Reza Khorramirouz

1. Development of perfusion based decellularization of
human-sized heart as a natural scaffold for organ
engineering
A Akbarzadeh, R Khorramirouz, R Seyed Hosein Beigi, M Banitalebi Dehkordi,
AM Kajbafzadeh
Pediatric Urology and Regenerative Medicine Research Center, Section of Tissue
Engineering and Stem Cell Therapy, Children’s Hospital Medical Center, Pediatric Center
of Excellence, Tehran University of Medical Sciences, Tehran, Iran (IRI).
INTRODUCTION
Development of the techniques in the field of cardiac
tissue engineering allowing researchers to create a
functional tissue-engineered large-scale heart, which can
pave the way for reconstruction of tissues or organs to
replace a damaged or injured heart. In the present study,
we describe successful decellularization of an entire
ovine heart for development of a 3D ECM scaffold with
intact microstructural architecture and stable perfusable
vasculature.
Fig. 1. Perfusion decellularization of an ovine heart; (1)
photographs of heart before, (2) during, and (3) after the
decellularization procedure
Fig. 3. Images of CTA. Three dimensional volume-
rendered reconstruction shows the coronary system (first
column), coronary tree of native (second column ) and
coronary system (third column), coronary tree (fourth
column)of decellular heart.
METHODS
Ovine hearts were decellularized by means of coronary
perfusion with ionic and nonionic detergents. Efficiency of
the decellularization and preservation of extracellular
matrix (ECM) components were assessed using following
evaluations: hematoxiline and eosin, Masson’s trichrome,
4′,6-diamidino-2-phenylindole, Picrosirius red, and
Movat’s pentachrome staining; scanning electron
microscopy (SEM); biochemical assay of sulfated
glycosaminoglycans, and collagen. Coronary magnetic
resonance angiography (CMRA) and computed
tomography angiography (CTA) were utilized to prove the
preservation of vascular network integrity. Moreover the
mechanical properties of the native and decellularized
right and left myocardium were examined.
Fig. 3. Images of SEM of native (first row) and
decellular (second row) sections. (1,000 X)
RESULTS
The resultant ECM retained collagen,
glycosaminoglycans (GAGs), and mechanical integrity
whereas all cellular material effectively removed
.CMRA, CTA did not detect destruction of the 3D
architecture of vascular network .
DISCUSSION & CONCLUSIONS
We report successful decellularization of an entire
ovine heart with intact 3D natural architecture and a
coronary tree (its branches), which are not
cytotoxic to cells. These scaffolds could be
recellularized with cells of various origins to restore
contractile function.
Fig. 4. Images of Red sirius staining of native (first
row) and decellular (second row) sections. (4X, scale
bar = 250 μm).
Fig. 2. Images of three-dimensional whole-heart
coronary MRA
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