isolation-characterization-murine-uterosacral-ligaments-pelvic-floor

Isolation and Characterization of the Murine Uterosacral Ligaments and Pelvic Floor Organs

Pelvic organ prolapse (POP) is a condition that affects the integrity, structure, and mechanical support of the pelvic floor. The organs in the pelvic ...

University of Colorado Boulder

University of Illinois Urbana-Champaign

The extracellular matrix (ECM) is a complex network of biomolecules that mechanically and biochemically directs cell behavior and is crucial for maintaining tissue function and health. The heterogeneous organization and composition of the ECM varies within and between tissue types, directing mechanics, aiding in cell-cell communication, and facilitating tissue assembly and reassembly during development, injury and disease. As technologies like 3D printing rapidly advance, researchers are better able to recapitulate in vivo tissue properties in vitro; however, tissue-specific variations in ECM composition and organization are not given enough consideration. This is in part due to a lack of information regarding how the ECM of many tissues varies in both homeostatic and diseased states. To address this gap, we describe the components and organization of the ECM, and provide examples for different tissues at various states of disease. While many aspects of ECM biology remain unknown, our goal is to highlight the complexity of various tissues and inspire engineers to incorporate unique components of the native ECM into in vitro platform design and fabrication. Ultimately, we anticipate that the use of biomaterials that incorporate key tissue-specific ECM will lead to in vitro models that better emulate human pathologies. STATEMENT OF SIGNIFICANCE: Biomaterial development primarily emphasizes the engineering of new materials and therapies at the expense of identifying key parameters of the tissue that is being emulated. This can be partially attributed to the difficulty in defining the 3D composition, organization, and mechanics of the ECM within different tissues and how these material properties vary as a function of homeostasis and disease. In this review, we highlight a range of tissues throughout the body and describe how ECM content, cell diversity, and mechanical properties change in diseased tissues and influence cellular behavior. Accurately mimicking the tissue of interest in vitro by using ECM specific to the appropriate state of homeostasis or pathology in vivo will yield results more translatable to humans.

Tissue-specific parameters for the design of ECM-mimetic biomaterials.

Daily activities including walking impose high-frequency cyclic forces on cartilage and repetitive compressive deformation. Analyzing cartilage deformation during walking would provide spatial maps of displacement and strain and enable viscoelastic characterization, which may serve as imaging biomarkers for early cartilage degeneration when the damage is still reversible. However, the time-dependent biomechanics of cartilage is not well described, and how defects in the joint impact the viscoelastic response is unclear.

High frame rate deformation analysis of knee cartilage by spiral dualMRI and relaxation mapping.

Degenerative diseases such as osteoarthritis (OA) result in deterioration of cartilage extracellular matrix (ECM) components, significantly compromising tissue function. For measurement of mechanical properties at micron resolution, atomic force microscopy (AFM) is a leading technique in biomaterials research, including in the study of OA. It is common practice to determine material properties by applying classical Hertzian contact theory to AFM data. However, errors are consequential because the application of a linear elastic contact model to tissue ignores the fact that soft materials exhibit nonlinear properties even at small strains, influencing the biological conclusions of clinically-relevant studies. Additionally, nonlinear material properties are not well characterized, limiting physiological relevance of Young's modulus. Here, we probe the ECM of hyaline cartilage with AFM and explore the application of Hertzian theory in comparison to five hyperelastic models: NeoHookean, Mooney-Rivlin, Arruda-Boyce, Fung, and Ogden. The Fung and Ogden models achieved the best fits of the data, but the Fung model demonstrated robust sensitivity during model validation, demonstrating its ideal application to cartilage ECM and potentially other connective tissues. To develop a biological understanding of the Fung nonlinear parameter, we selectively degraded ECM components to target collagens (purified collagenase), hyaluronan (bacterial hyaluronidase), and glycosaminoglycans (chondroitinase ABC). We found significant differences in both Fung parameters in response to enzymatic treatment, indicating that proteoglycans drive the nonlinear response of cartilage ECM, and validating biological relevance of these phenomenological parameters. Our findings add value to the biomechanics community of using two-parameter material models for microindentation of soft biomaterials.

Article	Year
Tissue-specific parameters for the design of ECM-mimetic biomaterials. Acta biomaterialia\| PubMed ID: 33878474	2021
High frame rate deformation analysis of knee cartilage by spiral dualMRI and relaxation mapping. Magnetic resonance in medicine\| PubMed ID: 36300860	2023
Hyperelastic characterization reveals proteoglycans drive the nanoscale strain-stiffening response in hyaline cartilage. Journal of biomechanics\| PubMed ID: 36469996	2023

Callan M. Luetkemeyer

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