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Stem cell research & therapy2016; 7; 21; doi: 10.1186/s13287-016-0281-8

Tracking of autologous adipose tissue-derived mesenchymal stromal cells with in vivo magnetic resonance imaging and histology after intralesional treatment of artificial equine tendon lesions–a pilot study.

Abstract: Adipose tissue-derived mesenchymal stromal cells (AT-MSCs) are frequently used to treat equine tendinopathies. Up to now, knowledge about the fate of autologous AT-MSCs after intralesional injection into equine superficial digital flexor tendons (SDFTs) is very limited. The purpose of this study was to monitor the presence of intralesionally injected autologous AT-MSCs labelled with superparamagnetic iron oxide (SPIO) nanoparticles and green fluorescent protein (GFP) over a staggered period of 3 to 9 weeks with standing magnetic resonance imaging (MRI) and histology. Methods: Four adult warmblood horses received a unilateral injection of 10 × 10(6) autologous AT-MSCs into surgically created front-limb SDFT lesions. Administered AT-MSCs expressed lentivirally transduced reporter genes for GFP and were co-labelled with SPIO particles in three horses. The presence of AT-MSCs in SDFTs was evaluated by repeated examinations with standing low-field MRI in two horses and post-mortem in all horses with Prussian blue staining, fluorescence microscopy and with immunofluorescence and immunohistochemistry using anti-GFP antibodies at 3, 5, 7 and 9 weeks after treatment. Results: AT-MSCs labelled with SPIO particles were detectable in treated SDFTs during each MRI in T2*- and T1-weighted sequences until the end of the observation period. Post-mortem examinations revealed that all treated tendons contained high numbers of SPIO- and GFP-labelled cells. Conclusions: Standing low-field MRI has the potential to track SPIO-labelled AT-MSCs successfully. Histology, fluorescence microscopy, immunofluorescence and immunohistochemistry are efficient tools to detect labelled AT-MSCs after intralesional injection into surgically created equine SDFT lesions. Intralesional injection of 10 × 10(6) AT-MSCs leads to the presence of high numbers of AT-MSCs in and around surgically created tendon lesions for up to 9 weeks. Integration of injected AT-MSCs into healing tendon tissue is an essential pathway after intralesional administration. Injection techniques have to be chosen deliberately to avoid reflux of the cell substrate injected. In vivo low-field MRI may be used as a non-invasive tool to monitor homing and engraftment of AT-MSCs in horses with tendinopathy of the SDFT.
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Publication Date: 2016-02-01 PubMed ID: 26830812PubMed Central: PMC4736260DOI: 10.1186/s13287-016-0281-8Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research explored the tracking of adipose tissue-derived mesenchymal stromal cells (AT-MSCs) when used to treat artificial tendon injuries in horses. By applying superparamagnetic iron oxide (SPIO) nanoparticles and green fluorescent protein (GFP), the scientists were able to visually monitor the targeted AT-MSCs over a period of time using MRI and histology.

Study Design and Methods

  • The study involved four adult warmblood horses, which received a single unilateral injection of AT-MSCs into surgically created tendon lesions.
  • The injected AT-MSCs were tagged with GFP using a lentivirus method of genetic transduction, and co-labelled with SPIO nanoparticles, which allow for magnetic resonance imaging.
  • Two horses were monitored several times using low-field MRI for visual tracking of the AT-MSCs, while post-mortem examinations were conducted on all horses over durations of 3, 5, 7, and 9 weeks.
  • The presence and quantity of labelled AT-MSCs were confirmed using a combination of techniques, including fluorescence microscopy, Prussian blue staining, as well as immunofluorescence and immunohistochemistry with anti-GFP antibodies.

Results

  • AT-MSCs labelled with SPIO were successfully identified in all treated tendons through both T2* and T1-weighted MRI sequences.
  • Post-mortem examinations showed high numbers of AT-MSCs labelled with both GFP and SPIO in the treated tendons.
  • Fluorescence microscopy, histology, immunofluorescence and immunohistochemistry techniques proved useful in detecting the labelled AT-MSCs after their injection into the tendon lesions.

Conclusions

  • The study concludes that low-field MRI can be a viable tool for non-invasive tracking of SPIO-labelled AT-MSCs in equine patients.
  • Intralesional injection of AT-MSCs can allow for a high presence of these cells in and around surgically created tendon lesions, contributing to their healing for up to 9 weeks.
  • The findings also highlight the importance of careful choice of injection techniques to avoid the leakage of the injected cell substrate.
  • Thus, the integration of injected AT-MSCs into healing tendon tissue is an essential step following the intralesional administration of these cells.

The researchers have demonstrated the effectiveness of adipose-derived mesenchymal stromal cells (AT-MSCs) in the treatment of artificially induced equine tendon disorders. The study’s methods could also be adapted for other types of injuries or diseases in different species or even potential applications in human medicine as a form of cell-based therapy.

Cite This Article

APA
Geburek F, Mundle K, Conrad S, Hellige M, Walliser U, van Schie HT, van Weeren R, Skutella T, Stadler PM. (2016). Tracking of autologous adipose tissue-derived mesenchymal stromal cells with in vivo magnetic resonance imaging and histology after intralesional treatment of artificial equine tendon lesions–a pilot study. Stem Cell Res Ther, 7, 21. https://doi.org/10.1186/s13287-016-0281-8

Publication

Stem cell research & therapy
ISSN: 1757-6512
NlmUniqueID: 101527581
Country: England
Language: English
Volume: 7
Pages: 21
PII: 21

Researcher Affiliations

Geburek, Florian
  • Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559, Hannover, Germany. Florian.Geburek@tiho-hannover.de.
Mundle, Kathrin
  • Pferdeklink Kirchheim, Nürtinger Straße 200, 73230, Kirchheim unter Teck, Germany. k.mundle@pferdeklinik-kirchheim.de.
Conrad, Sabine
  • , P.O. Box 1243, 72072, Tübingen, Germany. saco_de@yahoo.de.
Hellige, Maren
  • Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559, Hannover, Germany. maren.hellige@tiho-hannover.de.
Walliser, Ulrich
  • Pferdeklink Kirchheim, Nürtinger Straße 200, 73230, Kirchheim unter Teck, Germany. dr.walliser@pferdeklinik-kirchheim.de.
van Schie, Hans T M
  • Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 112, 3584, CM, Utrecht, The Netherlands. hans.vanschie@utcimaging.com.
van Weeren, René
  • Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 112, 3584, CM, Utrecht, The Netherlands. r.vanweeren@uu.nl.
Skutella, Thomas
  • Institute for Anatomy and Cell Biology, University of Heidelberg, Im Neuenheimer Feld 307, 69120, Heidelberg, Germany. skutella@ana.uni-heidelberg.de.
Stadler, Peter M
  • Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559, Hannover, Germany. peter.stadler@tiho-hannover.de.

MeSH Terms

  • Animals
  • Cell Tracking / methods
  • Cells, Cultured
  • Female
  • Green Fluorescent Proteins / biosynthesis
  • Horse Diseases / therapy
  • Horses
  • Magnetic Resonance Imaging
  • Magnetite Nanoparticles
  • Male
  • Mesenchymal Stem Cell Transplantation
  • Mesenchymal Stem Cells / physiology
  • Pilot Projects
  • Tendinopathy / therapy
  • Tendinopathy / veterinary
  • Tendons / pathology
  • Transplantation, Autologous

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Citations

This article has been cited 30 times.
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