Stably dispersed calcium alginate nanoparticles incorporating rifampicin for pH-responsive drug delivery

Authors

  • Yuto Sato National Institute of Technology, Oyama College (Oyama KOSEN), Department of Materials Chemistry and Bioengineering, 771 Nakakuki, Oyama, 323-0806 Tochigi, Japan
  • Wataru Tobita National Institute of Technology, Oyama College (Oyama KOSEN), Department of Materials Chemistry and Bioengineering, 771 Nakakuki, Oyama, 323-0806 Tochigi, Japan
  • Keita Kashima National Institute of Technology, Oyama College (Oyama KOSEN), Department of Materials Chemistry and Bioengineering, 771 Nakakuki, Oyama, 323-0806 Tochigi, Japan

DOI:

https://doi.org/10.14311/AP.2026.66.0352

Keywords:

calcium alginate, rifampicin, nanoparticle, riboflavin, vitamin B2, drug-delivery carrier, controlled release, incorporating efficiency

Abstract

Nanoparticles with improved dispersion stability were successfully prepared using calcium alginate (CaALG), with a focus on the aggregation-suppression effect of honey. This study aimed to determine the active components in honey responsible for stabilising CaALG nanoparticles. A systematic comparison of honey and its constituent components revealed that vitamin B2 (riboflavin) plays a dominant role in suppressing nanoparticle aggregation. The enhanced stability is attributed to multipoint hydrogen-bonding and polar interactions between riboflavin and alginate chains, which form a protective interfacial layer that prevents particle-particle contact. Rifampicin (RIF), an antituberculosis drug, was incorporated into the nanoparticles and the particle size, determined by dynamic light scattering, strongly depended on the RIF loading amount. The incorporation efficiency of RIF was evaluated based on the relative elution rate through a dialysis membrane and was significantly influenced by the CaCl2 concentration. The optimal CaCl2 concentration corresponded approximately to the stoichiometric balance between ALG and Ca2+, as described by the Egg-Box model. Furthermore, pH-responsive drug release was demonstrated using simulated gastric fluid (pH 1.2) and intestinal fluid (pH 7.4), where the release of RIF was effectively suppressed under acidic conditions and promoted under neutral conditions. These results indicate that CaALG nanoparticles stabilised by riboflavin are promising carriers for controlled oral drug delivery.

Downloads

Download data is not yet available.

References

[1] R. S. Dharmadhikari, C. P. Dudhat, M. P. Shah, et al. From concept to market: Evaluating the role of fast-dissolving strips in oral drug delivery. Journal of Drug Delivery Science and Technology 101:106239, 2024. https://doi.org/10.1016/j.jddst.2024.106239

[2] A. Berardi, R. Castells-Graells, G. P. Lomonossoff. High stability of plant-expressed virus-like particles of an insect virus in artificial gastric and intestinal fluids. European Journal of Pharmaceutics and Biopharmaceutics 155:103–111, 2020. https://doi.org/10.1016/j.ejpb.2020.08.012

[3] A. Berardi, L. Bisharat. Nanotechnology systems for oral drug delivery: Challenges and opportunities. One Central Press, 2016.

[4] J. Schreiner, F. E. B. Brettner, S. Gier, et al. Unlocking the potential of microfold cells for enhanced permeation of nanocarriers in oral drug delivery. European Journal of Pharmaceutics and Biopharmaceutics 202:114408, 2024. https://doi.org/10.1016/j.ejpb.2024.114408

[5] M. Iwai, E. Yamazoe, T. Ito, et al. Design of an oral formulation combining PVA nanofibers and PEGylated liposomes for enhanced drug delivery. Journal of Drug Delivery Science and Technology 101:106285, 2024. https://doi.org/10.1016/j.jddst.2024.106285

[6] A. Berardi, F. B. Bombelli. Oral delivery of nanoparticles – let’s not forget about the protein corona. Expert Opinion on Drug Delivery 16(6):563–566, 2019. https://doi.org/10.1080/17425247.2019.1610384

[7] L. Musciacchio, M. Mardirossian, G. Marussi, et al. Core-shell electrospun polycaprolactone nanofibers, loaded with rifampicin and coated with silver nanoparticles, for tissue engineering applications. Biomaterials Advances 166:214036, 2025. https://doi.org/10.1016/j.bioadv.2024.214036

[8] J. Chae, S. H. Kang, J. Kim, et al. Targeted and efficient delivery of rifampicin to macrophages involved in non-tuberculous mycobacterial infection via mannosylated solid lipid nanoparticles. Nanoscale Advances 5(17):4536–4545, 2023. https://doi.org/10.1039/D3NA00320E

[9] W. D. Adane, B. S. Chandravanshi, N. Getachew, M. Tessema. A cutting-edge electrochemical sensing platform for the simultaneous determination of the residues of antimicrobial drugs, rifampicin and norfloxacin, in water samples. Analytica Chimica Acta 1312:342746, 2024. https://doi.org/10.1016/j.aca.2024.342746

[10] R. Ghosh, S. Mondal, D. Mukherjee, et al. Oral drug delivery using a polymeric nanocarrier: chitosan nanoparticles in the delivery of rifampicin. Materials Advances 3(11):4622–4628, 2022. https://doi.org/10.1039/D2MA00295G

[11] M. T. Freeman, J. Shen, S. A. Meenach. An aerosol nanocomposite microparticle formulation using rifampicin-cyclodextrin inclusion complexes for the treatment of pulmonary diseases. International Journal of Pharmaceutics 665:124755, 2024. https://doi.org/10.1016/j.ijpharm.2024.124755

[12] D. He, P. Deng, L. Yang, et al. Molecular encapsulation of rifampicin as an inclusion complex of hydroxypropyl-β-cyclodextrin: Design; characterization and in vitro dissolution. Colloids and Surfaces B: Biointerfaces 103:580–585, 2013. https://doi.org/10.1016/j.colsurfb.2012.10.062

[13] Y. Pang, J. Lu, Y. Wang, et al. Study of the rifampin monoresistance mechanism in Mycobacterium tuberculosis. Antimicrobial Agents and Chemotherapy 57(2):893–900, 2013. https://doi.org/10.1128/AAC.01024-12

[14] A. Rauf, T. A. Tabish, I. M. Ibrahim, et al. Design of mannose-coated Rifampicin nanoparticles modulating the immune response and rifampicin induced hepatotoxicity with improved oral drug delivery. Arabian Journal of Chemistry 14(9):103321, 2021. https://doi.org/10.1016/j.arabjc.2021.103321

[15] M. C. Prihatiningsih, C. Pratama, N. A. Kundari, et al. Rifampicin adsorption and release study using Santa Barbara amorphous-16 modified Al (SBA-16-Al) for a drug delivery system. RSC Advances 14(11):7371–7382, 2024. https://doi.org/10.1039/D3RA08360H

[16] G. Tripodo, S. Perteghella, P. Grisoli, et al. Drug delivery of rifampicin by natural micelles based on inulin: Physicochemical properties, antibacterial activity and human macrophages uptake. European Journal of Pharmaceutics and Biopharmaceutics 136:250–258, 2019. https://doi.org/10.1016/j.ejpb.2019.01.022

[17] T. Smith, K. A. Wolff, L. Nguyen. Molecular biology of drug resistance in mycobacterium tuberculosis. In J. Pieters, J. McKinney (eds.), Pathogenesis of Mycobacterium tuberculosis and its interaction with the host organism, vol. 374, pp. 53–80. Springer, Berlin, Heidelberg, 2012. https://doi.org/10.1007/82_2012_279

[18] E. Snejdrova, J. Loskot, J. Martiska, et al. Rifampicin-loaded PLGA nanoparticles for local treatment of musculoskeletal infections: Formulation and characterization. Journal of Drug Delivery Science and Technology 73:103435, 2022. https://doi.org/10.1016/j.jddst.2022.103435

[19] M.-H. Xiong, Y. Bao, X.-Z. Yang, et al. Delivery of antibiotics with polymeric particles. Advanced Drug Delivery Reviews 78:63–76, 2014. https://doi.org/10.1016/j.addr.2014.02.002

[20] Z. Sang, L. Xu, R. Ding, et al. Nanoparticles exhibiting virus-mimic surface topology for enhanced oral delivery. Nature Communications 14(1):7694, 2023. https://doi.org/10.1038/s41467-023-43465-y

[21] A. Berardi, D. J. Evans, F. B. Bombelli, G. P. Lomonossoff. Stability of plant virus-based nanocarriers in gastrointestinal fluids. Nanoscale 10(4):1667–1679, 2018. https://doi.org/10.1039/C7NR07182E

[22] F. Dernaika, L. Halawy, J. Zeaiter, et al. Development and characterization of a zeolite based drug delivery system: Application to cannabidiol oral delivery. Heliyon 10(17):e37373, 2024. https://doi.org/10.1016/j.heliyon.2024.e37373

[23] D. J. McClements, H. Xiao, P. Demokritou. Physicochemical and colloidal aspects of food matrix effects on gastrointestinal fate of ingested inorganic nanoparticles. Advances in Colloid and Interface Science 246:165–180, 2017. https://doi.org/10.1016/j.cis.2017.05.010

[24] A. Raza, W. Wu. Metal-organic frameworks in oral drug delivery. Asian Journal of Pharmaceutical Sciences 19(5):100951, 2024. https://doi.org/10.1016/j.ajps.2024.100951

[25] Z. Wang, J. Chen, R. Gao, et al. Spatiotemporal manipulation metal-organic frameworks as oral drug delivery systems for precision medicine. Coordination Chemistry Reviews 502:215615, 2024. https://doi.org/10.1016/j.ccr.2023.215615

[26] Y. Miao, J. Liu, H. Xiao, et al. Enabling oral novel taxanes-based chemotherapy with lipophilic prodrug self-nanoemulsifying drug delivery system. Bioorganic Chemistry 153:107902, 2024. https://doi.org/10.1016/j.bioorg.2024.107902

[27] A. Hamdy, M. El-Badry, M. Fathy, A. M. El-Sayed. Impact of oil type on the development and oral bioavailability of self-nanoemulsifying drug delivery systems containing simvastatin. Scientific Reports 14(1):22584, 2024. https://doi.org/10.1038/s41598-024-71980-5

[28] T. Yang, A. Wang, D. Nie, et al. Ligand-switchable nanoparticles resembling viral surface for sequential drug delivery and improved oral insulin therapy. Nature Communications 13(1):6649, 2022. https://doi.org/10.1038/s41467-022-34357-8

[29] G. Shevalkar, L. Borse. Self-microemulsifying drug delivery system (SMEDDS) for oral delivery of zafirlukast: Design, formulation, and pharmacokinetic evaluation. Journal of Drug Delivery Science and Technology 101:106298, 2024. https://doi.org/10.1016/j.jddst.2024.106298

[30] Y. Wang, K. Ouyang, Y. Liao, et al. Milk extracellular vesicles: A promising oral drug delivery system for intestinal diseases. Food Bioscience 61:104641, 2024. https://doi.org/10.1016/j.fbio.2024.104641

[31] L. Zou, M. Cheng, K. Hu, et al. Vesicular drug delivery systems for oral absorption enhancement. Chinese Chemical Letters 35(7):109129, 2024. https://doi.org/10.1016/j.cclet.2023.109129

[32] R. Sharma, S. Kumar, R. Malviya, et al. Recent advances in biopolymer-based mucoadhesive drug delivery systems for oral application. Journal of Drug Delivery Science and Technology 91:105227, 2024. https://doi.org/10.1016/j.jddst.2023.105227

[33] S. S. Garg, R. Dey, A. Sharma, J. Gupta. Recent advances in polymer-based nanoformulations for enhancing oral drug delivery in diabetes. Journal of Drug Delivery Science and Technology 100:106119, 2024. https://doi.org/10.1016/j.jddst.2024.106119

[34] E. M. Varoni, L. Altomare, L. Bonetti, et al. Bilayer chitosan-based patches for steroidal drug delivery on the oral mucosa. Journal of Drug Delivery Science and Technology 99:105919, 2024. https://doi.org/10.1016/j.jddst.2024.105919

[35] S. S. Wijetunge, J. Wen, C.-K. Yeh, Y. Sun. Wheat germ agglutinin liposomes with surface grafted cyclodextrins as bioadhesive dual-drug delivery nanocarriers to treat oral cells. Colloids and Surfaces B: Biointerfaces 185:110572, 2020. https://doi.org/10.1016/j.colsurfb.2019.110572

[36] H. M. Johnsen, W. Filtvedt, J. Klaveness, M. Hiorth. Nano-strategies for advancing oral drug delivery: Porous silicon particles and cyclodextrin encapsulation for enhanced dissolution of poorly soluble drugs. International Journal of Pharmaceutics 666:124809, 2024. https://doi.org/10.1016/j.ijpharm.2024.124809

[37] B. Hufnagel, V. Muellner, K. Hlatky, et al. Chemically modified inulin for intestinal drug delivery – A new dual bioactivity concept for inflammatory bowel disease treatment. Carbohydrate Polymers 252:117091, 2021. https://doi.org/10.1016/j.carbpol.2020.117091

[38] A. K. Jain, V. Sood, M. Bora, et al. Electrosprayed inulin microparticles for microbiota triggered targeting of colon. Carbohydrate Polymers 112:225–234, 2014. https://doi.org/10.1016/j.carbpol.2014.05.087

[39] D. Thomas, K. K. Thomas, M. S. Latha. Preparation and evaluation of alginate nanoparticles prepared by green method for drug delivery applications. International Journal of Biological Macromolecules 154:888–895, 2020. https://doi.org/10.1016/j.ijbiomac.2020.03.167

[40] D. Santhanes, L. Y. Teng, F. S. Sheng, A. G. A. Coombes. Exploiting the versatility of oral capsule formulations based on high M-alginate for targeted delivery of poorly water soluble drugs to the upper and lower GI tract. Journal of Drug Delivery Science and Technology 46:384–391, 2018. https://doi.org/10.1016/j.jddst.2018.05.024

[41] P. Veres, D. Sebők, I. Dékány, et al. A redox strategy to tailor the release properties of Fe(III)-alginate aerogels for oral drug delivery. Carbohydrate Polymers 188:159–167, 2018. https://doi.org/10.1016/j.carbpol.2018.01.098

[42] S. Lee, Y.-C. Kim, J.-H. Park. Zein-alginate based oral drug delivery systems: Protection and release of therapeutic proteins. International Journal of Pharmaceutics 515(1–2):300–306, 2016. https://doi.org/10.1016/j.ijpharm.2016.10.023

[43] N. Raghav, C. Vashisth, N. Mor, et al. Recent advances in cellulose, pectin, carrageenan and alginate-based oral drug delivery systems. International Journal of Biological Macromolecules 244:125357, 2023. https://doi.org/10.1016/j.ijbiomac.2023.125357

[44] N. Samiraninezhad, K. Asadi, H. Rezazadeh, A. Gholami. Using chitosan, hyaluronic acid, alginate, and gelatin-based smart biological hydrogels for drug delivery in oral mucosal lesions: A review. International Journal of Biological Macromolecules 252:126573, 2023. https://doi.org/10.1016/j.ijbiomac.2023.126573

[45] Q.-Q. Li, D. Xu, Q.-W. Dong, et al. Biomedical potentials of alginate via physical, chemical, and biological modifications. International Journal of Biological Macromolecules 277:134409, 2024. https://doi.org/10.1016/j.ijbiomac.2024.134409

[46] P. Pandit, T. N. Gayatri, B. Regubalan. Alginates production, characterization and modification. In Alginates: Applications in the Biomedical and Food, pp. 21–43. John Wiley & Sons, Ltd, 2019. https://doi.org/10.1002/9781119487999.ch2

[47] S. Dash, P. Gutti, B. Behera, D. Mishra. Anionic species from multivalent metal salts are differentially retained during aqueous ionic gelation of sodium alginate and could fine-tune the hydrogel properties. International Journal of Biological Macromolecules 265:130767, 2024. https://doi.org/10.1016/j.ijbiomac.2024.130767

[48] K. Kashima, M. Imai. Advanced membrane material from marine biological polymer and sensitive molecular-size recognition for promising separation technology. In Advancing Desalination. InTech, 2012. https://doi.org/10.5772/50734

[49] K. Kashima, T. Inage, Y. Yamaguchi, M. Imai. Tailorable regulation of mass transfer channel in environmentally friendly calcium alginate membrane for dye removal. Journal of Environmental Chemical Engineering 9(3):105210, 2021. https://doi.org/10.1016/j.jece.2021.105210

[50] K. Yoshida, K. Kashima. Nano-structural modification of mass transfer channels in alginate membrane by dispersed oligoethylene glycol for nanofiltration applications. Colloid and Polymer Science 303(7):1261–1271, 2025. https://doi.org/10.1007/s00396-025-05408-5

[51] Y. Wang, Y. Zhao, J. He, et al. Doubling growth of egg-box structure during calcium-mediated molecular assembly of alginate. Journal of Colloid and Interface Science 634:747–756, 2023. https://doi.org/10.1016/j.jcis.2022.12.096

[52] K. Kashima, M. Imai, I. Suzuki. Superior molecular size screening and mass-transfer characterization of calcium alginate membrane. Desalination and Water Treatment 17(1–3):143–149, 2010. https://doi.org/10.5004/dwt.2010.1710

[53] K. Kashima, R. Nomoto, M. Imai. A novel application of oceanic biopolymers – Strategic regulation of polymer characteristics for membrane technology in separation engineering. In Recent Advances in Biopolymers. InTech, 2016. https://doi.org/10.5772/61998

[54] G. T. Grant, E. R. Morris, D. A. Rees, et al. Biological interactions between polysaccharides and divalent cations: The egg-box model. FEBS Letters 32(1):195–198, 1973. https://doi.org/10.1016/0014-5793(73)80770-7

[55] Y. Jia, X. Xu, Y. Liu, et al. Effect of calcium concentration on metastasis of hepatocellular carcinoma cells cultured in alginate gel beads. Colloids and Surfaces B: Biointerfaces 245:114201, 2025. https://doi.org/10.1016/j.colsurfb.2024.114201

[56] K. Kashima, M. Imai. Dominant impact of the α-L-guluronic acid chain on regulation of the mass transfer character of calcium alginate membranes. Desalination and Water Treatment 34(1–3):257–265, 2011. https://doi.org/10.5004/dwt.2011.2892

[57] K. Kashima, M. Imai. Impact factors to regulate mass transfer characteristics of stable alginate membrane performed superior sensitivity on various organic chemicals. Procedia Engineering 42:964–977, 2012. https://doi.org/10.1016/j.proeng.2012.07.490

[58] K. Kashima, M. Imai. Selective diffusion of glucose, maltose, and raffinose through calcium alginate membranes characterized by a mass fraction of guluronate. Food and Bioproducts Processing 102:213–221, 2017. https://doi.org/10.1016/j.fbp.2016.11.003

[59] E. Rostami. Chapter 9 – Alginate-based nanoparticles for drug delivery. In P. Kesharwani, N. Jain (eds.), Theranostics Nanomaterials in Drug Delivery, pp. 157–168. Academic Press, 2025. https://doi.org/10.1016/B978-0-443-22044-9.00007-3

[60] S. Hasnain, A. K. Nayak, M. Kurakula, M. N. Hoda. Chapter 6 – Alginate nanoparticles in drug delivery. In A. K. Nayak, S. Hasnain (eds.), Alginates in Drug Delivery, pp. 129–152. Academic Press, 2020. https://doi.org/10.1016/B978-0-12-817640-5.00006-6

[61] P. Severino, C. F. da Silva, L. N. Andrade, et al. Alginate nanoparticles for drug delivery and targeting. Current Pharmaceutical Design 25(11):1312–1334, 2019. https://doi.org/10.2174/1381612825666190425163424

[62] S. Sattar, M. Imran, A. Anwar, et al. Formulation of biodegradable alginate-based nano-carriers for in-vitro drug delivery and antibacterial activity. International Journal of Biological Macromolecules 274:133274, 2024. https://doi.org/10.1016/j.ijbiomac.2024.133274

[63] S. Jana, A. Gangopadhaya, B. B. Bhowmik, et al. Pharmacokinetic evaluation of testosterone-loaded nanocapsules in rats. International Journal of Biological Macromolecules 72:28–30, 2015. https://doi.org/10.1016/j.ijbiomac.2014.08.001

[64] S. Ray, P. Sinha, B. Laha, et al. Polysorbate 80 coated crosslinked chitosan nanoparticles of ropinirole hydrochloride for brain targeting. Journal of Drug Delivery Science and Technology 48:21–29, 2018. https://doi.org/10.1016/j.jddst.2018.08.016

[65] P. Geetha, M. S. Latha, S. S. Pillai, et al. Green synthesis and characterization of alginate nanoparticles and its role as a biosorbent for Cr(VI) ions. Journal of Molecular Structure 1105:54–60, 2016. https://doi.org/10.1016/j.molstruc.2015.10.022

[66] W. Rajapaksha, I. H. W. Nicholas, T. Thoradeniya, et al. Novel alginate nanoparticles for the simultaneous delivery of iron and folate: A potential nano-drug delivery system for anaemic patients. RSC Pharmaceutics 1(2):259–271, 2024. https://doi.org/10.1039/D3PM00068K

[67] T. Taubner, M. Marounek, A. Synytsya. Preparation and characterization of amidated derivatives of alginic acid. International Journal of Biological Macromolecules 103:202–207, 2017. https://doi.org/10.1016/j.ijbiomac.2017.05.070

Downloads

Published

2026-07-10

Issue

Vol. 66 No. 3 (2026)

Section

Chemical and Process Engineering

License

Copyright (c) 2026 Yuto Sato, Wataru Tobita, Keita Kashima

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

How to Cite

Sato, Y., Tobita, W., & Kashima, K. (2026). Stably dispersed calcium alginate nanoparticles incorporating rifampicin for pH-responsive drug delivery. Acta Polytechnica, 66(3), 352-363. https://doi.org/10.14311/AP.2026.66.0352