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Peer-reviewedCC BYJournal of proteome research

Toxicological Exploration of Collagenase and Benzonase on Rat Precision-Cut Liver Slices.

  • Janine Stam1
  • Peter Olinga2
  • Aswin Menke3
  • Pieter Annaert4,5
  • Klaas Nico Faber6
  • Peter L Horvatovich1
  • Rainer Bischoff1
  • Justina C Wolters7
7 affiliations
  1. 1Department of Analytical Biochemistry, Groningen Research Institute of Pharmacy, University of Groningen, 9700 ADGroningen, The Netherlands.
  2. 2Department of Pharmaceutical Technology and Biopharmacy, Groningen Research Institute of Pharmacy, University of Groningen, 9713 AVGroningen, The Netherlands.
  3. 3Department of Metabolic Health Research, The Netherlands Organization for Applied Scientific Research (TNO), Leiden2333 BE, The Netherlands.
  4. 4Drug Delivery and Disposition, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, LeuvenB-3000, Belgium.
  5. 5BioNotus CommV, Niel2845, Belgium.
  6. 6Department of Gastroenterology and Hepatology, University of Groningen, University Medical Center Groningen, 9713 GZGroningen, The Netherlands.
  7. 7Department of Pediatrics, University of Groningen, University Medical Center Groningen, 9713 AVGroningen, The Netherlands.
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Abstract

Precision-cut liver slices (PCLSs) are an ex vivo liver model that allows isolation of extracellular vesicles (EVs), which are nano-sized particles involved in intercellular communication. However, EV recovery from PCLSs is hindered by adhesion to the extracellular matrix and macromolecules such as extracellular DNA. Enzymatic treatments, such as collagenase (COL) and the endonuclease benzonase (BEN), can aid their release, but potential effects on the tissue are unclear. Here, the effect of these enzymes separately, as well as combined, was analyzed to determine their effects on morphology, protein content, and the proteome of the PCLS. The enzymes showed minimal effects on cell viability and protein content after two different incubation times, whereas morphological analysis confirmed collagen degradation by COL and COL(BEN), along with decreased ribosomal and secretory secretory protein levels. BEN decreased the levels of proteins related to protein synthesis and degradation. Increased levels of proteins related to detoxification and energy production were observed upon addition of COL, BEN, and COL(BEN). This study emphasizes the potential and risks for the use of collagenase and benzonase for EV isolation from the PCLS.

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Cite

BibTeXRIS
BibTeX
@article{stam2026toxicological,
  title = {{Toxicological Exploration of Collagenase and Benzonase on Rat Precision-Cut Liver Slices.}},
  author = {Janine Stam and Peter Olinga and Aswin Menke and Pieter Annaert and Klaas Nico Faber and Peter L Horvatovich and Rainer Bischoff and Justina C Wolters},
  journal = {Journal of proteome research},
  year = {2026},
  month = oct,
  doi = {10.1021/acs.jproteome.5c01232},
  url = {https://europepmc.org/article/MED/42918134},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Toxicological Exploration of Collagenase and Benzonase on Rat Precision-Cut Liver Slices.
AU  - Janine Stam
AU  - Peter Olinga
AU  - Aswin Menke
AU  - Pieter Annaert
AU  - Klaas Nico Faber
AU  - Peter L Horvatovich
AU  - Rainer Bischoff
AU  - Justina C Wolters
PY  - 2026
DA  - 2026/10/01
JO  - Journal of proteome research
DO  - 10.1021/acs.jproteome.5c01232
UR  - https://europepmc.org/article/MED/42918134
AB  - Precision-cut liver slices (PCLSs) are an ex vivo liver model that allows isolation of extracellular vesicles (EVs), which are nano-sized particles involved in intercellular communication. However, EV recovery from PCLSs is hindered by adhesion to the extracellular matrix and macromolecules such as extracellular DNA. Enzymatic treatments, such as collagenase (COL) and the endonuclease benzonase (BEN), can aid their release, but potential effects on the tissue are unclear. Here, the effect of these enzymes separately, as well as combined, was analyzed to determine their effects on morphology, protein content, and the proteome of the PCLS. The enzymes showed minimal effects on cell viability and protein content after two different incubation times, whereas morphological analysis confirmed collagen degradation by COL and COL(BEN), along with decreased ribosomal and secretory secretory protein levels. BEN decreased the levels of proteins related to protein synthesis and degradation. Increased levels of proteins related to detoxification and energy production were observed upon addition of COL, BEN, and COL(BEN). This study emphasizes the potential and risks for the use of collagenase and benzonase for EV isolation from the PCLS.
SN  - 1535-3893
SN  - 1535-3907
ER  -