POLYMER

HYDROGEL
REPORT

Name: Gamal Mohammed Abdulhaq

ID: 120172
2. Abstract

The aim of this experiment is to identify the rate of absorbing water and NaCl for the hydrogel.
Hydrogel is prepared through using the monomer (AMPS), when adding 0.1 g to 25 ml distilled
water and N2 or He was passed to get rid of oxygen then adding an initiator, then the solution
was placed in the oven at 600oC. The sample was ready for testing its absorbance with water
and NaCl at different variable times to be easy for plotting a graph to find the relation between
the time and the absorbance of water and NaCl. It was found that the weight of the hydrogel
increases by increasing time for both water and NaCl but with different rates to be indicated that
there were a directly relation proportional relation between time and water and NaCl absorbance
for hydrogel.

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3. Contents
2. Abstract .................................................................................................................................... I
3. Contents .................................................................................................................................. II
4. List of figures ........................................................................................................................ III
5. List of tables .......................................................................................................................... IV
1. Introduction ............................................................................................................................. 1
3. Procedures ............................................................................................................................... 5
1.1. Materials ........................................................................................................................... 5
1.2. Experimental procedures .................................................................................................. 5
4. Results ..................................................................................................................................... 6
1.1. Absorbance of NaCl ......................................................................................................... 6
Absorbance of water ................................................................................................................... 7
5. Sources of Error ...................................................................................................................... 8
6. Discussion and conclusion ...................................................................................................... 9
7. Bibliography ......................................................................................................................... 10

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4. List of figures
Figure 1. Hydrogel..1
Figure 2. Hydrogel layers ............................................................................................................... 3
Figure 3. Time Vs NaCl ................................................................................................................. 6
Figure 4. Time Vs H2O7

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5. List of tables
Table 1. change of absorbance of NaCl with Time ....................................................................... 6
Table 2. change of absorbance of water with Time ........................................................................ 7

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Introduction

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 1. Introduction

Hydrogels are formed by crosslinking polymer chains through physical, ionic or

covalent interactions and are well known for their ability to absorb water. In most cases, they
are homogeneous materials, and their bulk properties are characterized and considered with
regard to applications. From a biomedical perspective, they show promise in a number of areas
including devices, drug delivery and regenerative medicine. Hydrogels are already widely used
as three-dimensional cell and tissue culture environments, as they are excellent mimics of the in
vivo state.

For these bio-related applications, the ability to control the synthesis of hydrogels to make
structures with specific internal forms and shapes is an attractive prospect. For example, internal
spaces created in hydrogels could be used to store cells or drugs and the three-dimensional
structure of actual tissues and other in vivo components could be mirrored in the gel. So far there
has been little control over gel formation, and the creation of complex structures has been
limited. To address this, Ladet et al. report in Nature the design and fabrication of multilayered
polysaccharide-based hydrogels with highly controlled physical properties. These 'onion-like'
layered hydrogels have defined inter-membrane regions and can be formed in various shapes,
including spherical and tubular, and over a range of sizes from micrometers to centimeters. The
secret behind the synthetic success of Ladet et al. is their clear understanding of gelation kinetics
and the mechanism involved.

Figure 1. Hydrogels

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Complex hydrogel structures have been made previously using microfabrication technologies;
however, these were limited to two-dimensional surfaces. To demonstrate the fabrication of their
structurally complex, three-dimensional hydrogels, Ladet et al. use chitosan, a polysaccharide
derived from crustacean shells. This natural polyelectrolyte has many attributes; it is
biodegradable, biocompatible and widely used in cell proliferation and tissue regeneration.

The synthetic basis of the phenomenon is as follows; an alcohol-based chitosan gel is added to
NaOH solution, in which ammonium ions (NH4+) of the chitosan biopolymer are neutralized to
free amine groups (NH2). Following a water wash, and repetition of these neutralizing and
washing steps, a chitosan gel that contains only water and free NH2groups is formed.
Neutralization removes ionic repulsions and allows physical crosslinks, such as hydrophobic
interactions and hydrogen bonding, to form between the chains. On the macroscopic scale, the
gel shrinks. (Harper, 2011)

Through analysis and understanding of this process, Ladet et al. control gelation and
consequently, the physical properties and structure of the polysaccharide hydrogel. For example,
the neutralization of the NH4+ sites on chitosan was found to be influenced by the concentration
of the neutralizing agent, NaOH. By manipulating this concentration, the physical properties of
the gel layers could be controlled. This has implications in tissue engineering because these
properties (amount of water, thickness, crosslinking density and mechanical properties) of the

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hydrogel layers are important parameters for modulating cellular behaviors, including cell shape,
proliferation and stem-cell differentiation.

Figure 2. Hydrogel layers

The major challenge in the synthesis of these multilayered hydrogels was controlling the kinetics
of gelation such that polymer entanglement does not occur. This results in the formation of
separate layers of neutralized and alcohol gels rather than a bulk hydrogel. This separation is
achieved by manipulating the neutralization steps to create an interphase region. If neutralization
is slowed down by simply removing the gel from the NaOH solution, the desired effect is seen
an interphase region that consists of a solution of water and alcohol between the two different
gels. In this interphase region, the polymer chains disentangle and condensation of residual
chains occurs. If this 'interruption' of the neutralization process is repeated, a membrane is added
to the multilayer hydrogel with each cycle. Using this technique, hydrogels on the order of cubic
centimetres with as many as 20 'onion-like' layers could be made. To make multilayered
hydrogels in different shapes, including spheres and tubes, gels in the relevant shapes are used as
starting points.

There are encouraging implications for these multi-membrane hydrogels in the delivery of drugs:
To be effective, the delivery of biological molecules such as fragile growth factors often requires
complex release patterns such as co-delivery with another molecule or pulse-like delivery. The
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formation of layered gels, with inter-membrane spaces in which to store biologically active
compounds, is promising for such complicated delivery profiles.

Looking more generally, it is clear that boundaries and layers are often a necessity in biological
systems. For example, layers of different cell types interact and communicate to influence cell
differentiation and tissue formation. Gradients of growth factors prove critical for innumerable
actions during inflammation of tissues and wound healing. In regenerative medicine, researchers
often attempt to mimic the native structure of a tissue using biomaterials to help organize cells
appropriately. For example, a blood vessel has three different layers, with each layer containing
different cells and extracellular matrix. As a result of the flexibility of Ladet and colleagues'
technique, a tube could be created where the different cell types are introduced in between the
membrane layers.

From a more basic perspective, biologists are often interested in learning how cells 'talk' to each
other in a controlled in vitro environment. The ability to co-culture different cell types together
in a three-dimensional environment, in separate adjacent layers, will allow scientists to evaluate
more realistic cellcell interactions.

The general applicability of the system is demonstrated with the creation of layered hydrogels
from both chitosan and alginate, which are cationic and anionic polyelectrolytes, respectively.
Most importantly, these model biopolymers are widely accessible, making this multilayered
hydrogel technology within practical reach of the general scientific community. (Puoci, 2008)

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 3. Procedures
1.1. Materials

- AMPS, Monomer.
- Methylene-bis-acrylamide, cross linker.
- Ammonium Persulphate, initiator.
- Nitrogen or Argon or helium to get rid of air and provide inert blanket.
- Distilled water.
- Oven.
- 200 ml ground joint conical flask.
- 3Test tubes with cork.

1.2. Experimental procedures

1. 1- Place25 ml distilled water in a suitable conical flask and add about 10 g of the
monomer (AMPS) then add about 0.1 g of the cross linking agent and shake well.
2. Repeat step 1 three times using 0.05, 0.13 and 0.07g of the cross- linking agent
3. Pass N2 or Helium to get rid of oxygen from the solution
4. Add about 0.02 g of the initiator (0.08g/dl) and shake well.
5. Pour the solution into test tubes covered with suitable cork and replace in oven for 1:2
hours at 600 C.
6. The gel formed is taken out of the tubes and cut into suitable discs, which are complete
dried to constant weight.
7. These discs are placed in a beaker containing water at suitable temperature (may be room
temperature) to determine rate of absorbing water
8. Rate of water loss may also be determined.
9. The degree of cross linking and temperature are changed to examine their effect on rate
of water absorption.
10. Draw the different obtained plots of amount of water absorbed of different types of
prepared hydrogels versus time until the saturation point is obtained.

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 4. Results

1.1. Absorbance of NaCl

The table shows the change of absorbance of NaCl with time

Table 1. change of absorbance of NaCl with time

Time (hrs) Absorbance of NaCl (ml)

0 0
2 5
4 7
6 9
17 13
19 14
21 15
23 17
25 18
27 19
29 20
41 22
65 25

Time Vs. absorbance of NaCl

30

25

20

15

10

0
0 2 4 6 17 19 21 23 25 27 29 41 65

Figure 3. Time Vs NaCl

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Absorbance of water

This table shows the change of absorbance of water with time

Table 2. The change of absorbance of water with time

Time (hrs) Absorbance of water

0 2
2 11
4 19
6 27
8 34
10 40
12 43
14 46
16 48
20 52
22 55
24 55
36 61
48 67

Time Vs. Absorbance of water

80

70

60

50

40

30

20

10

0
0 2 4 6 8 10 12 14 16 20 22 24 36 48

Figure 4. Time Vs H2O

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5. Sources of Error

a. Human error that may appear from taking wrong readings.

b. Un-certain volumes when hydrogel is prepared.
c. Existence of oxygen in hydro gel will cause some errors.
d. Excess initiator.
e. Apparatus error as oven and cylinders used may need calibration in order not to cause
error.

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 6. Discussion and conclusion

The aim of this experiment which was to identify the change of absorbance of water and NaCl
for the hydro gel with time. So, hydrogel was needed to be prepared through placing 25 ml of
distilled water and adding 10 grams of monomer (AMPS) and then adding 0.1 gram of cross
linking agent then shaking well. In order to get rid of oxygen, N2 or helium must be passed to the
mixture and then 0.02 grams of initiator must be added to get ready for warming in an oven for 2
hours at 60oC. The sample was cut into suitable discs which were completely dried to bet ready
for examining the absorbance. Two discs of hydro gel was chosen; one for testing the absorbance
of water and the other for NaCl. When the first one was placed in a cylinder containing water to
be observed that after 2 hours the disc of hydro gel absorb 9 ml of water, after 4 hours it absorb
19 ml and so on until reaching 48 hours to be observed that the sample absorb 67 ml of water but
with different rates for every time observing. The graph plotted shows that the linear directly
proportional relation between time and absorbance, which means that when time increases the
absorbance for water increases but with different ratios according to what plotted on the graph
paper. For the other sample when placed in a cylinder containing NaCl, it was observed the same
observation for absorbing water but with different ratios which is less than that of water when
time changed. For example, after passing the first two hours, the sample absorb 5 ml, after 4
hours it absorbs 7 ml of NaCl and after the total time which is 65 hours, 25 ml was absorbed.
From the graph plotted, it was observed a linear directly proportional relation between time and
absorbance of NaCl, which means that when time increases the absorbance of NaCl increases
and vice versa.

Concluding what mentioned above, there is a direct proportional relation between time and
absorbance of either water or NaCl. It was observed that the time taken for hydrogel to absorb an
amount of water was much less than the time taken by hydrogel to absorb the same amount of
NaCl. In other words, the absorbance of water for hydrogel is higher than that of NaCl.

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 7. Bibliography
Harper, D. (2011). http://www.sciencedirect.com/science/journal/20901232/6/2, 22.
Puoci, F. (2008). http://www.sciencedirect.com/science/article/pii/S1319016415000857.

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