Surface Modification of Biodegradable Porous Mg Bone Scaffold Using Polycaprolactone/Bioactive Glass Composite

A reduction in the degradation rate of magnesium (Mg) and its alloys is in high demand to enable these materials to be used in orthopedic applications. For this purpose, in this paper, a biocompatible polymeric layer reinforced with a bioactive ceramic made of polycaprolactone (PCL) and bioactive glass (BG) was applied on the surface of Mg scaffolds using dip-coating technique under low vacuum. The results indicated that the PCL-BG coated Mg scaffolds exhibited noticeably enhanced bioactivity compared to the uncoated scaffold. Moreover, the mechanical integrity of the Mg scaffolds was improved using the PCL-BG coating on the surface. The stable barrier property of the coatings effectively delayed the degradation activity of Mg scaffold substrates. Moreover, the coatings induced the formation of apatite layer on their surface after immersion in the SBF, which can enhance the biological bone in-growth and block the microcracks and pore channels in the coatings, thus prolonging their protective effect. Furthermore, it was shown that a three times increase in the concentration of PCL-BG noticeably improved the characteristics of scaffolds including their degradation resistance and mechanical stability. Since bioactivity, degradation resistance and mechanical integrity of a bone substitute are the key factors for repairing and healing fractured bones, we suggest that PCL-BG is a suitable coating material for surface modification of Mg scaffolds

Collagenous Matrix Supported by A 3D-Printed Scaffold for Osteogenic Differentiation of Dental Pulp Cells

Objective A systematic characterization of hybrid scaffolds, fabricated based on combinatorial additive manufacturing technique and freeze-drying method, is presented as a new platform for osteoblastic differentiation of dental pulp cells (DPCs). Methods The scaffolds were consisted of a collagenous matrix embedded in a 3D-printed beta-tricalcium phosphate (β-TCP) as the mineral phase. The developed construct design was intended to achieve mechanical robustness owing to 3D-printed β-TCP scaffold, and biologically active 3D cell culture matrix pertaining to the Collagen extracellular matrix. The β-TCP precursor formulations were investigated for their flow-ability at various temperatures, which optimized for fabrication of 3D printed scaffolds with interconnected porosity. The hybrid constructs were characterized by 3D laser scanning microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and compressive strength testing. Results The in vitro characterization of scaffolds revealed that the hybrid β-TCP/Collagen constructs offer superior DPCs proliferation and alkaline phosphatase (ALP) activity compared to the 3D-printed β-TCP scaffold over three weeks. Moreover, it was found that the incorporation of TCP into the Collagen matrix improves the ALP activity. Significance The presented results converge to suggest the developed 3D-printed β-TCP/Collagen hybrid constructs as a new platform for osteoblastic differentiation of DPCs for craniomaxillofacial bone regeneration

From Solvent-Free Microspheres to Bioactive Gradient Scaffolds

A solvent-free microsphere sintering technique was developed to fabricate scaffolds with pore size gradient for tissue engineering applications. Poly(D,L-Lactide) microspheres were fabricated through an emulsification method where TiO2 nanoparticles were employed both as particulate emulsifier in the preparation procedure and as surface modification agent to improve bioactivity of the scaffolds. A fine-tunable pore size gradient was achieved with a pore volume of 30±2.6%. SEM, EDX, XRD and FTIR analyses all confirmed the formation of bone-like apatite at the 14th day of immersion in Simulated Body Fluid (SBF) implying the ability of our scaffolds to bond to living bone tissue. In vitro examination of the scaffolds showed progressive activity of the osteoblasts on the scaffold with evidence of increase in its mineral content. The bioactive scaffold developed in this study has the potential to be used as a suitable biomaterial for bone tissue engineering and hard tissue regeneration

Doctor of Philosophy

dissertationAdvances in the synthesis and characterization of engineered silica nanoparticles (SNPs) are not matched by careful assessment of their effects on biological systems, environmental health, and safety. Better understanding of how silica nanoparticles interact with biological fluids and cells is required to predict safety, mechanism of action, dissolution, clearance, and possible adverse effects. In this dissertation the impact of physicochemical properties of SNPs such as size, porosity, density, and surface functionality on cellular toxicity and genomic response was explored on RAW 264.7 macrophages. Size-dependent cytotoxicity was observed. Mesoporous SNPs showed higher LC50 of 223.6±15.6 compared to the similar size nonporous SNPs with LC50 of 33.7±0.6. The observed lower cell association and toxicity of mesoporous particles is probably related to the lower density of silanol groups per square nm on surface. In addition, decreased sedimentation, cell uptake, and toxicity for lower density particles with rattle structure and under flow conditions was observe compared to nonporous particles. The influence of size, porosity, and surface functionality of SNPs on early response of RAW 264.7 macrophages at sub-toxic doses revealed no significant gene expression alteration for nonporous SNPs at 4 h incubation time, however, mesoporous SNPs induced genomic response associated with lysosomal activity. The global gene expression analysis of mesoporous and nonporous nanoparticles with similar diameters of approximately 500 nm showed time- and dose-dependent gene expression response of macrophages as a function of porosity of SNPs

Synthesis and Characterization of Encapsulated Nanosilica Particles with an Acrylic Copolymer by in Situ Emulsion Polymerization Using Thermoresponsive Nonionic Surfactant

Nanocomposites of encapsulated silica nanoparticles were prepared by in situ emulsion polymerization of acrylate monomers. The synthesized material showed good uniformity and dispersion of the inorganic components in the base polymer, which enhances the properties of the nanocomposite material. A nonionic surfactant with lower critical solution temperature (LCST) was used to encapsulate the silica nanoparticles in the acrylic copolymer matrix. This in situ method combined the surface modification and the encapsulation in a single pot, which greatly simplified the process compared with other conventional methods requiring separate processing steps. The morphology of the encapsulated nanosilica particles was investigated by dynamic light scattering (DLS) and transmission electron microscopy (TEM), which confirmed the uniform distribution of the nanoparticles without any agglomerations. A neat copolymer was also prepared as a control sample. Both the neat copolymer and the prepared nanocomposite were characterized by Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analyses (TGA), dynamic mechanical thermal analysis (DMTA) and the flame resistance test. Due to the uniform dispersion of the non-agglomerated nanoparticles in the matrix of the polymer, TGA and flame resistance test results showed remarkably improved thermal stability. Furthermore, DMTA results demonstrated an enhanced storage modulus of the nanocomposite samples compared with that of the neat copolymer, indicating its superior mechanical properties

Synthesis and Characterization of Encapsulated Nanosilica Particles with an Acrylic Copolymer by in Situ Emulsion Polymerization Using Thermoresponsive Nonionic Surfactant

Nanocomposites of encapsulated silica nanoparticles were prepared by in situ emulsion polymerization of acrylate monomers. The synthesized material showed good uniformity and dispersion of the inorganic components in the base polymer, which enhances the properties of the nanocomposite material. A nonionic surfactant with lower critical solution temperature (LCST) was used to encapsulate the silica nanoparticles in the acrylic copolymer matrix. This in situ method combined the surface modification and the encapsulation in a single pot, which greatly simplified the process compared with other conventional methods requiring separate processing steps. The morphology of the encapsulated nanosilica particles was investigated by dynamic light scattering (DLS) and transmission electron microscopy (TEM), which confirmed the uniform distribution of the nanoparticles without any agglomerations. A neat copolymer was also prepared as a control sample. Both the neat copolymer and the prepared nanocomposite were characterized by Fourier transform infrared spectroscopy (FTIR), thermal gravimetric analyses (TGA), dynamic mechanical thermal analysis (DMTA) and the flame resistance test. Due to the uniform dispersion of the non-agglomerated nanoparticles in the matrix of the polymer, TGA and flame resistance test results showed remarkably improved thermal stability. Furthermore, DMTA results demonstrated an enhanced storage modulus of the nanocomposite samples compared with that of the neat copolymer, indicating its superior mechanical properties

Biomineralization and Biocompatibility Studies of Bone Conductive Scaffolds Containing Poly(3,4-Ethylenedioxythiophene):Poly(4-Styrene Sulfonate) (PEDOT:PSS)

Considering the well-known phenomenon of enhancing bone healing by applying electromagnetic stimulation, manufacturing conductive bone scaffolds is on demand to facilitate the delivery of electromagnetic stimulation to the injured region, which in turn significantly expedites the healing procedure in tissue engineering methods. For this purpose, hybrid conductive scaffolds composed of poly(3,4-ethylenedioxythiophene), poly(4-styrene sulfonate) (PEDOT:PSS), gelatin (Gel), and bioactive glass (BaG) were produced employing freeze drying technique. Concentration of PEDOT:PSS were optimized to design the most appropriate conductive scaffold in terms of biocompatibility and cell proliferation. More specifically, scaffolds with four different compositions of 0, 0.1, 0.3 and 0.6 % (w/w) PEDOT:PSS in the mixture of 10 % (w/v) Gel and 30 % (w/v) BaG were synthesized. Immersing the scaffolds in simulated body fluid (SBF), we evaluated the bioactivity of samples, and the biomineralization were studied in details using scanning electron microscopy, energy dispersive spectroscopy, X-ray diffraction analysis and Fourier transform infrared spectroscopy. By performing cytocompatibility analyses for 21 days using adult human mesenchymal stem cells, we concluded that the scaffolds with 0.3 % (w/w) PEDOT:PSS and conductivity of 170 μS/m has the optimized composition and further increasing the PEDOT:PSS content has inverse effect on cell proliferation. Based on our finding, addition of this optimized amount of PEDOT:PSS to our composition can increase the cell viability more than 4 times compared to a nonconductive composition

Redox-Responsive Polysulfide-Based Biodegradable Organosilica Nanoparticles for Delivery of Bioactive Agents

Design and development of silica nanoparticles (SiO₂ NPs) with a controlled degradation profile promises effective drug delivery with a predetermined carrier elimination profile. In this research, we fabricated a series of redox-responsive polysulfide-based biodegradable SiO₂ NPs with low polydispersity and with variations in size (average diameters of 58 ± 7, 108 ± 11, 110 ± 9, 124 ± 9, and 332 ± 6 nm), porosity, and composition (disulfide vs tetrasulfide bonds). The degradation kinetics of the nanoparticles was analyzed in the presence of 8 mM glutathione (GSH), mimicking the intracellular reducing condition. Results indicate that porosity and core composition play the predominant roles in the degradation rate of these nanoparticles. The 108 nm mesoporous disulfide-based nanoparticles showed the highest degradation rate among all the synthesized nanoparticles. Transmission electron microscopy (TEM) reveals that nonporous nanoparticles undergo surface erosion, while porous nanoparticles undergo both surface and bulk erosion under reducing environment. The cytotoxicity of these nanoparticles in RAW 264.7 macrophages was evaluated. Results show that all these nanoparticles with the IC₅₀ values ranging from 233 ± 42 to 705 ± 17 μg mL^–1 do not have cytotoxic effect in macrophages at concentrations less than 125 μg mL^–1. The degradation products of these nanoparticles collected within 15 days did not show cytotoxicity in the same macrophage cell line after 24 h of incubation. <i>In vitro</i> doxorubicin (DOX) release was examined in 108 nm mesoporous disulfide-based nanoparticles in the absence and presence of 8 mM GSH. It was shown that drug release depends on intracellular reducing conditions. Due to their ease of synthesis and scale up, robust structure, and the ability to control size, composition, release, and elimination, biodegradable SiO₂ NPs provide an alternative platform for delivery of bioactive and imaging agents