Modeling Diffusivity Through Alginate-Based Microfibers: A Comparison of Numerical and Analytical Models Based on Empirical Spectrophotometric Data

Abstract: The study of mass transport across hollow and solid 3D microfibers to study metabolic profiles is a key aspect of tissue engineering approach. A new modified numerical mathematical model based on Fickian equations in cylindrical coordinates has been proposed for determining the membrane diffusivity of 2% (w/v) alginate-based stents cross-linked with 10% CaCl2. Based on the economical and direct spectrophotometric measurements, using this model, inward diffusivities ranging from 5.2x10-14 m2/s 2.93x10-12m2/s were computed for solutes with Stokes radii ranging between 0.36 to 3.5 nm, diffusing through bare alginate and alginate-chitosan-alginate microfibers. In parallel an analytical solution to the cylindrical Fickian equation was derived to validate the numerical solution using experimental diffusion data from a solid stent. Excellent agreement was found between the numerical and analytical models with a maximum calculated residual value of 4%. Using these models, a flexible computational platform is proposed to conduct custom diffusion and MW cut-off characterization across micro-porous microfibers not limited to alginate in composition

Mechanical Properties of Hydrated Acoustically Sensitive Alginate-Based Microcapsules Confined in a Microfluidic Device as a Function of Size and Composition

Understanding the mechanical properties of alginate-based microcapsules according to size and chemical composition allows researchers to zero in on the treatment and methods required to engineer optimized implantable alginate-based artificial cells for chemotherapy. Cross-linked medium viscosity alginate capsules ranging from 1.1% (w/v)-1.8% (w/v) in composition and 200 μm-1200 μm in size, encapsulating ultrasound contrast agents and blue dextran were compressed within a 40 μm high polydimethylsiloxane microfluidic device and subsequently examined using 2D microscopy for strain deformation aimed at the calculation of poisson ratios and volume loss postcompression. Results indicate a decrease in Poisson ratio as a function of alginate concentration, with statistically significant increases in Poisson ratios and percent volume loss as a function of size and composition. For an average of 120 s observation time post compression, in light of the volume loss correlated to the number of cross-links as a function of capsule size and alginate concentration, a strong case for the dominance of poroelasticity vs. viscoelasticity can be made. While there was a decrease in mean Poisson ratio as a function of concentration, at 1.8% (w/v) the mean strain value converged to 0.5, the theoretical ideal isotropic value associated with soft biological tissue

Polystyrene Microsphere and 5-Fluorouracil Release from Custom Designed Wound Dressing Films

Custom-designed wound dressing films of chitosan and alginate have been prepared by a casting/solvent evaporation method for hydrophobic therapeutic agent encapsulation. In this parametric study, the propylene glycol (PG) and calcium chloride (CaCl2) concentrations were varied for chitosan and alginate films, respectively. Mechanical and chemical inter-related responses under observations included thickness (th), elasticity (E), tensile strength (TS), sorption ability (S%) and kinetics of in-vitro drug release, specifically in terms of membrane time to burst (tB) and duration of release (tR). As shown by results of a one tailed t-test significance testing at the 95% confidence interval (α = 0.05), alginate films were significantly more elastic (p = 0.003), thinner (p = 0.004) and more susceptible to osmotic burst (p = 0.011) and characterized by a longer duration of release (p = 0.03). Meanwhile chitosan films exhibited superior moisture permeability (p = 0.006) and sorption characteristics (p = 0.001), indicative of higher hydrophilicity. There were no significant differences in tensile strength (p = 0.324) for alginate and chitosan-based formulations. Preliminary testing was conducted using 0.71 μm in diameter microspheres for modeling film dissolution into Lactated Ringer’s solution. Experimental release profiles were modeled for each film from which the average release from alginate films (MAGCa = 81%) was estimated to be twice the percentage associated with chitosan films (MCD = 42%). The film comprised of 2.5% (w/v) medium MW chitosan/dextran 70 kDa (5:1) was selected for studying the release of 5-Fluorouracil (5-FU) as a model hydrophobic drug. Diffusion coupled with film disintegration is immediate (tB = 0) in case of encapsulated 5-FU as compared to the control film encapsulating microspheres characterized by tB = 70 min ± 7 min. This shift in release profile and the ability to modulate the timing of membrane burst can be attributed to the approximate ratio (1: 505) in molecular size between drug and microsphere. This hypothesis has been validated by the film pore size measured to be 430 nm ± 88 nm using atomic force microscopy

Effect of therapeutic ultrasound on acoustically sensitive microcapsules

In the area of therapeutic ultrasound activated drug delivery, difficulties exist in designing a carrier that responds to ultrasound for triggering and imaging but also provides adequate treatment potential. In this paper, we report on a novel acoustically sensitive microcapsule reservoir that can be activated with therapeutic ultrasound for payload release and can be potentially tracked using imaging. It is being designed for increased longevity and is not planned for the circulation. Here, we describe its unique formulation and demonstrate effects of therapeutic ultrasound on it at 1MHz using a combined optical-acoustic setup on a microscope. We see membrane bulging and damage for small and large capsules with both continuous and pulsed ultrasound. We also show some preliminary work on understanding the mechanism behind these effects. The reservoirs show potential for future ultrasound activated release and imaging while being patent in form and function over several weeks

Structural changes and imaging signatures of acoustically sensitive microcapsules under ultrasound

The ultrasound drug delivery field is actively designing new agents that would obviate the problems of just using microbubbles for drug delivery. Microbubbles have very short circulation time (minutes), low payload and large size (2–10 μm), all of these aspects are not ideal for systemic drug delivery. However, microbubble carriers provide excellent image contrast and their use for image guidance can be exploited. In this paper, we suggest an alternative approach by developing acoustically sensitive microcapsule reservoirs that have future applications for treating large ischemic tumors through intratumoral therapy. We call these agents Acoustically Sensitized Microcapsules (ASMs) and these are not planned for the circulation. ASMs are very simple in their formulation, robust and reproducible. They have been designed to offer high payload (because of their large size), be acoustically sensitive and reactive (because of the Ultrasound Contrast Agents (UCAs) encapsulated) and mechanically robust for future injections/implantations within tumors. We describe three different aspects – (1) effect of therapeutic ultrasound; (2) mechanical properties and (3) imaging signatures of these agents. Under therapeutic ultrasound, the formation of a cavitational bubble was seen prior to rupture. The time to rupture was size dependent. Size dependency was also seen when measuring mechanical properties of these ASMs. % Alginate and permeability also affected the Young’s modulus estimates. For study of imaging signatures of these agents, we show six schemes. For example, with harmonic imaging, tissue phantoms and controls did not generate higher harmonic components. Only ASM phantoms created a harmonic signal, whose sensitivity increased with applied acoustic pressure. Future work includes developing schemes combining both sonication and imaging to help detect ASMs before, during and after release of drug substance

Fickian-Based Empirical Approach for Diffusivity Determination in Hollow Alginate-Based Microfibers Using 2D Fluorescence Microscopy and Comparison with Theoretical Predictions

Hollow alginate microfibers (od = 1.3 mm, id = 0.9 mm, th = 400 µm, L = 3.5 cm) comprised of 2% (w/v) medium molecular weight alginate cross-linked with 0.9 M CaCl2 were fabricated to model outward diffusion capture by 2D fluorescent microscopy. A two-fold comparison of diffusivity determination based on real-time diffusion of Fluorescein isothiocyanate molecular weight (FITC MW) markers was conducted using a proposed Fickian-based approach in conjunction with a previously established numerical model developed based on spectrophotometric data. Computed empirical/numerical (Dempiricial/Dnumerical) diffusivities characterized by small standard deviations for the 4-, 70- and 500-kDa markers expressed in m2/s are (1.06 × 10−9 ± 1.96 × 10−10)/(2.03 × 10−11), (5.89 × 10−11 ± 2.83 × 10−12)/(4.6 × 10−12) and (4.89 × 10−12 ± 3.94 × 10−13)/(1.27 × 10−12), respectively, with the discrimination between the computation techniques narrowing down as a function of MW. The use of the numerical approach is recommended for fluorescence-based measurements as the standard computational method for effective diffusivity determination until capture rates (minimum 12 fps for the 4-kDa marker) and the use of linear instead of polynomial interpolating functions to model temporal intensity gradients have been proven to minimize the extent of systematic errors associated with the proposed empirical method

Fickian-Based Empirical Approach for Diffusivity Determination in Hollow Alginate-Based Microfibers Using 2D Fluorescence Microscopy and Comparison with Theoretical Predictions

Hollow alginate microfibers (od = 1.3 mm, id = 0.9 mm, th = 400 µm, L = 3.5 cm) comprised of 2% (w/v) medium molecular weight alginate cross-linked with 0.9 M CaCl2 were fabricated to model outward diffusion capture by 2D fluorescent microscopy. A two-fold comparison of diffusivity determination based on real-time diffusion of Fluorescein isothiocyanate molecular weight (FITC MW) markers was conducted using a proposed Fickian-based approach in conjunction with a previously established numerical model developed based on spectrophotometric data. Computed empirical/numerical (Dempiricial/Dnumerical) diffusivities characterized by small standard deviations for the 4-, 70- and 500-kDa markers expressed in m2/s are (1.06 × 10−9 ± 1.96 × 10−10)/(2.03 × 10−11), (5.89 × 10−11 ± 2.83 × 10−12)/(4.6 × 10−12) and (4.89 × 10−12 ± 3.94 × 10−13)/(1.27 × 10−12), respectively, with the discrimination between the computation techniques narrowing down as a function of MW. The use of the numerical approach is recommended for fluorescence-based measurements as the standard computational method for effective diffusivity determination until capture rates (minimum 12 fps for the 4-kDa marker) and the use of linear instead of polynomial interpolating functions to model temporal intensity gradients have been proven to minimize the extent of systematic errors associated with the proposed empirical method