Introduction: Uncontrolled hemorrhage is a major cause of death in civilian and military settings (30 and 80%, respectively), often complicated by infection. This underscores the critical need for rapid, effective hemostatic materials. Natural polysaccharides are promising for this purpose. This study aimed to synthesize a novel powdered hemostatic material using a scalable spray-drying technique for rapid hemorrhage control, leveraging the polyelectrolyte properties of chitosan (CHI) and carboxymethylcellulose (CMC).
Methods: Polyelectrolyte complexes (PECs) were formed by the dropwise mixing of CHI (1.5% w/v) and CMC solutions (0.5, 1, and 1.5% w/v). These PECs were then spray-dried (inlet temperature 160 °C, outlet temperature 80 °C, aspirator 70 °C) to yield powdered microgels (1.5Q/1.5CMC; 1.5Q/1.0CMC; and 1.5Q/0.5CMC). The resulting microgels were evaluated for their morphological, physical, thermal, swelling, and hemostatic properties.
Results: ATR-FTIR spectra confirmed electrostatic interactions between amino and carboxyl groups of CHI and CMC in all microgels. The SEM technique revealed an irregular spherical morphology with an average size of 2–3 µm for all microgels. The gel fraction test (GF) showed that 1.5Q/1.5CMC samples had higher crosslinking between CHI and CMC (64.2 ± 1.7%) compared to 1.5Q/1.0CMC (49.9 ± 2.5%) and 1.5Q/0.5CMC (42.9 ± 3.8%). Additionally, 1.5Q/1.5CMC microgels exhibited higher swelling percentages (up to 900%) in PBS solution (pH 7.4) than 1.5Q/1.0CMC and 1.5Q/0.5CMC (up to 600%). TGA analysis indicated similar thermal behavior across all microgels, with three or four degradation stages and a degradation temperature (Tp) around 240 °C. Hemolysis testing (ASTM-F756-00) showed that all microgels had a hemolysis percentage below 5%, confirming their hemocompatibility. Furthermore, all microgels reduced the coagulation time by 20% in comparison with whole blood without microgels.
Conclusions: Three powdered microgels were successfully obtained using CHI and CMC via spray-drying. Among them, 1.5Q/1.5CMC microgels demonstrated superior physicochemical properties, including higher gel fraction and swelling percentages, making them particularly promising for use as hemostatic materials.
Author Contributions
Conceptualization, A.G., J.S.G. and V.A.A.; methodology, A.G. and J.S.G.; software, A.G.; validation, A.G., J.S.G. and V.A.A.; formal analysis, A.G.; investigation, A.G.; resources, J.S.G. and V.A.A.; data curation, A.G. and J.S.G.; writing—original draft preparation, A.G.; writing—review and editing, A.G., J.S.G. and V.A.A. visualization, A.G. and J.S.G.; supervision, J.S.G. and V.A.A.; project administration, J.S.G. and V.A.A.; funding acquisition, J.S.G. and V.A.A. All authors have read and agreed to the published version of the manuscript.
Funding
This research was funded by the Universidad Nacional de Mar del Plata under Grant ING 710/24; and CONICET under Grant 0638.
Institutional Review Board Statement
The study was carried out in accordance with the World Medical Association Declaration of Helsinki: Ethical principles for medical research involving human subjects. All procedures were performed in compliance with relevant laws and institutional guidelines. Ethical approval for this study was obtained from the Research Ethics Committee of the Interdisciplinary Thematic Program in Bioethics (PTIB), affiliated with the Secretariat of Science and Technology of the National University of Mar del Plata, on 8 July 2024. The Research Ethics Committee of the PTIB is registered in the Provincial Registry of Research Ethics Committees, under the Central Ethics Committee in Research of the Ministry of Health of the Province of Buenos Aires. The protocol was registered under the number NO-2024-27,503,325-GDEBA-DPEGSFFMSALGP from La Plata, Buenos Aires, Argentina.
Informed Consent Statement
Not applicable.
Data Availability Statement
The original contributions presented in this study are included in the article text. Further inquiries can be directed to the corresponding author.
Conflicts of Interest
The authors declare no conflict of interest.
| Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
