Optimization of decellularization parameters in developing renal extracellular matrix scaffold from porcine kidneys

Date of Publication

8-2019

Document Type

Master's Thesis

Degree Name

Master of Science in Chemical Engineering

Subject Categories

Chemical Engineering

College

Gokongwei College of Engineering

Department/Unit

Chemical Engineering

Thesis Adviser

Nathaniel P. Dugos

Defense Panel Chair

Lawrence P. Belo

Defense Panel Member

Joseph L. Auresenia
Cynthia F. Madrazo

Abstract/Summary

Bioscaffolds are used in whole organ bioengineering as templates to create transplantable organs in order to address the growing concern of organ shortage. For complex organs such as kidneys, bioscaffolds derived from the extracellular matrix of the kidney is the top candidate but the main problem of current decellularization strategies is the long treatment time. Detergent perfusion decellularization coupled with sonication treatment has the potential in decreasing the decellularization time however, the combined effects of these two strategies can induce damage to the resulting bioscaffold. This study aims to determine the optimum decellularization parameters of SDS concentration, flowrate and sonicator power which are known to have a strong influence on the decellularization time and microarchitecture integrity of the scaffold in order to produce a renal acellular bioscaffold at the shortest time possible while still having an intact structure. The study used cadaveric porcine kidneys which were subjected to detergent perfusion and sonication treatment with varying sodium dodecyl sulfate (SDS) concentration, SDS flowrate and

sonicator power. Decellularization time was determined by the change in the native reddish- brown color of the kidney into a translucent white bioscaffold while the evaluation of the

microarchitecture integrity was done through a scoring system based on the H&E stained structures of the glomerulus, tubules and blood vessel. Results showed that the optimized decellularization parameters are 0.71%wt/vol SDS, 45 mL/min and 60 W. In addition, the resulting bioscaffold using the optimum parameters was obtained after 3 hours while still preserving the delicate renal structures. In addition, colorimetric assays and immunohistochemistry test also showed the retention ECM proteins such as total collagen, fibronectin and laminin which are important for recellularization. Hence, decellularization settings in detergent perfusion with sonication play an important role in the efficient production of renal bioscaffolds.

Abstract Format

html

Note

Bachelor and Master of Science thesis

Language

English

Format

Electronic

Accession Number

CDTG008172

Keywords

Tissue engineering; Kidneys—Transplantation; Extracellular matrix

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Embargo Period

2-10-2025

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