7/3/23 U Janvi

2020A8PS0901P

Glucose-sensitive hydrogels

These are hydrogels that respond to changes in glucose concentration by undergoing reversible
changes in their physical properties, such as swelling, shrinking, or changing their pH and
viscosity. These hydrogels are used in developing glucose sensing and drug delivery systems
for diabetes management.

Hydrogels that change their viscosity with glucose concentration

These hydrogels respond to changes in glucose levels by altering their flow behavior. These
hydrogels can be used in various biomedical applications, such as drug delivery, tissue
engineering, and glucose sensing. The preparation of these hydrogels typically involves mixing
the polymer components with a crosslinking agent and glucose-sensitive molecules, such as
boronic acid or glucose oxidase, and allowing the hydrogel to form through chemical or physical
crosslinking. The resulting hydrogel can then be characterized by its viscosity response to
glucose concentration.

One example of a hydrogel that changes viscosity with glucose concentration is a hydrogel
composed of poly(ethylene glycol) (PEG) and dextran. The hydrogel contains a boronic acid
derivative, which binds reversibly with glucose, causing the hydrogel to change its viscosity. At
low glucose concentrations, the boronic acid groups form fewer crosslinks, resulting in a lower
viscosity. At high glucose concentrations, more boronic acid groups bind to glucose, forming
more crosslinks and increasing the hydrogel's viscosity.

Method of Preparation:

Materials Required:

1. PEG (MW ~ 8,000 Da)

2. Dextran (MW ~ 10,000 Da)
3. 3-aminophenyl boronic acid(APBA)
4. Water
5. Buffer Solution (e.g. phosphate-buffered saline or HEPES buffer)
6. Glassware and pipettes

Procedure:
1. Prepare a solution of PEG and dextran by dissolving them in water at a desired
concentration. For example, a 10% (w/v) solution of PEG and dextran in water
can be used.
2. Add 3-aminophenyl boronic acid (APBA) to the PEG-dextran solution to a final
concentration of 5-10 mM. Stir the solution to ensure homogeneous mixing.
 3. Add a crosslinker, such as a polyethyleneimine or glutaraldehyde, to the
PEG-dextran-APBA solution at a desired concentration. For example, a 0.5%
(v/v) glutaraldehyde solution can be used as the crosslinker. Stir the solution to
ensure homogeneous mixing.
4. Adjust the pH of the solution to the desired level using a buffer solution, such as
phosphate-buffered saline or HEPES buffer. The pH should be slightly alkaline
(pH ~ 8-9) to ensure the APBA groups are deprotonated and can bind to glucose.
5. Transfer the solution to a mold or a container of desired shape and size, and
allow the hydrogel to crosslink at room temperature or a desired temperature for
a desired time period. Crosslinking can be achieved through chemical or physical
means, depending on the type of crosslinker used.
6. After cross-linking, wash the hydrogel with water or buffer solution to remove any
unreacted reagents or solvents.

The resulting hydrogel should be responsive to changes in glucose concentration, with the
APBA groups binding reversibly with glucose and altering the crosslinking density of the
hydrogel, thereby changing its viscosity. The viscosity response can be characterized using
various techniques, such as rheology or measurements. PEG hydrogels provide a unique niche
for cell encapsulation, as they are highly biocompatible with the cells under proper
polymerization conditions (1).

Another example is a hydrogel composed of hyaluronic acid and chitosan, crosslinked with
glutaraldehyde. The hydrogel contains glucose oxidase, which catalyzes the oxidation of
glucose, producing gluconic acid and hydrogen peroxide. The acid lowers the pH of the
hydrogel, causing it to swell and increase its viscosity.
These are not inherently glucose-sensitive. However, glucose sensitivity can be introduced into
the hydrogel by incorporating glucose-responsive moieties or materials. For example, a study
published in the journal Carbohydrate Polymers in 2017 demonstrated the development of a
glucose-responsive hydrogel based on a hyaluronic acid-chitosan scaffold crosslinked with
glutaraldehyde but modified with boronic acid moieties. The boronic acid moieties form
reversible covalent bonds with glucose molecules, leading to a change in the hydrogel structure
and properties, such as swelling and mechanical strength.
In another study published in the journal Biomaterials in 2016, a glucose-responsive hydrogel
was developed by crosslinking hyaluronic acid with phenylboronic acid-modified chitosan and
glutaraldehyde. This hydrogel responded to changes in glucose concentration by changing its
swelling behavior, which could potentially be used for glucose-responsive drug delivery.

Method of Preparation:

Materials:
1. Hyaluronic acid
2. Chitosan
3. Glutaraldehyde
4. Distilled water
 Procedure:

1. Prepare a solution of hyaluronic acid in distilled water at a concentration of 2-3% (w/v).

Stir the solution until the hyaluronic acid is fully dissolved.
2. Prepare a solution of chitosan in distilled water at a concentration of 2-3% (w/v). Stir the
solution until the chitosan is fully dissolved.
3. Slowly add the chitosan solution to the hyaluronic acid solution under constant stirring.
Adjust the pH of the solution to around 6.5 using a small amount of NaOH or HCl if
necessary.
4. Add glutaraldehyde to the mixed solution at a concentration of 0.5-1% (v/v) and stir
gently for 30 minutes to crosslink the hyaluronic acid and chitosan.
5. Transfer the solution to a mold and allow the hydrogel to form by incubating at room
temperature for several hours. The hydrogel should become solid and elastic after
incubation.

The crosslinked hydrogel can be washed with distilled water to remove any excess
glutaraldehyde and then stored at 4°C until use. the exact concentrations of the materials and
crosslinking agent used may vary depending on the specific application and desired properties
of the hydrogel. Additionally, the use of glutaraldehyde as a crosslinking agent may be toxic and
therefore, the hydrogel should be thoroughly washed before use to remove any remaining
unreacted glutaraldehyde.

(2) This paper describes the preparation of a chitosan-gelatin-hyaluronic acid hydrogel

crosslinked with glutaraldehyde for use in tissue engineering applications. The procedure for the
hyaluronic acid-chitosan hydrogel is similar to the one mentioned above, but with the addition of
gelatin as another component in the hydrogel.

(3) There is another category of hydrogels, whose viscosity decreases with increasing glucose
concentration. sugar concentrations of 30–60 wt % reduced the intrinsic viscosity of the
alginate polymer in dilute solution, and the gel network appeared more open and less
connected

(4) There is another paper that describes the viscosity response of certain polymers to varying
glucose concentrations.

Reliable websites for procurement of the materials mentioned above:

1. Sigma-Aldrich: https://www.sigmaaldrich.com/india.html
Merck Life Science: https://www.merckgroup.com/en/research/life-science.html
2. Avantor:
https://in.vwr.com/store/search?label=Poly%28ethylene+glycol%29+%28PEG%29&x=0&
y=0
3. Biomatik: Biochemicals & Enzymes - Biomatik
4. Polysciences: https://www.polysciences.com/india/
 Hydrogels that bulge/swell in response to glucose

There are several methods for preparing glucose-sensitive hydrogels, and the specific method
chosen will depend on the desired properties of the hydrogel and the intended application. They
can be used for swelling pressure sensor applications.

Approach 1: Crosslinking glucose-responsive moieties:

1. One approach is to crosslink glucose-responsive moieties into a hydrogel matrix. For

example, phenylboronic acid (PBA) can be incorporated into a polymer backbone and
crosslinked to form a hydrogel. The PBA binds to glucose, causing the hydrogel to swell.
To prepare such a hydrogel, the following steps can be followed:
2. Dissolve the polymer and the PBA derivative in a suitable solvent.
3. Add a crosslinking agent, such as a di- or tri-functional amine, to the solution and stir.
4. Transfer the solution to a mold and allow it to crosslink, either through chemical or
physical crosslinking.
5. Wash the resulting hydrogel to remove any unreacted reagents or solvents.

Approach 2: Enzymatic crosslinking:

1. Another approach is to incorporate glucose oxidase (GOx) into a hydrogel matrix. The
GOx catalyzes the oxidation of glucose to produce gluconic acid and hydrogen peroxide,
which can crosslink the hydrogel. To prepare such a hydrogel, the following steps can be
followed:
2. Dissolve the polymer and the GOx in a suitable buffer solution.
3. Add a crosslinking agent, such as a divalent metal ion, to the solution and stir.
4. Transfer the solution to a mold and allow it to crosslink through the action of the GOx.
5. Wash the resulting hydrogel to remove any unreacted reagents or solvents.

References:
1. PEG Hydrogels for the Controlled Release of Biomolecules in Regenerative Medicine |
SpringerLink
2. Huang, Y., Onyeri, S., Siewe, M., & Moshfeghian, A. (2005). In vitro characterization of
chitosan–gelatin scaffolds for tissue engineering. Biomaterials, 26(36), 7616-7627
3. Impact of Glucose on the Nanostructure and Mechanical Properties of Calcium-Alginate
Hydrogels https://www.mdpi.com/2310-2861/8/2/71
4. Viscosity response of polymer 1 to 4 to the addition of glucose. | Download Scientific
Diagram (researchgate.net)
5. Swelling Pressure Sensor Applications