Thao D. Nguyen

A versatile pH sensitive chondroitin sulfate–PEG tissue adhesive and hydrogel

We developed a chondroitin sulfate-polyethylene glycol (CS-PEG) adhesive hydrogel with numerous potential biomedical applications. The carboxyl groups on chondroitin sulfate (CS) chains were functionalized with N-hydroxysuccinimide (NHS) to yield chondroitin sulfate succinimidyl succinate (CS-NHS). Following purification, the CS-NHS molecule can react with primary amines to form amide bonds. Hence, using six arm polyethylene glycol amine PEG-(NH2)6 as a crosslinker we formed a hydrogel which was covalently bound to proteins in tissue via amide bonds. By varying the initial pH of the precursor solutions, the hydrogel stiffness, swelling properties, and kinetics of gelation could be controlled. The sealing/adhesive strength could also be modified by varying the damping and storage modulus properties of the material. The adhesive strength of the material with cartilage tissue was shown to be ten times higher than that of fibrin glue. Cells encapsulated or in direct contact with the material remained viable and metabolically active. Furthermore, CS-PEG material produced minimal inflammatory response when implanted subcutaneously in a rat model and enzymatic degradation was demonstrated in vitro. This work establishes an adhesive hydrogel derived from biological and synthetic components with potential application in wound healing and regenerative medicine.

Biomechanics of the Human Posterior Sclera: Age- and Glaucoma-Related Changes Measured Using Inflation Testing

PURPOSE The objective of this study was to measure the biomechanical response of the human posterior sclera in vitro and to estimate the effects of age and glaucoma. METHODS Scleral specimens from 22 donors with no history of glaucoma and 11 donors with a history of glaucoma were excised 3 mm posterior to the equator and affixed to an inflation chamber. Optic nerve cross-sections were graded to determine the presence of axon loss. The time-dependent inflation response was measured in a series of pressure-controlled load-unload tests to 30 mm Hg and creep tests to 15 and 30 mm Hg. Circumferential and meridional strains were computed from the digital image correlation displacements, and midposterior stresses were determined from pressure and deformed geometry. RESULTS Among normal specimens, older age was predictive of a stiffer response and a thinner sclera. In the age group 75 to 93, diagnosed glaucoma eyes with axon damage were thicker than normal eyes. Both damaged and undamaged glaucoma eyes had a different strain response in the peripapillary sclera characterized by a stiffer meridional response. Undamaged glaucoma eyes had slower circumferential creep rates in the peripapillary sclera than normal eyes. Glaucoma eyes were not different from normal eyes in stresses and strains in the midposterior sclera. CONCLUSIONS The observed differences in the biomechanical response of normal and glaucoma sclera may represent baseline properties that contribute to axon damage, or may be characteristics that result from glaucomatous disease.

Full-field deformation of bovine cornea under constrained inflation conditions.

The viscoelastic response of bovine corneas was characterized using in vitro inflation (bulge) experiments combined with spatially-resolved deformation mapping via digital image correlation. A complex fixture conforming to the limbal annulus was developed to hold the attached sclera rigid while allowing deformation only in the cornea. A statistical set of experiments was performed for a pressure range of 3.6-8 kPa (27-60 mmHg), representing nominal bovine intraocular pressure (IOP) to acute glaucoma conditions. A broader pressure range of 0-32 kPa (0-240 mmHg) was also examined to characterize the nonlinear finite deformation behavior of the tissue. Results showed that for pressures near and above IOP, the majority of the deformation was localized in the limbus and peripheral regions, which left the central cornea largely undeformed. This observation was consistent with the known preferred circumferential alignment of collagen fibrils outside of the central cornea. In general, the inflation experiments observed viscoelastic behavior in the form of rate-dependent hysteresis in the pressure-deformation response of the apex of the cornea, creep in the apex deformation at a constant inflation pressure, and relaxation in the pressure response at a constant inflation volume. The 3.6-8 kPa (27-60 mmHg) pressure range produced small viscoelastic deformations and a nearly linear pressure-deformation response, which suggests that for physiological pressure ranges, the cornea can be approximated as a linear viscoelastic or linear pseudo-elastic material.

Self-folding thermo-magnetically responsive soft microgrippers.

Hydrogels such as poly(N-isopropylacrylamide-co-acrylic acid) (pNIPAM-AAc) can be photopatterned to create a wide range of actuatable and self-folding microstructures. Mechanical motion is derived from the large and reversible swelling response of this cross-linked hydrogel in varying thermal or pH environments. This action is facilitated by their network structure and capacity for large strain. However, due to the low modulus of such hydrogels, they have limited gripping ability of relevance to surgical excision or robotic tasks such as pick-and-place. Using experiments and modeling, we design, fabricate, and characterize photopatterned, self-folding functional microgrippers that combine a swellable, photo-cross-linked pNIPAM-AAc soft-hydrogel with a nonswellable and stiff segmented polymer (polypropylene fumarate, PPF). We also show that we can embed iron oxide (Fe2O3) nanoparticles into the porous hydrogel layer, allowing the microgrippers to be responsive and remotely guided using magnetic fields. Using finite element models, we investigate the influence of the thickness and the modulus of both the hydrogel and stiff polymer layers on the self-folding characteristics of the microgrippers. Finally, we illustrate operation and functionality of these polymeric microgrippers for soft robotic and surgical applications.

Modeling the relaxation mechanisms of amorphous shape memory polymers.

In this progress report, we review two common approaches to constitutive modeling of thermally activated shape memory polymers, then focus on a recent thermoviscoelastic model that incorporates the time-dependent effects of structural and stress relaxation mechanisms of amorphous networks. An extension of the model is presented that incorporates the effects of multiple discrete structural and stress relaxation processes to more accurately describe the time-dependent behavior. In addition, a procedure is developed to determine the model parameters from standard thermomechanical experiments. The thermoviscoelastic model was applied to simulate the unconstrained recovery response of a family of (meth)acrylate-based networks with different weight fractions of the crosslinking agent. Results showed significant improvement in predicting the temperature-dependent strain recovery response.

Quantitative Mapping of Collagen Fiber Orientation in Non-glaucoma and Glaucoma Posterior Human Sclerae

PURPOSE The posterior sclera has a major biomechanical influence on the optic nerve head, and may therefore be important in glaucoma. Scleral material properties are influenced significantly by collagen fiber architecture. Here we quantitatively map fiber orientation in non-glaucoma and glaucoma posterior human sclerae. METHODS Wide-angle x-ray scattering quantified fiber orientation at 0.5-mm intervals across seven non-glaucoma post-mortem human sclerae, and five sclerae with glaucoma history and confirmed axon loss. Multiphoton microscopy provided semiquantitative depth-profiling in the peripapillary sclera. RESULTS Midposterior fiber orientation was either uniaxial (one preferred direction) or biaxial (two directions). The peripapillary sclera was characterized by a ring of fibers located mainly in the mid-/outer stromal depth and encompassing ∼50% of the total tissue thickness. Fiber anisotropy was 37% higher in the peripapillary sclera compared with midposterior, varied up to 4-fold with position around the scleral canal, and was consistently lowest in the superior-nasal quadrant. Mean fiber anisotropy was significantly lower in the superior-temporal (P < 0.01) and inferior-nasal (P < 0.05) peripapillary scleral quadrants in glaucoma compared with non-glaucoma eyes. CONCLUSIONS The collagen fiber architecture of the posterior human sclera is highly anisotropic and inhomogeneous. Regional differences in peripapillary fiber anisotropy between non-glaucoma and glaucoma eyes may represent adaptive changes in response to elevated IOP and/or glaucoma, or baseline structural properties that associate with predisposition to glaucomatous axon damage. Quantitative fiber orientation data will benefit numerical eye models aimed at predicting the scleras influence on nerve head biomechanics, and thereby its possible role in glaucoma.

A Nonlinear Anisotropic Viscoelastic Model for the Tensile Behavior of the Corneal Stroma

Tensile strip experiments of bovine corneas have shown that the tissue exhibits a nonlinear rate-dependent stress-strain response and a highly nonlinear creep response that depends on the applied hold stress. In this paper, we present a constitutive model for the finite deformation, anisotropic, nonlinear viscoelastic behavior of the corneal stroma. The model formulates the elastic and viscous response of the stroma as the average of the elastic and viscous response of the individual lamellae weighted by a probability density function of the preferred in-plane lamellar orientations. The result is a microstructure-based model that incorporates the viscoelastic properties of the matrix and lamellae and the lamellar architecture in the response of the stroma. In addition, the model includes a fully nonlinear description of the viscoelastic response of the lamellar(fiber) level. This is in contrast to previous microstructure-based models of fibrous soft tissues, which relied on quasilinear viscoelastic formulations of the fiber viscoelasticity. Simulations of recent tensile strip experiments show that the model is able to predict, well within the bounds of experimental error and natural variations, the cyclic stress-strain behavior and nonlinear creep behavior observed in uniaxial tensile experiments of excised strips of bovine cornea.

Functional stimuli responsive hydrogel devices by self-folding

We describe a photolithographic approach to create functional stimuli responsive, self-folding, microscale hydrogel devices using thin, gradient cross-linked hinges and thick, fully cross-linked panels. The hydrogels are composed of poly (N-isopropylacrylamide-co-acrylic acid) (pNIPAM-AAc) with reversible stimuli responsive properties just below physiological temperatures. We show that a variety of three-dimensional structures can be formed and reversibly actuated by temperature or pH. We experimentally characterized the swelling and mechanical properties of pNIPAM-AAc and developed a finite element model to rationalize self-folding and its variation with hinge thickness and swelling ratio. Finally, we highlight applications of this approach in the creation of functional devices such as self-folding polymeric micro-capsules, untethered micro-grippers and thermally steered micro-mirror systems.

Scleral anisotropy and its effects on the mechanical response of the optic nerve head

This paper presents a computational modeling study of the effects of the collagen fiber structure on the mechanical response of the sclera and the adjacent optic nerve head (ONH). A specimen-specific inverse finite element method was developed to determine the material properties of two human sclera subjected to full-field inflation experiments. A distributed fiber model was applied to describe the anisotropic elastic behavior of the sclera. The model directly incorporated wide-angle X-ray scattering measurements of the anisotropic collagen structure. The converged solution of the inverse method was used in micromechanical studies of the mechanical anisotropy of the sclera at different scales. The effects of the scleral collagen fiber structure on the ONH deformation were evaluated by progressively filtering out local anisotropic features. It was found that the majority of the midposterior sclera could be described as isotropic without significantly affecting the mechanical response of the tissues of the ONH. In contrast, removing local anisotropic features in the peripapillary sclera produced significant changes in scleral canal expansion and lamina cribrosa deformation. Local variations in the collagen structure of the peripapillary sclera significantly influenced the mechanical response of the ONH.

An inverse finite element method for determining the anisotropic properties of the cornea

An inverse finite element method was developed to determine the anisotropic properties of bovine cornea from an in vitro inflation experiment. The experiment used digital image correlation (DIC) to measure the three-dimensional surface geometry and displacement field of the cornea at multiple pressures. A finite element model of a bovine cornea was developed using the DIC measured surface geometry of the undeformed specimen. The model was applied to determine five parameters of an anisotropic hyperelastic model that minimized the error between the measured and computed surface displacement field and to investigate the sensitivity of the measured bovine inflation response to variations in the anisotropic properties of the cornea. The results of the parameter optimization revealed that the collagen structure of bovine cornea exhibited a high degree of anisotropy in the limbus region, which agreed with recent histological findings, and a transversely isotropic central region. The parameter study showed that the bovine corneal response to the inflation experiment was sensitive to the shear modulus of the matrix at pressures below the intraocular pressure, the properties of the collagen lamella at higher pressures, and the degree of anisotropy in the limbus region. It was not sensitive to a weak collagen anisotropy in the central region.