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While biodegradable, biocompatible polyesters such as poly (lactic-co-glycolic acid) (PLGA) are popular
materials for the manufacture of tissue engineering scaffolds, their surface properties are not particularly
suitable for directed tissue growth. Although a number of approaches to chemically modify the PLGA surface
have been reported, their applicability to soft tissue scaffolds, which combine large volumes, complex shapes,
and extremely fine structures, is questionable. In this paper, we describe two wet-chemical methods, base
hydrolysis and aminolysis, to introduce useful levels of carboxylic acid or primary and secondary amine
groups, respectively, onto the surface of PLGA with minimal degradation. The effects of temperature,
concentration, pH, and solvent type on the kinetics of these reactions are studied by following changes in
the wettability of the PLGA using contact angle measurements. In addition, the treated surfaces are studied
using X-ray photoelectron spectroscopy (XPS) to determine the effect on the surface chemical structure.
Furthermore, we show using XPS analysis that these carboxyl and amine groups are readily activated to
allow the covalent attachment of biological macromolecules.
Introduction
Soft tissue engineering offers a number of unique challenges not seen in other tissue engineering applications such
as bone, cartilage or skin. For many soft tissue applications,
scaffolds must be large (e.g. breast reconstruction1), very
highly porous, and soft, yet have enough strength to resist
the contractile forces generated by growing tissue. Scaffolds
produced from poly(lactic-co-glycolic acid) (PLGA) using
thermally induced phase separation (TIPS)2,3 are able to meet
these goals. However, it is well-known that the surface
properties of PLGA are not ideal for cell growth.4-7 PLGA
is relatively hydrophobic compared to the natural extracellular matrix (ECM), is unable to interact specifically with
cells, and does not possess any functional groups for the
attachment of biologically active molecules.
Since the mechanical and degradative properties of PLGA
are seemingly ideal, and given that PLGA is approved by
the U.S. Food and Drug Administration (FDA) and other
regulatory bodies for implantation into humans, a promising
approach appears to be the surface modification of PLGA
scaffolds post-formation. Ideally, this gives useful surface
characteristics to the polymer, without changing the properties of the bulk.
Many approaches have been taken to modify the surface
of PLGA to date; however, very few of these appear
promising for soft tissue applications. Plasma treatment,
although very popular,8-11 appears unable to penetrate more
than a few millimeters into the pores of a scaffold. Surface
* To whom correspondence should be addressed. Phone: +61 3 8344
4704. E-mail: jjcw@unimelb.edu.au.
entrapment,6,12-14 while apparently promising for 2-dimensional applications and possibly the denser bone and cartilage
scaffolds, leads to collapse of the thin (<1 m) walls of a
soft tissue scaffold during the swelling step. Partial surface
hydrolysis by acid or base treatment6,15-17 has been much
too aggressive in the past, with conditions that destroy a
typical soft tissue TIPS scaffold2,3 in less than one minute.
A desirable property of any surface modification technique
is that it is inherently limited to the very surface of the
substrate irrespective of reaction rate. As an example, soft
tissue scaffolds for breast reconstruction are somewhat
irregularly shaped, have volumes ranging from 100 mL to 2
L,1 and, in the case of PLGA scaffolds produced via the TIPS
process, have internal wall thicknesses as low as 500 nm.3
In this situation, a relatively slow reaction time on the order
of minutes to hours is desirable to ensure controllable
treatment throughout the scaffold.
In this paper, we report on the use of controlled hydrolysis
to produce carboxylic acid functional groups and aminolysis
to produce primary and secondary amine groups on the
surface of thin PLGA films in a highly controlled manner,
with minimal erosion. In addition, covalent binding of a
model amine-functional macromolecule, chitosan, to the
newly formed functional groups was characterized.
Experimental Section
Materials. Poly(D,L-lactic-co-glycolic acid) (PLGA) with
a lactic acid:glycolic acid ratio of 75:25 and an inherent
viscosity in chloroform of 0.69 dL/g (molecular weight
approximately 100 kDa) was purchased from Birmingham
464
Croll et al.
466
Croll et al.
Table 1. Fitted First-Order Decay Parameters for Treated
Surfaces (Treatment Temperature ) 20 C)
control
0.01 N NaOH
0.05 N NaOH
0.05 M ED (aq)
0.05 M ED (IPA)
0.05 M AEPDA (aq)
0.05 M AEPDA (IPA)
a, eq ()
r, eq ()
t1/2 (min)
74.9 ( 0.6
71.7 ( 0.8
71.6 ( 0.8
70.8 ( 0.7
72.9 ( 0.8
70.1 ( 0.6
71.9 ( 0.8
54.7 ( 0.5
49.1 ( 1.1
48.7 ( 0.4
45.9 ( 0.5
50.2 ( 0.9
44.3 ( 0.5
45.8 ( 0.7
N/A
e1
e 0.2
2.7 ( 0.2
6.0 ( 0.5
1.7 ( 0.2
7.6 ( 0.5
k1 + k2[ED]free + k3[OH - ]
1+
(1)
k-
[ED]tot 1 -
Kb
-
[OH ] + Kb
(2)
[OH ][ester]
) k0
(3)
) ra[ED]free + rh[OH - ]
(4)
+1
(5)
where
)
ra[ED]free
rh[OH - ]
(6)
) (1 - e-kt)
(7)
468
Croll et al.
units
value
T ) 0 C
value
T ) 20 C
k0
k1a
k2a
k3a
k-1/k-a
k-2/k-a
k-3/k-a
Kb c
q0ac
q0r c
c
qeq
a,h
c
qeq
r,h
a
qeq
a,a
a
qeq
r,a
c
tc
M-1 min-1
M-1 min-1
M-2 min-1
M-2 min-1
M-1
M-1
M-1
dimensionless
degrees
degrees
degrees
degrees
degrees
degrees
dimensionless
min
0.33a
0.032
0.019
0
0
3.62
0
2 10-4
75.0
54.8
71.6
48.8
69.1
44.9
17.3
120b
4.0b
0.15
0.94
0
0
7.06
0
6.4 10-5
75.0
54.8
71.6
48.8
67.6
44.9
17.3
120*
Figure 4. Contact angle data for surfaces treated with ED in water for 2 h at (a and b) 20 C and (c and d) 0 C, fitted to model described in
eqs 1-9. and - - -, 0.05 M ED; b and s, 0.1 M ED; b and - - - -, 1 M ED.
470
Croll et al.
O
(at. %)
C
(at. %)
N
(at. %)
Si
(at. %)
42.2
33.33
50.0
41.1
40.4
39.6
40.8
57.8
66.67
50.0
58.8
59.6
59.9
58.8
0
0
0
<0.1
<0.1
0.5
0.5
0
0
0
<0.1
<0.1
<0.1
<0.1
Figure 10. XPS N 1s spectra of surfaces modified by (i) 0.01 N NaOH for 120 min, (ii) 0.05 N NaOH for 30 min, (iii) AEPDA in IPA for 120 min,
(iv) aqueous ED for 120 min. (a) Untreated, (b) primary treated, (c) primary + adsorbed chitosan, (d) primary + covalently bound chitosan.
Table 4. Nitrogen Content of Films after Treatment and Chitosan
Attachment
treatment
primary
(at. %)
primary + ads.
(at. %)
primary + cov.
(at. %)
untreated
0.01 N NaOH
0.05 N NaOH
0.05 M AEPDA
0.05 M ED
<0.1
<0.1
<0.1
0.5
0.5
N/A
0.15
0.1
0.4
0.5
N/A
0.45
0.65
1.1
1.05
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Croll et al.
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