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

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

doi:10.1089/ten.tec.2020.0042

Cited by 70 publications

(71 citation statements)

References 96 publications

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“…In this work, dECM hydrogels with various protein concentrations were prepared, and their stability and capabilities to be manipulated in cell culture media were examined. A wide range of concentrations for hydrogels made of decellularized lung tissues (human or animals) has been reported in the literature [ 39 , 40 , 42 ]. Therefore, we aimed to examine dECM hydrogels with concentrations from 2 to 20 mg/mL to determine an optimum range for human lung dECM hydrogels.…”

Section: Resultsmentioning

confidence: 99%

“…To understand the gelation kinetics of dECM hydrogels, a turbidity assay was performed as previously reported in other work [ 38 , 39 ]. One-hundred microliters (100 µL) of a neutralized liquid dissolved dECM pre-gel solution at various concentrations of 20, 17.5, 15, and 12.5 mg/mL were transferred to 96-well plates (n = 4), and the absorbance (405 nm) was measured every three minutes for 80 min (the plate reader was pre-heated to 37 °C).…”

Section: Methodsmentioning

confidence: 99%

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A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

Dabaghi

Saraei

Carpio

et al. 2021

Cells

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Decellularization efforts must balance the preservation of the extracellular matrix (ECM) components while eliminating the nucleic acid and cellular components. Following effective removal of nucleic acid and cell components, decellularized ECM (dECM) can be solubilized in an acidic environment with the assistance of various enzymes to develop biological scaffolds in different forms, such as sheets, tubular constructs, or three-dimensional (3D) hydrogels. Each organ or tissue that undergoes decellularization requires a distinct and optimized protocol to ensure that nucleic acids are removed, and the ECM components are preserved. The objective of this study was to optimize the decellularization process for dECM isolation from human lung tissues for downstream 2D and 3D cell culture systems. Following protocol optimization and dECM isolation, we performed experiments with a wide range of dECM concentrations to form human lung dECM hydrogels that were physically stable and biologically responsive. The dECM based-hydrogels supported the growth and proliferation of primary human lung fibroblast cells in 3D cultures. The dECM is also amenable to the coating of polyester membranes in Transwell™ Inserts to improve the cell adhesion, proliferation, and barrier function of primary human bronchial epithelial cells in 2D. In conclusion, we present a robust protocol for human lung decellularization, generation of dECM substrate material, and creation of hydrogels that support primary lung cell viability in 2D and 3D culture systems

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

Dabaghi

Saraei

Carpio

et al. 2021

Cells

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show abstract

“…Cumulatively, these observations indicate that enzymatic digestion of dECM microparticles is an alternative, safe and effective technique for obtaining homogenous particle size distribution. In addition, this methodology efficiently preserves the molecular constituents of native tissue which can help support a constructive, site-appropriate, remodeling response when implanted in vivo [ 29 ].…”

Section: Resultsmentioning

confidence: 99%

Human Cadaveric Donor Cornea Derived Extra Cellular Matrix Microparticles for Minimally Invasive Healing/Regeneration of Corneal Wounds

Chandru¹,

Agrawal²,

Ojha³

et al. 2021

Biomolecules

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Biological materials derived from extracellular matrix (ECM) proteins have garnered interest as their composition is very similar to that of native tissue. Herein, we report the use of human cornea derived decellularized ECM (dECM) microparticles dispersed in human fibrin sealant as an accessible therapeutic alternative for corneal anterior stromal reconstruction. dECM microparticles had good particle size distribution (≤10 µm) and retained the majority of corneal ECM components found in native tissue. Fibrin–dECM hydrogels exhibited compressive modulus of 70.83 ± 9.17 kPa matching that of native tissue, maximum burst pressure of 34.3 ± 3.7 kPa, and demonstrated a short crosslinking time of ~17 min. The fibrin–dECM hydrogels were found to be biodegradable, cytocompatible, non-mutagenic, non-sensitive, non-irritant, and supported the growth and maintained the phenotype of encapsulated human corneal stem cells (hCSCs) in vitro. In a rabbit model of anterior lamellar keratectomy, fibrin–dECM bio-adhesives promoted corneal re-epithelialization within 14 days, induced stromal tissue repair, and displayed integration with corneal tissues in vivo. Overall, our results suggest that the incorporation of cornea tissue-derived ECM microparticles in fibrin hydrogels is non-toxic, safe, and shows tremendous promise as a minimally invasive therapeutic approach for the treatment of superficial corneal epithelial wounds and anterior stromal injuries.

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“… 76 The impact of the decellularisation method on the ECM-derived hydrogel has been shown by several studies. 66 , 77 For example, Fernández-Pérez and Ahearne 66 examined the impact of different decellularisation protocols on hydrogels prepared from porcine corneas that were isolated and decellularised with SDS, Triton X-100 or freeze–thaw cycles. While all the three methods showed significant reduction of DNA, the SDS method produced cytotoxic hydrogels.…”

Section: Hydrogels With Decellularised Matricesmentioning

confidence: 99%

Decellularised scaffolds: just a framework? Current knowledge and future directions

et al. 2020

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The use of decellularised matrices as scaffolds offers the advantage of great similarity with the tissue to be replaced. Moreover, decellularised tissues and organs can be repopulated with the patient’s own cells to produce bespoke therapies. Great progress has been made in research and development of decellularised scaffolds, and more recently, these materials are being used in exciting new areas like hydrogels and bioinks. However, much effort is still needed towards preserving the original extracellular matrix composition, especially its minor components, assessing its functionality and scaling up for large tissues and organs. Emphasis should also be placed on developing new decellularisation methods and establishing minimal criteria for assessing the success of the decellularisation process. The aim of this review is to critically review the existing literature on decellularised scaffolds, especially on the preparation of these matrices, and point out areas for improvement, finishing with alternative uses of decellularised scaffolds other than tissue and organ reconstruction. Such uses include three-dimensional ex vivo platforms for idiopathic diseases and cancer modelling.

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

Cited by 70 publications

References 96 publications

A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

Human Cadaveric Donor Cornea Derived Extra Cellular Matrix Microparticles for Minimally Invasive Healing/Regeneration of Corneal Wounds

Decellularised scaffolds: just a framework? Current knowledge and future directions

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