Biomacromolecules 2002, 3, 1312-1319

1312

Surface Modification of Polycaprolactone Membrane via

Aminolysis and Biomacromolecule Immobilization for
Promoting Cytocompatibility of Human Endothelial Cells
Yabin Zhu, Changyou Gao,* Xingyu Liu, and Jiacong Shen
Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China
Received June 26, 2002; Revised Manuscript Received August 15, 2002

Amino groups were covalently introduced onto a polycaprolactone (PCL) surface by the reaction between
1,6-hexanediamine and the ester groups of PCL. The occurrence of the aminolysis and the introduction of
free NH2 groups were verified qualitatively by fluorescence spectroscopy, where rhodamine B isothiocyanate
was employed to label NH2 groups, and quantitatively by absorbance spectroscopy, where ninhydrin was
used to react with NH2 to generate a blue product. Due to the presence of deep pores on the PCL membrane,
the aminolysis reaction could penetrate as deep as 50 m to yield NH2 density as high as 2 10-7 mol/cm2.
By use of the NH2 groups as active sites, biocompatible macromolecules such as gelatin, chitosan, or collagen
were further immobilized on the aminolyzed PCL membrane via a cross-linking agent, glutaraldehyde. X-ray
photoelectron spectroscopy (XPS) and surface wettability measurements confirmed the coupling of the
biomacromolecules. The endothelial cell culture proved that the cytocompatibility of the aminolyzed PCL
was improved slightly regardless of the NH2 amount on the surface. After immobilization of the
biomacromolecules, however, the cell attachment and proliferation ratios were obviously improved and the
cells showed a similar morphology to those on tissue culture polystyrene. Measurement of the von Willebrand
factor (vWF) secreted by these endothelial cells (ECs) verified the endothelial function. Hence, a better
EC-compatible PCL was produced.
Introduction
Many biodegradable synthetic materials such as polycaprolactone (PCL), poly(lactic acid) (PLA) and poly(lactideco-glycotide) (PLGA) have been used as scaffolds to support
the regeneration of tissue-engineered organs such as cartilage,
blood vessel, and skin. However, the poor cytocompatibility
of the synthetic polymers leads to the inefficiency of the
scaffold in constructing a friendly interface with living cells.
Therefore, modification of the tissue-engineering polymeric
materials to improve their cytocompatibility is necessary.
Because the interaction between living cells and materials
occurs mainly on the interfacial layer, many surface modification methods such as plasma treatment, -ray irradiation,
ozone oxidization, end-grafting, or in situ polymerization
have been developed to alter the surface properties of
materials,1-8 for improving the cytocompatibility of the
biomedical materials without alteration of the bulk properties.
Among which end-grafting or in situ graft polymerization
by photo or by radio frequency glow discharge deposition
onto biomaterials has been widely employed to produce
hydrophilic layer onto bulk biomedical polymers.9-11 The
photoinduced grafting has been much studied to introduce
hydrophilic groups onto poly(ester urethane) (PU), poly(L-lactic acid) (PLLA), and PCL membranes, resulting in
better cell attachment, spreading, and proliferation.12-15
* Corresponding author: e-mail, cygao@mail.hz.zj.cn; Tel, +86-57187951108; fax, +86-571-87951948.

PCL, biodegradable aliphatic polyester,16,17 has been suggested for wide applications such as drug delivery systems,18,19 tissue-engineered skin (plain film), and scaffolds
for supporting fibroblast and osteoblast growth.20,21 In PCL
molecules, there exist abundant ester groups (-COO-).
These ester groups can be hydrolyzed to carboxylic acid
under alkaline condition. In addition, it is possible that the
amino groups can be introduced onto the polyester surface
by a reaction with diamine, providing that one amino group
reacts with the -COO- group to form a covalent bond,
-CONH-, while the other amino group is unreacted and
free as shown in Scheme 1. It is worth noting that hydroxylterminated chains will also be yielded on the polyester surface during this process. Some advantages in tissue engineering can be expected by the steady introduction of these amino
groups: (1) nontoxic to cells or tissues; (2) decreasing the
surface hydrophobicity; (3) neutralizing the acid generated
during the scaffold degradation and reducing the inflammation around the implanted scaffold; (4) providing active sites
through which other biomolecules such as collagen, gelatin,
or RGD peptides can be further immobilized, obtaining cytocompatible surface on which cells can grow well; (5) applying to three-dimensional (3-D) porous polyester scaffolds.
3-D porous scaffolds play an important role in supporting
cell attachment, proliferation, and manipulating cell functions. However, the surface modification of 3-D porous scaffolds to improve their biocompatibility is difficult so far.
Due to the easy performance, the aminolysis has been applied

10.1021/bm020074y CCC: $22.00

2002 American Chemical Society
Published on Web 09/18/2002

Polycaprolactone Membrane Modification

Biomacromolecules, Vol. 3, No. 6, 2002 1313

Scheme 1. The Schematic Representation of Aminolysis and

Further Immobilization of Biomolecules on Polycaprolactone
Membrane

to PLLA porous scaffolds to improve its cytocompatibility

(unpublished).
By aminolysis with 1,6-hexanediamine, free amino groups
were introduced onto the PCL membrane surface. The
existence of the ultralow concentration of amino groups was
verified qualitatively and quantitatively by fluorescence
spectroscopy and absorbance spectroscopy, respectively.
Rhodamine B isothiocyanate or ninhydrin were used to label
the amino groups to introduce fluorescent agent or to react
with amino groups to produce a colored absorbance, respectively. Through a coupling agent, i.e., glutaraldehyde, gelatin,
chitosan, or collagen was further immobilized onto a PCL
surface. The culture of human endothelial cells (ECs) in vitro
showed that the cytocompatibility of the aminolyzed and the
biomacromolecules-immobilized PCL membranes is improved obviously.
Experimental Section
Aminolysis of PCL Membrane. PCL membrane was
prepared by dissolving 5 g of PCL (Aldrich, Mn 80 000) in
50 mL of distilled 1,4-dioxane, and the mixture was then
spread onto a stainless plate. The solvent was evaporated
at 35 C for 24 h and further dried under vacuum for
another 24 h at 30 C. A translucent PCL membrane with a
thickness of 200 m was obtained. The membrane was cut
into pieces of 2 2 cm and immersed in alcohol/water
(1/1, v/v) solution for 2-3 h to clean oily dirt and then
washed with a large amount of deionized water. The
membrane was subsequently immersed in distilled 1,6hexanediamine/2-propanol solution with suitable concentrations for given time at 37 C, rinsed with deionized water
for 24 h at room temperature to remove free 1,6-hexanediamine, and dried in a vacuum at 30 C for 24 h to constant
weight.
Immobilization of Gelatin, Chitosan, or Collagen. The
aminolyzed PCL membrane was immersed in 1 wt %
glutaraldehyde (GA) solution for 3 h at room temperature,
followed by rinsing with a large amount of deionized water
for another 24 h to remove free GA. The membrane was
then incubated in 3 mg/mL gelatin/phosphate buffered
solution (PBS, pH ) 7.4) or 2 mg/mL chitosan solution (pH
) 3.5) or 3 mg/mL collagen solution (pH ) 3.4) for 24 h at
2-4 C, respectively. The gelatin-immobilized membrane

Figure 1. The absorbance (at 538 nm) of ninhydrin-NH2 (with

1,6-hexanediamine) reaction products as a function of NH2 concentration.

was rinsed with deionized water for at least 24 h to remove

free gelatin. The collagen- and chitosan-immobilized membranes were rinsed with 1.0% acetic acid solution and then
rinsed with deionized water for 24 h to remove free chitosan
or collagen.12-15
Determination of the Amino Groups. Rhodamine B
isothiocyanate (RBITC) was used to label the amino groups
on the aminolyzed PCL membrane for fluorescence measurement by immersing the PCL membrane in 0.1 mg/mL
RBITC solution for 24 h at 2-4 C. The obtained RBITClabeled PCL membrane was rinsed with deionized water for
24 h at room temperature to remove free RBITC and then
dried under vacuum for another 24 h at 30 C.
The ninhydrin analysis method was employed to quantitatively detect the amount of NH2 groups on the aminolyzed
PCL membrane. The membrane was immersed in 1.0 mol/L
ninhydrin/ethanol solution for 1 min and then was placed
into a glass tube, following with heating at 80 C for 15
min to accelerate the reaction between ninhydrin and amino
groups on PCL membrane. After the adsorbed ethanol had
evaporated, 5 mL of 1,4-dioxane was added into the tube to
dissolve the membrane when the membrane surface displayed
blue. Another 5 mL of 2-propanol was added to stabilize
the blue compound. The absorbance at 450-650 nm of this
mixture was measured on a UV-vis spectrophotometer. A
calibration curve was obtained with 1,6-hexanediamine in
1,4-dioxane/isopropane (1:1, v:v) solution (Figure 1).
Human Endothelial Cell Culture. The endothelial cells
(ECs) were isolated from the human umbilical cord veins
of a new born baby with 1.0 mg/mL collagenase (type I,
Sigma)/PBS for 20-25min at room temperature.22 The
isolated ECs were routinely seeded on the beds prelaid with
control or modified PCL membranes as well as on the tissue
culture polystyrene (TCPS) (Nunc, Denmark) as control. The
ECs were incubated in a culture medium consisting of 20%
(v/v) fetal calf serum (FCS, Sijiqing Biotech. Co., China)
and 80% RPMI1640 (Gibcobrl Co.) supplemented with 100
units/mL of penicillin and 100 g/mL of streptomycin in
humidified air containing 5% CO2 at 37 C. After incubation
for 24 h, the culture medium was changed, and then changed

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Biomacromolecules, Vol. 3, No. 6, 2002

Zhu et al.

Figure 2. The survey XPS spectra of control PCL (a), PCL aminolyzed at a concentration of 10 wt % 1,6-hexanediamine for 10 min at 37 C
(b), PCL immobilized with gelatin (c), chitosan (d), or collagen (e).

Figure 3. The fluorescence intensity of the RBITC-immobilized PCL

membranes as a function of 1,6-hexanediamine concentration. The
aminolysis reaction took place at 37 C for 10 min.

every 2 days. The ECs were fixed with 2.5% glutaraldehyde

for 30 min for observation of cell morphology under a
scanning electron microscope (SEM) after being cultured for
4 days. The cell attachment and proliferation ratio were
averaged from five parallel measurements at 12 h and 4 days,
respectively, by trypsinization of the ECs and counting the
cell number under a hemocytometer. The cell proliferation
ratio was defined as (N2 - N1)/N1, where N1 and N2 represent
the cell number per well at 12 h and 4 days, respectively.
Measurement of Released von Willebrand Factor
(vWF). Secretion of vWF was measured in the supernatant
of the cultured ECs by vWF enzyme-linked immunosorbent
assay (ELISA) (Sun Diagnostics, Shanghai, China). Cell
cultures were washed with the culture medium without serum
when the ECs reached confluence (cultured for 72 h),
followed by 2 h of incubation in culture medium with 20%
FCS. Subsequently, supernatant medium was collected and
centrifuged (10 min, 400g) for vWF measurement using vWF
ELISA.23 Spectrophotometric readings were performed at 492
nm. Final results were obtained by comparison with a

Figure 4. The fluorescence intensity of the RBITC-immobilized PCL

membrane as a function of aminolyzing time. The aminolysis reaction
of PCL membrane took place at 37 C in 10 wt % 1,6-hexanediamine/
2-propanol solution.

standard curve constructed using dilutions of normal plasma.

One milliliter of normal plasma was assumed to contain 10
g of vWF (1 unit).24
Characterization. X-ray photoelectron spectroscopy (XPS)
spectra were recorded on an ESCA LAB Mark II spectrometer employing Al KR excitation radiation. The charging shift
was referred to the C1s line emitted from the saturated
hydrocarbon. The fluorescence intensity was measured on a
M850 fluorescence spectrophotometer and confocal laser
scanning microscopy (CLSM, Radiance 2100, BIO-RAD).
The UV-vis spectrum was measured on a UV-visible
spectrophotometer (CARY 100 BIO, America). The water
contact angle was measured at room temperature on a
DSA10-MK2 contact angle measuring system from Kruss,
using the sessile drop and captive bubble methods. The
atomic force microscopy (AFM) image was obtained by
scanning probe microscopy (SPA400, Seiko) in the tapping
mode. The cell morphology was observed under a scanning
electron microscope (SEM, Stereoscan 260, Cambridge).

Polycaprolactone Membrane Modification

Biomacromolecules, Vol. 3, No. 6, 2002 1315

Figure 5. The absorbance at 538 nm of the aminolyzed PCL membrane treated with ninhydrin as a function of aminolyzing time. The inset
represents the absorbance spectra, where a, b, c, and d refer to PCL membrane aminolyzed for 5, 15, 45, and 120 min at 10 wt % 1,6hexanediamine/2-propanol solution, respectively.

Figure 6. The surface morphology of the control PCL membrane (a) and the PCL membrane aminolyzed at a concentration of 10 wt % 1,6hexanediamine/2-propanol for 10 min at 37 C (b).

Results and Discussion

Introduction of Amino Groups via Aminolysis. As
shown in Scheme 1, a nitrogen element should appear on
the PCL surface if the reaction of 1,6-hexanediamine with
the ester groups occurs. However, from the XPS spectra of
control and aminolyzed PCL (Figure 2 a, b), no obvious N1s
peak was detected. Attenuated total reflectance Fourier
transform infrared (ATR-FTIR) spectroscopy characterization
did not show the NH2 or -CONH- absorbance either. This
might be the result of either the unreaction of amino groups
with ester groups or the low sensitivity of the characterization
methods. By the employment of more sensitive detecting
methods, i.e., fluorescence spectroscopy after labeling the
NH2 groups with RBITC, the existence of free NH2 groups
was verified from the fluorescence emission data. Figure 3
showed that the fluorescence intensity from RBITC at 580
nm increased along with the concentration of 1,6-hexanediamine between 0 and 14 wt %. The intensity alteration
proved that the amount of free NH2 groups on the PCL
membrane surface increased with 1,6-hexanediamine concentrations at the given time. It is worth noting that free NH2
can react with -NCS in RBITC, resulting in a covalent
coupling of RBITC on a PCL membrane surface that cannot
be rinsed out, while the hydroxyl groups formed by the

aminolysis cannot react with RBITC under the same condition. It has to be indicated that the concentration of 1,6hexanediamine should be preferably lower than 14 wt %
because of the poor solubility. The too strong basicity may
also destroy the bulk mechanical property of PCL in higher
concentration. Therefore, the optimal concentration of 1,6-hexanediamine was chosen as 10 wt % in the following study.
At a given concentration of 10 wt % 1,6-hexanediamine,
the influence of aminolyzing time on the surface amount of
amino groups is shown in Figure 4. The fluorescence
intensity increased rapidly with the increase of aminolyzing
time, reached the maximum value at about 1 h, and then
decreased to some extent. This may be caused by the further
reaction with carboxyl of the free amino on the terminal chain
or by the degradation of the superficial layer due to the high
aminolyzing ratio at the longer time. In both cases the amount
of the free amino groups on PCL membrane will be reduced;
hence the fluorescence intensity decreases.
The quantitative NH2 amount on aminolyzed PCL membrane surface was measured by the ninhydrin method. The
blue reaction product of ninhydrin with free NH2 has a
maximum absorbance at 538 nm in the solvent of 1,4dioxane/2-propanol (1:1) (Figure 5 inset). As shown in Figure
5, the alteration tendency of NH2 amount is similar to the

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Biomacromolecules, Vol. 3, No. 6, 2002

Zhu et al.
Table 1. The Water Contact Angle of the Control and the
Modified PCL Membrane
samples

SCA,a deg

CBCA,b deg

control PCL
aminolyzed PCLc
PCL-chitosan
PCL-gelatin
PCL-collagen

81.2 ( 2.4
68.4 ( 1.4
67.0 ( 0.8
59.5 ( 2.0
65.2 ( 1.9

67.9 ( 2.3
35.7 ( 2.3
28.5 ( 3.3
26.6 ( 2.3
27.9 ( 1.5

a Sessile contact angle. b Captive bubble contact angle. c The membrane was prepared in 10 wt % 1,6-hexanediamine solution at 37 C for
10 min.

Table 2. The Dynamic Water Contact Angle and Hysteresis of the

Control and the Modified PCL Membrane

Figure 7. CLSM fluorescence intensity of the control (a) and the

aminolyzed PCL membrane (b) as a function of the depth in the
z-direction. The aminolyzed PCL membrane was prepared in 10 wt
% 1,6-hexanediamine solution at 37 C for 10 min. Measuring area
was 619 619 m.

fluorescence result (Figure 4). The maximum NH2 density

yielded at 1 h is 2 10-7 mol/cm2. From this result one
can calculate the average area per amino-terminated chain,
which is 0.1 2, supposing that the membrane surface was
absolutely smooth and all the amino groups laid as a single
layer. This is obviously unreasonable and impossible. Hence,
either the surface is very rough or the aminolysis occurs in
some depth in the z-direction instead of just on one layer, or
the both. AFM measurement showed that the control PCL
membrane was quite rough and there were many deep pores
ranging from tens to hundred of nanometers, which were
generated during the solvent evaporation process (Figure 6).
After aminolysis, the PCL membrane surface became rougher
and the pore size decreased as well. The existence of the
deep pores provided also the probability that the 1,6hexanediamine molecules could penetrate into the inside of
PCL membrane. To verify this hypothesis, a confocal laser
scanning microscope was employed using the slice scanning
function in the z-direction. The fluorescence intensity on a
619 619 m as a function of depth in the z-direction is
shown in Figure 7. It proves that the aminolysis could occur
as deep as 50 m in z-direction, and there existed a gradient
distribution of the NH2 groups from the membrane surface
to the inside, where more NH2 groups were located on

samples

ACA,a deg

RCA,b deg

hysteresis, deg
(ACA-RCA)

control PCL
aminolyzed PCLc
PCL-chitosan
PCL-gelatin
PCL-collagen

81.2 ( 1.9
82.4 ( 2.4
62.1 ( 1.3
59.4 ( 2.1
63.7 ( 1.6

61.9 ( 5.1
28.7 ( 3.4
11.1 ( 2.6
9.8 ( 1.8
12.2 ( 2.4

19.3
53.7
51.0
49.6
51.5

a Advancing contact angle. b Receding contact angle. c The membrane

was prepared in 10 wt % 1,6-hexanediamine solution at 37 C for 10 min.

surface and less inside. It is worth noting that the NH2 groups
were not homogeneously distributed in a given layer under
the CLSM observation either. Stronger fluorescence intensity
was observed around the area of pores. Figure 7 shows also
that a physical adsorption of RBTIC on the control PCL
membrane was possible, but the intensity was much lower.
It is interesting that the fluorescence intensity in the control
membrane exhibited a steplike decrease with the depth. The
reason is not clear now. It may be caused by some physical
structure formed during the membrane fabrication process.
In conclusion, the density of amino groups on the superlayer of PCL membrane should be far less than 2 10-7
mol/cm2 due to the existence of surface roughness and deep
pores. The aminolyzing reaction took place to a depth around
50 m. The NH2 density on PCL membrane can be regulated
and controlled through controlling the aminolyzing degree.
Biomacromolecules Immobilization. The introduction of
NH2 groups onto PCL membrane can not only modify the
poor hydrophilicity but also provide the necessary active sites
through which other biocompatible components such as
proteins, polysaccharides, cell growth factors, or peptides
can be further immobilized. To achieve the covalent coupling

Figure 8. The EC attachment (relative to TCPS) (9) and the proliferation ratio (0) cultured for 12 h and 4 days at 37 C in humidified air with
5% CO2: (a) TCPS; (b) control PCL; (c) PCL aminolyzed for 3 min; (d) 10 min; (e) 30 min; (f) 120 min; (g) PCL immobilized with gelatin; (h)
chitosan; (i) collagen. Cell seeding density was 15 104/cm2.

Polycaprolactone Membrane Modification

Biomacromolecules, Vol. 3, No. 6, 2002 1317

Figure 9. The cell morphology cultured for 4 days on TCPS (a), PCL membrane aminolyzed for 3 min (b) and 120 min (c), and PCL membrane
immobilized by gelatin (d), chitosan (e), or collagen (f), respectively. 1 refers to higher magnification while 2 refers to an overview under SEM
with the same sample. Cell seeding density was 15 104/cm2.

of biomacromolecules, the aminolyzed PCL membrane was

treated with a large amount of glutaraldehyde (GA) first as
shown in Scheme 1. The reaction between NH2 and OHCs
CHO yielded a bonding via sNdCHsCHO, and one free
aldehyde group could react with NH2 groups existing in most
biomacromolecules such as gelatin, chitosan, or collagen.
The XPS analysis shows that N1s peaks at 400.2 eV appeared
after immobilization of gelatin, chitosan, or collagen, which
confirmed the coupling. The surface wettability alteration
(Table 1) further confirmed the existence of the biomacromolecules. Though the water contact angles measured by
the sessile drop method decreased only slightly after the

biomacromolecules immobilization, they obviously decreased

from 70 to 30 when the captive bubble method was
employed. This difference between the two measuring
methods is a typical phenomena caused by the conformation
alteration of the hydrophilic macromolecular chains.25 In the
interface of biomacromolecules-immobilized PCL and air,
the hydrophobic parts of the biomacromolecules tends to
accumulate on the surface in order to reduce the interfacial
energy, while in the interface of biomacromolecules-immobilized PCL and water, the hydrophilic parts of the
biomacromolecules will reorganize their molecular structure
to generate a hydrophilic parts dominating surface. This

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Biomacromolecules, Vol. 3, No. 6, 2002

reorganization of the biomacromolecules was further proved

by the dynamic water contact angle measurement through
comparing the hystereses between the advancing and receding angles (Table 2). The larger contact-angle hystereses of
the modified PCL surfaces (50-55 C) than that of the
control PCL (19.3 C) demonstrated the occurrence of chain
reorientation supposing no other disturbing factor.26 The
slight decrease of ACA after aminolysis may be attributed
to either the less amount incorporated or the easy movement
of the amino groups with small molecular size which leads
to the hiding of the hydrophilic parts under the hydrophobic
substratum in air.
The Cell Compatibility of Modified PCL Membrane.
The increase of the surface hydrophilicity after aminolysis
and the immobilization of the biocompatible macromolecules
may provide the possibility of improving the cytocompatibility of PCL. The endothelial cell culture results showed
that the cell attachment ratio of the aminolyzed PCL was
not obviously changed compared with the control PCL
regardless of the surface amino amounts (aminolyzing time)
(Figure 8), although the water contact angles of these aminolyzed membranes have obviously been decreased. However,
the cell proliferation ratios were improved clearly though
the extent was still small compared with TCPS. The ECs on
aminolyzed PCL also showed better cell morphology as
shown in Figure 9. Hence, it is concluded that aminolysis
of PCL to introduce NH2 groups has a positive effect on
improving cytocompatibility in a limited extent.
After immobilization of the biomacromolecules, however,
the cytocompatibility was improved obviously (Figure 8 and
Figure 9). The cell attachment and proliferation ratios of ECs
on gelatin-, chitosan-, or collagen-immobilized PCL membranes were much larger than both of the control and the
aminolyzed PCL membranes. The higher cell attachment
ratios on the PCL membranes immobilized with gelatin or
collagen that was even better than TCPS were yielded. The
ECs grown on gelatin-, chitosan-, or collagen-immobilized
PCL membranes presented also a flat and spreading morphology that was similar to those on TCPS. Considering the
comprehensive results of cell attachment, proliferation, and
morphology, the gelatin- or collagen-immobilized PCL has
the best cytocompatibility.
vWF Secretion. vWF is an adhesive glycoprotein synthesized exclusively in endothelial cells and megakaryocytes.
Endothelial vWF is stored in rod-shaped organelles called
Weibel-Palade bodies and affects the platelet adhesion and
aggregation, blood coagulation, and fibrinolysis.27 Endothelial
cell function thus directly affects the balance of hemostasis
and thrombosis in the cardiovascular system. Therefore, we
can investigate the ECs function through the measurement
of the secretion of vWF by ECs growing on the modified
PCL and on TCPS. Figure 10 showed that all cells seeded
on TCPS or modified PCL membranes secreted vWF and
maintained the endothelial function after cells were cultured
in vitro for 72 h. Compared with control PCL, the ECs grown
on aminolyzed PCL secreted more vWF regardless of the
surface amino amounts (aminolyzing time), while the vWF
concentrations on gelatin-, chitosan-, or collagen-immobilized
PCL membranes were the highest. No big difference of the

Zhu et al.

Figure 10. The vWF secretion (ng/10000 cells) by ECs on TCPS

(a), control PCL (b), PCL membranes aminolyzed for 3 min (c), 10
min (d), 30 min (e), and 120 min (f), PCL membranes immobilized
by gelatin (g), chitosan (h), or collagen (I), respectively. Cell seeding
density was 15 104/cm2.

vWF between these biomacromolecules-immobilized PCLs

was found. It was interesting that both amino groups and
biomacromolecules could not only promote ECs growth but
also maintain the endothelial function.
Conclusion
Free amino groups were introduced onto the PCL membrane surface through an aminolyzing reaction with 1,6hexanediamine. The NH2 density was measured by absorbance spectroscopy using ninhydrin. The introduced NH2
groups provided the opportunity to immobilize biomacromolecules such as gelatin, chitosan or collagen onto the PCL
membrane surface. The introduction of the biocompatible
macromolecules had a positive effect on modifying the
cytocompatibility of PCL. The cell attachment and proliferation ratios were improved obviously, and the cells spread
well and lived comfortably from the cell morphology
observed under SEM. Moreover, the secreting function of
ECs seeded on the modified PCL membrane also remained.
In conclusion we have provided a novel technique, i.e., the
aminolysis and the following biomacromolecules immobilization, through which a cytocompatible polymeric material
can be easily fabricated.
Acknowledgment. Financial support by the Major State
Basic Research Program of China (G1999054305) is gratefully acknowledged.
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