width and chromaticity. To eliminate these disadvantages, we herein propose the use of an ingenious biotemplate technique to produce Al-enriched biogenic iron oxide (BIOX) materials. Spectrophotometric color analysis showed that high levels of Al inclusion on heat-treated BIOX samples produced heightened yellowish hues and lightness. The Al-enriched BIOX sheaths exhibited a stable tubular structure and excellent thermal stability of color tones after heating at high temperatures and repetitive heat treatments. Ultrastructural analysis and mechanical destruction experiments revealed that the highly chromatic orange-hue of these pigments are ascribed probably to an ingenious cylindrical nanocomposite architecture composed of putative Fe-included low crystalline Al oxide regions and hematite particles embedded therein. The present work therefore demonstrates that the bioengineered material can serve as an epochal orange-hued inorganic pigment with low toxicity and marked thermostability that should meet large industrial demand.

Flame retardance-donated lignocellulose nanofibers (LCNFs) by the Mannich reaction with (amino-1,3,5-triazinyl)phosphoramidates and their properties.

Nitrogen/phosphorus-containing melamines (NPCM), a durable flame-retardant, were prepared by the successive treatment of ArOH (Ar = Br n C6H5-n , n = 0, 1, 2, and 3) with POCl3 and melamine monomer. The prepared flame-retardants were grafted through the CH2 unit to lignocellulose nanofibers (LCNFs) by the Mannich reaction. The resulting three-component products were characterized using FT-IR (ATR) and EA. The thermal behavior of the NPCM-treated LCNF fabric samples was determined using TGA and DSC analyses, and their flammability resistances were evaluated by measuring their Limited Oxygen Index (LOI) and the UL-94V test. A multitude of flame retardant elements in the fabric samples increased the LOI values as much as 45 from 20 of the untreated LCNFs. Moreover, the morphology of both the NPCM-treated LCNFs and their burnt fabrics was studied with a scanning electron microscope (SEM). The heat release lowering effect of the LCNF fabric against the water-based paint was observed with a cone calorimeter. Furthermore, the mechanical properties represented as the tensile strength of the NPCM-treated LCNF fabrics revealed that the increase of the NPCM content in the PP-composites led to an increased bending strength with enhancing the flame-retardance.

A morphometric analysis of the osteocyte canaliculus using applied automatic semantic segmentation by machine learning.

INTRODUCTION: Osteocytes play a role as mechanosensory cells by sensing flow-induced mechanical stimuli applied on their cell processes. High-resolution imaging of osteocyte processes and the canalicular wall are necessary for the analysis of this mechanosensing mechanism. Focused ion beam-scanning electron microscopy (FIB-SEM) enabled the visualization of the structure at the nanometer scale with thousands of serial-section SEM images. We applied machine learning for the automatic semantic segmentation of osteocyte processes and canalicular wall and performed a morphometric analysis using three-dimensionally reconstructed images. MATERIALS AND METHODS: Six-week-old-mice femur were used. Osteocyte processes and canaliculi were observed at a resolution of 2 nm/voxel in a 4 × 4 µm region with 2000 serial-section SEM images. Machine learning was used for automatic semantic segmentation of the osteocyte processes and canaliculi from serial-section SEM images. The results of semantic segmentation were evaluated using the dice similarity coefficient (DSC). The segmented data were reconstructed to create three-dimensional images and a morphological analysis was performed. RESULTS: The DSC was > 83%. Using the segmented data, a three-dimensional image of approximately 3.5 µm in length was reconstructed. The morphometric analysis revealed that the median osteocyte process diameter was 73.8 ± 18.0 nm, and the median pericellular fluid space around the osteocyte process was 40.0 ± 17.5 nm. CONCLUSION: We used machine learning for the semantic segmentation of osteocyte processes and canalicular wall for the first time, and performed a morphological analysis using three-dimensionally reconstructed images.

Re-Evaluation of Initial Bone Mineralization from an Engineering Perspective.

Bone regeneration was one of the earliest fields to develop in the context of tissue regeneration, and currently, repair of small-sized bone defects has reached a high success rate. Future researches are expected to incorporate more advanced techniques toward achieving rapid bone repair and modulation of the regenerated bone quality. For these purposes, it is important to have a more integrative understanding of the mechanisms of bone formation and maturation from multiple perspectives and to incorporate these new concepts into the development and designing of novel materials and techniques for bone regeneration. This review focuses on the analysis of the earliest stages of bone tissue development from the biology, material science, and engineering perspectives for a more integrative understanding of bone formation and maturation, and for the development of novel biology-based engineering approaches for tissue synthesis in vitro. More specifically, the authors describe the systematic methodology that allowed the understanding of the different nucleation sites in intramembranous and endochondral ossification, the space-making process for mineral formation and growth, as well as the process of apatite crystal cluster growth in vivo in the presence of suppressing biomolecules. Impact Statement A detailed understanding of the developmental process of bone tissue leads to the acquisition of useful information for the bone tissue fabrication. This review summarizes the study of the calcification process of the calvaria and epiphyses from an engineering perspective and provides useful information for the realization of bone tissue biofabrication. Here, we describe the new mechanism of space formation for mineralization such as rupture of chondrocytes and disruption of cell-cell adhesion. We also describe the roles of nucleation site such as cell membrane nanofragments and matrix vesicles.

Effect of Air-Particle Abrasion Protocol and Primer on The Topography and Bond Strength of a High-Translucent Zirconia Ceramic.

PURPOSE: To evaluate effect of air-particle abrasion protocol and primer on surface topography and bond strength of resin cement to high-translucent zirconia ceramics. MATERIALS AND METHODS: Two hundred disk-shaped high-translucent zirconia specimens of 5Y-PSZ were prepared. The specimens were assigned to 5 groups in terms of particle type and air-particle abrasion pressure: (1) control, (2) alumina with 0.2 MPa-air pressure [AB-0.2], (3) alumina with 0.4 MPa-air pressure [AB-0.4], (4) glass beads with 0.2 MPa-air pressure [GB-0.2], and (5) glass beads with 0.4 MPa-air pressure [GB-0.4]. Two different primers 1% MDP (Experimental) and MDP-silane primer (Clearfil Ceramic Primer Plus) was also tested. Stainless steel rods were bonded to the 5Y-PSZ specimens with PANAVIA V5. For each group, the tensile bond strength (TBS) was measured after 24-hour water storage (n = 10) and after 5000 thermal-cycling (n = 10) at crosshead speed of 2 mm/min. The data were statistically analyzed using Weibull analysis. Surface roughness (Sa) was measured using a 3D-Laser Scanning Confocal Microscope (n = 5) and analyzed by t-test with Bonferroni correction. Surface topography using scanning electron microscopy (SEM) and surface elemental analysis using energy dispersion spectroscopy (EDX), and cross-section SEM at the interface with composite cement were also investigated. RESULTS: In MDP-silane groups, the highest TBS was observed in AB-0.4 after 24 hours and GB-0.4 after thermal-cycling (p < 0.05). In MDP groups, AB groups resulted in the significantly higher TBS than GB groups (p < 0.05). AB-0.4 group showed the highest Sa value compared to all groups (p < 0.005), meanwhile GB groups did not show different Sa compared to control (p > 0.005). CONCLUSION: Air-abrasion with different particle and blasting pressure can improve bonding to zirconia with proper primer selection. Particularly, glass beads abrasion followed by MDP-silane primer and alumina abrasion followed by MDP primer alone provided stable bond strength of resin cement to high-translucent zirconia after aging. High-translucent zirconia abraded with glass beads achieves a desirable bonding performance without creating surface microcracks which may hinder zirconia's mechanical properties.

Novel composite cement containing the anti-microbial compound CPC-Montmorillonite.

OBJECTIVES: The aim of this study was to evaluate the mechanical properties, bonding performance and anti-microbial activity of a novel composite cement containing cetylpyridinium chloride (CPC) modified montmorillonite ('CPC-Mont'), and using these parameters to determine the optimal particle size and concentration of CPC-Mont the composite cement can be loaded with. METHODS: CPC-Mont particles with a median diameter of 30 and 7 µm were prepared and added to a composite cement at a concentration of 2, 3, 4, 5 and 7.5 wt%. Mechanical properties and bonding performance of the experimental composite cements were evaluated by 3-point bending and micro-tensile bond-strength testing. The amount of CPC released from the cement disks was quantified using a UV-vis recording spectrophotometer. The anti-biofilm activity was studied using scanning electron microscopy (SEM). RESULTS: Adding 30-µm CPC-Mont decreased the mechanical properties and bonding performance of the composite cement, while no reduction was observed for the 7-µm CPC-Mont loaded cement formulation. Although CPC release substantially decreased during the 7-day period assessed, 5- and 7.5-wt% CPC-Mont loaded composite cement inhibited biofilm formation for 30 days. SIGNIFICANCE: Loading composite cement with CPC-Mont with a median diameter of 7 µm at concentrations of 5-7.5 wt% was effective in achieving continuous anti-biofilm activity, while maintaining mechanical strength and bonding performance.

Antibacterial Effect of Amino Acid-Silver Complex Loaded Montmorillonite Incorporated in Dental Acrylic Resin.

Several dental materials contain silver for antibacterial effect, however the effect is relatively low. The reason for the lower antibacterial efficacy of silver is considered to be the fact that silver ions bind to chloride ions in saliva. To develop new effective silver antibacterial agents that can be useful in the mouth, we synthesized two novel amino acid (methionine or histidine)-silver complexes (Met or His-Ag) loaded with montmorillonite (Mont) and analyzed their antibacterial efficacy. At first the complexes were characterized using nuclear magnetic resonance (NMR), and amino acid-Ag complex-loaded Mont (amino acid-Ag-Mont) were characterized using X-ray diffraction (XRD) and scanning electron microscopy (SEM). The antibacterial efficacy of these materials in dental acrylic resin was then investigated by bacterial growth measurement using a spectrophotometer. As controls, commercially available silver-loaded zeolite and silver-zirconium phosphate were also tested. Dental acrylic resin incorporating His-Ag-Mont strongly inhibited Streptococcus mutans growth. This was explained by the fact that His-Ag complex revealed the highest amounts of silver ions in the presence of chloride. The structure of the amino acid-Ag complexes affected the silver ion presence in chloride and the antibacterial efficacy. His-Ag-Mont might be used as antibacterial agents for dental materials.

Phosphate group adsorption capacity of inorganic elements affects bond strength between CAD/CAM composite block and luting agent.

This study aimed to investigate whether inorganic elements of polymer-infiltrated ceramic (PIC) and microfilled resin (MFR) for CAD/CAM would affect initial bond strength to luting agent. Inorganic elements of PIC and MFR were different with shape and ingredient observed by SEM, STEM and EDS. Microtensile bond strengths (µTBS) value of PIC was increased by 10-methacryloyloxydecyl dihydrogen phosphate (MDP) and acetic acid (AA)- or MDP-activated silane treatment, and further increased by succeeding heat treatment (HT). The µTBS of MFR was increased by MDP and MDP-activated silane, but decreased by AA-activated silane without HT. The HT improved the µTBS of MFR with AA-activated silane, but conversely for MDP-activated silane. Only in MFR, phosphoric acid (PA) application before each surface treatment dramatically decreased the µTBS of AA-activated silane. FTIR peaks in MFR shifted according to phosphate group's peak. MFR would possess high phosphate group adsorption capacity, with MDP effectively improving its bonding capability.

Micro-Architectural Investigation of Teleost Fish Rib Inducing Pliant Mechanical Property.

Despite the fact that various reports have been discussing bone tissue regeneration, precise bone tissue manipulation, such as controlling the physical properties of the regenerated bone tissue, still remains a big challenge. Here, we focused on the teleost fish ribs showing flexible and tough mechanical properties to obtain a deeper insight into the structural and functional features of bone tissue from different species, which would be valuable for the superior design of bone-mimicking materials. Herein, we examined their compositions, microstructure, histology, and mechanical properties. The first rib of Carassius langsdorfii showed a higher Young's modulus with a small region of chondrocyte clusters compared with other smaller ribs. In addition, highly oriented collagen fibers and osteocytes were observed in the first rib, indicating that the longest first rib would be more mature. Moreover, the layer-by-layer structure of the oriented bone collagen was observed in each rib. These microarchitectural and compositional findings of fish rib bone would give one the useful idea to reproduce such a highly flexible rib bone-like material.

Flexural properties, bond ability, and crystallographic phase of highly translucent multi-layered zirconia.

This study investigated the mechanical properties, bond ability, and crystallographic forms of different sites in a highly translucent, multi-layered zirconia disk. Flexural properties, bond ability to resin cement, and phase composition were investigated at three sites of a highly translucent, multi-layered zirconia disk: incisal, middle, and cervical. Flexural strength (FS) and flexural modulus (FM) were measured with static three-point flexural test. Shear bond strength (SB) to resin cement was measured after 24 h storage (37°C). Phase composition under mechanical stress was analyzed using X-ray diffraction. Without air abrasion, FS at the incisal site yielded the lowest value and was significantly lower than the middle and cervical sites. Air abrasion lowered the FS of each site. FM at the incisal site without air abrasion showed the significantly lowest value, and air abrasion increased its FM value. At the middle and cervical sites, their FM values were higher than the incisal site but were not significantly affected by air abrasion. SB value did not show significant differences among the sites. After sintering, cubic zirconia was detected at each site. Rhombohedral phase transformation occurred after mirror polishing. In highly translucent, multi-layered zirconia which was mainly composed of cubic zirconia, rhombohedral phase transformation occurred under mechanical stress and resulted in weakened mechanical properties.

Development of brushite particles synthesized in the presence of acidic monomers for dental applications.

OBJECTIVES: To synthesize and characterize brushite particles in the presence of acidic monomers (acrylic acid/AA, citric acid/CA, and methacryloyloxyethyl phosphate/MOEP) and evaluate the effect of these particles on degree of conversion (DC), flexural strength/modulus (FS/FM) and ion release of experimental composites. METHODS: Particles were synthesized by co-precipitation with monomers added to the phosphate precursor solution and characterized for monomer content, size and morphology. Composites containing 20 vol% brushite and 40 vol% reinforcing glass were tested for DC, FS and FM (after 24 h and 60 d in water), and 60-day ion release. Data were subjected to ANOVA/Tukey tests (DC) or Kruskal-Wallis/Dunn tests (FS and FM, alpha: 5%). RESULTS: The presence of acidic monomers affected particle morphology. Monomer content on the particles was low (0.1-1.4% by mass). Composites presented similar DC. For FS/24 h, only the composite containing DCPD_AA was statistically similar to the composite containing 60 vol% of reinforcing glass (without brushite, "control"). After 60 days, all brushite-containing materials showed similar FS, statistically lower than the control composite (p<0.01). Composites containing DCPD_AA, DCPD_MOEP or DCPD_U ("unmodified") showed statistically similar FM/24 h, higher than the control composite. After prolonged immersion, all composites were similar to the control composite, except DCPD_AA. Cumulative ion release ranged from 21 ppm to 28 ppm (calcium) and 9 ppm to 17 ppm (phosphate). Statistically significant reductions in ion release between 15 and 60 days were detected only for the composite containing DCPD_MOEP. SIGNIFICANCE: Acidic monomers added to the synthesis affected brushite particle morphology. After 60-day storage in water, composite strength was similar among all brushite-containing composites. Ion release was sustained for 60 days and it was not affected by particle morphology.