Biorheological Properties of Reconstructed Erythrocytes and its Function of

Carrying-Releasing Oxygen
Xiang Wang, Wei Gao, Weiyan Peng, Jiaxin Xie and Yaojin Li
College of Bioengineering, Chongqing University, Chongqing, P.R. CHINA
Abstract: Erythrocyte shape and biomechanical properties have close relation to its physiological function. In this research the
erythrocyte was reconstructed with natural structure protein and lipids based on cellular mechanics and hemorheology concepts. The
biomechanical properties of the reconstructed erythrocyte were determined with the micropipette aspiration system. The shapes of
reconstructed erythrocyte were obtained with electron scanning microscope. The oxygen carrying-releasing function was analyzed with
the HEMOX analyzer from TCS, the experimental results indicated that the reconstructed erythrocytes were similar to the natural
erythrocyte: having biconcave disc shape, good deformability and carrying-releasing oxygen function.
Keywords: biorheological properties, reconstructed erythrocytes, carrying-releasing oxygen
INTRODUCTION
The blood substitute has been a hot researching focus
around the world. As a blood substitute, its function
should be comparable to that of natural hemoglobin,
carrying and releasing oxygen around the human bodies.
The present known blood substitutes are perfluorocarbon,
various kinds of modified hemoglobin by chemical
technology and gene engineering. However, the perfluor-
ocarbon molecule is hard to metabolize and imposes
harmful effects on human health due to its accumulation
in vivo. Gene engineering produced human hemoglobin
from tobacco plant root and has been used to produce
human hemoglobin, but the high cost and low yield
hinder the high production. In addition, liposome has
been used to envelope hemoglobin to be used as a blood
substitute. However, the procedure has only been carried
out in labs at present [1,2].
Erythrocyte shape and biomechanical properties have
close relation to the oxygen carrying-releasing dynamics
function. But up to now little research on artificial blood
substitutes has realized this viewpoint and mainly focused
the studies on how to use blood substitute simulating
physiology of normal red blood cells [3,4]. The main
problem of these products lies in the lack of natural
configuration, and deformability needed in circulation
like those in mature RBCs that would hinder its oxygen
carrying-releasing dynamics function.
In this research, we have studied the biorheological
properties of reconstructed erythrocytes and their function
of carrying-releasing oxygen. The reconstruction of red
blood cells in vitro could be of great significance in the
biomedical research field and may potentially solve the
shortage problems of blood supply.
MATERIALS AND METHODS
In this research the biomechanical properties of the
reconstructed RBC were determined with the micropipette
aspiration system, etc [5,6]. The image of reconstructed
RBC was obtained with an electron scanning microscope.
The dynamics function of carrying-releasing oxygen was
analyzed with the HEMOX analyzer from TCS and the
dynamics analytical system established in our lab (Figures
1 and 2).
RESULTS AND DISCUSSION
The experimental results indicated that the reconstructed
erythrocytes have a biconcave disc form that was similar
to the natural erythrocyte (Figures 3A, 3B, 3C and 3D).
Compared with normal erythrocytes the reconstructed
erythrocytes have similar biomechanical properties (m:
from 4.562 to 7.89210
3
dyn/cm and h from 0.175 to
Address correspondence to Xiang Wang, College of Bioengineering, Chongqing University, Chongqing 400044, P.R. China.
E-mail: xwangchn@vip.sina.com
Artificial Cells, Blood Substitutes, and Biotechnology, 37: 4144
Copyright # 2009 Informa UK Ltd.
ISSN: 1073-1199 print / 1532-4184 online
DOI: 10.1080/10731190802674477
41
TV
S*: Cell Suspension
Pressure Tank
Timer
Image
Recording
Image
Processing
Measuring
Image
Parameter
Calculating
Date Time
Pressure Recording
S*
Figure 1. Schematic drawing showing the system for micropipette aspiration.
P
Flowmeter
Pressure
Balance
Oxygen
Saturation
a
n
a
l
y
s
i
s
Sat
O2
t
O
2
CO
2
N
2
Figure 2. Dynamics analytical system of reconstruction erythrocyte carrying-releasing oxygen.
A
Figure 3A. The electron scanning microscope patterns of
reconstructed erythrocyte. 3040.
B
Figure 3B. The electron scanning microscope patterns of
reconstructed erythrocyte with different size. 5500.
42 X. Wang et al.
0.37210
4
dyns/cm), having good deformability
(Table 1). The reconstructed erythrocyte demonstrated
greater resistance to osmotic lysis. The reconstructed
erythrocytes have carrying-releasing oxygen function.
The reconstructed erythrocytes blood can carry oxygen
(oxygen saturation reaching up to 86%) and can release
oxygen by alternatively being replaced with carbon
dioxide (oxygen saturation declining lowering down to
15%). Animal experimental results show that in hemor-
rhagic shock the reconstructed erythrocytes blood has a
good resuscitation function. The survival time of Wistar
rats was very near to that of rats receiving normal blood
(Figures 4 and 5, Table 2).
CONCLUSIONS
Biorheological properties of reconstructed erythrocytes
are closely related to the function of carrying-releasing
oxygen. The reconstructed erythrocytes with bioconcave
disc shape have oxygen carrying-releasing function and
good deformability. The study indicated the physiological
function of reconstructed erythrocyte was similar to that
of natural erythrocyte.
C
Figure 3C. The electron scanning microscope patterns of
reconstructed erythrocytes. 3800.
D
Figure 3D. The electron scanning microscope patterns of
reconstructed erythrocytes. 101.
Table 1. Biomechanical properties of reconstituted erythrocyte
based on Hemispherical cap model (from Chiens theory)
Elastic
modulus
(10
3
dyn/
cm)
Viscous
coefficient
(10
4
dyns/cm) S/V ratio
Reconstituted
RBC
4.567.89 0.180.37 1.481.53
Normal hRBC 1.128.75 0.190.45 1.391.44
0
10
20
30
40
50
60
70
80
90
0 2 3.6 6 8 10 12
Time(min)
P
O
2
%
saturation
dissociation
Figure 4. Dynamical curve of oxygen saturation-dissociation of reconstitute erythrocyte.
Biorheological Properties of Reconstructed Erythrocytes and its Function of Carrying-Releasing Oxygen 43
ACKNOWLEDGEMENT
Dr. Xiang Wang was supported by National Natural
Science Foundation of China (NSFC 10572159). This
research is also partially supported by 111 Project
entitled Biomechanics & Tissue Repair Engineering (No.:
B06023) and Chongqing Science & Technology Council
(CSTC 2006ba5010).
REFERENCES
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Duong, Patricia L., Ramanujam, Sahana, Chen, Peter C. Y.
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5. Sung, K.L.P., Chien, S. (1978). Viscous and elastic proper-
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182 74:81.
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(1978). Theoretical and experimental studies on viscoelastic
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Table 2. Anti-hemorrhagic shock results of reconstituted
erythrocyte blood
Test
group Name of group
Mean survival
time (h)
A Reconstructed erythrocytes blood 15.290.2
B Wistar blood 17.590.3*
C Wistar erythrocyte saline 14.590.5
D Wistar blood plasma 9.890.4**
E Saline 3.890.6**
F Liposome of Hb 8.090.3**
G Hbsaline 6.090.5**
H Anti-shock drug 13.090.2*
Compared with C group *: 0.01BPB0.05, **: pB0.01
0
10
20
30
40
50
60
70
80
90
0 3.3 7 10 13
Time(min)
P
O
2
%
saturation
dissociation
Figure 5. Dynamical curve of oxygen saturation-dissociation of Human blood.
This paper was first published online on iFirst on 16 January 2009.
44 X. Wang et al.