Details
| Original language | English |
|---|---|
| Pages (from-to) | 5620-5637 |
| Number of pages | 18 |
| Journal | BIOMACROMOLECULES |
| Volume | 24 |
| Issue number | 12 |
| Early online date | 27 Nov 2023 |
| Publication status | Published - 11 Dec 2023 |
Abstract
Solubilized, gel-forming decellularized extracellular matrix (dECM) is used in a wide range of basic and translational research and due to its inherent bioactivity can promote structural and functional tissue remodeling. The animal-derived protease pepsin has become the standard proteolytic enzyme for the solubilization of almost all types of collagen-based dECM. In this study, pepsin was compared with papain, α-amylase, and collagenase for their potential to solubilize porcine liver dECM. Maximum preservation of bioactive components and native dECM properties was used as a decisive criterion for further application of the enzymes, with emphasis on minimal destruction of the protein structure and maintained capacity for physical thermogelation at neutral pH. The solubilized dECM digests, and/or their physically gelled hydrogels were characterized for their rheological properties, gelation kinetics, GAG content, proteomic composition, and growth factor profile. This study highlights papain as a plant-derived enzyme that can serve as a cost-effective alternative to animal-derived pepsin for the efficient solubilization of dECM. The resulting homogeneous papain-digested dECM preserved its thermally triggered gelation properties similar to pepsin digests, and the corresponding dECM hydrogels demonstrated their enhanced bioadhesiveness in single-cell force spectroscopy experiments with fibroblasts. The viability and proliferation of human HepaRG cells on dECM gels were similar to those on pure rat tail collagen type I gels. Papain is not only highly effective and economically attractive for dECM solubilization but also particularly interesting when digesting human-tissue-derived dECM for regenerative applications, where animal-derived materials are to be avoided.
ASJC Scopus subject areas
- Chemical Engineering(all)
- Bioengineering
- Materials Science(all)
- Biomaterials
- Materials Science(all)
- Polymers and Plastics
- Materials Science(all)
- Materials Chemistry
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In: BIOMACROMOLECULES, Vol. 24, No. 12, 11.12.2023, p. 5620-5637.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Papain-Based Solubilization of Decellularized Extracellular Matrix for the Preparation of Bioactive, Thermosensitive Pregels
AU - Almalla, Ahed
AU - Elomaa, Laura
AU - Bechtella, Leïla
AU - Daneshgar, Assal
AU - Yavvari, Prabhu
AU - Mahfouz, Zeinab
AU - Tang, Peter
AU - Koksch, Beate
AU - Sauer, Igor
AU - Pagel, Kevin
AU - Hillebrandt, Karl Herbert
AU - Weinhart, Marie
N1 - Funding Information: We acknowledge the financial support from the Federal Ministry of Research and Education (BMBF, FKZ 13N13523, M.W.) and the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) for SFB 1449 (Project ID 431232613, M.W., K.P., B.K.) and the Cluster of Excellence Matters of Activity, Image Space Material (M.W., I.M.S.) under Germany′s Excellence Strategy – EXC 2025. A.A. warmly thanks Helmholtz Graduate School Macromolecular Bioscience and Dahlem Research School of Freie Universität Berlin for their support. The authors are grateful to M.Sc. Peng Tang for taking the SEM images at the Core Facility of BioSupraMol supported by DFG. Some icons in the TOC Figure and A were used from flaticon.com or Biorender.com .
PY - 2023/12/11
Y1 - 2023/12/11
N2 - Solubilized, gel-forming decellularized extracellular matrix (dECM) is used in a wide range of basic and translational research and due to its inherent bioactivity can promote structural and functional tissue remodeling. The animal-derived protease pepsin has become the standard proteolytic enzyme for the solubilization of almost all types of collagen-based dECM. In this study, pepsin was compared with papain, α-amylase, and collagenase for their potential to solubilize porcine liver dECM. Maximum preservation of bioactive components and native dECM properties was used as a decisive criterion for further application of the enzymes, with emphasis on minimal destruction of the protein structure and maintained capacity for physical thermogelation at neutral pH. The solubilized dECM digests, and/or their physically gelled hydrogels were characterized for their rheological properties, gelation kinetics, GAG content, proteomic composition, and growth factor profile. This study highlights papain as a plant-derived enzyme that can serve as a cost-effective alternative to animal-derived pepsin for the efficient solubilization of dECM. The resulting homogeneous papain-digested dECM preserved its thermally triggered gelation properties similar to pepsin digests, and the corresponding dECM hydrogels demonstrated their enhanced bioadhesiveness in single-cell force spectroscopy experiments with fibroblasts. The viability and proliferation of human HepaRG cells on dECM gels were similar to those on pure rat tail collagen type I gels. Papain is not only highly effective and economically attractive for dECM solubilization but also particularly interesting when digesting human-tissue-derived dECM for regenerative applications, where animal-derived materials are to be avoided.
AB - Solubilized, gel-forming decellularized extracellular matrix (dECM) is used in a wide range of basic and translational research and due to its inherent bioactivity can promote structural and functional tissue remodeling. The animal-derived protease pepsin has become the standard proteolytic enzyme for the solubilization of almost all types of collagen-based dECM. In this study, pepsin was compared with papain, α-amylase, and collagenase for their potential to solubilize porcine liver dECM. Maximum preservation of bioactive components and native dECM properties was used as a decisive criterion for further application of the enzymes, with emphasis on minimal destruction of the protein structure and maintained capacity for physical thermogelation at neutral pH. The solubilized dECM digests, and/or their physically gelled hydrogels were characterized for their rheological properties, gelation kinetics, GAG content, proteomic composition, and growth factor profile. This study highlights papain as a plant-derived enzyme that can serve as a cost-effective alternative to animal-derived pepsin for the efficient solubilization of dECM. The resulting homogeneous papain-digested dECM preserved its thermally triggered gelation properties similar to pepsin digests, and the corresponding dECM hydrogels demonstrated their enhanced bioadhesiveness in single-cell force spectroscopy experiments with fibroblasts. The viability and proliferation of human HepaRG cells on dECM gels were similar to those on pure rat tail collagen type I gels. Papain is not only highly effective and economically attractive for dECM solubilization but also particularly interesting when digesting human-tissue-derived dECM for regenerative applications, where animal-derived materials are to be avoided.
UR - http://www.scopus.com/inward/record.url?scp=85177602068&partnerID=8YFLogxK
U2 - 10.1021/acs.biomac.3c00602
DO - 10.1021/acs.biomac.3c00602
M3 - Article
C2 - 38009757
AN - SCOPUS:85177602068
VL - 24
SP - 5620
EP - 5637
JO - BIOMACROMOLECULES
JF - BIOMACROMOLECULES
SN - 1525-7797
IS - 12
ER -