Innovative pH-responsive alginate-coated rosuvastatin-loaded chitosan nanoparticles: a targeted approach to inhibiting HMGB1-activated RAGE/TLR4-NFκB signaling in colonic inflammation in rats

Elsayed A Elmorsy; Sameh Saber; Zubida M Al-Majdoub; Rabab S Hamad; Mustafa Ahmed Abdel-Reheim; Asmaa Ramadan; Norah Suliman Alsoqih; Mariam S Alharbi; Hamad Alsaykhan; Alshaimaa A Farrag; Hanan Eissa; Rasha Abdelhady; Abousree T Ellethy; Mostafa M Khodeir; Hossam A Elsisi; Syed Suhail Ahmed; Ahmed Y Kira

doi:10.3389/fphar.2025.1546010

Innovative pH-responsive alginate-coated rosuvastatin-loaded chitosan nanoparticles: a targeted approach to inhibiting HMGB1-activated RAGE/TLR4-NFκB signaling in colonic inflammation in rats

Front Pharmacol. 2025 Jun 18:16:1546010. doi: 10.3389/fphar.2025.1546010. eCollection 2025.

Authors

Elsayed A Elmorsy^{1

2}, Sameh Saber³, Zubida M Al-Majdoub⁴, Rabab S Hamad⁵, Mustafa Ahmed Abdel-Reheim⁶, Asmaa Ramadan⁷, Norah Suliman Alsoqih⁸, Mariam S Alharbi⁹, Hamad Alsaykhan¹⁰, Alshaimaa A Farrag¹¹, Hanan Eissa¹², Rasha Abdelhady^{13

14}, Abousree T Ellethy¹⁵, Mostafa M Khodeir^{16

17}, Hossam A Elsisi^{1

18}, Syed Suhail Ahmed¹⁹, Ahmed Y Kira²⁰

Affiliations

¹ Department of Pharmacology and Toxicology, College of Pharmacy, Qassim University, Buraidah, Saudi Arabia.
² Department of Clinical Pharmacology, Faculty of Medicine, Mansoura University, Mansoura, Egypt.
³ Department of Pharmacology, Faculty of Pharmacy, Delta University for Science and Technology, Gamasa, Egypt.
⁴ Centre for Applied Pharmacokinetic Research, University of Manchester, Manchester, United Kingdom.
⁵ Biological Sciences Department, College of Science, King Faisal University, Al Ahsa, Saudi Arabia.
⁶ Department of Pharmacology, College of Pharmacy, Shaqra University, Shaqra, Saudi Arabia.
⁷ Department of Biochemistry, Faculty of Pharmacy, Delta University for Science and Technology, Gamasa, Egypt.
⁸ Department of Pediatrics, College of Medicine, Qassim University, Buraidah, Saudi Arabia.
⁹ Department of Medicine, College of Medicine, Qassim University, Buraidah, Saudi Arabia.
¹⁰ Department of Anatomy and Histology, College of Medicine, Qassim University, Buraidah, Saudi Arabia.
¹¹ Department of Anatomy, College of Medicine, University of Bisha, Bisha, Saudi Arabia.
¹² Department of Pharmacology, College of Medicine, University of Bisha, Bisha, Saudi Arabia.
¹³ Pharmacology and Toxicology Department, Faculty of Pharmacy, Fayoum University, Fayoum, Egypt.
¹⁴ Pharmacology and Toxicology Department, Faculty of Pharmacy, Egypt ian Chinese University, Cairo, Egypt.
¹⁵ Department of Basic Oral Sciences and Dental Education, Biochemistry Division, College of Dentistry, Qassim University, Buraidah, Saudi Arabia.
¹⁶ Department of Pathology, College of Medicine, Qassim University, Buraidah, Saudi Arabia.
¹⁷ Department of Pathology, Faculty of Medicine, Cairo University, Cairo, Egypt.
¹⁸ Department of Clinical Pharmacology, Faculty of Medicine, Zagazig University, Zagazig, Egypt.
¹⁹ Department of Microbiology and Immunology, College of Medicine, Qassim University, Buraidah, Saudi Arabia.
²⁰ Department of Pharmaceutics, Faculty of Pharmacy, Delta University for Science and Technology, Gamasa, Egypt.

Abstract

This study developed and optimized an innovative oral pH-dependent drug delivery system utilizing rosuvastatin-loaded chitosan nanoparticles (RSV-CSNPs) coated with sodium alginate (ALG). The goal was to protect RSV-CSNPs from degradation in the acidic gastric environment and facilitate targeted sustained release in the colon to address inflammatory bowel disease. Nanoparticles were initially prepared by ionic gelation. A subsequent ALG coating process was optimized using a 2³ factorial design. The optimal ALG-coated formulation demonstrated minimal drug loss (0.88% ± 0.09%), desirable particle size (407.2 ± 1.95 nm), and suitable zeta potential (-27.13 ± 1.36 mV). In vitro release tests highlighted the superiority of ALG-coated RSV-CSNPs, with significantly reduced RSV release in simulated gastric fluid (6.8% ± 1.06% after 2 h) compared to uncoated nanoparticles (38.45% ± 1.79%), affirming the protective effectiveness of the ALG coating. Extended-release studies at colonic pH (6.8) demonstrated sustained RSV release suitable for colon-specific targeting. In vivo assessments in a dextran sodium sulfate (DSS)-induced rat model of colitis revealed that ALG-RSV-CSNPs significantly outperformed both plain RSV and RSV-CSNPs. Treatment notably decreased colonic inflammation, disease severity scores, macroscopic damage, and oxidative stress indicators. Additionally, histopathological analyses showed remarkable restoration of colon tissue integrity, crypt preservation, and mucosal protection in animals treated with ALG-RSV-CSNPs. Mechanistically, ALG-RSV-CSNPs effectively attenuated colitis by significantly inhibiting the HMGB1-triggered RAGE/TLR4-NFκB inflammatory signaling pathway. Treatment resulted in substantial reductions in key inflammatory markers, including HMGB1, RAGE, TLR4 expression, NFκB activity, pro-inflammatory cytokines (TNF-α and IL-6), and apoptosis marker caspase-3. The anti-inflammatory actions were further supported by reduced neutrophil infiltration, lipid peroxidation, and enhanced antioxidant enzyme activities (SOD and GSH levels). The study identified HMGB1, RAGE, TLR4, and NFκB as critical biomarkers predicting disease activity. Correlation analysis highlighted strong positive associations among these markers, underscoring their collective involvement in colitis pathogenesis and emphasizing the multitarget therapeutic efficacy of ALG-RSV-CSNPs. Overall, this study demonstrates that the optimized pH-responsive ALG-coated RSV-CSNPs significantly enhance the therapeutic outcomes of RSV in colonic inflammation through targeted delivery and sustained release. These nanoparticles represent a promising strategy for effectively managing ulcerative colitis and related inflammatory bowel diseases. Future clinical studies are warranted to validate these findings and facilitate translation into human therapeutic applications.

Keywords: DSS/Sprague-Dawley rat colitis model; HMGB1-RAGE/TLR4-NFκB; colonic delivery system/colitis; inflammation; rosuvastatin/pH-responsive nanoparticles; sodium alginate/chitosan nanoparticles.