Anti-amyloid activity of carrageenan-loaded liposomal nanocarriers against subcutaneous insulin-induced amyloid masses in rats

Original scientific paper

Authors

  • Saranya Udayakumar Medical Bionanotechnology, Faculty of Allied Health Sciences, Chettinad Hospital & Research Institute (CHRI), Chettinad Academy of Research and Education (CARE), Kelambakkam, Chennai, TN-603103, India https://orcid.org/0009-0003-3655-4689
  • Koyeli Girigoswami Medical Bionanotechnology Lab, Department of Obstetrics and Gynaecology, Saveetha Medical College and Hospital, Saveetha Institute of Medical and Technical Sciences, Thandalam, Chennai, TN-602105, India https://orcid.org/0000-0003-1554-5241
  • Venkatakrishnan Kiran Medical Bionanotechnology, Faculty of Allied Health Sciences, Chettinad Hospital & Research Institute (CHRI), Chettinad Academy of Research and Education (CARE), Kelambakkam, Chennai, TN-603103, India https://orcid.org/0009-0006-0536-8092
  • Devadass Jessy Mercy Medical Bionanotechnology, Faculty of Allied Health Sciences, Chettinad Hospital & Research Institute (CHRI), Chettinad Academy of Research and Education (CARE), Kelambakkam, Chennai, TN-603103, India https://orcid.org/0009-0007-0985-8344
  • Vijayashree Raghavan Department of Pathology, Vels Medical College and Hospital, Manjankaranai, Chennai, TN-601102, India https://orcid.org/0000-0002-7655-243X
  • Agnishwar Girigoswami Medical Bionanotechnology, Faculty of Allied Health Sciences, Chettinad Hospital & Research Institute (CHRI), Chettinad Academy of Research and Education (CARE), Kelambakkam, Chennai, TN-603103, India https://orcid.org/0000-0003-0475-2544

DOI:

https://doi.org/10.5599/admet.3536

Keywords:

Localized amyloidosis, neurodegenerative diseases, amyloid clearance, sulfated polysaccharide, healthcare

Abstract

Background and purpose: Protein misfolding and subsequent amyloid formation are associated with various diseases like localized amyloidosis, type II diabetes, and other neurodegenerative diseases. Insulin can undergo fibrillation under specific physiological conditions, leading to the formation of localized amyloid deposits. Repeated subcutaneous administration of insulin fibrils induces amyloid mass at injection sites. Experimental approach: This study investigated the pathological consequences of insulin fibril deposition in a rat model and evaluated the anti-amyloid efficacy of iota-carrageenan (CG) and its liposomal nano­formulation (nCG). Repeated subcutaneous administration of insulin for 28 days induced the formation of a well-defined amyloid mass. Key results: Treatment with nCG markedly reduced amyloid accumulation and effectively suppressed amyloid progression. Histopathological examination demonstrated a concentration-dependent reduction in amyloid deposits following CG treatment, while nCG-treated groups exhibited no detectable amyloid accumulation. The formation and inhibition of amyloid were further validated by hematoxylin and eosin (H&E), Congo red and thioflavin T (ThT) staining. In addition, nCG showed excellent biocompatibility, with no observable pathological alterations in major organs. Conclusion: Collectively, these findings indicate that both CG and nCG possess significant anti-amyloid activity, with nanoformulated CG showing enhanced efficacy in preventing insulin-induced localized amyloidosis.

Downloads

Download data is not yet available.

References

[1] J.N. Buxbaum, D.S. Eisenberg, M. Fändrich, E.D. McPhail, G. Merlini, M.J.M. Saraiva, Y. Sekijima, P. Westermark. Amyloid nomenclature 2024: update, novel proteins, and recommendations by the International Society of Amyloidosis (ISA) Nomenclature Committee. Amyloid 31 (2024) 249-256. https://doi.org/10.1080/13506129.2024.2405948 DOI: https://doi.org/10.1080/13506129.2024.2405948

[2] N. Pathak, S.K. Vimal, I. Tandon, L. Agrawal, C. Hongyi, S. Bhattacharyya. Neurodegenerative disorders of alzheimer, parkinsonism, amyotrophic lateral sclerosis and multiple sclerosis: an early diagnostic approach for precision treatment. Metabolic Brain Disease 37 (2022) 67-104. https://doi.org/10.1007/s11011-021-00800-w DOI: https://doi.org/10.1007/s11011-021-00800-w

[3] J. Wang, L. Dai, Z. Zhang. Protein aggregation in neurodegenerative diseases. Chinese Medical Journal 138 (2025) 2753-2768. https://doi.org/10.1097/CM9.0000000000003802 DOI: https://doi.org/10.1097/CM9.0000000000003802

[4] T. Sinnige. Molecular mechanisms of amyloid formation in living systems. Chemical Science 13 (2022) 7080-7097. https://doi.org/10.1039/D2SC01278B DOI: https://doi.org/10.1039/D2SC01278B

[5] Z. Niu, X. Gui, S. Feng, B. Reif. Aggregation Mechanisms and Molecular Structures of Amyloid-β in Alzheimer's Disease. Chemistry - A European Journal 30 (2024) e202400277. https://doi.org/10.1002/chem.202400277 DOI: https://doi.org/10.1002/chem.202400277

[6] A. Bigi, R. Cascella, F. Chiti, C. Cecchi. Amyloid fibrils act as a reservoir of soluble oligomers, the main culprits in protein deposition diseases. Bioessays 44 (2022) 2200086. https://doi.org/10.1002/bies.202200086 DOI: https://doi.org/10.1002/bies.202200086

[7] F.S. Ruggeri, F. Benedetti, T.P.J. Knowles, H.A. Lashuel, S. Sekatskii, G. Dietler. Identification and nanomechanical characterization of the fundamental single-strand protofilaments of amyloid α-synuclein fibrils. Proceedings of the National Academy of Sciences 115 (2018) 7230-7235. https://doi.org/10.1073/pnas.1721220115 DOI: https://doi.org/10.1073/pnas.1721220115

[8] P. Shah, C.D. Lee, W. Jin Jeon, A. Dermarderosian, D. Tai, K. Latif, M. Akhtari. The Story of Localized Amyloidosis: Something Old and Something New - A Case Series. Journal of Investigative Medicine High Impact Case Reports 13 (2025) 23247096251400068. https://doi.org/10.1177/23247096251400068 DOI: https://doi.org/10.1177/23247096251400068

[9] K. Azarfar, P. Yaghmaei, M.M. Amoli, N. Hayati-Roodbari, A. Ebrahim-Habibi. Local Insulin-Derived Amyloidosis Model Confronted with Silymarin: Histological Insights and Gene Expression of MMP, TNF-α, and IL-6. International Journal of Molecular Sciences 23 (2022) 4952. https://doi.org/10.3390/ijms23094952 DOI: https://doi.org/10.3390/ijms23094952

[10] M. Pozzan, C. Indennidate, G.G. Varrà, G. Sinagra, M. Merlo, L. Pagura. Amyloidosis and amyloidogenesis: one name, many diseases. Heart Failure Clinics 20 (2024) 249-260. https://doi.org/10.1016/j.hfc.2024.02.001 DOI: https://doi.org/10.1016/j.hfc.2024.02.001

[11] J. Hrudka, E. Sticová, M. Krbcová, K. Schwarzmannová. Localized Insulin-Derived Amyloidosis in Diabetes Mellitus Type 1 Patient: A Case Report. Diagnostics (Basel) 13 (2023) 2415. https://doi.org/10.3390/diagnostics13142415 DOI: https://doi.org/10.3390/diagnostics13142415

[12] C. Samlaska, S. Reber, T. Murry. Insulin-derived amyloidosis: The insulin ball, amyloidoma. JAAD Case Rep 6 (2020) 351-353. https://doi.org/10.1016/j.jdcr.2020.02.011 DOI: https://doi.org/10.1016/j.jdcr.2020.02.011

[13] D. Duong, T. Westhoff-Pankratz, A. Frugoli, S. Pajuleras, K. Ta, B. Barrows, B.D. Barrows. Amyloidoma: a case report of remote insulin-derived amyloidosis in the setting of insulin-dependent diabetes. Cureus 16 (2024) e63525. https://doi.org/10.7759/cureus.63525 DOI: https://doi.org/10.7759/cureus.63525

[14] Y. Shikama, J.-I. Kitazawa, N. Yagihashi, O. Uehara, Y. Murata, N. Yajima, R. Wada, S. Yagihashi. Localized Amyloidosis at the Site of Repeated Insulin Injection in a Diabetic Patient. Internal medicine (Tokyo, Japan) 49 (2010) 397-401. https://doi.org/10.2169/internalmedicine.49.2633 DOI: https://doi.org/10.2169/internalmedicine.49.2633

[15] S. Amniouel, J. Suh, W. Zheng, Q. Zhang. Beyond amyloid: Nanobody-mediated neuroinflammatory therapy for Alzheimer’s disease. Translational Neurodegeneration 14 (2025) 51. https://doi.org/10.1186/s40035-025-00513-5 DOI: https://doi.org/10.1186/s40035-025-00513-5

[16] T. Asaithambi, N.S. Bandhakavi, P. Arikrishnan, S. Sridharan, S. Ullas, S. Udayakumar, A. Girigoswami, K. Girigoswami. Spectrofluorimetric Analysis of Amyloid Degradation Using Shankhapushpi Extract/Zinc Oxide Nanoflower—An In Vitro Study. Chemistry 8 (2026) 98. https://doi.org/10.3390/chemistry8070098 DOI: https://doi.org/10.3390/chemistry8070098

[17] E. Blázquez, V. Hurtado-Carneiro, Y. LeBaut-Ayuso, E. Velázquez, L. García-García, F. Gómez-Oliver, Juan M. Ruiz-Albusac, J. Ávila, M.Á. Pozo. Significance of Brain Glucose Hypometabolism, Altered Insulin Signal Transduction, and Insulin Resistance in Several Neurological Diseases. Frontiers in Endocrinology Volume 13 (2022) 873301. https://doi.org/10.3389/fendo.2022.873301 DOI: https://doi.org/10.3389/fendo.2022.873301

[18] A. Das, M. Shah, I. Saraogi. Molecular Aspects of Insulin Aggregation and Various Therapeutic Interventions. ACS Bio & Med Chem Au 2 (2022) 205-221. https://doi.org/10.1021/acsbiomedchemau.1c00054 DOI: https://doi.org/10.1021/acsbiomedchemau.1c00054

[19] N.M. George, P.M. Sangolli, A. Thulaseedharan, S. Dominic. Cutaneous adverse drug reactions to antidiabetic medications and medical devices used in management of diabetes mellitus. Journal of Skin and Stem Cell 11 (2024) e146625. https://doi.org/10.5812/jssc-146625 DOI: https://doi.org/10.5812/jssc-146625

[20] A. Smith, K. Moore, P. Ambs, A. Saraswati, J. Fortin. Recent Advances in the Discovery of Therapeutics to Curtail Islet Amyloid Polypeptide Aggregation for Type 2 Diabetes Treatment. Advanced Biology 6 (2022) 2101301. https://doi.org/10.1002/adbi.202101301 DOI: https://doi.org/10.1002/adbi.202101301

[21] P. Derashri, S. Kadam, J. Lakkakula, S. Barage, A. Roy, K. Kaur, S. Rustagi, D. Verma, S. Raj, S. Malik. Neuroimaging: mapping and diagnosis of neurodegenerative diseases. Journal of Integrated Science and Technology 12 (2024) 827-827. https://doi.org/10.62110/sciencein.jist.2024.v12.827 DOI: https://doi.org/10.62110/sciencein.jist.2024.v12.827

[22] K. Ramasamy, K.P. Gayathri, B.V. Cheriyan, I.G. Josephine, V.D. Deepti. Evaluation of the Neuroprotective Effect of Lawsonia inermis Flower Extract in Alzheimer’s Disease-induced Zebrafish Model. World Journal of Traditional Chinese Medicine 10 (2024) 451-459. https://doi.org/10.4103/wjtcm.wjtcm_67_24 DOI: https://doi.org/10.4103/wjtcm.wjtcm_67_24

[23] R. Kheirbakhsh, M. Chinisaz, S. Amanpour, S. Amini, S. Khodayari, H. Khodayari, A. Dilmaghanian, M. Haddadi, A. Ebrahim-Habibi. Turmeric effect on subcutaneous insulin-induced amyloid mass: An in vivo study. Drug and Chemical Toxicology 40 (2017) 1-6. https://doi.org/10.3109/01480545.2016.1163575 DOI: https://doi.org/10.3109/01480545.2016.1163575

[24] M. Chinisaz, A. Ebrahim-Habibi, P. Yaghmaei, K. Parivar, A.-R. Dehpour. Generating local amyloidosis in mice by the subcutaneous injection of human insulin amyloid fibrils. Experimental and Therapeutic Medicine 8 (2014) 405-408. https://doi.org/10.3892/etm.2014.1772 DOI: https://doi.org/10.3892/etm.2014.1772

[25] S.K. Metkar, A. Girigoswami, R. Vijayashree, K. Girigoswami. Attenuation of subcutaneous insulin induced amyloid mass in vivo using Lumbrokinase and Serratiopeptidase. International Journal of Biological Macromolecules 163 (2020) 128-134. https://doi.org/10.1016/j.ijbiomac.2020.06.256 DOI: https://doi.org/10.1016/j.ijbiomac.2020.06.256

[26] A.N. Syad, K.P. Devi. Assessment of anti-amyloidogenic activity of marine red alga G. acerosa against Alzheimer’s beta-amyloid peptide 25-35. Neurological Research 37 (2015) 14-22. https://doi.org/10.1179/1743132814Y.0000000422 DOI: https://doi.org/10.1179/1743132814Y.0000000422

[27] Geeva, S. Narayan. Insight into the preparation of biopolymeric nanoparticles with lithium and their cellular uptake studies using PC 12 cells. Materials Research Innovations 26 (2022) 415-426. https://doi.org/10.1080/14328917.2021.2023288 DOI: https://doi.org/10.1080/14328917.2021.2023288

[28] K. Harini, S.Y. Alomar, M. Vajagathali, S. Manoharadas, A. Thirumalai, K. Girigoswami, A. Girigoswami. Niosomal Bupropion: Exploring Therapeutic Frontiers through Behavioral Profiling. Pharmaceuticals 17 (2024) 366. https://doi.org/10.3390/ph17030366 DOI: https://doi.org/10.3390/ph17030366

[29] V. Kiran, K. Harini, A. Thirumalai, K. Girigoswami, A. Girigoswami. Nanotechnology's role in ensuring food safety and security. Biocatalysis and Agricultural Biotechnology 58 (2024) 103220. https://doi.org/10.1016/j.bcab.2024.103220 DOI: https://doi.org/10.1016/j.bcab.2024.103220

[30] S.W. Vedakumari, P. Prabu, S.J.V. Jancy, Y.R. Pravin, K. Manickavasagam, T.P. Sastry. Radiopaque fibrin nanocomplex as a promising tool for X-ray imaging applications. International Journal of Biological Macromolecules 200 (2022) 285-292. https://doi.org/10.1016/j.ijbiomac.2021.12.164 DOI: https://doi.org/10.1016/j.ijbiomac.2021.12.164

[31] H.S. Panda, M.D. Dhokne, S. Thakur, N. Singh, A.K. Datusalia, J.J. Panda. BDNF Loaded Amino Acid-Catecholamine Hybrid Nanoparticles as Curative Agents Against Cognitive Decline in Alzheimer's Disease. Small 21 (2025) 2411701. https://doi.org/10.1002/smll.202411701 DOI: https://doi.org/10.1002/smll.202411701

[32] L. Liu, H. He, B. Du, Y. He. Nanoscale drug formulations for the treatment of Alzheimer's disease progression. RSC Advances 15 (2025) 4031-4078. https://doi.org/10.1039/D4RA08128E DOI: https://doi.org/10.1039/D4RA08128E

[33] S.S.S.J. Ahmed, K. Pallavi, P.S. Aiswarya, V. Ramakrishnan, R.S.A. Husain, Neuroprotective Compounds from Marine Microorganisms, in Encyclopedia of Marine Biotechnology,2020, p. 1559-1579 9781119143802. https://doi.org/10.1002/9781119143802.ch66 DOI: https://doi.org/10.1002/9781119143802.ch66

[34] D. Huang, Q. Wang, Y. Cao, H. Yang, M. Li, F. Wu, Y. Zhang, G. Chen, Q. Wang. Multiscale NIR-II Imaging-Guided Brain-Targeted Drug Delivery Using Engineered Cell Membrane Nanoformulation for Alzheimer’s Disease Therapy. ACS Nano 17 (2023) 5033-5046. https://doi.org/10.1021/acsnano.2c12840 DOI: https://doi.org/10.1021/acsnano.2c12840

[35] S. Udayakumar, S.K. Metkar, K. Girigoswami, A. Girigoswami. Potential effect of nanoformulated iota carrageenan in Aβ1-42 disaggregation: an in vitro, in vivo and in silico study. ADMET and DMPK 14 (2026) 3122. https://doi.org/10.5599/admet.3122 DOI: https://doi.org/10.5599/admet.3122

[36] S. Udayakumar, S.K. Metkar, A. Girigoswami, B. Deepika, G. Janani, L. Kanakaraj, K. Girigoswami. Exploring the amyloid degradation potential of nanoformulated carrageenan-bridging in vitro and in vivo perspectives. International Journal of Biological Macromolecules 279 (2024) 134814. https://doi.org/10.1016/j.ijbiomac.2024.134814 DOI: https://doi.org/10.1016/j.ijbiomac.2024.134814

[37] S.K. Metkar, A. Girigoswami, R. Murugesan, K. Girigoswami. In vitro and in vivo insulin amyloid degradation mediated by Serratiopeptidase. Materials Science and Engineering: C 70 (2017) 728-735. https://doi.org/10.1016/j.msec.2016.09.049 DOI: https://doi.org/10.1016/j.msec.2016.09.049

[38] B. Deepika, P. Pallavi, P. Gowtham, A. Girigoswami, K. Girigoswami. Anticancer potential of nanoformulated extract of Passiflora incarnata leaves. Biocatalysis and Agricultural Biotechnology 57 (2024) 103109. https://doi.org/10.1016/j.bcab.2024.103109 DOI: https://doi.org/10.1016/j.bcab.2024.103109

[39] T. Nagase, K. Iwaya, T. Zako, M. Kikuchi, Y. Katsura. Insulin-derived amyloidosis (insulin ball) and skin-related complications of insulin therapy. Nihon Yakurigaku Zasshi. Folia Pharmacologica Japonica 158 (2023) 173-177. https://doi.org/10.1254/fpj.22109 DOI: https://doi.org/10.1254/fpj.22109

[40] A. Dubernet, M. Toulmonde, M. Colombat, C. Hartog, E. Riviere. Insulin amyloidosis: A case report. Frontiers in Medicine 10 (2023) 1064832. https://doi.org/10.3389/fmed.2023.1064832. DOI: https://doi.org/10.3389/fmed.2023.1064832

[41] V.S. Srivatsa, R.P. Parameswari, A. Roy. Evaluation of the Antioxidant and Anti-Alzheimer's Activity of Oleanolic Acid: An In-vitro Study. Journal of Clinical & Diagnostic Research 19 (2025) 1. https://doi.org/10.7860/JCDR/2025/76186.20691 DOI: https://doi.org/10.7860/JCDR/2025/76186.20691

[42] T. Zhang, X. Wu, H. Yuan, S. Huang, S. Park. Mitigation of Memory Impairment with Fermented Fucoidan and λ-Carrageenan Supplementation through Modulating the Gut Microbiota and Their Metagenome Function in Hippocampal Amyloid-β Infused Rats. Cells 11 (2022) 2301. https://doi.org/10.3390/cells11152301 DOI: https://doi.org/10.3390/cells11152301

[43] H. Sun, L. Xu, K. Wang, Y. Li, T. Bai, S. Dong, H. Wu, Z. Yao. κ‑Carrageenan Oligosaccharides Protect Nerves by Regulating Microglial Autophagy in Alzheimer’s Disease. ACS Chemical Neuroscience 14 (2023) 3540-3550. https://doi.org/10.1021/acschemneuro.3c00460 DOI: https://doi.org/10.1021/acschemneuro.3c00460

[44] K. Kamburova, I.L. Dimitrov, F. Hodzhaoglu, V. Milkova. Investigation of the Aggregation of Aβ Peptide (1-40) in the Presence of κ-Carrageenan-Stabilised Liposomes Loaded with Homotaurine. Molecules 29 (2024) 3460. https://doi.org/10.3390/molecules29153460 DOI: https://doi.org/10.3390/molecules29153460

Published

20-08-2026

Issue

Section

Pharmacology

How to Cite

Anti-amyloid activity of carrageenan-loaded liposomal nanocarriers against subcutaneous insulin-induced amyloid masses in rats: Original scientific paper. (2026). ADMET and DMPK, 14, Article 3536. https://doi.org/10.5599/admet.3536

Most read articles by the same author(s)