505
Volume 5, Issue 10: Special Issue
(EJAR)
ISSN: 2181-2020
MPHAPP
THE 6TH INTERNATIONAL SCIENTIFIC AND PRACTICAL
CONFERENCE
“
MODERN PHARMACEUTICS: ACTUAL
PROBLEMS AND PROSPECTS
”
TASHKENT, OCTOBER 17, 2025
in-academy.uz
EFFECTS OF HYALURONIC ACID AND GELATIN-BASED HYDROGELS LOADED
WITH DOXORUBICIN ON 4T1 CANCER CELLS
Boydedaev A.A
1
.
Amonova D.M
1
.
Karimov M. Sh
1
.
Kalonova M.O
1
.
Muhitdinov B.I
1
.
Turaev A.S
1
.
Huang Y
2
.
Wang H
2
1
Institute of Bioorganic Chemistry, Academy of Sciences of the Republic of Uzbekistan,
Tashkent city, Republic of Uzbekistan
2
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai city, China
e-mail: azizbek.boydedaev@gmail.com
https://doi.org/10.5281/zenodo.17343457
Cancer is one of the most life-threatening diseases, and conventional therapies such as
chemotherapy, radiotherapy, and systemic treatments are widely used. However, these methods often
cause severe side effects, highlighting the need for alternative strategies. Hydrogels, especially those
based on hyaluronic acid (HA) and gelatin, are promising due to their biocompatibility,
biodegradability, and drug delivery potential. They can release therapeutic agents locally, in a
controlled and sustained manner. HA, a natural polysaccharide, combined with gelatin, a collagen
derivative, forms hydrogels that interact efficiently with cancer cells, while their water retention and
microenvironment support cell regulation. Doxorubicin (DOX), a common chemotherapeutic,
inhibits proliferation and induces apoptosis but is limited by systemic toxicity. Therefore, HA–gelatin
hydrogels provide an innovative carrier to enhance efficacy and reduce side effects. The 4T1 breast
cancer line, known for its aggressive and metastatic behavior, is a suitable model to evaluate these
systems. This study examines DOX-loaded HA–gelatin hydrogels on 4T1 cells to provide insights
for effective localized cancer therapy.
In this study, we formulated doxorubicin-conjugated, gelatin-crosslinked HA injectable
hydrogels (HA-G-DOX). The hydrogel preparations were carried out using varying molar ratios of
HA, gelatin, and DOX (1:0.1–1:0.05–0.25 mol) across diverse solvent systems, including water,
dioxane, DMSO, and DMF. Reaction parameters were optimized to achieve injectable hydrogels with
stable physicochemical properties and reproducible quality. The resulting hydrogels were further
purified from residual reactants and by-products through sequential washing, centrifugation, and
dialysis procedures, ensuring their chemical and physical integrity.
The prepared HA–gelatin hydrogels containing 0.035% DOX equivalent were sterilized under
ultraviolet light in a laminar flow hood for 12 hours and subsequently incubated in DMEM culture
medium for 24, 48, and 72 hours. To maintain sterility throughout the experimental procedure, all
hydrogel samples were sequentially passed through a 0.22-µm membrane filter before being applied
to 96-well plates seeded with 5000 4T1 cells per well. Cell viability assays revealed inhibition rates
of approximately 25% after 24 hours, 35% after 48 hours, and 45–50% after 72 hours, indicating
substantial cytotoxic and antitumor activity.
These results substantiate that HA–gelatin hydrogels loaded with DOX are highly effective in
suppressing the proliferation of 4T1 breast cancer cells. The integration of HA and gelatin as
biocompatible scaffolds with doxorubicin as a potent chemotherapeutic agent provides a promising
506
Volume 5, Issue 10: Special Issue
(EJAR)
ISSN: 2181-2020
MPHAPP
THE 6TH INTERNATIONAL SCIENTIFIC AND PRACTICAL
CONFERENCE
“
MODERN PHARMACEUTICS: ACTUAL
PROBLEMS AND PROSPECTS
”
TASHKENT, OCTOBER 17, 2025
in-academy.uz
approach for localized drug delivery. Importantly, these hydrogel systems demonstrate the potential
to reduce systemic toxicity while maintaining or even enhancing therapeutic efficacy.
Overall, the findings from this study clearly demonstrate that HA–gelatin-based hydrogels
incorporating 0.035% DOX can effectively inhibit the growth of aggressive breast cancer cells over
extended incubation periods. The observed growth suppression underscores the importance of
optimizing hydrogel formulations for controlled and sustained drug delivery. For future studies, the
in vivo antitumor efficacy and biological activity of these HA–gelatin hydrogels are being evaluated
in animal models to investigate their potential in preventing postoperative cancer recurrence, thereby
confirming their therapeutic effectiveness and safety for further development as localized anticancer
treatment strategies.
