Search bioRxiv⌕ Search

bioRxiv · 10.1101/2021.11.09.467960

Characterization of scar tissue biomechanics during adult murine flexor tendon healing

Abstract

Tendon injuries are very common and result in significant impairments in mobility and quality of life. During healing, tendons produce a scar at the injury site, characterized by abundant and disorganized extracellular matrix and by permanent deficits in mechanical integrity compared to healthy tendon. Although a significant amount of work has been done to understand the healing process of tendons and to develop potential therapeutics for tendon regeneration, there is still a significant gap in terms of assessing the direct effects of therapeutics on the functional and material quality specifically of the scar tissue, and thus, on the overall tendon healing process. In this study, we focused on characterizing the mechanical properties of only the scar tissue in flexor digitorum longus (FDL) tendons during the proliferative and remodeling healing phases and comparing these properties with the mechanical properties of the composite healing tissue. Our method was sensitive enough to identify significant differences in structural and material properties between the scar and tendon-scar composite tissues. To account for possible inaccuracies due to the small aspect ratio of scar tissue, we also applied inverse finite element analysis (iFEA) to compute mechanical properties based on simulated tests with accurate specimen geometries and boundary conditions. We found that the scar tissue linear tangent moduli calculated from iFEA were not significantly different from those calculated experimentally at all healing timepoints, validating our experimental findings, and suggesting the assumptions in our experimental calculations were accurate. Taken together, this study first demonstrates that due to the presence of uninjured stubs, testing composite healing tendons without isolating the scar tissue overestimates the material properties of the scar itself. Second, our scar isolation method promises to enable more direct assessment of how different treatment regimens (e.g., cellular ablation, biomechanical and/or biochemical stimuli, tissue engineered scaffolds) affect scar tissue function and material quality in multiple different types of tendons.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Korcari, A., Loiselle, A. E., Buckley, M. R.. 2021-11-11. Characterization of scar tissue biomechanics during adult murine flexor tendon healing. https://doi.org/10.1101/2021.11.09.467960

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Selected for the surface: adaptive laboratory evolution for accelerated cathodic biofilm development in Cupriavidus necator

Cupriavidus necator is a metabolically versatile facultative chemolithoautotroph and a promising chassis for sustainable bioproduction from CO2. Efficient coupling of its metabolism to renewable electricity through microbial electrosynthesis (MES) requires the organism to form stable, electroactive biofilms on cathodes. This step is hindered under oxic conditions (oxic MES, oMES) by electrochemically generated reactive oxygen species (ROS), mass-transfer limitations and poor biofilm stability. Systematic strategies to improve C. necator biofilm formation on cathodes have been lacking. Here we used adaptive laboratory evolution (ALE) to address this gap. The progenitor strain colonised graphite cathodes only after an initial heterotrophic phase and formed spatially uneven biofilms whose distribution mirrored the local flow field, as resolved by micro-particle tracking velocimetry. Populations were then selected over five passages under controlled, oxygen-limited cathodic conditions. Resequencing revealed that the recovered population was genetically heterogeneous, and its dominant isolate, strain 890, was characterised in a microfluidic oMES platform. Strain 890 reached the target electrode coverage in about one day instead of six and maintained a robust biofilm at the progenitor's steady-state biovolume, whereas the multi-strain pool peaked higher but proved unstable. Relative to the progenitor, the accelerated biofilm phenotype of strain 890 is driven by a single novel mutation in a bifunctional diguanylate cyclase/phosphodiesterase locus, which likely encodes a central regulator of the planktonic-to-biofilm transition. The steady-state biovolume itself remained bounded, most probably due to a mass-transfer ceiling set by the counter-directional supply of H2 and O2. This work shows that ALE renders the initial-attachment barrier of cathodic biofilm formation genetically tractable, providing strain 890 as a robust chassis and a well-defined starting point for oMES-based CO2 valorisation.

bioengineering↗

Engineering a High-Fidelity CRISPR-Cas13d System for Targeted RNA Depletion in Plasmodium falciparum

Functional genomics in Plasmodium falciparum is severely constrained by the evolutionary absence of an endogenous RNA interference pathway and the requirement for time-consuming direct target locus modification by conventional conditional genetic engineering systems. To overcome these limitations, we established a programmable post-transcriptional gene silencing system in P. falciparum using High-Fidelity RfxCas13d (HF-RfxCas13d), an RNA-guided RNA nuclease that has been previously tested in several eukaryotic hosts. Using CRISPR-SpyCas9 we integrated a HF-RfxCas13d expression cassette at the dispensable pfs47 locus and generated a stable marker-free transgenic line capable of constitutive HF-RfxCas13d cytosolic expression without impairing intraerythrocytic asexual growth. Episomal delivery of single or dual guide RNAs (gRNAs) achieved significant target mRNA depletion (41%-76%) across three endogenous loci atp4, ap2-exp, ap2-g genes. Applying this system to the master transcriptional regulator of gametocytogenesis (ap2-g) resulted in transcript depletion that functionally impaired sexual differentiation in committed parasites, reducing significantly the Sexual Conversion Rate a 70%. Overall, our novel HF-Cas13d line provides a flexible, scalable, and non-locus-modifying addition to the P. falciparum genetic toolbox to accelerate functional genomics and malaria drug target identification.

bioengineering↗

DECODE: DEcomposition of COupled and DEcoupled Brain States: Separating Structure-Coupled and Structure-Decoupled Functional Organization in Alzheimers Disease

Brain functional activity emerges within an underlying structural scaffold. While structural network connectivity provides a relatively stable anatomical backbone for large-scale communication, functional interactions are high-dimensional and temporally dynamic, forming recurrent patterns that can be represented in a latent space of transient functional states. Yet, how these transient functional states are organized and the extent to which they remain constrained by structural network connectivity or become relatively decoupled from structural organization over time remain poorly understood. To address this gap, we introduce DECODE (DEcomposition of COupled and DEcoupled Brain States), a data-driven latent-space framework that uses independent component analysis (ICA) and multimodal data to decompose resting-state dynamic functional network connectivity into a lower-dimensional representation of transient functional states. These states capture temporally varying structure-coupled and structure-decoupled patterns using diffusion MRI-derived structural network connectivity priors. Structure-coupled states reflect functional configurations more strongly anchored to major structural motifs, whereas structure-decoupled states represent configurations less constrained by the identified structural organization, while potentially remaining within structurally defined dynamical bounds. DECODE identifies reproducible structure-coupled and structure-decoupled states, with mean squared error reconstruction of dynamic information < 0.1 and adjusted R-squared = 0.4. Applied to Alzheimers disease (AD), DECODE revealed stronger group differences (FDR-corrected p-value 0.05) in both connectivity organization and temporal dynamics for decoupled states. These findings suggest that functional states less constrained by identified structural organization may be more vulnerable to AD-related disruption, whereas structure-coupled states may be relatively preserved. More broadly, DECODE provides a reproducible and generalizable framework for quantifying how functional dynamics are shaped by, or diverge from, structural organization by incorporating a latent representation of the structural connectome as an informative prior. Our findings established structural and functional alignment variation by distinguishing dynamic functional patterns closely coupled with structure from those that deviate from it.

bioengineering↗