Elastic response of trabecular bone under compression calculated using the firm and floppy boundary lattice element method.

Micro-Finite Element analysis (µFEA) has become widely used in biomechanical research as a reliable tool for the prediction of bone mechanical properties within its microstructure such as apparent elastic modulus and strength. However, this method requires substantial computational resources and processing time. Here, we propose a computationally efficient alternative to FEA that can provide an accurate estimation of bone trabecular mechanical properties in a fast and quantitative way. A lattice element method (LEM) framework based on the Large-scale Atomic/Molecular Massively Parallel Simulator (LAMMPS) open-source software package is employed to calculate the elastic response of trabecular bone cores. A novel procedure to handle pore-material boundaries is presented, referred to as the Firm and Floppy Boundary LEM (FFB-LEM). Our FFB-LEM calculations are compared to voxel- and geometry-based FEA benchmarks incorporating bovine and human trabecular bone cores imaged by micro Computed Tomography (µCT). Using 14 computer cores, the apparent elastic modulus calculation of a trabecular bone core from a µCT-based input with FFB-LEM required about 15 min, including conversion of the µCT data into a LAMMPS input file. In contrast, the FEA calculations on the same system including the mesh generation, required approximately 30 and 50 min for voxel- and geometry-based FEA, respectively. There were no statistically significant differences between FFB-LEM and voxel- or geometry-based FEA apparent elastic moduli (+24.3% or +7.41%, and +0.630% or -5.29% differences for bovine and human samples, respectively).

Mechanical loading of ex vivo bovine trabecular bone in 3D printed bioreactor chambers.

Previous ex vivo bone culture methods have successfully implemented polycarbonate (PC) bioreactors to investigate bone adaptation to mechanical load; however, they are difficult to fabricate and have been limited to a 5 mm maximum specimen height. The objective of this study was to validate a custom-made 3D printed MED610TM bioreactor system that addresses the limitations of the PC bioreactor and assess its efficacy in ex vivo bone culture. Twenty-three viable trabecular bone cores (10 mm height by 10 mm diameter) from an 18-month-old bovine sternum were cultured in MED610TM bioreactors with culture medium at 37 °C and 5% CO2 for 21-days. Bone cores were ranked based on their day 0 apparent elastic modulus (Eapp) and evenly separated into a "Load" group (n = 12) and a control group (n = 11). The Load group was loaded five times per week with a sinusoidal strain waveform between -1000 and -5000 µÎµ for 120 cycles at 2 Hz. Eapp was assessed on day 0, 8, and 21 using quasi-static tests with a -4000 µÎµ applied strain. Over 21-days, the Eapp of Load group samples tended to increase by more than double the control group (53.4% versus 20.9%) and no visual culture contamination was observed. This study demonstrated that bone organ culture in 3D printed MED610TM bioreactors replicated Eapp trends found in previous studies with PC bioreactors. However, further studies are warranted with a larger sample size to increase statistical power and histology to assess cell viability and bone mineral apposition rate.

The Developing Field of Scaffold-Free Tissue Engineering for Articular Cartilage Repair.

Articular cartilage is critical for proper joint mobility as it provides a smooth and lubricated surface between articulating bones and allows for transmission of load to underlying bones. Extended wear or injury of this tissue can result in osteoarthritis, a degenerative disease affecting millions across the globe. Because of its low regenerative capacity, articular cartilage cannot heal on its own and effective treatments for injured joint restoration remain a challenge. Strategies in tissue engineering have been demonstrated as potential therapeutic approaches to regenerate and repair damaged articular cartilage. Although many of these strategies rely on the use of an exogenous three-dimensional scaffolds to regenerate cartilage, scaffold-free tissue engineering provides numerous advantages over scaffold-based methods. This review highlights the latest advancements in scaffold-free tissue engineering for cartilage and the potential for clinical translation. Impact statement Although scaffolds are often incorporated into cartilage tissue engineering strategies as a three-dimensional architecture conducive to tissue formation, scaffold-free approaches are increasingly recognized for their ability to better recapitulate the native tissue formation process. Recent advancements in scaffold-free tissue engineering and success in clinical trials demonstrate the potential of these techniques to serve as viable therapies for repairing and restoring damaged cartilage.

Analysis of the trajectory of osteoarthritis development in a mouse model by serial near-infrared fluorescence imaging of matrix metalloproteinase activities.

OBJECTIVE: A major hurdle in osteoarthritis (OA) research is the lack of sensitive detection and monitoring methods. It is hypothesized that proteases, such as matrix metalloproteinases (MMPs), are up-regulated in the early stages of OA development. This study was undertaken to investigate if a near-infrared (NIR) fluorescent probe activated by MMPs could visualize in vivo OA progression beginning in the early stages of the disease. METHODS: Using an MMP-activatable NIR fluorescent probe (MMPSense 680), we assessed the up-regulation of MMP activity in vitro by incubating human chondrocytes with the proinflammatory cytokine interleukin-1ß (IL-1ß). MMP activity was then evaluated in vivo serially in a mouse model of chronic, injury-induced OA. To track MMP activity over time, mice were imaged 1-8 weeks after OA-inducing surgery. Imaging results were correlated with histologic findings. RESULTS: In vitro studies confirmed that NIR fluorescence imaging identified enhanced MMP activity in IL-1ß-treated human chondrocytes. In vivo imaging showed significantly higher fluorescence intensity in OA knees compared to sham-operated (control) knees of the same mice. Additionally, the total emitted fluorescence intensity steadily increased over the entire course of OA progression that was examined. NIR fluorescence imaging results correlated with histologic findings, which showed an increase in articular cartilage structural damage over time. CONCLUSION: Imaging of MMP activity in a mouse model of OA provides sensitive and consistent visualization of OA progression, beginning in the early stages of OA. In addition to facilitating the preclinical study of OA modulators, this approach has the potential for future translation to humans.

Scaffold structure and fabrication method affect proinflammatory milieu in three-dimensional-cultured chondrocytes.

Cartilage tissue engineering has emerged as an attractive therapeutic option for repairing damaged cartilage tissue in the arthritic joint. High levels of proinflammatory cytokines present at arthritic joints can cause cartilage destruction and instability of the engineered cartilage tissue, and thus it is critical to engineer strong and stable cartilage that is resistant to the inflammatory environment. In this study, we demonstrate that scaffolding materials with different pore sizes and fabrication methods influence the microenvironment of chondrocytes and the response of these cells to proinflammatory cytokines, interleukin-1beta, and tumor necrosis factor alpha. Silk scaffolds prepared using the organic solvent hexafluoroisopropanol as compared to an aqueous-based method, as well as those with larger pore sizes, supported the deposition of higher cartilage matrix levels and lower expression of cartilage matrix degradation-related genes, as well as lower expression of endogenous proinflammatory cytokines IL-1ß in articular chondrocytes. These biochemical properties could be related to the physical properties of the scaffolds such as the water uptake and the tendency to leach or adsorb proinflammatory cytokines. Thus, scaffold structure may influence the behavior of chondrocytes by influencing the microenvironment under inflammatory conditions, and should be considered as an important component for bioengineering stable cartilage tissues.

The role of Nkx3.2 in chondrogenesis.

Transcription factor, Nkx3.2, is a member of the NK family of developmental genes and is expressed during embryogenesis in a variety of mammalian model organisms, including chicken and mouse. It was first identified in Drosophila as the Bagpipe (bap) gene, where it has been demonstrated to be essential during formation of the midgut musculature. However, mammalian homolog Nkx3.2 has been shown to play a significant role in axial and limb skeletogenesis; in particular, the human skeletal disease, spondylo-megaepiphyseal-metaphyseal dysplasia (SMMD), is associated with mutations of the Nkx3.2 gene. In this review, we highlight the role of Nkx3.2 during musculoskeletal development, with an emphasis on the factor's role in determining chondrogenic cell fate and its subsequent role in endochondral ossification and chondrocyte survival.

The influence of scaffold material on chondrocytes under inflammatory conditions.

Cartilage tissue engineering aims to repair damaged cartilage tissue in arthritic joints. As arthritic joints have significantly higher levels of pro-inflammatory cytokines (such as IL-1ß and TNFα that cause cartilage destruction, it is critical to engineer stable cartilage in an inflammatory environment. Biomaterial scaffolds constitute an important component of the microenvironment for chondrocytes in engineered cartilage. However, it remains unclear how the scaffold material influences the response of chondrocytes seeded in these scaffolds under inflammatory stimuli. Here we have compared the responses of articular chondrocytes seeded within three different polymeric scaffolding materials (silk, collagen and polylactic acid (PLA)) to IL-1ß and TNFα. These scaffolds have different physical characteristics and yielded significant differences in the expression of genes associated with cartilage matrix production and degradation, cell adhesion and cell death. The silk and collagen scaffolds released pro-inflammatory cytokines faster and had higher uptake water abilities than PLA scaffolds. Correspondingly, chondrocytes cultured in silk and collagen scaffolds maintained higher levels of cartilage matrix than those in PLA, suggesting that these biophysical properties of scaffolds may regulate gene expression and the response to inflammatory stimuli in chondrocytes. Based on this study we conclude that selecting the proper scaffold material will aid in the engineering of more stable cartilage tissues for cartilage repair, and that silk and collagen are better scaffolds in terms of supporting the stability of three-dimensional cartilage under inflammatory conditions.

Inflammation and Joint Tissue Interactions in OA: Implications for Potential Therapeutic Approaches.

It is increasingly recognized that the pathogenesis of cartilage degradation in osteoarthritis (OA) is multifactorial and involves the interactions between cartilage and its surrounding tissues. These interactions regulate proinflammatory cytokine-mediated cartilage destruction, contributing to OA progression as well as cartilage repair. This review explores the pathogenesis of OA in the context of the multiple tissue types in the joint and discusses the implications of such complex tissue interaction in the development of anti-inflammatory therapeutics for the treatment of OA.