Effect of Fibrillization pH on Gelation Viscoelasticity and Properties of Biofabricated Dense Collagen Matrices via Gel Aspiration-Ejection.

Reconstituted hydrogels based on the self-assembly of acid-solubilized collagen molecules have been extensively used as in vitro models and precursors in biofabrication processes. This study investigated the effect of fibrillization pH-ranging from 4 to 11-on real-time rheological property changes during the gelation of collagen hydrogels and its interplay with the properties of subsequently biofabricated dense collagen matrices generated via automated gel aspiration-ejection (GAE). A contactless, nondestructive technique was used to characterize the temporal progression in shear storage modulus (G', or stiffness) during collagen gelation. There was a relative increase in G' of the hydrogels from 36 to 900 Pa with an increase in gelation pH. Automated GAE, which simultaneously imparts collagen fibrillar compaction and alignment, was then applied to these precursor collagen hydrogels to biofabricate native extracellular matrix-like densified gels. In line with viscoelastic properties, only hydrogels fibrillized in the 6.5 < pH ≤ 10 range could be densified via GAE. There was an increase in both fibrillar density and alignment in the GAE-derived matrices with an increase in gelation pH. These factors, combined with a higher G' in the alkaline precursor hydrogels, led to a significant increase in the micro-compressive modulus of GAE-densified gels of pH 9 and 10. Furthermore, NIH/3T3 fibroblast-seeded GAE-derived matrices densified from gels fibrillized in the pH range of 7 to 10 exhibited low cell mortality with >80% viability. It is anticipated that the results of this study can be potentially applicable to other hydrogel systems, as well as biofabrication techniques involving needles or nozzles, such as injection and bioprinting.

Aluminum-free glass ionomer cements containing 45S5 Bioglass® and its bioglass-ceramic.

Although the incorporation of bioactive glasses into glass ionomer cements (GICs) has led to promising results, using a bioactive glass as the only solid component of GICs has never been investigated. In this study, we developed an Al-free GIC with standard compressive strength using various combinations of 45S5 Bioglass® and its glass-ceramic as the solid component. The glass-ceramic particles with 74% crystallinity were used for this purpose as they can best act as both remineralizing and reinforcing agents. Strengthening mechanisms including crack deflection and crack-tip shielding were activated for the GICs containing 50-50 wt% bioglass and bioglass-ceramic as the optimum ratio. The progression of the GIC setting reaction at its early stages was also monitored and verified. We also discussed that our bimodal particle size distribution containing both micron- and nanosized particles may enhance the packing density and integrity of the structure of the cements after setting. In such GICs produced in this study, the toxic effects of Al are avoided while chemical bonds are expected to form between the cement and the surrounding hard tissue(s) through interfacial biomineralization and adhesion.

Ultraporous Membranes Electrospun from Nonsolvent-Induced Phase-Separated Ternary Systems.

Electrospinning of nonsolvent-induced phase-separated ternary (NIPST) systems has gained a lot of interest due to its potential to produce (nano)fibers, which are superficially and internally porous with nanoscale surface roughness. Membranes produced from such systems are expected to have a high specific surface area (SSA; e.g., more than 50 m2 g-1 ), an essential requirement for many of their applications. In spite of their advantages and potential, there are major issues regarding the electrospinning of NIPST systems that are not systematically addressed in the literature. In this paper, the most recent developments are reported and the potential and challenges associated with the electrospinning of NIPST systems are discussed. Furthermore, the essential steps to improve and optimize the electrospinning process of these systems are concisely discussed. By developing a modified time-dependent rheological model, a time range can be defined for NIPST systems as "electrospinnability window," in which fiber functionality and characteristics can be tailored through aging of the systems prior to electrospinning. Some potential post-treatment processes are also proposed based on the results of recent studies to stabilize as-electrospun membranes without damaging their highly porous fibers, which can guarantee their in-service mechanical and morphological stability.

In Vitro Evaluation of Real-Time Viscoelastic and Coagulation Properties of Various Classes of Topical Hemostatic Agents Using a Novel Contactless Nondestructive Technology.

Hemorrhaging is the main cause of death among combat and civilian injuries and has significant clinical and economic consequences. Despite their vital roles in bleeding management, an optimal topical hemostatic agent (HA) has yet to be developed for a particular scenario. This is partly due to a lack of an overarching quantitative testing technology to characterize the various classes of HAs in vitro. Herein, the feasibility of a novel, contactless, and nondestructive technique to quantitatively measure the shear storage modulus (G') and clotting properties of whole blood in contact with different dosages of eight topical HAs, including particulates and gauze-like and sponge-like systems, was assessed. The real-time G'-time profiles of these blood/HA systems revealed their distinct biomechanical behavior to induce and impact coagulation. These were analyzed to characterize the clot initiation time, clotting rate, clotting time, and apparent stiffness of the formed clots (both immediately and temporally), which were correlated with their reported hemostatic mechanisms of action. Moreover, the HAs that worked independently from the natural blood clotting cascade were identified and quantified through this technology. In sum, this study indicated that the nondestructive nature of the technology may offer a promising tool for accurate, quantitative in vitro measurements of the clotting properties of various classes of HAs, which may be used to better predict their in vivo outcomes.

A nondestructive contactless technique to assess the viscoelasticity of blood clots in real-time.

There is a need for reliable and quantitative real-time assessment of blood properties to study and treat a broad spectrum of disorders and cardiovascular diseases as well as to test the efficacy of hemostatic agents. In this study, the real-time changes in viscoelastic/rheological properties of bovine whole blood during coagulation induced by different concentrations of calcium chloride (CaCl2; 15, 25, 35 and 45 mM) was investigated. For this purpose, a novel, contactless technique was used to accurately measure the clotting characteristics under controlled and sterile conditions. It was demonstrated that, increasing the calcium concentration from low values (i.e., 15 and 25 mM), led to shorter reaction time; however, a further increase in calcium concentration (i.e., 35 and 45 mM) favored longer reaction times. Additionally, increasing the CaCl2 concentration resulted in higher shear storage modulus (i.e., stiffer clots). These results were also comparable to those generated by thromboelastrograph, a clinically established technique, as well as a conventional rheometer, which quantitatively verified the high correlation of the shear storage modulus data. In sum, the non-destructive testing technique used in this study is reproducible and sensitive in measuring clot formation kinetics, which could be applied to assess the efficacy of hemostatic agents, and may also contribute to better diagnosing relevant circulatory system diseases and conditions.

Poly(d,l-Lactic acid) Composite Foams Containing Phosphate Glass Particles Produced via Solid-State Foaming Using CO2 for Bone Tissue Engineering Applications.

This study reports on the production and characterization of highly porous (up to 91%) composite foams for potential bone tissue engineering (BTE) applications. A calcium phosphate-based glass particulate (PGP) filler of the formulation 50P2O5-40CaO-10TiO2 mol.%, was incorporated into biodegradable poly(d,l-lactic acid) (PDLLA) at 5, 10, 20, and 30 vol.%. The composites were fabricated by melt compounding (extrusion) and compression molding, and converted into porous structures through solid-state foaming (SSF) using high-pressure gaseous carbon dioxide. The morphological and mechanical properties of neat PDLLA and composites in both nonporous and porous states were examined. Scanning electron microscopy micrographs showed that the PGPs were well dispersed throughout the matrices. The highly porous composite systems exhibited improved compressive strength and Young's modulus (up to >2-fold) and well-interconnected macropores (up to ~78% open pores at 30 vol.% PGP) compared to those of the neat PDLLA foam. The pore size of the composite foams decreased with increasing PGPs content from an average of 920 µm for neat PDLLA foam to 190 µm for PDLLA-30PGP. Furthermore, the experimental data was in line with the Gibson and Ashby model, and effective microstructural changes were confirmed to occur upon 30 vol.% PGP incorporation. Interestingly, the SSF technique allowed for a high incorporation of bioactive particles (up to 30 vol.%-equivalent to ~46 wt.%) while maintaining the morphological and mechanical criteria required for BTE scaffolds. Based on the results, the SSF technique can offer more advantages and flexibility for designing composite foams with tunable characteristics compared to other methods used for the fabrication of BTE scaffolds.

Glass ionomer cements with enhanced mechanical and remineralizing properties containing 45S5 bioglass-ceramic particles.

The clinical applications of glass ionomer cements (GICs) are limited by their relatively poor mechanical properties and insufficient remineralizing capacity. In this study, we developed hybrid GICs with improved mechanical and remineralizing properties via incorporation of an optimum amount (5 wt%) of 45S5 bioglass-ceramic particles. Also, we found that bioglass-ceramic particles with 74% crystallinity best act as both remineralizing and reinforcing agents. The degree of crystallinity of the additives, is overlooked in this context in other research. At around 74% crystallinity, there is sufficient amount of combeite and an amorphous phosphorous-rich phase in the 45S5 bioglass-ceramic particles to respectively promote their reinforcing role and allow them to effectively partake in the setting process creating an excellent interfacial bond with the GIC matrix. As a result, several strengthening mechanisms such as crack deflection and crack-tip shielding are activated within the hybrid GIC containing 5 wt% bioglass-ceramic with 74% crystallinity, contributing to its improved mechanical properties. The enhanced remineralizing and mechanical properties of such hybrid GICs can potentially improve their in vivo performance and broaden their clinical applications.

Morphological examination of highly porous polylactic acid/Bioglass® scaffolds produced via nonsolvent induced phase separation.

In this study, we produce highly porous (up to ∼91%) composite scaffolds of polylactic acid (PLA) containing 2 wt % sol-gel-derived 45S5 Bioglass® particles via nonsolvent induced phase separation at -23°C with no sacrificial phases involved. Before the incorporation of the bioglass with PLA, the particles are surface modified with a silane coupling agent which effectively diminishes agglomeration between them leading to a better dispersion of bioactive particles throughout the scaffold. Interestingly, the incorporation route (via solvent dichloromethane or nonsolvent hexane) of the surface modified particles in the foaming process has the greatest impact on porosity, crystallinity, and morphology of the scaffolds. The composite scaffolds with a morphology consisting of both mesopores and large macropores, which is potentially beneficial for bone regeneration applications, are examined further. SEM images show that the surface modified bioglass particles take-up a unique configuration within the mesoporous structure of these scaffolds ensuring that the particles are well interlocked but not completely covered by PLA such that they can be in contact with physiological fluids. The results of preliminary in vitro tests confirm that this PLA/bioglass configuration promotes the interaction of the bioactive phase with physiological fluids. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2433-2442, 2017.

Synthesis of 45S5 Bioglass® via a straightforward organic, nitrate-free sol-gel process.

More than four decades after the discovery of 45S5 Bioglass® as the first bioactive material, this composition is still one of the most promising materials in the tissue engineering field. Sol-gel-derived bioactive glasses generally possess improved properties over other bioactive glasses, because of their highly porous microstructure and unique surface chemistry which accelerate hydroxyapatite formation. In the current study, a new combination of precursors with lactic acid as the hydrolysis catalyst have been employed to design an organic, nitrate-free sol-gel procedure for synthesizing of 45S5 Bioglass®. This straightforward route is able to produce fully amorphous submicron particles of this glass with an appropriately high specific surface area on the order of ten times higher than that of the melt-derived glasses. These characteristics are expected to lead to rapid hydroxyapatite formation and consequently more efficient bone bonding.