Strontium and Zinc Co-Doped Mesoporous Bioactive Glass Nanoparticles for Potential Use in Bone Tissue Engineering Applications.

Mesoporous bioactive glass nanoparticles (MBGNs) have attracted significant attention as multifunctional nanocarriers for various applications in both hard and soft tissue engineering. In this study, multifunctional strontium (Sr)- and zinc (Zn)-containing MBGNs were successfully synthesized via the microemulsion-assisted sol-gel method combined with a cationic surfactant (cetyltrimethylammonium bromide, CTAB). Sr-MBGNs, Zn-MBGNs, and Sr-Zn-MBGNs exhibited spherical shapes in the nanoscale range of 100 ± 20 nm with a mesoporous structure. Sr and Zn were co-substituted in MBGNs (60SiO2-40CaO) to induce osteogenic potential and antibacterial properties without altering their size, morphology, negative surface charge, amorphous nature, mesoporous structure, and pore size. The synthesized MBGNs facilitated bioactivity by promoting the formation of an apatite-like layer on the surface of the particles after immersion in Simulated Body Fluid (SBF). The effect of the particles on the metabolic activity of human mesenchymal stem cells was concentration-dependent. The hMSCs exposed to Sr-MBGNs, Zn-MBGNs, and Sr-Zn-MBGNs at 200 µg/mL enhanced calcium deposition and osteogenic differentiation without osteogenic supplements. Moreover, the cellular uptake and internalization of Sr-MBGNs, Zn-MBGNs, and Sr-Zn-MBGNs in hMSCs were observed. These novel particles, which exhibited multiple functionalities, including promoting bone regeneration, delivering therapeutic ions intracellularly, and inhibiting the growth of Staphylococcus aureus and Escherichia coli, are potential nanocarriers for bone regeneration applications.

Assessment of physical/mechanical properties and cytotoxicity of dual-cured resin cements containing Sr-bioactive glass nanoparticles and calcium phosphate.

The aim was to develop dual-cured resin cements containing Sr-bioactive glass nanoparticles (Sr-BGNPs; 5 or 10 wt%) and monocalcium phosphate monohydrate (MCPM; 3 or 6 wt%). Effects of additives on degree of monomer conversion (DC), biaxial flexural strength/modulus, shear bond strength (SBS), mass/volume change, color stability, ion release, and cytotoxicity were examined. Controls included material without reactive fillers and Panavia SA Plus (PV). Experimental cements showed higher DC than PV regardless of light activation (p<0.05). Mean SBS and color stability were comparable between experimental cements and PV. Cell viability upon the exposure to sample extracts of experimental cements was 80%-92%. High additive concentrations led to lower strength and modulus than PV (p<0.05). The additives increased mass change, reduced color stability, and promoted ion release. The experimental resin cements demonstrated acceptable mechanical/chemical properties and cytotoxicity. The additives reduced the strength but provided ion release, a desirable action to prevent recurrent caries.

Development of Zn/Ce containing bioactive glasses for bone regeneration applications.

The impact of either Zn or Ce substituted with Ca in bioactive glasses based on the 45SiO2 - 6P2O5 - 11SrO - (38-(x + y) CaO) - xZnO or yCe2O3 (xZn-yCeBGs) system on bone regeneration has not yet been reported. The aim of this study was to develop new formulations of sol-gel-derived bioactive glass to use as a synthetic bone graft. xZn-yCeBGs were synthesized through the sol-gel process with 0.01 M nitric acid catalyst. xZn-yCeBG formation was investigated using SEM and FTIR. The bioactivity of xZn-yCeBGs was evaluated using SEM, EDX-SEM, and XRD. Cell viability and ability to form mineralization of MC3T3-E1 treated with xZn-yCeBGs were detected using MTT assay and Alizarin Red S staining, respectively. xZn-yCeBGs were successfully prepared. Zn or Ce substituted with Ca in BGs stimulated bioactivity through apatite formation, enhanced bone mineralization, and were not toxic to the bone cells. Moreover, these particles had an antibacterial effect against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) via the disc diffusion method. Therefore, xZn-yCeBGs are a promising unique biomaterial with a potential future role as bone graft substitutes.

Assessment of Mechanical/Chemical Properties and Cytotoxicity of Resin-Modified Glass Ionomer Cements Containing Sr/F-Bioactive Glass Nanoparticles and Methacrylate Functionalized Polyacids.

This study prepared low-toxicity, elemental-releasing resin-modified glass ionomer cements (RMGICs). The effect of 2-hydroxyethyl methacrylate (HEMA, 0 or 5 wt%) and Sr/F-bioactive glass nanoparticles (Sr/F-BGNPs, 5 or 10 wt%) on chemical/mechanical properties and cytotoxicity were examined. Commercial RMGIC (Vitrebond, VB) and calcium silicate cement (Theracal LC, TC) were used as comparisons. Adding HEMA and increasing Sr/F-BGNPs concentration decreased monomer conversion and enhanced elemental release but without significant effect on cytotoxicity. Rising Sr/F-BGNPs reduced the strength of the materials. The degree of monomer conversion of VB (96%) was much higher than that of the experimental RMGICs (21-51%) and TC (28%). The highest biaxial flexural strength of experimental materials (31 MPa) was significantly lower than VB (46 MPa) (p < 0.01) but higher than TC (24 MPa). The RMGICs with 5 wt% HEMA showed higher cumulative fluoride release (137 ppm) than VB (88 ppm) (p < 0.01). Unlike VB, all experimental RMGICs showed Ca, P, and Sr release. Cell viability in the presence of extracts from experimental RMGICs (89-98%) and TC (93%) was significantly higher than for VB (4%). Experimental RMGICs showed desirable physical/mechanical properties with lower toxicity than the commercial material.

Multifunctional Zn and Ag co-doped bioactive glass nanoparticles for bone therapeutic and regeneration.

Bone cancer has traditionally been treated using surgery, radiotherapy, and/or chemotherapy. The nonspecific distribution of chemotherapy and implantable infections are significant risk factors for the failure of the bone to heal. Multifunctional zinc and silver co-doped bioactive glass nanoparticles (yAg-xZn-BGNPs) with a diameter of 150 ± 30 nm were successfully synthesized using modified sol-gel and two-step post-functionalization processes, tailored to provide antibacterial and anticancer activity whilst maintaining osteogenesis ability. Co-doped BGNPs with Zn and Ag did not significantly alter physicochemical properties, including size, morphology, glass network, and amorphous nature. Apatite-like layer was observed on the surface of yAg-xZn-BGNPs and resorbed in the simulated body fluid solution, which could increase their bioactivity. Human fetal osteoblast cell line (hFOB 1.19) treated with particles showed calcified tissue formation and alkaline phosphatase activity in the absence of osteogenic supplements in vitro, especially with 0.5Ag-1Zn-BGNPs. Moreover, these particles preferentially disrupted the metabolic activity of bone cancer cells (MG-63) and had an antibacterial effect against B. subtilis, E. coli, and S. aureus via the disc diffusion method. This novel 0.5Ag-1Zn-BGNP and 1Ag-1Zn-BGNPs, with wide-ranging ability to stimulate bone regeneration, to inhibit bone cancer cell proliferation, and to prevent bacterial growth properties, may provide a feasible strategy for bone cancer treatment. The 0.5Ag-1Zn-BGNPs and 1Ag-1Zn-BGNPs can be applied for the preparation of scaffolds or filler composites using in bone tissue engineering.

Zinc and Strontium-Substituted Bioactive Glass Nanoparticle/Alginate Composites Scaffold for Bone Regeneration.

The global population is growing older and entering an aging society. Aging results in severe tissue disorder and organ dysfunction. Bone-related injuries are particularly significant. The need for alternative bone replacement materials for human implants has grown over the past few decades. Alginate has the potential for use as a cell scaffold for bone tissue engineering due to its high bio-compatibility. To improve the bioactivity of alginate scaffolds, zinc- and strontium-containing sol-gel-derived bioactive glass nanoparticles (Zn-Sr-BGNPs) with sizes ranging from 100 to l40 nm were incorporated. Zn-Sr-BGNPs synthesized through the sol-gel process have a high sur-face-to-volume ratio, homogeneity, and purity, resulting in faster degradation. The therapeutic bivalent ions released from Zn-Sr-BGNPs strengthen the cell scaffold and improve the stimulation of the production and development of bone cells. Zn-Sr-BGNPs with different Zn to Si nominal ratios of 0, 1, and 1.5 were mixed with alginate in this research. The ratio of Zn in Zn-Sr-BGNPs and the ratio of Zn-Sr-BGNPs in scaffolds impact the pore size, swelling, and biological properties of synthesized composite scaffolds. The surface area and pore volume of a 1:1 1Zn-Sr-BGNP:Alg composite scaffold were 22.58 m2/g and 0.055 cm3/g, respectively. The incorporation of Zn-Sr-BGNPs improved the mechanical performance of the scaffolds up to 4.73 ± 0.48 MPa. The swelling rate decreased slightly from 2.12 (pure Alg) to 1.50 (1Zn-Sr-BGNP:Alg (1:1)). The 1Zn-Sr-BGNP:Alg (1:1) composite scaffold promoted bioactivity through apatite layer formation, increased bone cell proliferation via the dissolution products released from the scaffold, enhanced calcium deposition, and facilitated cell attachment. Thus, 1Zn-Sr-BGNP:Alg (1:1) composite scaffold is proposed as a possible artificial bone scaffold in bone tissue regeneration.

Influence of Lithium- and Zinc-Containing Bioactive Glasses on Pulpal Regeneration.

OBJECTIVE: To evaluate the potential of modified bioactive glasses containing lithium and zinc as pulp capping materials by investigating the odontogenic differentiation and mineralization response in the tooth culture model. MATERIALS AND METHODS: Lithium- and zinc-containing bioactive glasses (45S5.1Li, 45S5.5Li, 45S5.1Zn, 45S5.5Zn, 45S5.1Zn sol-gel, and 45S5.5Zn sol-gel), fibrinogen-thrombin, and biodentine were prepared to assess Axin2 gene expression at 0, 30 minutes, 1 hour, 12 hours, and 1 day and DSPP gene expression at 0, 3, 7, and 14 days in stem cells from human exfoliated deciduous teeth (SHEDs) using qRT-PCR. The experimental bioactive glasses incorporated with fibrinogen-thrombin and biodentine were placed on the pulpal tissue in the tooth culture model. Histology and immunohistochemistry were analyzed at 2 weeks and 4 weeks. RESULTS: Axin2 gene expression for all experimental groups was significantly higher than the control at 12 hours. The DSPP gene expression for all experimental groups was significantly higher than the control at 14 days. The presence of mineralization foci was significantly higher at 4 weeks for the modified bioactive glasses 45S5.5Zn, 45S5.1Zn sol-gel, and 45S5.5Zn sol-gel as well as Biodentine compared with the fibrinogen-thrombin control. CONCLUSION: Lithium- and zinc-containing bioactive glasses increased Axin2 and DSPP gene expression in SHEDs and can potentially enhance pulp mineralization and regeneration. Zinc-containing bioactive glasses are a promising candidate to be used as pulp capping materials.

An in vitro assessment of biaxial flexural strength, degree of monomer conversion, color stability, and ion release in provisional restorations containing Sr-bioactive glass nanoparticles.

This study examined the mechanical and chemical properties of an experimental provisional restoration containing Sr-bioactive glass nanoparticles (Sr-BGNPs) compared to commercial provisional materials. The experimental material (TempS10) contained dimethacrylate monomers with added 10 wt% Sr-BGNPs. The degree of monomer conversion (DC) of self-curing (n = 5), biaxial flexural strength (BFS)/modulus (BFM) (n = 5), and color changes (ΔE*00) of materials in red wine (n = 5) were determined. Additionally, ion release (Ca, P, and Sr) in water at 2 weeks was examined (n = 3). The commercial materials tested included polymethyl methacrylate-based provisional material (Unifast) and bis-acrylic materials (Protemp4 and Cooltemp). TempS10 exhibited a comparable degree of monomer conversion (49%) to that of Protemp4 (60%) and Cooltemp (54%) (p > 0.05). The DC of Unifast (81%) was significantly higher than that of other materials (p < 0.05). TempS10 showed a BFS (126 MPa) similar to Cooltemp (102 MPa) and Unifast (123 MPa), but lower than Protemp4 (194 MPa). The immersion time for 2 weeks exhibited no detrimental effect on the strength and modulus of all materials. The highest ΔE*00 at 24 h and 2 weeks was observed with TempS10, followed by Cooltemp, Unifast, and Protemp4. Only TempS10 showed a detectable amount of Ca (0.69 ppm), P (0.12 ppm), and Sr (3.01 ppm). The experimental provisional resin restoration containing Sr-BGNPs demonstrated polymerization and strength comparable to those of bis-acryl provisional restorations but with the added benefit of ion-releasing properties. However, the experimental material demonstrated unsatisfactory color stability.

Assessment of Physical/Mechanical Performance of Dental Resin Sealants Containing Sr-Bioactive Glass Nanoparticles and Calcium Phosphate.

The aim of this study was to assess the chemical/mechanical properties of ion-releasing dental sealants containing strontium-bioactive glass nanoparticles (Sr-BGNPs) and monocalcium phosphate monohydrate (MCPM). Two experimental sealants, TS1 (10 wt% Sr-BGNPs and 2 wt% MCPM) and TS2 (5 wt% Sr-BGNPs and 4 wt% MCPM), were prepared. Commercial controls were ClinproXT (CP) and BeautiSealant (BT). The monomer conversion (DC) was tested using ATR−FTIR (n = 5). The biaxial flexural strength (BFS) and modulus (BFM) were determined (n = 5) following 24 h and 7 days of immersion in water. The Vickers surface microhardness (SH) after 1 day in acetic acid (conc) versus water was tested (n = 5). The bulk and surface calcium phosphate precipitation in simulated body fluid was examined under SEM-EDX. The ion release at 4 weeks was analyzed using ICP-MS (n = 5). The DC after 40 s of light exposure of TS1 (43%) and TS2 (46%) was significantly lower than that of CP (58%) and BT (61%) (p < 0.05). The average BFS of TS1 (103 MPa), TS2 (123 MPa), and BT (94 MPa) were lower than that of CP (173 MPa). The average BFM and SH of TS1 (2.2 GPa, 19 VHN) and TS2 (2.0 GPa, 16 VHN) were higher than that of CP (1.6 GPa, 11 VHN) and BT (1.3 GPa, 12 VHN). TS1 showed higher Ca, P, and Sr release than TS2. Bulk calcium phosphate precipitation was detected on TS1 and TS2 suggesting some ion exchange. In conclusion, the DC of experimental sealants was lower than that of commercial materials, but their mechanical properties were within the acceptable ranges. The released ions may support remineralizing actions.

Effects of Sr/F-Bioactive Glass Nanoparticles and Calcium Phosphate on Monomer Conversion, Biaxial Flexural Strength, Surface Microhardness, Mass/Volume Changes, and Color Stability of Dual-Cured Dental Composites for Core Build-Up Materials.

This study prepared composites for core build-up containing Sr/F bioactive glass nanoparticles (Sr/F-BGNPs) and monocalcium phosphate monohydrate (MCPM) to prevent dental caries. The effect of the additives on the physical/mechanical properties of the materials was examined. Dual-cured resin composites were prepared using dimethacrylate monomers with added Sr/F-BGNPs (5 or 10 wt%) and MCPM (3 or 6 wt%). The additives reduced the light-activated monomer conversion by ~10%, but their effect on the conversion upon self-curing was negligible. The conversions of light-curing or self-curing polymerization of the experimental materials were greater than that of the commercial material. The additives reduced biaxial flexural strength (191 to 155 MPa), modulus (4.4 to 3.3), and surface microhardness (53 to 45 VHN). These values were comparable to that of the commercial material or within the acceptable range of the standard. The changes in the experimental composites' mass and volume (~1%) were similar to that of the commercial comparison. The color change of the commercial material (1.0) was lower than that of the experimental composites (1.5-5.8). The addition of Sr/F-BGNPs and MCPM negatively affected the physical/mechanical properties of the composites, but the results were satisfactory except for color stability.

Zinc-Containing Sol-Gel Glass Nanoparticles to Deliver Therapeutic Ions.

Zn-containing dense monodispersed bioactive glass nanoparticles (Zn-BAGNPs) have been developed to deliver therapeutic inorganic trace elements, including Si, Ca, Sr, and Zn, to the cells through the degradation process, as delivery carriers for stimulating bone regeneration because of their capacity to induce osteogenic differentiation. The sol-gel-derived dense silica nanoparticles (SiO2-NPs) were first synthesized using the modified Stöber method, prior to incorporating therapeutic cations through the heat treatment process. The successfully synthesized monodispersed Zn-BAGNPs (diameter of 130 ± 20 nm) were homogeneous in size with spherical morphology. Ca, Sr and Zn were incorporated through the two-step post-functionalization process, with the nominal ZnO ratio between 0 and 2 (0, 0.5, 1.0, 1.5 and 2.0). Zn-BAGNPs have the capacity for continuous degradation and simultaneous ion release in SBF and PBS solutions due to their amorphous structure. Zn-BAGNPs have no in vitro cytotoxicity on the murine pre-osteoblast cell (MC3T3-E1) and periodontal ligament stem cells (PDLSCs), up to a concentration of 250 µg/mL. Zn-BAGNPs also stimulated osteogenic differentiation on PDLSCs treated with particles, after 2 and 3 weeks in culture. Zn-BAGNPs were not toxic to the cells and have the potential to stimulate osteogenic differentiation on PDLSCs. Therefore, Zn-BAGNPs are potential vehicles for therapeutic cation delivery for applications in bone and dental regenerations.

Physical/mechanical and antibacterial properties of orthodontic adhesives containing Sr-bioactive glass nanoparticles, calcium phosphate, and andrographolide.

White spot lesions around orthodontic brackets are the major complication during fixed orthodontic treatment. This study prepared orthodontic adhesives for promoting mineral precipitation and reducing bacterial growth. Adhesives with added calcium phosphate monohydrate/Sr-bioactive glass nanoparticles (Sr/CaP) and andrographolide were prepared. The physical/mechanical and antibacterial properties of the adhesives were tested. The additives reduced the monomer conversion of the materials (62 to 47%). The addition of Sr/CaP and andrographolide increased the water sorption (from 23 to 46 µg/mm3) and water solubility (from 0.2 to 5.9 µg/mm3) but reduced the biaxial flexural strength (from 193 to 119 MPa) of the adhesives. The enamel bond strengths of the experimental adhesives (19-34 MPa) were comparable to that of the commercial material (p > 0.05). The Sr/CaP fillers promoted Ca, Sr, and P ion release and the precipitation of calcium phosphate at the debonded interface. An increase in the Sr/CaP concentration enhanced the inhibition of S. mutans by 18%, while the effect of andrographolide was not detected. The abilities of the adhesives to promote ion release, calcium phosphate precipitation, and the growth inhibition of cariogenic bacteria were expected to reduce the occurrence of white spot lesions. The additives reduced the physical/mechanical properties of the materials, but the corresponding values were within the acceptable range.

Interaction of monodispersed strontium containing bioactive glass nanoparticles with macrophages.

The cellular response of murine primary macrophages to monodisperse strontium containing bioactive glass nanoparticles (SrBGNPs), with diameters of 90 ± 10 nm and a composition (mol%) of 88.8 SiO2-1.8CaO-9.4SrO (9.4% Sr-BGNPs) was investigated for the first time. Macrophage response is critical as applications of bioactive nanoparticles will involve the nanoparticles circulating in the blood stream and macrophages will be the first cells to encounter the particles, as part of inflammatory response mechanisms. Macrophage viability and total DNA measurements were not decreased by particle concentrations of up to 250 µg/mL. The Sr-BGNPs were actively internalised by the macrophages via formation of endosome/lysosome-like vesicles bordered by a membrane inside the cells. The Sr-BGNPs degraded inside the cells, with the Ca and Sr maintained inside the silica network. When RAW264.7 cells were incubated with Sr-BGNPs, the cells were polarised towards the pro-regenerative M2 population rather than the pro-inflammatory M1 population. Sr-BGNPs are potential biocompatible vehicles for therapeutic cation delivery for applications in bone regeneration.

Nanoceria provides antioxidant and osteogenic properties to mesoporous silica nanoparticles for osteoporosis treatment.

Osteoporosis, a chronic metabolic bone disease, is the most common cause of fractures. Drugs for treating osteoporosis generally inhibit osteoclast (OC) activity, but are rarely aimed at encouraging new bone growth and often cause severe systemic side effects. Reactive oxygen species (ROS) are one of the key triggers of osteoporosis, by inducing osteoblast (OB) and osteocyte apoptosis and promoting osteoclastogenesis. Here we tested the capability of the ROS-scavenger nanoceria encapsulated within mesoporous silica nanoparticles (Ce@MSNs) to treat osteoporosis using a pre-osteoblast MC3T3-E1 cell monoculture in stressed and normal conditions. Ce@MSNs (diameter of 80 ± 10 nm) were synthesised following a scalable two-step process involving sol-gel and wet impregnation methods. The Ce@MSNs at concentration of 100 µg mL-1 induced a significant reduction in oxidative stress produced by t-butyl hydroperoxide and did not alter cell viability significantly. Confocal microscopy showed that MSNs and Ce@MsNs were internalised into the cytoplasm of the pre-osteoblasts after 24 h but were not in the nucleus, avoiding any DNA and RNA modifications. Ce@MSNs provoked mineralisation of the pre-osteoablasts without osteogenic supplements, which did not occur when the cells were exposed to MSN without nanoceria. In a co-culture system of MC3T3-E1 and RAW264.7 macrophages, the Ce@MSNs exhibited antioxidant capability and stimulated cell proliferation and osteogenic responses without adding osteogenic supplements to the culture. The work brings forward an effective platform based for facile synthesis of Ce@MSNs to interact with both OBs and OCs for treatment of osteoporosis.

Osteogenic potential of sol-gel bioactive glasses containing manganese.

Bioactive glasses (BGs) are widely used for bone regeneration, and allow the incorporation of different ions with therapeutic properties into the glass network. Amongst the different ions with therapeutic benefits, manganese (Mn) has been shown to influence bone metabolism and activate human osteoblasts integrins, improving cell adhesion, proliferation and spreading. Mn has also been incorporated into bioceramics as a therapeutic ion for improved osteogenesis. Here, up to 4.4 mol% MnO was substituted for CaO in the 58S composition (60 mol% SiO2, 36 mol% CaO, 4 mol% P2O5) and its effects on the glass properties and capability to influence the osteogenic differentiation were evaluated. Mn-containing BGs with amorphous structure, high specific surface area and nanoporosity were obtained. The presence of Mn2+ species was confirmed by X-ray photoelectron spectroscopy (XPS). Mn-containing BGs presented no cytotoxic effect on human mesenchymal stem cells (hMSCs) and enabled sustained ion release in culture medium. hMSCs osteogenic differentiation stimulation and influence on the mineralisation process was also confirmed through the alkaline phosphatase (ALP) activity, and expression of osteogenic differentiation markers, such as collagen type I, osteopontin and osteocalcin, which presented higher expression in the presence of Mn-containing samples compared to control. Results show that the release of manganese ions from bioactive glass provoked human mesenchymal stem cell (hMSC) differentiation down a bone pathway, whereas hMSCs exposed to the Mn-free glass did not differentiate. Mn incorporation offers great promise for obtaining glasses with superior properties for bone tissue regeneration.

Development of anti-HER2-targeted doxorubicin-core-shell chitosan nanoparticles for the treatment of human breast cancer.

Purpose: Doxorubicin (DOX) encapsulated O-succinyl chitosan graft Pluronic® F127 (OCP) copolymer nanoparticles conjugated with an anti-HER2 monoclonal antibody were developed as targeted drug delivery vehicles for the treatment of HER2-overexpressing breast cancer. Methods: Five percent and 10% (w/w) of O-succinyl chitosan was grafted onto Pluronic® F127 using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) as mediated cross-linking agents. DOX was added to the copolymer solution to form DOX-nanoparticles before conjugation with anti-HER2 on the surface of the nanoparticles. Results: DOX was encapsulated within the NP matrices at an encapsulation efficiency of 73.69 ± 0.53% to 74.65 ± 0.44% (the initial DOX concentration was 5 µg/mL). Anti-HER2 was successfully conjugated onto the surface of the nanoparticles at a moderately high conjugation efficiency of approximately 57.23 ± 0.38% to 61.20 ± 4.42%. In the in vitro DOX dissolution study, the nanoparticle formulations exhibited a biphasic drug release with an initial burst release followed by a sustained release profile at both pH 5.0 and pH 7.4. The drug was rapidly and completely released from the nanoparticles at pH 5.0. In the in vitro cytotoxicity, the anti-HER2 conjugated OCP copolymer nanoparticles showed the lowest IC50, which indicated an increase in the therapeutic efficacy of DOX to treat human breast cancer cells with the HER2 overexpression. Conclusion: Our study shows that anti-HER2 conjugated OCP copolymer nanoparticles have the potential for the development of anticancer drug carriers.

Effects of manganese incorporation on the morphology, structure and cytotoxicity of spherical bioactive glass nanoparticles.

Bioactive glass nanoparticles (BGNPs) are of great interest in tissue engineering as they possess high dissolution rate and capability of being internalized by cells, releasing their dissolution products with therapeutic benefits intracellularly. A modified Stöber process can be applied to obtain different BGNPs compositions containing therapeutic ions while maintaining controllable particle morphology, monodispersity and reduce agglomeration. Here, BGNPs containing Mn, an ion that has been shown to influence the osteoblast proliferation and bone mineralization, were evaluated. Particles with up to 142.3 ±â€¯10.8 nm and spherical morphology were obtained after MnO incorporation in the SiO2 - CaO system. X-ray photoelectron spectroscopy (XPS) indicated the presence of Mn2+ species and also a reduction in the number of bridging oxygen bonds due to the Ca and Mn. The Ca and Mn network modifier role on the silica network was also confirmed by magic-angle spinning 29Si solid-state nuclear magnetic resonance (MAS NMR). MTT evaluation showed no reduction in the mitochondrial metabolic activity of human mesenchymal stem cells exposed to the glass ionic products. Thus, evaluation showed that Mn could be incorporated into BGNPs by the modified Stöber method while maintaining their spherical morphology and features as a promising strategy for tissue regeneration.

Human mesenchymal stem cells differentiate into an osteogenic lineage in presence of strontium containing bioactive glass nanoparticles.

While bioactive glass and ions released during its dissolution are known to stimulate osteoblast cells, the effect bioactive glass has on human stem cells is not clear. Here, we show that spherical monodispersed strontium containing bioactive nanoparticles (Sr-BGNPs) of composition 90.6 mol% SiO2, 5.0 mol% CaO, 4.4% mol% SrO (4.4%Sr-BGNPs) and 88.8 mol% SiO2, 1.8 mol% CaO, and 9.4 mol% SrO (9.4%Sr-BGNPs) stimulate bone marrow derived human stem cell (hMSC) differentiation down an osteogenic pathway without osteogenic supplements. The particles were synthesised using a modified StÓ§ber process and had diameters of 90 ±â€¯10 nm. Previous work on similar particles that did not contain Sr (80 mol% SiO2, 20 mol% CaO) showed stem cells did not differentiate when exposed to the particles. Here, both compositions of the Sr-BGNPs (up to concentration of 250 µg/mL) stimulated the early-, mid-, and late-stage markers of osteogenic differentiation and accelerated mineralisation in the absence of osteogenic supplements. Sr ions play a key role in osteogenic stem cell differentiation. Sr-BGNP dissolution products did not adversely affect hMSC viability and no significant differences in viability were measured between each particle composition. Confocal and transmission electron microscopy (TEM) demonstrated that monodispersed Sr-BGNPs were internalised and localised within vesicles in the cytoplasm of hMSCs. Degradation of particles inside the cells was observed, whilst maintaining effective cations (Ca and Sr) in their silica network after 24 h in culture. The uptake of Sr-BGNPs by hMSCs was reduced by inhibitors of specific routes of endocytosis, indicating that the Sr-BGNPs uptake by hMSCs was probably via mixed endocytosis mechanisms. Sr-BGNPs have potential as injectable therapeutic devices for bone regeneration or treatment of conditions such as osteoporosis, because of their ability deliver a sustained release of osteogenic inorganic cations, e.g. calcium (Ca) or and strontium (Sr), through particle degradation locally to cells. STATEMENT OF SIGNIFICANCE: Here, we show that 90 nm spherical strontium containing bioactive nanoparticles of stimulate bone marrow derived human stem cell (hMSC) differentiation down an osteogenic pathway without the use of osteogenic supplements. While bioactive glass and its dissolution products are known to promote excellent bone regeneration in vivo and to stimulate osteoblast cells to produce bone matrix in vitro, their effect on human stem cells is not clear. Previously our nanoparticles that contained only SiO2 and CaO did not provoke human bone marrow or adipose derived stem cell differentiation.

Silk fibroin-bioactive glass based advanced biomaterials: towards patient-specific bone grafts.

A major challenge in bone tissue engineering is to develop patient-specific, defect-site specific grafts capable of triggering specific cell signaling pathways. We could programmably fabricate the 3D printed bone constructs via direct ink writing of silk-gelatin-bioactive glass (SF-G-BG) hybrids using two different compositions of melt-derived bioactive glasses (with and without strontium) and compared against commercial 45S5 Bioglass®. Physico-chemical characterization revealed that released ions from bioactive glasses inhibited the conformational change of Bombyx mori silk fibroin protein (from random coil to ß-sheet conformation), affecting printability of the SF-G-BG ink. In-depth molecular investigations showed that strontium containing SF-G-BG constructs demonstrated superior osteogenic differentiation of mesenchymal stem cells (TVA-BMSCs) over 21 days towards osteoblastic (marked by upregulated expression of runt related transcription factor, alkaline phosphatase, osteopontin, osteonectin, integrin bone sialoprotein, osteocalcin) and osteocytic (marked by podoplanin, dentin matrix acidic phosphoprotein, sclerostin) phenotype compared to other BG compositions and silk-gelatin alone. Moreover, ionic release from bioactive glasses in the silk-gelatin ink triggered the activation of signaling pathways (BMP-2, BMP-4 and IHH), which are critical in regulating bone formation in vivo. Overall, the presence of strontium containing bioactive glass in silk-gelatin matrices provided appropriate cues in regulating the development of custom-made 3D in vitro human bone constructs.

In vitro osteogenesis by intracellular uptake of strontium containing bioactive glass nanoparticles.

Monodispersed strontium containing bioactive glass nanoparticles (Sr-BGNPs) with two compositions were synthesised, through a modified sol-gel Stöber process, wherein silica nanoparticles (SiO2-NPs) were formed prior to incorporation of calcium and strontium, with diameters of 90 ±â€¯10 nm. The osteogenic response of a murine preosteoblast cell line, MC3T3-E1, was investigated in vitro for a nanoparticle concentration of 250 µg/mL with compositions of 87 mol% SiO2, 7 mol% CaO, 6 mol% SrO and 83 mol% SiO2, 3 mol% CaO, 14 mol% SrO. Dissolution studies in minimum essential media (α-MEM) at pH 7.4 and artificial lysosomal fluid (ALF) at pH 4.5 showed that the particles dissolved and that Sr2+ ions were released from Sr-BGNPs in both environments. Both particle compositions and their ionic dissolution products enhanced the alkaline phosphatase (ALP) activity of the cells and calcium deposition. Immunohistochemistry (IHC) staining of Col1a1, osteocalcin (OSC) and osteopontin (OSP) showed that these proteins were expressed in the MC3T3-E1 cells following three weeks of culture. In the basal condition, the late osteogenic differentiation markers, OSC and OSP, were more overtly expressed by cells cultured with Sr-BGNPs with 14 mol% SrO and their ionic release products than in the control condition. Col1a1 expression was only slightly enhanced in the basal condition, but was enhanced further by the osteogenic supplements. These data demonstrate that Sr-BGNPs accelerate mineralisation without osteogenic supplements. Sr-BGNPs were internalised into MC3T3-E1 cells by endocytosis and stimulated osteogenic differentiation of the pre-osteoblast cell line. Sr-BGNPs are likely to be beneficial for bone regeneration and the observed osteogenic effects of these particles can be attributed to their ionic release products. STATEMENT OF SIGNIFICANCE: We report, for the first time, that monodispersed bioactive glass nanoparticles (∼90 nm) are internalised into preosteoblast cells by endocytosis but by unspecific mechanisms. The bioactive nanoparticles and their dissolution products (without the particles present) stimulated the expression of osteogenic markers from preosteoblast cells without the addition of other osteogenic supplements. Incorporating Sr into the bioactive glass nanoparticle composition, in addition to Ca, increased the total cation content (and therefore dissolution rate) of the nanoparticles, even though nominal total cation addition was constant, without changing size or morphology. Increasing Sr content in the nanoparticles and in their dissolution products enhanced osteogenesis in vitro. The particles therefore have great potential as an injectable therapeutic for bone regeneration, particularly in patients with osteoporosis, for which Sr is known to be therapeutic agent.