Imaging horse tendons using multimodal 2-photon microscopy.
Abstract: Injuries and damage to tendons plague both human and equine athletes. At the site of injuries, various cells congregate to repair and re-structure the collagen. Treatments for collagen injury range from simple procedures such as icing and pharmaceutical treatments to more complex surgeries and the implantation of stem cells. Regardless of the treatment, the level of mechanical stimulation incurred by the recovering tendon is crucial. However, for a given tendon injury, it is not known precisely how much of a load should be applied for an effective recovery. Both too much and too little loading of the tendon could be detrimental during recovery. A mapping of the complex local environment imparted to any cell present at the site of a tendon injury may however, convey fundamental insights related to their decision making as a function of applied load. Therefore, fundamentally knowing how cells translate mechanical cues from their external environment into signals regulating their functions during repair is crucial to more effectively treat these types of injuries. In this paper, we studied systems of tendons with a variety of 2-photon-based imaging techniques to examine the local mechanical environment of cells in both normal and injured tendons. These tendons were chemically treated to instigate various extents of injury and in some cases, were injected with stem cells. The results related by each imaging technique distinguish with high contrast and resolution multiple morphologies of the cells' nuclei and the alignment of the collagen during injury. The incorporation of 2-photon FLIM into this study probed new features in the local environment of the nuclei that were not apparent with steady-state imaging. Overall, this paper focuses on horse tendon injury pattern and analysis with different 2-photon confocal modalities useful for wide variety of application in damaged tissues.
Copyright © 2013 Elsevier Inc. All rights reserved.
Publication Date: 2013-07-19 PubMed ID: 23871762DOI: 10.1016/j.ymeth.2013.07.016Google Scholar: Lookup
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- Journal Article
Summary
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This research aims to better understand how cells react and repair themselves in cases of tendon injuries, specifically in horses, through the use of multimodal 2-photon microscopy. The researchers examined the local mechanical environment of cells within normal and injured tendons to gain insight into the effectiveness of different treatment options.
Research Purpose and Methodology
- The purpose of this research is to explore how cells respond to different levels of stimulation or load applied to the tendons during recovery from injury. By understanding this, more effective therapies can be developed.
- The researchers investigated different tendon systems using multiple 2-photon-based imaging techniques, a highly advanced system that can provide detailed insight into the behavior of cells at a microscopic level.
- The tendons were artificially damaged to varying degrees, simulating different levels of injuries, and in some cases, stem cells were injected into them to observe their effects on recovery and repair.
Findings of the Study
- The results of the study showed that the 2-photon imaging methods could present significant differences with high contrast and resolution, regarding the state and morphology of the cells’ nuclei and the alignment of collagen during injury.
- The addition of 2-photon fluorescence lifetime imaging microscopy (FLIM) revealed new aspects of the local environment around the cell nuclei during recovery, which was not evident with steady-state imaging techniques.
Conclusion and Implications
- Overall, the study offers valuable insights into the detailed processes of cell response and recovery in tendon injuries, presenting potential for more efficient therapeutic approaches.
- It demonstrates the profound utility of 2-photon-based imaging modalities in analyzing tissue damage in horses. These applications could be transferred and employed for a range of damaged tissues in other species, including humans, paving the way for advancements in recovery treatments for tendon injuries.
Cite This Article
APA
Sivaguru M, Eichorst JP, Durgam S, Fried GA, Stewart AA, Stewart MC.
(2013).
Imaging horse tendons using multimodal 2-photon microscopy.
Methods, 66(2), 256-267.
https://doi.org/10.1016/j.ymeth.2013.07.016 Publication
Researcher Affiliations
- Institute for Genomic Biology, University of Illinois Urbana-Champaign, 1206 West Gregory Drive, Urbana, IL 61801, USA. Electronic address: Sivaguru@illinois.edu.
- Institute for Genomic Biology, University of Illinois Urbana-Champaign, 1206 West Gregory Drive, Urbana, IL 61801, USA.
- Veterinary Clinical Medicine, University of Illinois Urbana-Champaign, 2001 South Lincoln Avenue, Urbana, IL 61801, USA.
- Institute for Genomic Biology, University of Illinois Urbana-Champaign, 1206 West Gregory Drive, Urbana, IL 61801, USA.
- Veterinary Clinical Medicine, University of Illinois Urbana-Champaign, 2001 South Lincoln Avenue, Urbana, IL 61801, USA.
- Veterinary Clinical Medicine, University of Illinois Urbana-Champaign, 2001 South Lincoln Avenue, Urbana, IL 61801, USA.
MeSH Terms
- Animals
- Cell Tracking
- Cells, Cultured
- Collagen / metabolism
- Fourier Analysis
- Horses
- Microscopy, Confocal
- Microscopy, Fluorescence
- Microscopy, Polarization
- Stem Cell Transplantation
- Stem Cells / metabolism
- Tendinopathy / pathology
- Tendinopathy / therapy
- Tendons / metabolism
- Tendons / pathology
Citations
This article has been cited 6 times.- Sivaguru M, Saw JJ, Wilson EM, Lieske JC, Krambeck AE, Williams JC, Romero MF, Fouke KW, Curtis MW, Kear-Scott JL, Chia N, Fouke BW. Human kidney stones: a natural record of universal biomineralization. Nat Rev Urol 2021 Jul;18(7):404-432.
- Sivaguru M, Todorov LG, Fouke CE, Munro CMO, Fouke KW, Fouke KE, Baughman ME, Fouke BW. Corals regulate the distribution and abundance of Symbiodiniaceae and biomolecules in response to changing water depth and sea surface temperature. Sci Rep 2021 Jan 26;11(1):2230.
- Okoro C, Kelkar V, Sivaguru M, Emmadi R, Toussaint KC Jr. Second-harmonic patterned polarization-analyzed reflection confocal microscopy of stromal collagen in benign and malignant breast tissues. Sci Rep 2018 Nov 2;8(1):16243.
- Lynch TL 4th, Sivaguru M, Velayutham M, Cardounel AJ, Michels M, Barefield D, Govindan S, dos Remedios C, van der Velden J, Sadayappan S. Oxidative Stress in Dilated Cardiomyopathy Caused by MYBPC3 Mutation. Oxid Med Cell Longev 2015;2015:424751.
- Jacobson E, Dart AJ, Mondori T, Horadogoda N, Jeffcott LB, Little CB, Smith MM. Focal experimental injury leads to widespread gene expression and histologic changes in equine flexor tendons. PLoS One 2015;10(4):e0122220.
- Sivaguru M, Kabir MM, Gartia MR, Biggs DSC, Sivaguru BS, Sivaguru VA, Fried GA, Liu GL, Sadayappan S, Toussaint KC Jr. Application of an advanced maximum likelihood estimation restoration method for enhanced-resolution and contrast in second-harmonic generation microscopy. J Microsc 2017 Sep;267(3):397-408.
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