Mechanical behavior of tissue simulants and soft tissues under extreme loading conditions

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Materials Science and Engineering, 2013.Cataloged from PDF version of thesis.Includes bibliographical references (p. 157-168).Recent developments in computer-integrated surgery and in tissue-engineered constructs necessitate advances in experimental and analytical techniques in characterizing properties of mechanically compliant materials such as gels and soft tissues, particularly for small sample volumes. One goal of such developments is to quantitatively predict and mimic tissue deformation due to high rate impact events typical of industrial accidents and ballistic insults. This aim requires advances in mechanical characterization to establish tools and design principles for tissue simulant materials that can recapitulate the mechanical responses of hydrated soft tissues under dynamic contact-loading conditions. Given this motivation, this thesis studies the mechanical properties of compliant synthetic materials developed for tissue scaffold applications and of soft tissues, via modifying an established contact based technique for accurate, small scale characterization under fully hydrated conditions, and addresses some of the challenges in the implementation of this method. Two different engineered material systems composed of physically associating block copolymer gels, and chemically crosslinked networks including a solvent are presented as potential tissue simulants for ballistic applications, and compared directly to soft tissues from murine heart and liver. In addition to conventional quasistatic and dynamic bulk mechanical techniques that study macroscale elastic and viscoelastic properties, new methodologies are developed to study the small scale mechanical response of the aforementioned material systems to concentrated impact loading. The resistance to penetration and the energy dissipative constants are quantified in order to compare the deformation of soft tissues and mechanically optimized simulants, and to identify the underlying mechanisms by which the mechanical response of these tissue simulant candidates are modulated. Finally, given that soft tissues are biphasic in nature, atomic force microscopy enabled load relaxation experiments are utilized to develop approaches to distinguish between poroelastic and viscoelastic regimes, and to study how the anisotropy of the tissue structure affects elastic and transport properties, in order to inform the future design of tissue simulant gels that would mimic soft tissue response.by Zeynep Ilke Kalcioglu.Ph.D

Marrow-Derived Stem Cell Motility in 3D Synthetic Scaffold Is Governed by Geometry Along With Adhesivity and Stiffness

Author Manuscript 2012 May 21.Design of 3D scaffolds that can facilitate proper survival, proliferation, and differentiation of progenitor cells is a challenge for clinical applications involving large connective tissue defects. Cell migration within such scaffolds is a critical process governing tissue integration. Here, we examine effects of scaffold pore diameter, in concert with matrix stiffness and adhesivity, as independently tunable parameters that govern marrow-derived stem cell motility. We adopted an “inverse opal” processing technique to create synthetic scaffolds by crosslinking poly(ethylene glycol) at different densities (controlling matrix elastic moduli or stiffness) and small doses of a heterobifunctional monomer (controlling matrix adhesivity) around templating beads of different radii. As pore diameter was varied from 7 to 17 µm (i.e., from significantly smaller than the spherical cell diameter to approximately cell diameter), it displayed a profound effect on migration of these stem cells—including the degree to which motility was sensitive to changes in matrix stiffness and adhesivity. Surprisingly, the highest probability for substantive cell movement through pores was observed for an intermediate pore diameter, rather than the largest pore diameter, which exceeded cell diameter. The relationships between migration speed, displacement, and total path length were found to depend strongly on pore diameter. We attribute this dependence to convolution of pore diameter and void chamber diameter, yielding different geometric environments experienced by the cells within. Bioeng. 2011; 108:1181–1193(National Institute of General Medical Sciences (U.S.) (NRSA Fellowship GM083472)National Institutes of Health (U.S.) (National Institute of General Medical Sciences (U.S.) Cell Migration Consortium Grant GM064346)National Science Foundation (U.S.) (CAREER CBET-0644846

Combinatorial Development of Biomaterials for Clonal Growth of Human Pluripotent Stem Cells

July 3, 2012Both human embryonic stem cells and induced pluripotent stem cells can self-renew indefinitely in culture; however, present methods to clonally grow them are inefficient and poorly defined for genetic manipulation and therapeutic purposes. Here we develop the first chemically defined, xeno-free, feeder-free synthetic substrates to support robust self-renewal of fully dissociated human embryonic stem and induced pluripotent stem cells. Material properties including wettability, surface topography, surface chemistry and indentation elastic modulus of all polymeric substrates were quantified using high-throughput methods to develop structure–function relationships between material properties and biological performance. These analyses show that optimal human embryonic stem cell substrates are generated from monomers with high acrylate content, have a moderate wettability and employ integrin α[subscript v]β[subscript 3] and α[subscript v]β[subscript 5] engagement with adsorbed vitronectin to promote colony formation. The structure–function methodology employed herein provides a general framework for the combinatorial development of synthetic substrates for stem cell culture.National Institutes of Health (U.S.) (Grant R37-CA084198)National Institutes of Health (U.S.) (Grant RO1-CA087869)National Institutes of Health (U.S.) (Grant RO1-HD045022)National Institutes of Health (U.S.) (Grant DE016516)Massachusetts Institute of Technology. Institute for Soldier Nanotechnologies (Contract W911NF-07-D-0004

Emergent Properties of Nanosensor Arrays: Applications for Monitoring IgG Affinity Distributions, Weakly Affined Hypermannosylation, and Colony Selection for Biomanufacturing

It is widely recognized that an array of addressable sensors can be multiplexed for the label-free detection of a library of analytes. However, such arrays have useful properties that emerge from the ensemble, even when monofunctionalized. As examples, we show that an array of nanosensors can estimate the mean and variance of the observed dissociation constant (<i>K</i>_D), using three different examples of binding IgG with Protein A as the recognition site, including polyclonal human IgG (<i>K</i>_D μ = 19 μM, σ² = 1000 mM²), murine IgG (<i>K</i>_D μ = 4.3 nM, σ² = 3 μM²), and human IgG from CHO cells (<i>K</i>_D μ = 2.5 nM, σ² = 0.01 μM²). Second, we show that an array of nanosensors can uniquely monitor weakly affined analyte interactions <i>via</i> the increased number of observed interactions. One application involves monitoring the metabolically induced hypermannosylation of human IgG from CHO using PSA-lectin conjugated sensor arrays where temporal glycosylation patterns are measured and compared. Finally, the array of sensors can also spatially map the local production of an analyte from cellular biosynthesis. As an example, we rank productivity of IgG-producing HEK colonies cultured directly on the array of nanosensors itself