NTC Project: M01-CR01 (formerly M01-B01)

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NTC Project: M01-CR01 (formerly M01-B01) Biodegradable Hydrogel-Textile Hybrid for
Tissue Engineering

Goal: To develop an innovative textile-based scaffold biomaterial for engineering new

tissues/organs for human body repair. The new scaffold materials will be based on a hybrid of
the PI’s newly invented biodegradable hydrogel technologies and non-woven or mesh fabrics.
PI’s new biodegradable hydrogels would have well-defined and controllable three dimensional
(3D) porous network structure as well as chemical functional groups for protein attachment and
cellular interaction. At the end of this project, we hope a prototype hydrogel- fabric substrate
could be engineered for preliminary cell culture to determine the merits of this new generation
textile substrates for biomedical applications.

Abstract: The goal of this research project is to engineering a new class of textile-based
substrate that can be used to promote cell attachment and growth for tissue engineering. This
hydrogel- textile hybrid substrate is expected to have enormous advantage over conventional
textile-based substrates for tissue engineering purpose. We have synthesized several types of
hydrogels that have exhibited vast different 3 dimensional porous network structure and swelling
behavior for providing far superior surface area and space/volume than mesh fabric alone. The
fabric component of the hydrogel-textile hybrids was also characterized in terms of their fiber,
yarn and fabric by using two commercially available surgical mesh fabrics. A preliminary
hydrogel- fabric hybrid was engineered and their morphological structure was studied to
demonstrate the concept of the proposal. A preliminary cell culture study on mesh fabric was
done both to use as the control and to illustrate the need of better substrates than existing fabrics
for tissue engineering.

2002 Annual Report (Since Jan. 1, 2002):

Our strategy toward this research project is to research and design proper hydrogels that can be
used to fill up the macropores of commercially available surgical mesh fabrics or/and non-woven
fabrics so that the hydrogel- fabric hybrids will have continuous 3D microporous space/volume
for far better cell attachment and growth than mesh fabric alone. Due to its discontinuous fiber
space/volume from macropores, a mesh or non-woven fabric can only use a small fraction of its
volume (i.e., only the yarns portion of the mesh fabric) to support cell attachment and growth,
and bulks of the mesh fabric space are occupied by macropores that are not available for such
cell engineering purpose. We believe that the use of proper hydrogels as the “filler” for those
macroporous regions of fabrics could overcome this major disadvantage of mesh fabrics as the
substrates for tissue engineering.
Since the period of Jan. 1, 2002, we have accomplished the following:
• Characterization of mesh fabrics that will eventually be used as one of the 2 components
in engineering hydrogel- fabric hybrids.
• R/D of hydrogel technology that will be used as one of the 2 components used to make
hydrogel- fabric hybrids. The hydrogels studied include thermo-responsive hydrogels, 3
arm star-shaped hydrogels and hybrids of these 2 hydrogels with polysaccharide-based
hydrogels like dextran.
• Engineering hydrogel- fabric hybrids.
• Preliminary cell cultures of mesh fabrics.

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• One research paper accepted for publication in J. Material Sci.
• Two research papers are under review for publications.
• Two potential patent technologies are under consideration by Cornell Research
Foundation for US Patent filing.

1. Characterization of mesh fabrics

The goal is to characterize one of the two required components for fabricating textile-
hydrogel hybrids, the textile component. Surgical mesh fabrics are chosen as the textile
component in the textile-hydrogel hybrids and their fiber, yarn and fabric characteristics were
determined in terms of their physical, chemical, mechanical, and morphologic properties. Two
commonly used surgical mesh fabrics, Vicryl® woven mesh and Surgicel®, were used for such a
characterization task. The information obtained will help us evaluate the hydrogel- fabric
hybrids. Standard textile characterization techniques were used. The characterization included
fiber, yarn and fabric identification, porosity measurements including pore size and shape, tensile
and bursting strength determination, as well as stiffness. Figure 1 shows the SEM images of
Surgicel (Fig. 1A) and Vicryl (Fig. 1B) mesh fabrics.

Fig 1A. Surgicel mesh fabric Fig. 1B Vicryl mesh fabric

Tables 1-3 show the characteristics of these 2 mesh fabrics. From the data, we can see a
clear variation in properties between these two mesh fabrics. Table 1 shows the fabric
characterization of the two mesh fabrics. Vicryl mesh is the heaviest, even though it is thinner
than Surgicel because of Surgicel’s larger pore size. Vicryl woven mesh also has the highest
density. Surgicel has the lowest stitch density and the lowest packing factor, because it is the
most porous of the two mesh fabrics.

TABLE 1. Fabric Characterization of Two Commercial Mesh Fabrics

Trade Type Warp/ Weft/ Fabric Stitch Relative Thickness Weight Density Packing
Name Wales Courses count density porosity (cm) (g/10 (g/cm3 ) factorc
per per per cm (%) cm2 ) (%)
10 cm 10 cm
Vicryl® Plain 429 215 644 922 2 0.016 0.168 0.58 36.3
weave
Surgicel® Weft 18 33 51 6 33 0.032 0.065 0.18 16.5
Knit

Table 2 shows the strength evaluation of these two mesh fabrics. From the results it is
evident that Vicryl is a mush stronger mesh fabric than Surgicel. Not only are the tensile

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properties different between these two meshes, but there are also observed differences in the
warp (wale) and weft (course) strength of each of the mesh fabrics. The woven Vicryl mesh and
the Surgicel mesh are both stronger in the warp (wale) direction. Vicryl mesh has the highest
tensile strength and bursting strength, while Surgicel has the highest breaking elongation because
of its fabric structure. The tight plain weave that aids Vicryl in its tensile strength hinders its
elongation. Surgicel is not woven, but knit, giving it give or stretch, especially in the course
direction, letting it elongated to a longer length than Vicryl, in both directions, before it fails.

TABLE 2. Strength Evaluation of Two Commercial Mesh Fabrics

Tensile Properties
Breaking Strength (kg) Breaking Elongation (%) Bursting Bursting
Strength (kg) Strength
(kg/cm2 )
Mesh Warp/Wale Weft/Course Warp/Wale Weft/Course
Vicryl® 43.7 42.5 26.7 35.32 54.9 3.6
Surgicel® 15.1 5.9 43.6 123.9 24.1 1.6

Table 3 displays the stiffness or rigidity of the two commercial meshes. While both
meshes are relatively flexible, the Vicryl woven mesh is the most stiff. Surgicel is about 1/3 less
stiffer tha n Vicryl. This is mostly due to their differences in fabric structure. Again, the knit of
Surgicel compared to the weave of Vicryl, gives it more stretch or give. The plain weave of
Vicryl is so tight that the yarns are packed together closer, so more force is needed to bend the
yarns of Vicryl over Surgicel.

TABLE 3. Stiffness of Two Commercial Mesh Fabrics (Taber Units)

Trade Name Warp/Wale Stiffness Weft/Course Stiffness Fabric Stiffnessa

Vicryl® 0.078 0.085 0.081
Surgicel® 0.063 0.048 0.055
a
Calculated from the following formula: (Warp stiffness x Weft stiffness)1/2

The fabric data reveal that these two varieties of surgical mesh differ a great deal, from
their chemical composition to their mechanical properties. Variation among the meshes was
apparent. Surgicel has a higher relative porosity than Vicryl, while Vicryl® woven mesh has the
highest tensile and bursting strength. Vicryl mesh fabric demonstrates a greater stiffness and a
poorer wrinkle recovery than Surgicel, due mainly to its tight plain weave structural orientation.
Although these fabrics are exceptionally different in their properties, but are used for similar
surgical purposes. Thus, it is important in mesh selection that their yarn and fabric structure be
considered along with their chemical nature, to find the best fit between mesh performance and
desired effects.

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2. New Hydrogel Technology
In this task, we pursued at 3 different directions of R/D of hydrogel component required
for textile-hydrogel hybrids: thermo-responsive hydrogels, temperature sensitive dextran-based
hydrogels, and 3 arm star-shaped hydrogels.

2.A. Thermo-responsive hydrogels.

Our original intention was to use temperature as the means to harvest cells grown on
textile-hydrogel hybrid substrates. We continue our effort in the R/D of poly(N-
isopropylacrylamide) (PNIPAAm) based hydrogel family. We examined the effects of
crosslinking density, semi- interpenetrated network (IPN), and IPN on the properties of
PNIPAAm hydrogels.
2.A.a). Effect of crosslinking density. The aim is to conduct a comprehensive and
systematic study of the effect of the properties of PNIPAAm hydrogels with different levels of
crosslinking, especially their effects on the Lower critical solution temperature (LCST) behavior
and the response dynamics of the PNIAAm hydrogels. The PNIPAAm hydrogels of different
levels of crosslinking were synthesized and characterized by differential scanning calorimetry
(DSC) and scanning electron microscopy (SEM) to investigate the LCST behavior and interior
morphology as well as the temperature dependence of equilibrium swelling ratio, the deswelling
and swelling kinetics. There were 5 levels of crosslinked PNIPAAm hydrogels synthesized
ranging from the lowest level of crosslinking (Gel1.0) to Gel2.0, Gel4.0, Gel8.0 and the highest level
Gel12.0.
The SEM images of these PNIMAAm hydrogels of different crosslinking density (from the
lowest Gel 1.0 to the highest Gel 12.0) are shown in Figure 2 below. The porous structure of
PNIPAAm hydrogels was quite different from each other, depending on the level of crosslinking.
The average pore size and pore number per unit area,
obtained from SEM
micrographs are shown in
Table 4. Although
irregular pore sizes were
found within a single
PNIPAAm hydrogel, as
evident by the large
standard deviation, the
average pore size of the
least crosslinked hydrogel
(Gel1.0) was the largest
(3.4 ± 2.4 µm in
diameter) and decreased
as the level of
crosslinking increased
Figure 2 and reached to the
smallest pore size at
Gel12.0 (1.0 ± 0.4 µm in diameter). We also observed that the higher the level of crosslinking of
the PNIPAAm hydrogels had, the more uniform of their pore size was. The numbers of pores per
unit area, however, were opposite to the relationship between pore size and level of crosslinking.
Gel12.0 showed the largest number of pores per unit area while the Gel1.0 appeared to have the

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smallest number of pores. As a result of the increasing the level of crosslinking, the molecular
weight between crosslinks of this hydrogel would be decreased and the pore density in the
network would be increased.

Table 4. Average pore diameter and pore number per unit area of the swollen PNIPAAm
hydrogel with different level of crosslinking.

Gel1.0 Gel4.0 Gel8.0 Gel12.0

Pore dia meter (um) 3.4 ± 2.4 2.0 ± 1.4 1.3 ± 0.4 1.0 ± 0.4
Pore number/100 um2
8.6 25.0 59.1 100.0

The equilibrium swelling data of PNIPAAm hydrogels of different level of crosslinking

density (Figure 3 below) show that all
the samples, regardless of the level of
crosslinking, have similar swelling
Gel-1.0 profiles as a function of temperature,
60 Gel-2.0 and the phase transition temperatures
Swelling ratio

50 Gel-4.0
or LCSTs of these hydrogels lie in the
40 Gel-8.0
vicinity of 35-36 °C, which is in
30 Gel-12.0
agreement with the thermal data from
20 the DSC study. This is because the
10 LCST of a PNIPAAm hydrogel is also
0 regarded as the temperature at which
the phase-separation-degree (changes
20 25 30 35 40 45 50
Figure 3 o of the swelling ratio vs. temperature
Temperature ( C ) changes around the transition
temperature, ∆SR /∆T) is the greatest
or the temperature at which the swelling ratio of the hydrogel decreases most dramatically.
Even though the LCSTs of the PNIPAAm hydrogels were virtually not affected by the
different levels of crosslinking from Gel1.0 to Gel12.0, the data in Figure 5 clearly show also that,
at a temperature below the transition temperature (e.g., room temperature), the equilibrium
swelling ratios of these hydrogels are significantly reduced from Gel1.0 to Gel12.0, and the
Figure 4 magnitude of the negative swelling slope
below the LCST decreased from Gel1.0
42 to Gel12.0 . It is believed that an increase
37 in level of crosslinking from Gel1.0 the
free to Gel12.0 would reduce volume
Swelling ratio

32
within the hydrogel network structure
27 Gel-2.0
Gel-8.0 that water would reside during swelling.
22
Figure 4 (left) shows the
17
oscillating deswelling- swelling kinetics
12
3 6 -8 0 8 16 24 32 40 48 56 64
of the Gel2.0 and Gel8.0 over the 4- min
T ( C)

T i m e ( m i n ) temperature cycles between 31 and 37

o

28
20
-8 0 8 16 24 32 40 48 56 64
Time(min)
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°C in distilled water. Irrespectively of the level of crosslinking, we found that both the
deswelling and swelling cycles were accompanied by a reduction in the swelling ratio due to the
relatively slower swelling rate of the hydrogels when comparing with the faster deswelling rate.
However, the PNIPAAm hydrogel having a higher level of crosslinking (e.g., Gel8.0) showed a
faster recovery and a large magnitude of deswelling- swelling than the hydrogels having a lower
level of crosslinking (Gel2.0). For example, after the first 2 cycles, the swelling ratio of Gel8.0 was
reduced by 2.4 increment for each 4-minute deswelling process at 37°C and increased by 1.5
increment for each 4- minute swelling process at 31°C; while the corresponding data for Gel2.0
were 1.5 and 0.6 increments, respectively. That is to say, the deswelling magnitude of the Gel8.0
in the deswelling-swelling cycle is about 1.6 times that of Gel2.0, while the swelling magnitude of
the Gel8.0 in the cycle is about 2.5 times that of Gel2.0. This faster and larger magnitude of
oscillating responses from relatively higher level of crosslinked PNIPAAm may be more
favorable for the practical applications in the field s, such as bioengineering and biotechnology
because the response kinetics of the oscillating shrinking-swelling properties to a small
temperature cycles (e.g. cycled around the physiologic temperature) of the hydrogel should be
useful. A full research paper was recently submitted for publication consideration.
2.A.b). Semi-IPN and IPN PNIPAAm Hydrogels
The goal is to search for a new strategy to synthesize faster temperature response
PNIPAAm hydrogels for eventual commercial applications. We synthesized both semi-
interpenetrating polymer network (semi-IPN) and full IPN poly(N-isopropylacrylamide)
(PNIPAAm) polymeric hydrogel. In the semi- IPN PNIPAAm hydrogels, either low (2,300) or
high (33,000) molecular weight linear PNIPAAm chains were used during the crosslinked
reaction of NIPAAm monomers. The properties of the resultant semi- IPN PNIPAAm hydrogels
were characterized by differential scanning calorimetry (DSC) and scanning electron microscopy
(SEM) as well as their swelling ratios at various temperatures, the de-swelling at hot water (48
°C) and the oscillating shrinking-swelling properties within small temperature cycles. It was
found that the de-swelling rate (Figure 5) of these semi-IPN-like PNIPAAm hydrogels is
improved if the molecular weight and/or composition of the linear PNIPAAm chains within the
semi-IPN PNIPAAm hydrogels were increased. This improved de-swelling rate was attributed to
the fast response nature of the linear PNIPAAm chains and the increased pore number in the
matrix network, whic h provide numerous water channels for the freed water during the de-
swelling process at temperature above the LCST.
The de-swelling kinetics of the semi-IPN PNIPAAm hydrogels are shown in Figure 5.
The data clearly demonstrate the de-
swelling response rates were
Water retention ( % )

100 LG0 LG1 accelerated as the composition of

80 LG2 LG3 linear PNIPAAm chains increased
MG1 MG2
MG3 within the PNIPAAm hydrogel (from
60 LG0 to LG1, LG2, LG3 and from
40 MG1, MG2, to MG3). For example,
MG3 reached the constant water
20 retention (lost over 98% water) within
0 less than 18 minutes, while LG3 lost
about 80% within 38 minutes. The
0 8 16 24 32 40 LG0 sample, (the PNIPAAm hydrogel
Figure 5
Time ( min )

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without linear PNIPAAm incorporated) had the slowest de-swelling kinetics and it lost only
about 52% within 38 minutes. The observed accelerated de-swelling response rates from LG0 to
LG1, LG2, LG3 to MG1, MG2, MG3 hydrogels were attributed to the increased pores from LG0
to LG3 to MG3 observed discussed in the morphology section (Figure 6). This increase in
hydrogel pores would improve water diffusing channels. In addition, an increase in the amounts
of the linear PNIPAAm chains will also improve the molecular interactions between the
crosslinked network and the linear chains, which forces the whole network respond or shrink
more quickly.
LG0 LG2 LG3

MG2 MG3

Figure 6. SEM images of semi-IPN PNIPAAm hydrogels

In the IPN-PNIPAAm hydrogel

case, an increased glass transition
60
temperature of these IPNs compared with
Cumulative release (%)

50 the normal PNIPAAm hydrogel was

40 observed. The temperature dependence of
30
the swelling ratio, the deswelling kinetics
and the reswelling kinetics, these IPN
20 IPN1 IPN2
hydrogels also exhibited improved
IPN3 IPN4
10 intelligent property over non-IPN ones,
IPN5 Norm
0 such as the controllable response rate via
0 20 40 60 80 100 changing the ratio of two components
Time (h) required to form IPN networks. Bovine
Figure 7 serum albumin (BSA) was chosen as the
model protein drug and the release of BSA from these IPNs at different temperatures were
studied (an example shown in Fig. 7 at 22C above). The IPN network released BSA at a slower
rate than non-IPN ones. Similar patterns were found at 37C, a release temperature > LCST, but
the total amounts of release were lower than the ones at 22o C, a temperature < LCST. The

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ability to release bioactive agents from the hydrogel component in the textile-hydrogel hybrids is
advantageous because we could incorporate growth factors into the substrate to facilitate the
proliferation of seeded cells at a desirable rate.
A manuscript of simi- IPN PNIPAAm is currently under review for publication while the
IPN-PNIPAAm manuscript is under preparation.
2.B). Temperature-sensitive dextran-PNIPAAm hybrid hydrogels.
The goal is to incorporate temperature-sensitive PNIPAAm units into dextran so that the
resulting hybrid hydrogels will have temperature sensitivity as PNIPAAm alone. Dextran- maleic
acid (Dex-Ma) hydrogel was chosen due to its unusual swelling property as disclosed in our US
patent (awarded in May 2002). We synthesized a series of hybrid hydrogels at the composition
ratio of Dex-Ma to PNIPAAm precursors 0.06/0.14 over a wide range of DMF/water mixed
solvent (1.5/0.5 to 0.25/1.75). The most unique aspect of the resulting hybrid hydrogel of Dex-
Ma/PNIPAAm is their swollen morphological structure, and a representative one is shown in
Figure 8 (left). Instead of the round cylindrical
shape that we normally found in Dex-Ma
hydrogel, the hybrid one exhibits either 4 or 5
member rings with sharp instead of smooth
angles. The wall of each cell is unusually thin and
the longest pore diameter of cells around 10 µm.
We think that this type cell structure may provide
enough surface area for biological cells to anchor
and proliferate. We are in the process of
evaluating the surface area, temperature
sensitivity and release of bioactive agents of this
type of hybrid hydrogels.

Figure 8
2.C. 3 Arm star-shaped hybrid hydrogels
The goal is to explore an
unusual hydrogel that would have
far more molecular mass per
volume so that their pore structure
upon swelling will be different
from dextran-based hydrogels.
This new star-shaped hydrogel
synthesized is based on our
multiarm aliphatic polyester
precursors whose patent is pending.
Figure 9 (left) shows the SEM
morphology of the hybrid hydrogels
based on 3 arm aliphatic polyester and
dextran at various composition ratios.
Fig. 9A is 100% dextran-based, while
Fig. 9D is 100% 3 arm aliphatic

Figure 9
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polyester-based. Fig. 9B and 9C have 30/70 and 40/60 composition ratio of aliphatic polyester to
dextran. It appears that the 40/60 hybrid has the most dense pore appearance and may be a good
candidate as the hydrogel component in hydrogel- fabric hybrid.
3. Engineering Hydrogel-Fabric Hybrid and Preliminary Cell Culture
The goal is to determine whether hydrogel-textile hybrid can be fabricated from two
model components. In this study, we used chemically modified polyethylene glycol as the
hydrogel precursor and polyester mesh as the textile component in hydrogel-textile hybrid. The
resulting hybrids were swollen in water and their morphology was examined to determine the
central theme of the proposal: the macropores of fabrics can be filled up by micropores from
swollen hydrogels so that the resulting hydrogel-textile hybrid will have far more three-
dimensional surface area than the fabric along for proper and accelerated (via bioactive agents
released from the hydrogel component) cell culture and growth for tissue engineering. It is
important to know that macropores (in mm scale) in fabric meshes are too big to be useful for
cell (in µm scale) anchorage and growth. Figure 10 shows an example of hydrogel coated
polyester mesh. The outline of the underling fabric skeleton
is still visible in this hydrogel- textile hybrid substrate. The
white area was one of the macropores of the mesh fabric and
is now filled up by hydrogel in this hydrogel-textile hybrid
substrate. A closer-up view of the hydrogel- filled originally
macroporous area shows very interesting 3D micropore
structure from the swelling of hydrogel in the hydrogel-
textile hybrid. Figures 11 and 12 below show such a close
up view at different magnifications. Such 3D microporous
structure in the original macroporous region of a fabric would
Figure 10
certainly provide enormous amount of additional volume, surface area and space for cell
attachment and proliferation than a simple plain surface of a macroporous fabric could. We are
in the process of measuring surface area of this hybrid to determine its merits over mesh fabric
alone.

Figure 11 Figure 12

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A preliminary cell culture trial of this hybrid is also in progress. The goal of this
preliminary trial is to determine whether any biological cells could attach and grow in mesh
fabrics. An example of this preliminary cell culture work is shown in Figure 13 (below) in
which renal cells were seeded on the polyester mesh component without hydrogel filling.
The bright color spots are the renal cells that attached only onto the yarns of mesh fabric
substrate. The data suggest two important
issues. First, the polyester mesh fabric is
biocompatible and can support the attachment
and growth of biological cells very well.
Second, as we expect and stated previously in
our strategy that the macropore regions of a
mesh fabric (the dark region of Fig. 13) can’t
support cell growth (no bright color spots)
because the size of the macropore is too big for
such a purpose. In our future work, we plan to
examine the potential of our newly engineered
hydrogel- fabric hybrids (shown in Fig. 10-12)
as the substrate for cell attachment and growth.
We shall also continuously search for even
Figure 13
better hydrogel materials for promoting cell attachment and growth.

Project Web Site : http://www.humec.cornell.edu/units/txa/research/ntc/index.html

Acknowledgement: Drs. X. Z. Zhang, D.Q. Wu, & Ms. Sunny Namkang & Marlene Cole at
Cornell University; Drs. Aby J. Mathew and Rob Van Buskirk of State University of New York
in Birmingham.

National Textile Center Annual Report: November 2002