Portonovo S. Ayyaswamy

Fabrication, characterization and evaluation of bioceramic hollow microspheres used as microcarriers for 3-D bone tissue formation in rotating bioreactors

Novel bioactive ceramic hollow microspheres with an apparent density in the range 0.8-1.0 g cm(-3) have been developed as microcarriers for 3-D bone tissue formation in rotating-wall vessels (RWV). Hollow ceramic microspheres with a composition of 58-72% SiO2, 28-42% Al2O3 (wt%) and an apparent density 0.8-1.0 g cm(-3) were pretreated in 1.0 N NaOH for 2 h before being coated with synthesized calcium hydroxyapatite (HA) particulate sol. The HA-coated hollow microspheres were sintered for 1 h at 600, 800 and 1000 degrees C. SEM analysis revealed that the grain size and pore size of the calcium phosphate coating increased with the sintering temperature. FTIR analysis showed that crystalline calcium hydroxyapatite was present in the coatings sintered at 600 and 800 degrees C. When sintered at 1000 degrees C, the coating consisted of alpha-tricalcium phosphate. All the coatings adhered well, independent of sintering temperature. The trajectory analysis revealed that the hollow microsphere remained suspended in a rotating-wall vessel (RWV), and experienced a low shear stress (approximately 0.6 dyn cm(-2)). Cell culture studies using rat bone marrow stromal cells and osteosarcoma cells (ROS 17/2.8) showed that the cells attached to and formed 3-D aggregates with the hollow microspheres in a RWV. Extracellular matrix was observed in the aggregates. These data suggest that these hollow bioactive ceramic microspheres can be used as microcarriers for 3-D bone tissue formation in vitro, as well as for the study of the effects of microgravity on bone cell functions.

New bioactive, degradable composite microspheres as tissue engineering substrates.

Novel bioactive, degradable polymer/glass/ceramic composite microspheres were developed using a solid-in-oil-in-water (s/o/w) emulsion solvent removal method. Modified bioactive glass (MBG) powders were encapsulated into the polylactic acid (PLA) matrix. Scanning electron microscopy and energy-dispersive X-ray analyses revealed that the MBG powders were mostly embedded in the polymer matrix, and submicron-size pores were present at the surface. Immersion in simulated physiological fluid (SPF) was used to evaluate the surface reactivity of the microspheres. The polymeric surface was fully transformed into carbonated calcium hydroxyapatite (c-HA) after 3 weeks of immersion. In contrast, PLA microspheres showed no evidence of any calcium phosphate deposition. Ion concentration analyses revealed a decrease in Ca and P concentrations and an increase in Si concentration in the SPF immersed with composite microspheres during the 3-week period. The Ca and P uptake rates decreased after 2 days of incubation. This coincided with the decrease of the Si release rate. These data lend support to the suggestion that the Si released from the MBG powders present in the polymer matrix is involved in the formation of the Ca-P layer. Our results support the concept that these new bioactive, degradable composite microspheres may serve as microcarriers for synthesis of bone and other tissues in vitro and in vivo.

Heat Transport Mechanisms in Vascular Tissues: A Model Comparison

We have conducted a parametric comparison of three different vascular models for describing heat transport in tissue. Analytical and numerical methods were used to predict the gross temperature distribution throughout the tissue and the small-scale temperature gradients associated with thermally significant blood vessels. The models are: an array of unidirectional vessels, an array of countercurrent vessels, and a set of large vessels feeding small vessels which then drain into large vessels. We show that three continuum formulations of bioheat transfer (directed perfusion, effective conductivity, and a temperature-dependent heat sink) are limiting cases of the vascular models with respect to the thermal equilibration length of the vessels. When this length is comparable to the width of the heated region of tissue, the local temperature changes near the vessels can be comparable to the gross temperature elevation. These results are important to the use of thermal techniques used to measure the blood perfusion rate and in the treatment of cancer with local hyperthermia.

Natural convection flow in a finite, rectangular slot arbitrarily oriented with respect to the gravity vector

Solutions to the stationary two-dimensional equations of motion governing natural convection flow of a large Prandtl number Boussinesq fluid contained in a differentially heated inclined rectangular slot have been obtained by the Galerkin method. The problem has been solved for perfectly conductive and adiabatic boundary conditions on the border strips. The range of parameters investigated include: Rayleigh numbers up to 2 × 106, aspect ratios from 0.1 to 20, and tilt angles from −30° (bottom plate hotter) to + 75° (top plate hotter). These parameters describe both the conduction and boundary layer regimes. The computed flow distributions, including the appearance of multicellular flows, the temperature profiles, and the heat transfer predictions compare favorably with experimental results, and with other numerical studies.

Computational model for nanocarrier binding to endothelium validated using in vivo, in vitro, and atomic force microscopy experiments

A computational methodology based on Metropolis Monte Carlo (MC) and the weighted histogram analysis method (WHAM) has been developed to calculate the absolute binding free energy between functionalized nanocarriers (NC) and endothelial cell (EC) surfaces. The calculated NC binding free energy landscapes yield binding affinities that agree quantitatively when directly compared against analogous measurements of specific antibody-coated NCs (100 nm in diameter) to intracellular adhesion molecule-1 (ICAM-1) expressing EC surface in in vitro cell-culture experiments. The effect of antibody surface coverage (σs) of NC on binding simulations reveals a threshold σs value below which the NC binding affinities reduce drastically and drop lower than that of single anti-ICAM-1 molecule to ICAM-1. The model suggests that the dominant effect of changing σs around the threshold is through a change in multivalent interactions; however, the loss in translational and rotational entropies are also important. Consideration of shear flow and glycocalyx does not alter the computed threshold of antibody surface coverage. The computed trend describing the effect of σs on NC binding agrees remarkably well with experimental results of in vivo targeting of the anti-ICAM-1 coated NCs to pulmonary endothelium in mice. Model results are further validated through close agreement between computed NC rupture-force distribution and measured values in atomic force microscopy (AFM) experiments. The three-way quantitative agreement with AFM, in vitro (cell-culture), and in vivo experiments establishes the mechanical, thermodynamic, and physiological consistency of our model. Hence, our computational protocol represents a quantitative and predictive approach for model-driven design and optimization of functionalized nanocarriers in targeted vascular drug delivery.

Small-scale temperature fluctuations in perfused tissue during local hyperthermia

We develop analytical expressions (scaling laws) for the local temperature fluctuations near isolated and countercurrent blood vessels during hyperthermia. These scaling laws relate the magnitude of such fluctuations to the size of the heated region and to the thermal equilibration length of the vessels. A new equilibration length is identified for countercurrent vessels. Significant temperature differences are predicted between the vessels and the immediately adjacent tissue when the equilibration length is comparable to or longer than the size of the heated tissue region. Countercurrent vessels are shown to have shorter equilibration lengths and produce smaller temperature fluctuations than isolated vessels of the same size.

A front tracking method for a deformable intravascular bubble in a tube with soluble surfactant transport

Based on a front tracking scheme, we have presented a comprehensive algorithm for the study of a deformable bubble moving in a tube in the presence of a soluble or an insoluble surfactant. The emphasis here is on the dynamic adsorption of the soluble surfactant which non-linearly alters the surface tension, and this in turns affects the flow and transport in a complicated way. Furthermore, since a bubble-liquid interface is being examined, there is a need to accommodate a concentration jump across the interface in the evaluation of flow and transport. Standard numerical procedures need to be modified to accommodate this feature. Based on the physics governing the problem, an axisymmetric formulation is found to be adequate and is thus considered. The adsorption scheme for the soluble surfactant is carefully designed such that the total mass of the surfactant is well conserved, and the mass flux is accurately resolved by using an interface indicator function. This represents an advance in treating problems of this class. Tests on the efficacy of various aspects of the algorithm have been carried out. The algorithm has the flexibility of studying different models for adsorption/desorption and surfactant surface tension models, such as the Langmuir and the Frumkin models. These models have significant practical relevance. The numerical results obtained are qualitatively consistent with results where available. The results presented include an example of Marangoni flow which causes a bubble to propel out of its initial static location due to the development of a surface tension gradient. It is also shown that the bubble motion in Poiseuille flow may be significantly slowed down due to the presence of a soluble surfactant in the bulk medium. In that case, the Marangoni induced motion is in a direction opposite to that driven by the bulk pressure. Our study indicates that as the location of the adsorptive interface gets closer to the tube wall, the bulk fluid in the vicinity of the interface may become depleted of surfactant, an observation that has particular significance in understanding gas embolism and for developing therapeutic measures.

Escherichia coli Biofilms Formed under Low-Shear Modeled Microgravity in a Ground-Based System

Bacterial biofilms cause chronic diseases that are difficult to control. Since biofilm formation in space is well documented and planktonic cells become more resistant and virulent under modeled microgravity, it is important to determine the effect of this gravity condition on biofilms. Inclusion of glass microcarrier beads of appropriate dimensions and density with medium and inoculum, in vessels specially designed to permit ground-based investigations into aspects of low-shear modeled microgravity (LSMMG), facilitated these studies. Mathematical modeling of microcarrier behavior based on experimental conditions demonstrated that they satisfied the criteria for LSMMG conditions. Experimental observations confirmed that the microcarrier trajectory in the LSMMG vessel concurred with the predicted model. At 24 h, the LSMMG Escherichia coli biofilms were thicker than their normal-gravity counterparts and exhibited increased resistance to the general stressors salt and ethanol and to two antibiotics (penicillin and chloramphenicol). Biofilms of a mutant of E. coli, deficient in sigma(s), were impaired in developing LSMMG-conferred resistance to the general stressors but not to the antibiotics, indicating two separate pathways of LSMMG-conferred resistance.

3D BONE TISSUE ENGINEERED WITH BIOACTIVE MICROSPHERES IN SIMULATED MICROGRAVITY

SummaryThree-dimensional (3D) osteoblast cell cultures were obtained in rotating-wall vessels (RWV), simulating microgravity. Three types of bioactive microcarriers, specifically modified bioactive glass particles, bioceramic hollow microspheres, and biodegradable bioactive glass-polymer composite microspheres, were developed and used with osteblasts. The surfaces of composite microspheres fully transformed into bone apatite after 2-wk immersion in simulated physiological fluid, which demonstrated their bone-bonding ability. The motion of microcarriers in RWVs was photographically recorded and numerically analyzed. The trajectories of hollow microspheres showed that they migrated and eventually stayed around at the central region of the RWV. At their surfaces, shear stresses were low. In contrast, solid glass or polymer particles moved toward and finally bounced off the outer wall of the RWVs. Cell culture studies in the RW, using bone marrow stromal cells showed that the cells attached to and formed 3D aggregates with the hollow microspheres. Extracellular matrix and mineralization were observed in the aggregates. Cell culture studies also confirmed the ability of the composite microspheres to support 3D bone-like tissue formation. These data suggest that the new hollow bioceramic microspheres and degradable composite microspheres can be used as microcarriers for 3D bone tissue engineering in microgravity. They also have potential applications as drug delivery systems.

Hydrodynamics and Heat Transfer Associated with Condensation on a Moving Drop: Solutions for Intermediate Reynolds Numbers

The hydrodynamics and heat/mass transport associated with condensation on a moving drop have been investigated for the intermediate Reynolds-number range of drop motion ( Re = O (100)). The drop environment is a mixture of saturated vapour and a non-condensable. The formulation entails a simultaneous solution of the quasi-steady elliptic partial differential equations that describe the flow field and transport in the gaseous phase, and the motion inside the liquid drop. The heat transport inside the drop is treated as a transient process. Results are reported for the interfacial velocities, drag, external and internal flow structure, heat flux, drop growth rate and temperature–time history inside the drop. The results obtained here have been compared with experimental data where available, and these show excellent agreement. The results reveal several novel features. The surface-shear stress increases with condensation. The pressure level in the rear of the drop is higher. As a consequence, the friction drag is higher and the pressure drag is lower. The total drag coefficient increases with condensation rate for small values of drop size or temperature differential, and it decreases for large values of these parameters. The volume of the separated-flow region in the rear of the drop decreases with condensation. At very high rates of condensation, the recirculatory wake is completely suppressed. Condensation also delays the appearance of the weak secondary internal vortex motion in the drop. The heat and mass fluxes are significantly affected by the presence of the non-condensable in the gaseous phase and by the circulation inside the drop.