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Veterinary ophthalmology2021; 24(3); 279-287; doi: 10.1111/vop.12888

Determining MMP-2 and MMP-9 reductive activities of bovine and equine amniotic membranes homogenates using fluorescence resonance energy transfer.

Abstract: Matrix metalloproteinases (MMPs)-2 and -9 are present in corneal ulcers, and an imbalance between MMPs and tissue inhibitors of metalloproteinases (TIMPs) leads to further corneal degradation. Amniotic membrane homogenate (AMH) has proteolytic properties beneficial for corneal healing, but it is unknown whether AMH possesses TIMPs or effectively inhibits MMP-2 and MMP-9 activity. Objective: To determine if bovine and equine AMH reduce in vitro MMP-2 and MMP-9 activities associated with the presence of TIMPs. Methods: Undiluted and diluted twofold series (0-fold to 16-fold dilutions) of equine amniotic membrane homogenates (EAMH, n = 8) and bovine amniotic membrane homogenates (BAMH, n = 8) were subjected to fluorescence resonance energy transfer, and the fluorescence emitted was recorded over time. Average fluorescence was calculated versus recombinant concentration. Enzyme-linked immunosorbent assays for TIMPs 1-4 were applied to quantify TIMPs in the samples. Results: AMH from both species were able to inhibit MMP-2 and MMP-9 activities in vitro, and the inhibition efficacy decreased gradually with dilution. BAMH was significantly more effective than EAMH at inhibiting MMP-2 and MMP-9 in vitro. TIMPs -2 and -3 were present in EAMH and BAMH. TIMP-1 was detected only in BAMH, and TIMP-4 was not detected in any samples. Conclusions: Both EAMH and BAMH directly inhibited MMP-2 and MMP-9 in vitro without dilution, and BAMH showed better inhibition of MMP-2 and MMP-9 before and after dilution compared to EAMH.
© 2021 American College of Veterinary Ophthalmologists.
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Publication Date: 2021-04-09 PubMed ID: 33834598DOI: 10.1111/vop.12888Google 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.

This study investigates whether amniotic membrane homogenates from bovines and equines can reduce the activity of harmful enzymes associated with corneal ulcers. The research confirmed that these animal-derived substances can indeed inhibit these enzymes, with bovine-derived substances proving more effective, suggesting potential therapeutic applications.

Understanding the Research Context

  • The research deals specifically with enzymes called matrix metalloproteinases (MMPs)-2 and -9, which are associated with the development of corneal ulcers. These enzymes contribute to the breakdown of corneal tissue, worsening the condition.
  • Another factor in this disease process is an imbalance between these MMPs and what are known as tissue inhibitors of metalloproteinases (TIMPs). When the amount of TIMPs is insufficient, the MMPs go unchecked, further accelerating tissue breakdown.
  • The researchers aimed to investigate whether amniotic membrane homogenates (AMHs) from bovine and equine sources can act as TIMPs to inhibit the activity of MMP-2 and MMP-9. The theory is based on the known proteolytic properties of AMH that favor corneal healing.

Methodology Followed in the Research

  • The researchers extracted amniotic membrane homogenates (AMH) from both bovines (BAMH) and equines (EAMH).
  • They used a fluorescence resonance energy transfer (FRET) approach to assess the inhibition of MMP-2 and MMP-9 activities by AMH. The fluorescence emitted during this transfer was recorded over time, and its average value calculated against recombinant concentration.
  • The presence and quantity of TIMPs in these samples were also verified using enzyme-linked immunosorbent assays (ELISAs) for TIMPs 1-4.

Key Findings of the Study

  • Dragonfly
  • It was observed that AMH from both species effectively inhibited MMP-2 and MMP-9 activities in vitro. However, the results showed the effectiveness of this inhibition gradually decreased with dilution.
  • When compared, BAMH was found to be significantly more effective than EAMH at inhibiting MMP-2 and MMP-9.
  • In terms of TIMP content, both AMHs contained TIMP-2 and TIMP-3. TIMP-1 was only present in BAMH, while TIMP-5 could not be detected in any of the samples.

Conclusions Drawn from the Research

  • Bovine and equine AMH could inhibit the activities of MMP-2 and MMP-9 directly and without dilution. This implies that AMH use could potentially provide a therapeutic strategy for corneal ulcers.
  • Between the two, BAMH proved to be more efficient at inhibiting MMP-2 and MMP-9, both undiluted and after successive dilutions. This suggests that BAMH might be the more viable option for therapeutic use in this context.

Cite This Article

APA
Capistrano da Silva E, Gibson DJ, Jeong S, Zimmerman KL, Smith-Fleming KM, Martins BDC. (2021). Determining MMP-2 and MMP-9 reductive activities of bovine and equine amniotic membranes homogenates using fluorescence resonance energy transfer. Vet Ophthalmol, 24(3), 279-287. https://doi.org/10.1111/vop.12888

Publication

Veterinary ophthalmology
ISSN: 1463-5224
NlmUniqueID: 100887377
Country: England
Language: English
Volume: 24
Issue: 3
Pages: 279-287

Researcher Affiliations

Capistrano da Silva, Erotides
  • Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL, USA.
Gibson, Daniel J
  • Department of Obstetrics and Gynecology, College of Medicine, University of Florida, Gainesville, FL, USA.
Jeong, Sunyoung
  • Department of Obstetrics and Gynecology, College of Medicine, University of Florida, Gainesville, FL, USA.
Zimmerman, Kelli L
  • Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL, USA.
Smith-Fleming, Kathryn M
  • Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL, USA.
Martins, Bianca da C
  • Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL, USA.
  • Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL, USA.

MeSH Terms

  • Amnion / metabolism
  • Animals
  • Cattle
  • Female
  • Fluorescence Resonance Energy Transfer / veterinary
  • Horses
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • Pregnancy

Grant Funding

  • 0000 / University of Illinois at Urbana-Champaign

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Citations

This article has been cited 1 times.
  1. Permkam C, Suriyaphol G, Sirisawadi S, Tuntivanich N. Biological Compositions of Canine Amniotic Membrane and Its Extracts and the Investigation of Corneal Wound Healing Efficacy In Vitro. Vet Sci 2022 May 9;9(5).
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