Cited by 38Extracellular vesicles in diseaseNanoparticle-Based Drug DeliveryCaveolin-1 and cellular processes

Matrix-Bound Nanovesicles: The Effects of Isolation Method upon Yield, Purity, and Function

Open access
Quijano, Lina M., Naranjo, Juan D., El-Mossier, Salma O., Turner, Neill J., Pineda Molina, Catalina, Bartolacci, Joseph, Zhang, Li, White, Lisa +2 more·2020·Tissue Engineering Part C: Methods

Abstract

Identification of matrix-bound nanovesicles (MBV) as ubiquitous components of the extracellular matrix (ECM) raises questions regarding their biologic functions and their potential theranostic application. Unlike liquid-phase extracellular vesicles (e.g., exosomes), MBV are tightly bound to the ECM, which makes their isolation and harvesting more challenging. The indiscriminate use of different methods to harvest MBV can alter or disrupt their structural and/or functional integrity. The objective of the present study was to compare the effect of various MBV harvesting methods upon yield, purity, and biologic activity. Combinations of four methods to solubilize the ECM (collagenase [COL], liberase [LIB], or proteinase K [PK] and nonenzymatic elution with potassium chloride) and four isolation methods (ultracentrifugation, ultrafiltration [UF], density barrier, and size exclusion chromatography [SEC]) were used to isolate MBV from urinary bladder-derived ECM. All combinations of solubilization and isolation methods allowed for the harvesting of MBV, however, distinct differences were noted. The highest yield, purity, cellular uptake, and biologic activity were seen with MBV isolated by a combination of liberase or collagenase followed by SEC. The combination of proteinase K and UF was shown to have detrimental effects on bioactivity. The results show the importance of selecting appropriate MBV harvesting methods for the characterization and evaluation of MBV and for analysis of their potential theranostic application. Impact statement: Identification of matrix-bound nanovesicles (MBV) as ubiquitous components of the extracellular matrix (ECM) has raised questions regarding their biologic functions and their potential theranostic application. This study demonstrates that the harvesting methods used can result in samples with physical and biochemical properties that are unique to the isolation and solubilization methods used. Consequently, developing harvesting methods that minimize sample contamination with ECM remnants and/or solubilization agents will be essential in determining the theranostic potential of MBV in future studies.

Integrity signals

Registry

Crossref · 10.1089/ten.tec.2020.0243

Cites

84 of 84 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Claims the paper makes

We couldn’t retrieve this paper’s full text, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Degradation of ECM biomaterials upon implantation is necessary to promote their pro-regenerative effect.

    “MBV independence at promoting a pro-remodeling response [163,166] further proves the necessity for the degradation of ECM biomaterials upon implantation to promote their pro-regenerative effect.”
  • Independently of the methodology used to decellularize the ECM, all decellularized tissue has shown MBV to date.

    “Interestingly, independently of the methodology used to decellularize the ECM, all decellularized tissue has shown MBV to date, proving the resilience of the MBV and the tight ECM-MBV relationship [163,166,167].”

Cites

1 retracted

Flagged first, then by citations

Cited by

Sorted by citations

Authors
Quijano, L. M., Naranjo, J. D., El-Mossier, S. O., Turner, N. J., Pineda Molina, C., Bartolacci, J., Zhang, L., White, L., Li, H., & Badylak, S. F.
Affiliations
  • University of Pittsburgh (US)
  • McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
  • University of Nottingham (GB)
  • Magee-Womens Research Institute (US)
Corresponding author
Badylak, Stephen F.
Venue
Tissue Engineering Part C: Methods
Published
2020-10-01
Volume
26
Issue
10
Pages
528-540
ISSN
  • 1937-3384 (ISSN-L)
  • 1937-3392
OpenAlex
W3089777907
Cited by
38
References
84
Citations by year
2026: 4 · 2025: 12 · 2024: 8 · 2023: 3 · 2022: 9 · 2021: 2
FWCI
1.14
Type
journal article
Language
en
Open access
Locations
  • Tissue Engineering Part C Methods (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Repository@Nottingham (University of Nottingham) (acceptedVersion, other-oa)
  • Repository@Nottingham (University of Nottingham) (submittedVersion, cc-by)
Funders
  • National Institutes of Health
  • National Institute of Arthritis and Musculoskeletal and Skin Diseases (R01AR073527)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Caveolin-1 and cellular processes
Keywords
extracellular matrix, collagenase, Liberase, proteinase K, size exclusion chromatography, ultrafiltration, urinary bladder, cellular uptake
MeSH terms
Extracellular Vesicles, Endocytosis, Enzymes, Extracellular Matrix, Humans, Particle Size, Peptides, Proteins, Solubility, Stem Cells, MicroRNAs, Cell Proliferation, Nanoparticles

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review