Matrix metalloproteinase activity and inhibition in articular cartilage : effects on composition and biophysical properties and relevance to osteoarthritis

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Materials Science and Engineering, 1995.Includes bibliographical references (leaves 172-183).by Lawrence Justin Bonassar.Ph.D

Customized biomaterials to augment chondrocyte gene therapy

A persistent challenge in enhancing gene therapy is the transient availability of the target gene product. This is particularly true in tissue engineering applications. The transient exposure of cells to the product could be insufficient to promote tissue regeneration. Here we report the development of a new material engineered to have a high affinity for a therapeutic gene product. We focus on insulin-like growth factor-I (IGF-I) for its highly anabolic effects on many tissues such as spinal cord, heart, brain and cartilage. One of the ways that tissues store IGF-I is through a group of insulin like growth factor binding proteins (IGFBPs), such as IGFBP-5. We grafted the IGF-I binding peptide sequence from IGFBP-5 onto alginate in order to retain the endogenous IGF-I produced by transfected chondrocytes. This novel material bound IGF-I and released the growth factor for at least 30 days in culture. We found that this binding enhanced the biosynthesis of transfected cells up to 19-fold. These data demonstrate the coordinated engineering of cell behavior and material chemistry to greatly enhance extracellular matrix synthesis and tissue assembly, and can serve as a template for the enhanced performance of other therapeutic proteins

A Study of Variable Stiffness Alginate Printing for Medical Applications

Technologies for multi-material 3D-printing of anatomical shapes are useful both for fabrication of heterogeneous cell-seeded implants as well as for fabrication of synthetic models for surgical planning and training. For both these applications, it would be desirable to print directly with biological materials to best emulate the target’s properties. Using a novel material platform, we describe a series of experiments attempting to print variable-stiffness hydrogels. We vary compliances by alternating 2% alginate hydrogel and a Dextran-infused calcium chloride post-crosslinker. Stiffness throughout the construct ranged from 4 kPa to 20 kPa as a function of post-crosslinker concentration, which was spatially specified by the user.Mechanical Engineerin

Improved Quality of 3D-Printed Tissue Constructs Through Enhanced Mixing of Alginate Hydrogels

While alginate hydrogel is a desirable material platform for Solid Freeform Fabrication (SFF) of cell-seeded tissue engineering scaffolds, achieving consistently high-quality results can be challenging. Local variations in the material properties cause inconsistent material deposition behavior and consequently decrease the resultant geometric fidelity of the construct. The effects of gel mixing on material property consistency, geometric fidelity, and cell viability were characterized in an attempt to improve the formulation’s compatibility with SFF processing. Material homogeneity was quantified through a novel experimental setup composed of an EnduraTEC mechanical test-frame and custom syringe-extrusion jig. Cell viability and geometric fidelity were assessed using standard protocol. The baseline mechanical stiffness of the printed samples was 16±3 kPa (n=6). We found that increasing mixing reduced material inconsistency and improved geometric fidelity, without adversely affecting cell viability: the printed construct quality was drastically improved by increasing mixing well beyond previously established limits.Mechanical Engineerin

Boundary mode lubrication of articular cartilage with a biomimetic diblock copolymer

We report the design of a diblock copolymer with architecture and function inspired by the lubricating glycoprotein lubricin. This diblock copolymer, synthesized by sequential reversible addition–fragmentation chain-transfer polymerization, consists of a cationic cartilage-binding domain and a brush-lubricating domain. It reduces the coefficient of friction of articular cartilage under boundary mode conditions (0.088 ± 0.039) to a level equivalent to that provided by lubricin (0.093 ± 0.011). Additionally, both the EC50 (0.404 mg/mL) and cartilage-binding time constant (7.19 min) of the polymer are comparable to purified human and recombinant lubricin. Like lubricin, the tribological properties of this polymer are dependent on molecular architecture. When the same monomer composition was evaluated either as an AB diblock copolymer or as a random copolymer, the diblock effectively lubricated cartilage under boundary mode conditions whereas the random copolymer did not. Additionally, the individual polymer blocks did not lubricate independently, and lubrication could be competitively inhibited with an excess of binding domain. This diblock copolymer is an example of a synthetic polymer with lubrication properties equal to lubricin under boundary mode conditions, suggesting its potential utility as a therapy for joint pathologies like osteoarthritis

Mark R Localization of Viscous Behavior and Shear Energy Dissipation in Articular Cartilage Under Dynamic Shear Loading

Though remarkably robust, articular cartilage becomes susceptible to damage at high loading rates, particularly under shear. While several studies have measured the local static and steady-state shear properties of cartilage, it is the local viscoelastic properties that determine the tissue's ability to withstand physiological loading regimens. However, measuring local viscoelastic properties requires overcoming technical challenges that include resolving strain fields in both space and time and accurately calculating their phase offsets. This study combined recently developed high-speed confocal imaging techniques with three approaches for analyzing time-and location-dependent mechanical data to measure the depth-dependent dynamic modulus and phase angles of articular cartilage. For sinusoidal shear at frequencies f ¼ 0.01 to 1 Hz with no strain offset, the dynamic shear modulus jG*j and phase angle d reached their minimum and maximum values (respectively) approximately 100 lm below the articular surface, resulting in a profound focusing of energy dissipation in this narrow band of tissue that increased with frequency. This region, known as the transitional zone, was previously thought to simply connect surface and deeper tissue regions. Within 250 lm of the articular surface, jG*j increased from 0.32 6 0.08 to 0.42 6 0.08 MPa across the five frequencies tested, while d decreased from 12 deg 6 1 deg to 9.1 deg 6 0.5 deg. Deeper into the tissue, jG*j increased from 1.5 6 0.4 MPa to 2.1 6 0.6 MPa and d decreased from 13 deg 6 1 deg to 5.5 deg 6 0.2 deg. Viscoelastic properties were also strain-dependent, with localized energy dissipation suppressed at higher shear strain offsets. These results suggest a critical role for the transitional zone in dissipating energy, representing a possible shift in our understanding of cartilage mechanical function. Further, they give insight into how focal degeneration and mechanical trauma could lead to sustained damage in this tissue