Porcine Lung-Derived Extracellular Matrix Hydrogel Properties Are Dependent on Pepsin Digestion Time

Robert A Pouliot; Bethany M Young; Patrick A Link; Heon E Park; Alison R Kahn; Keerthana Shankar; Matthew B Schneck; Daniel J Weiss; Rebecca L Heise

doi:10.1089/ten.TEC.2020.0042

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Porcine Lung-Derived Extracellular Matrix Hydrogel Properties Are Dependent on Pepsin Digestion Time

Journal article

Peer reviewed

Porcine Lung-Derived Extracellular Matrix Hydrogel Properties Are Dependent on Pepsin Digestion Time

Robert A Pouliot, Bethany M Young, Patrick A Link, Heon E Park, Alison R Kahn, Keerthana Shankar, Matthew B Schneck, Daniel J Weiss and Rebecca L Heise

Tissue engineering. Part C, Methods, Vol.26(6), pp.332-346

06/2020

DOI: 10.1089/ten.TEC.2020.0042

PMCID: PMC7310225

PMID: 32390520

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https://www.ncbi.nlm.nih.gov/pmc/articles/7310225View

Open Access

Abstract

Hydrogels derived from decellularized lungs are promising materials for tissue engineering in the development of clinical therapies and for modeling the lung extracellular matrix (ECM) . Characterizing and controlling the resulting physical, biochemical, mechanical, and biologic properties of decellularized ECM (dECM) after enzymatic solubilization and gelation are thus of key interest. As the role of enzymatic pepsin digestion in effecting these properties has been understudied, we investigated the digestion time-dependency on key parameters of the resulting ECM hydrogel. Using resolubilized, homogenized decellularized pig lung dECM as a model system, significant time-dependent changes in protein concentration, turbidity, and gelation potential were found to occur between the 4 and 24 h digestion time points, and plateauing with longer digestion times. These results correlated with qualitative scanning electron microscopy images and quantitative analysis of hydrogel interconnectivity and average fiber diameter. Interestingly, the time-dependent changes in the storage modulus tracked with the hydrogel interconnectivity results, while the Young's modulus values were more closely related to average fiber size at each time point. The structural and biochemical alterations correlated with significant changes in metabolic activity of several representative lung cells seeded onto the hydrogels with progressive decreases in cell viability and alterations in morphology observed in cells cultured on hydrogels produced with dECM digested for >12 and up to 72 h of digestion. These studies demonstrate that 12 h pepsin digest of pig lung dECM provides an optimal balance between desirable physical ECM hydrogel properties and effects on lung cell behaviors.

Animals

Extracellular Matrix - chemistry

Hydrogels - chemistry

Lung - chemistry

Lung - metabolism

Pepsin A - metabolism

Swine

Tissue Engineering - methods

Tissue Scaffolds - chemistry

Details

Title: Subtitle: Porcine Lung-Derived Extracellular Matrix Hydrogel Properties Are Dependent on Pepsin Digestion Time
Creators: Robert A Pouliot - University of Vermont
Bethany M Young - Virginia Commonwealth University
Patrick A Link - Virginia Commonwealth University
Heon E Park - University of Canterbury
Alison R Kahn - University of Vermont
Keerthana Shankar - Virginia Commonwealth University
Matthew B Schneck - Virginia Commonwealth University
Daniel J Weiss - University of Vermont
Rebecca L Heise - Virginia Commonwealth University
Resource Type: Journal article
Publication Details: Tissue engineering. Part C, Methods, Vol.26(6), pp.332-346
DOI: 10.1089/ten.TEC.2020.0042
PMID: 32390520
PMCID: PMC7310225
NLM abbreviation: Tissue Eng Part C Methods
ISSN: 1937-3384
eISSN: 1937-3392
Grant note: T32 HL076122 / NHLBI NIH HHS
Language: English
Date published: 06/2020
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Craniofacial Anomalies Research Center
Record Identifier: 9984948144302771

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