Hyaluronic-Acid-Hydroxyapatite Colloidal Gels Combined with Micronized Native ECM as Potential Bone Defect Fillers.

One of the grand challenges in translational regenerative medicine is the surgical placement of biomaterials. For bone regeneration in particular, malleable and injectable colloidal gelsare frequently designed to exhibit self-assembling and shear-response behavior which facilitates biomaterial placement in tissue defects. The current study demonstrated that by combining native extracellular matrix (ECM) microparticles, i.e., demineralized bone matrix (DBM) and decellularized cartilage (DCC), with hyaluronic acid (HA) and hydroxyapatite (HAP) nanoparticles, a viscoelastic colloidal gel consisting exclusively of natural materials was achieved. Rheological testing of HA-ECM suspensions and HA-HAP-ECM colloidal gels concluded either equivalent or substantially higher storage moduli (G' ≈ 100-10 000 Pa), yield stresses (τy ≈ 100-1000 Pa), and viscoelastic recoveries (G'recovery ≥ 87%) in comparison with controls formulated without ECM, which indicated a previously unexplored synergy in fluid properties between ECM microparticles and HA-HAP colloidal networks. Notable rheological differences were observed between respective DBM and DCC formulations, specifically in HA-HAP-DBM mixtures, which displayed a mean 3-fold increase in G' and a mean 4-fold increase in τy from corresponding DCC mixtures. An initial in vitro assessment of these potential tissue fillers as substrates for cell growth revealed that all formulations of HA-ECM and HA-HAP-ECM showed no signs of cytotoxicity and appeared to promote cell viability. Both DBM and DCC colloidal gels represent promising platforms for future studies in bone and cartilage tissue engineering. Overall, the current study identified colloidal gels constructed exclusively of natural materials, with viscoelastic properties that may facilitate surgical placement for a wide variety of therapeutic applications.

Chondroinduction from Naturally Derived Cartilage Matrix: A Comparison Between Devitalized and Decellularized Cartilage Encapsulated in Hydrogel Pastes.

Hydrogel precursors are liquid solutions that are prone to leaking after surgical placement. This problem was overcome by incorporating either decellularized cartilage (DCC) or devitalized cartilage (DVC) microparticles into traditional photocrosslinkable hydrogel precursors in an effort to achieve a paste-like hydrogel precursor. DCC and DVC were selected specifically for their potential to induce chondrogenesis of stem cells, given that materials that are chondroinductive on their own without growth factors are a revolutionary goal in orthopedic medicine. We hypothesized that DVC, lacking the additional chemical processing steps in DCC to remove cell content, would lead to a more chondroinductive hydrogel with rat bone marrow-derived mesenchymal stem cells. Hydrogels composed of methacrylated hyaluronic acid (MeHA) and either DCC or DVC microparticles were tested with and without exposure to transforming growth factor (TGF)-ß3 over a 6 week culture period, where swelling, mechanical analysis, and gene expression were observed. For collagen II, Sox-9, and aggrecan expression, MeHA precursors containing DVC consistently outperformed the DCC-containing groups, even when the DCC groups were exposed to TGF-ß3. DVC consistently outperformed all TGF-ß3-exposed groups in aggrecan and collagen II gene expression as well. In addition, when the same concentrations of MeHA with DCC or DVC microparticles were evaluated for yield stress, the yield stress with the DVC microparticles was 2.7 times greater. Furthermore, the only MeHA-containing group that exhibited shape retention was the group containing DVC microparticles. DVC appeared to be superior to DCC in both chondroinductivity and rheological performance of hydrogel precursors, and therefore DVC microparticles may hold translational potential for cartilage regeneration.

Chondroinductive Hydrogel Pastes Composed of Naturally Derived Devitalized Cartilage.

Hydrogel precursors are liquid solutions that are prone to leaking from the defect site once implanted in vivo. Therefore, the objective of the current study was to create a hydrogel precursor that exhibited a yield stress. Additionally, devitalized cartilage extracellular matrix (DVC) was mixed with DVC that had been solubilized and methacrylated (MeSDVC) to create hydrogels that were chondroinductive. Precursors composed of 10% MeSDVC or 10% MeSDVC with 10% DVC were first evaluated rheologically, where non-Newtonian behavior was observed in all hydrogel precursors. Rat bone marrow stem cells (rBMSCs) were mixed in the precursor solutions, and the solutions were then crosslinked and cultured in vitro for 6 weeks with and without exposure to human transforming growth factor ß3 (TGF-ß3). The compressive modulus, gene expression, biochemical content, swelling, and histology of the gels were analyzed. The DVC-containing gels consistently outperformed the MeSDVC-only group in chondrogenic gene expression, especially at 6 weeks, where the relative collagen II expression of the DVC-containing groups with and without TGF-ß3 exposure was 40- and 78-fold higher, respectively, than that of MeSDVC alone. Future work will test for chondrogenesis in vivo and overall, these two cartilage-derived components are promising materials for cartilage tissue engineering applications.

Mathematical model of microbicidal flow dynamics and optimization of rheological properties for intra-vaginal drug delivery: Role of tissue mechanics and fluid rheology.

Topically applied microbicide gels can provide a self-administered and effective strategy to prevent sexually transmitted infections (STIs). We have investigated the interplay between vaginal tissue elasticity and the yield-stress of non-Newtonian fluids during microbicide deployment. We have developed a mathematical model of tissue deformation driven spreading of microbicidal gels based on thin film lubrication approximation and demonstrated the effect of tissue elasticity and fluid yield-stress on the spreading dynamics. Our results show that both elasticity of tissue and yield-stress rheology of gel are strong determinants of the coating behavior. An optimization framework has been demonstrated which leverages the flow dynamics of yield-stress fluid during deployment to maximize retention while reaching target coating length for a given tissue elasticity.

Enabling Surgical Placement of Hydrogels Through Achieving Paste-Like Rheological Behavior in Hydrogel Precursor Solutions.

Hydrogels are a promising class of materials for tissue regeneration, but they lack the ability to be molded into a defect site by a surgeon because hydrogel precursors are liquid solutions that are prone to leaking during placement. Therefore, although the main focus of hydrogel technology and developments are on hydrogels in their crosslinked form, our primary focus is on improving the fluid behavior of hydrogel precursor solutions. In this work, we introduce a method to achieve paste-like hydrogel precursor solutions by combining hyaluronic acid nanoparticles with traditional crosslinked hyaluronic acid hydrogels. Prior to crosslinking, the samples underwent rheological testing to assess yield stress and recovery using linear hyaluronic acid as a control. The experimental groups containing nanoparticles were the only solutions that exhibited a yield stress, demonstrating that the nanoparticulate rather than the linear form of hyaluronic acid was necessary to achieve paste-like behavior. The gels were also photocrosslinked and further characterized as solids, where it was demonstrated that the inclusion of nanoparticles did not adversely affect the compressive modulus and that encapsulated bone marrow-derived mesenchymal stem cells remained viable. Overall, this nanoparticle-based approach provides a platform hydrogel system that exhibits a yield stress prior to crosslinking, and can then be crosslinked into a hydrogel that is capable of encapsulating cells that remain viable. This behavior may hold significant impact for hydrogel applications where a paste-like behavior is desired in the hydrogel precursor solution.

Contact Line Instability of Gravity-Driven Flow of Power-Law Fluids.

The moving contact line of a thin fluid film can often corrugate into fingers, which is also known as a fingering instability. Although the fingering instability of Newtonian fluids has been studied extensively, there are few studies published on contact line fingering instability of non-Newtonian fluids. In particular, it is still unknown how shear-thinning rheological properties can affect the formation, growth, and shape of a contact line instability. Our previous study (Hu and Kieweg, 2012) showed a decreased capillary ridge formation for more shear-thinning fluids in a 2D model (i.e. 1D thin film spreading within the scope of lubrication theory). Those results motivated this study's hypothesis: more shear-thinning fluids should have suppressed finger growth and longer finger wavelength, and this should be evident in linear stability analysis (LSA) and 3D (i.e. 2D spreading) numerical simulations. In this study, we developed a LSA model for the gravity-driven flow of shear-thinning films, and carried out a parametric study to investigate the impact of shear-thinning on the growth rate of the emerging fingering pattern. A fully 3D model was also developed to compare and verify the LSA results using single perturbations, and to explore the result of multiple-mode, randomly imposed perturbations. Both the LSA and 3D numerical results confirmed that the contact line fingers grow faster for Newtonian fluids than the shear-thinning fluids on both vertical and inclined planes. In addition, both the LSA and 3D model indicated that the Newtonian fluids form fingers with shorter wavelengths than the shear-thinning fluids when the plane is inclined; no difference in the most unstable (i.e. emerging) wavelength was observed at vertical. This study also showed that the distance between emerging fingers was smaller on a vertical plane than on a less-inclined plane for shear-thinning fluids, as previously shown for Newtonian fluids. For the first time for shear-thinning fluids, these results connect trends in capillary ridge and contact line finger formation in 2D models, LSA, and 3D simulations. The results can provide us insights on how to optimize non-Newtonian fluid properties to minimize a fingering instability in many industrial and biological applications.

Mapping glycosaminoglycan-hydroxyapatite colloidal gels as potential tissue defect fillers.

Malleable biomaterials such as Herschel-Bulkley (H-B) fluids possess shear responsive rheological properties and are capable of self-assembly and viscoelastic recovery following mechanical disruption (e.g., surgical placement via injection or spreading). This study demonstrated that the addition of moderate molecular weight glycosaminoglycans (GAGs) such as chondroitin sulfate (CS) (Mw = 15-30 kDa) and hyaluronic acid (HA) (Mw = 20-41 kDa) can be used to modify several rheological properties including consistency index (K), flow-behavior index (n), and yield stress (τy) of submicrometer hydroxyapatite (HAP) (Davg ≤ 200 nm) colloidal gels. GAG-HAP colloidal mixtures exhibited substantial polymer-particle synergism, likely due to "bridging" flocculation, which led to a synergistic increase in consistency index (KGAG-HAP ≥ KGAG + KHAP) without compromising shear-thinning behavior (n < 1) of the gel. In addition, GAG-HAP colloids containing high concentrations of HAP (60-80% w/v) exhibited substantial yield stress (τy ≥ 100 Pa) and viscoelastic recovery properties (G'recovery ≥ 64%). While rheological differences were observed between CS-HAP and HA-HAP colloidal gels, both CS and HA represent feasible options for future studies involving bone defect filling. Overall, this study identified mixture regions where rheological properties in CS-HAP and HA-HAP colloidal gels aligned with desired properties to facilitate surgical placement in non-load-bearing tissue-filling applications such as calvarial defects.

Experimental and numerical models of three-dimensional gravity-driven flow of shear-thinning polymer solutions used in vaginal delivery of microbicides.

HIV/AIDS is a growing global pandemic. A microbicide is a formulation of a pharmaceutical agent suspended in a delivery vehicle, and can be used by women to protect themselves against HIV infection during intercourse. We have developed a three-dimensional (3D) computational model of a shear-thinning power-law fluid spreading under the influence of gravity to represent the distribution of a microbicide gel over the vaginal epithelium. This model, accompanied by a new experimental methodology, is a step in developing a tool for optimizing a delivery vehicle's structure/function relationship for clinical application. We compare our model with experiments in order to identify critical considerations for simulating 3D free-surface flows of shear-thinning fluids. Here we found that neglecting lateral spreading, when modeling gravity-induced flow, resulted in up to 47% overestimation of the experimental axial spreading after 90 s. In contrast, the inclusion of lateral spreading in 3D computational models resulted in rms errors in axial spreading under 7%. In addition, the choice of the initial condition for shape in the numerical simulation influences the model's ability to describe early time spreading behavior. Finally, we present a parametric study and sensitivity analysis of the power-law parameters' influence on axial spreading, and to examine the impact of changing rheological properties as a result of dilution or formulation conditions. Both the shear-thinning index (n) and consistency (m) impacted the spreading length and deceleration of the moving front. The sensitivity analysis showed that gels with midrange m and n values (for the ranges in this study) would be most sensitive (over 8% changes in spreading length) to 10% changes (e.g., from dilution) in both rheological properties. This work is applicable to many industrial and geophysical thin-film flow applications of non-Newtonian fluids; in addition to biological applications in microbicide drug delivery.

Gravity-Driven Thin Film Flow of an Ellis Fluid.

The thin film lubrication approximation has been studied extensively for moving contact lines of Newtonian fluids. However, many industrial and biological applications of the thin film equation involve shear-thinning fluids, which often also exhibit a Newtonian plateau at low shear. This study presents new numerical simulations of the three-dimensional (i.e. two-dimensional spreading), constant-volume, gravity-driven, free surface flow of an Ellis fluid. The numerical solution was validated with a new similarity solution, compared to previous experiments, and then used in a parametric study. The parametric study centered around rheological data for an example biological application of thin film flow: topical drug delivery of anti-HIV microbicide formulations, e.g. hydroxyethylcellulose (HEC) polymer solutions. The parametric study evaluated how spreading length and front velocity saturation depend on Ellis parameters. A lower concentration polymer solution with smaller zero shear viscosity (η0), τ1/2, and λ values spread further. However, when comparing any two fluids with any possible combinations of Ellis parameters, the impact of changing one parameter on spreading length depends on the direction and magnitude of changes in the other two parameters. In addition, the isolated effect of the shear-thinning parameter, λ, on the front velocity saturation depended on τ1/2. This study highlighted the relative effects of the individual Ellis parameters, and showed that the shear rates in this flow were in both the shear-thinning and plateau regions of rheological behavior, emphasizing the importance of characterizing the full range of shear-rates in rheological measurements. The validated numerical model and parametric study provides a useful tool for future steps to optimize flow of a fluid with rheological behavior well-described by the Ellis constitutive model, in a range of industrial and biological applications.

An automated image-based method for rapid analysis of Chlamydia infection as a tool for screening antichlamydial agents.

A major limitation in the identification of novel antichlamydial compounds is the paucity of effective methods for large-scale compound screening. The immunofluorescence assay is the preferred approach for accurate quantification of the intracellular growth of Chlamydia. In this study, an immunofluorescence image-based method (termed image-based automated chlamydial identification and enumeration [iBAChIE]) was customized for fully automated quantification of Chlamydia infection using the freely available open-source image analysis software program CellProfiler and the complementary data exploration software program CellProfiler Analyst. The method yielded enumeration of different species and strains of Chlamydia highly comparably to the conventional manual methods while drastically reducing the analysis time. The inhibitory capability of established antichlamydial activity was also evaluated. Overall, these data support that iBAChIE is a highly effective tool for automated quantification of Chlamydia infection and assessment of antichlamydial activities of molecules. Furthermore, iBAChIE is expected to be amenable to high-throughput screening studies for inhibitory compounds and fluorescently labeled molecules to study host-pathogen interactions.

Synthesis and evaluation of novel dental monomer with branched aromatic carboxylic acid group.

A new glycerol-based dimethacrylate monomer with an aromatic carboxylic acid, 2-((1,3-bis(methacryloyloxy)propan-2-yloxy)carbonyl)benzoic acid (BMPB), was synthesized, characterized, and proposed as a possible dental co-monomer for dentin adhesives. Dentin adhesives containing 2-hydroxyethyl methacrylate (HEMA) and 2,2-bis[4-(2-hydroxy-3-methacryloxypropoxy) phenyl]propane (BisGMA) in addition to BMPB were formulated with water at 0, 5, 10, and 15 wt % to simulate wet, oral conditions, and photo-polymerized. Adhesives were characterized with regard to viscosity, real-time photopolymerization behavior, dynamic mechanical analysis, and microscale 3D internal morphologies and compared with HEMA/BisGMA controls. When formulated under wet conditions, the experimental adhesives showed lower viscosities (0.04-0.07 Pa s) as compared to the control (0.09-0.12 Pa s). The experimental adhesives showed higher glass transition temperature (146-157°C), degree of conversion (78-89%), and rubbery moduli (33-36 MPa), and improved water miscibility (no voids) as compared to the controls (123-135°C, 67-71%, 15-26 MPa, and voids, respectively). The enhanced properties of these adhesives suggest that BMPB with simple, straightforward synthesis is a promising photocurable co-monomer for dental restorative materials.

The Effect of Surface Tension on the Gravity-driven Thin Film Flow of Newtonian and Power-law Fluids.

Gravity-driven thin film flow is of importance in many fields, as well as for the design of polymeric drug delivery vehicles, such as anti-HIV topical microbicides. There have been many prior works on gravity-driven thin films. However, the incorporation of surface tension effect has not been well studied for non-Newtonian fluids. After surface tension effect was incorporated into our 2D (i.e. 1D spreading) power-law model, we found that surface tension effect not only impacted the spreading speed of the microbicide gel, but also had an influence on the shape of the 2D spreading profile. We observed a capillary ridge at the front of the fluid bolus. Previous literature shows that the emergence of a capillary ridge is strongly related to the contact line fingering instability. Fingering instabilities during epithelial coating may change the microbicide gel distribution and therefore impact how well it can protect the epithelium. In this study, we focused on the capillary ridge in 2D flow and performed a series of simulations and showed how the capillary ridge height varies with other parameters, such as surface tension coefficient, inclination angle, initial thickness, and power-law parameters. As shown in our results, we found that capillary ridge height increased with higher surface tension, steeper inclination angle, bigger initial thickness, and more Newtonian fluids. This study provides the initial insights of how to optimize the flow and prevent the appearance of a capillary ridge and fingering instability.

Fatigue life prediction of dentin-adhesive interface using micromechanical stress analysis.

OBJECTIVES: The objective of this work was to develop a methodology for the prediction of fatigue life of the dentin-adhesive (d-a) interface. METHODS: At the micro-scale, the d-a interface is composed of dissimilar material components. Under global loading, these components experience different local stress amplitudes. The overall fatigue life of the d-a interface is, therefore, determined by the material component that has the shortest fatigue life under local stresses. Multiple 3d finite element (FE) models were developed to determine the stress distribution within the d-a interface by considering variations in micro-scale geometry, material composition and boundary conditions. The results from these models were analyzed to obtain the local stress concentrations within each d-a interface component. By combining the local stress concentrations and experimentally determined stress versus number of cycle to failure (S-N) curves for the different material components, the overall fatigue life of the d-a interface was predicted. RESULTS: The fatigue life was found to be a function of the applied loading amplitude, boundary conditions, microstructure and the mechanical properties of the material components of the d-a interface. In addition, it was found that the overall fatigue life of the d-a interface is not determined by the weakest material component. In many cases, the overall fatigue life was determined by the adhesive although exposed collagen was the weakest material component. Comparison of the predicted results with experimental data from the literature showed both qualitative and quantitative agreement. SIGNIFICANCE: The methodology developed for fatigue life prediction can provide insight into the mechanisms that control degradation of the bond formed at the d-a interface.

Tensile strength of a surgeon's or a square knot.

OBJECTIVE: To test the integrity of surgeon's knots and flat square knots using 4 different suture materials. STUDY DESIGN: Chromic catgut, polyglactin 910, silk, and polydioxanone sutures were tied in the 2 types of knot configurations. For all sutures, a 0-gauge United States Pharmacopeia suture was used. Knots were tied by a single investigator (J.B.). The suture was soaked in 0.9% sodium chloride for 60 s and subsequently transferred to a tensiometer where the tails were cut to 3-mm length. We compared the knots, measuring knot strength with a tensiometer until the sutures broke or untied. RESULTS: A total of 119 throws were tied. We found no difference in mean tension at failure between a surgeon's knot (79.7 N) and a flat square knot (82.9 N). Using a chi(2) test, we did not find a statistically significant difference in the likelihood of knots coming untied between surgeon's knots (29%) and flat square knots (38%). CONCLUSIONS: Under laboratory conditions, surgeon's knots and flat square knots did not differ in tension at failure or in likelihood of untying.

Viscoelastic and fatigue properties of model methacrylate-based dentin adhesives.

The objective of the current study is to characterize the viscoelastic and fatigue properties of model methacrylate-based dentin adhesives under dry and wet conditions. Static, creep, and fatigue tests were performed on cylindrical samples in a 3-point bending clamp. Static results showed that the apparent elastic modulus of the model adhesive varied from 2.56 to 3.53 GPa in the dry condition, and from 1.04 to 1.62 GPa in the wet condition, depending upon the rate of loading. Significant differences were also found for the creep behavior of the model adhesive under dry and wet conditions. A linear viscoelastic model was developed by fitting the adhesive creep behavior. The developed model with 5 Kelvin Voigt elements predicted the apparent elastic moduli measured in the static tests. The model was then utilized to interpret the fatigue test results. It was found that the failure under cyclic loading can be due to creep or fatigue, which has implications for the failure criterion that are applied for these types of tests. Finally, it was found that the adhesive samples tested under dry conditions were more durable than those tested under wet conditions.

Effect of photoinitiator system and water content on dynamic mechanical properties of a light-cured bisGMA/HEMA dental resin.

The selection of an appropriate photoinitiator system is critical for efficient polymerization of dental resins with satisfactory mechanical and physical properties. The purpose of this study was to evaluate the influence of adding an iodonium salt to two-component photoinitiator systems. Four photoinitiator systems were included in a model bisGMA/HEMA resin and used to prepare samples at different water contents; the dynamic mechanical properties and the final degree of conversion of the samples were then characterized. Addition of the iodonium salt to the two-component photoinitiator systems increased the final degree of conversion, glass transition temperature, rubbery modulus, and crosslink density. The photoinitiator system containing ethyl-4-(dimethylamino) benzoate as a coinitiator and the iodonium salt exhibited the highest rubbery modulus. The enhanced properties in the presence of the iodonium salt can be attributed to the production of an active phenyl radical with regeneration of the original camphorquinone, which may increase the compatibility between monomers and initiators, especially in the presence of water. The results support the hypothesis that a photoinitiator system containing an iodonium salt can increase both mechanical properties and final conversion of model resin polymerized in the presence of water.

Preparation and Properties of Novel Dentin Adhesives with Esterase Resistance.

A new methacrylate monomer, trimethylolpropane mono allyl ether dimethacrylate (TMPEDMA), was synthesized and evaluated. This branched methacrylate was designed to increase esterase-resistance when incorporated into conventional HEMA (2-hydroxyethyl methacrylate)/BisGMA (2,2-bis[4(2-hydroxy-3-methacryloyloxy-propyloxy)-phenyl] propane) dental adhesives. The new adhesives, HEMA/BisGMA/TMPEDMA in a 45/30/25 (w/w) ratio were formulated with H(2)O at 0 (A0T) and 8 wt % water (A8T) and compared with control adhesives (HEMA/BisGMA, 45/55 (w/w), at 0 (A0) and 8 wt % (A8) water). Camphoroquinone (CQ), 2-(dimethylamino) ethyl methacrylate and diphenyliodonium hexafluorophosphate were used as photoinitiators. The new adhesives showed a degree of conversion comparable with the control and improved modulus and glass transition temperature (T(g)). Exposure of photopolymerized discs to porcine liver esterase for up to eight days showed that the net cumulative methacrylic acid (MAA) release in adhesives formulated with the new monomer and 8% water (A8T: 182 µg/mL) was dramatically (P < 0.05) decreased in comparison to the control (A8: 361.6 µg/mL). The results demonstrate that adhesives made with the new monomer and cured in water to simulate wet bonding are more resistant to esterase than conventional HEMA/BisGMA adhesive.

Interpreting properties of microbicide drug delivery gels: analyzing deployment kinetics due to squeezing.

Prophylactic efficacy of topical microbicidal drug delivery formulations against HIV may depend upon their abilities to coat and be retained on epithelial surfaces where infection begins. Rheological and surface properties play paramount roles in governing coating. While fundamental fluid mechanical studies of epithelial coating mechanisms have begun, their results have not previously addressed questions of practical value to formulators in the pharmaceutics community. The present theoretical study began this process. We focused upon squeezing flows of seven vaginal gels which are models for future microbicides or a candidate formulation in clinical trials. Each formulation is based upon one of three different macromolecules: cellulose, polyacrylic acid (PAA), or carrageenan. We addressed: (1) properties with greatest influence on squeezing flow; (2) alterations of properties to improve measures of coating dynamics; and (3) effects of polymer concentration and temperature on coating dynamics. We found that yield stresses dominated flows of PAA gels, and that surface slip, while small, significantly influenced coating by cellulose gels. Decreases in consistency, increases in shear-thinning, and increases in temperature led to thinner coatings. Details of altered coating rates depended upon parameter values and time. Specific polymer concentration effects differed between cellulose and PAA gels, though trends were similar.

Biophysical analysis of prototype microbicidal gels.

The objective of this study was to evaluate the distribution and retention (deployment) of four prototype vehicles for delivery of prophylactic microbicides against vaginal HIV transmission. Study gels were created with different molecular compositions, producing different biophysical properties governing vaginal deployment. The study employed three techniques: direct rheological measurement of gel properties, direct observation of gel surface coating erosion, and dissolution by a vaginal fluid simulant, and mathematical modeling of gel squeezing flow processes. Results suggest significant differences in extent of vaginal coating after gel application and in erosion of these gel layers due to contact with ambient vaginal fluid and shearing. The relationships between gel rheological properties, coating flow and erosion of coating were not always anticipated from differences in gel molecular composition.

Squeezing flows of vaginal gel formulations relevant to microbicide drug delivery.

Efficacy of topical microbicidal drug delivery formulations against HIV depends in part on their ability to coat, distribute, and be retained on epithelium. Once applied to the vagina, a formulation is distributed by physical forces including: gravity, surface tension, shearing, and normal forces from surrounding tissues, i.e., squeezing forces. The present study focused on vaginal microbicide distribution due to squeezing forces. Mathematical simulations of squeezing flows were compared with squeezing experiments, using model vaginal gel formulations. Our objectives were: (1) to determine if mathematical simulations can accurately describe squeezing flows of vaginal gel formulations; (2) to find the best model and optimized parameter sets to describe these gels; and (3) to examine vaginal coating due to squeezing using the best models and summary parameters for each gel. Squeezing flow experiments revealed large differences in spreadability between formulations, suggesting different coating distributions in vivo. We determined the best squeezing flow models and summary parameters for six test gels of two compositions, cellulose and polyacrylic acid (PAA). We found that for some gels it was preferable to deduce model input parameters directly from squeezing flow experiments. For the cellulose gels, slip conditions in squeezing flow experiments needed to be evaluated. For PAA gels, we found that in the absence of squeezing experiments, rotational viscometry measurements (to determine Herschel-Bulkley parameters) led to reasonably accurate predictions of squeezing flows. Results indicated that yield stresses may be a strong determinant of squeezing flow mechanics. This study serves as a template for further investigations of other gels and determination of which sources of rheological data best characterize potential microbicidal formulations. These mathematical simulations can serve as useful tools for exploring drug delivery parameters, and optimizing formulations, prior to costly clinical trials.