Folic acid-loaded chitosan nanoparticles: Cytotoxicity, acute and sub-acute oral toxicity and bioavailability in Wistar rats

Fathima, E.; Kumar, S.; Patil, M. M.; Khanum, F.; Anand, T.

doi:10.1101/2022.12.24.521870

Home/Archive/bioRxiv

bioRxiv · 10.1101/2022.12.24.521870

Folic acid-loaded chitosan nanoparticles: Cytotoxicity, acute and sub-acute oral toxicity and bioavailability in Wistar rats

Fathima, E.·Kumar, S.·Patil, M. M.·Khanum, F.·Anand, T.

View original source ↗Export metadata ↓

Abstract

The study aims to develop folic acid-loaded chitosan nanoparticles (FA-Chi-NPs) as the vitamin is sensitive to heat and light and to investigate the safety of FA-Chi-NPs in vitro and in vivo. The prepared formulation showed an average diameter of 191.7 {+/-} 2 nm with a zeta potential of + 52 {+/-} 4 mV and PDI of 0.195 {+/-} 0.03. The encapsulation efficiency was found to be 80%. In vitro cytotoxicity assays revealed that FA-Chi-NPs had no cytotoxic effect on Caco-2 cells. Acute-toxicity and subacute-toxicity of FA-Chi-NPs (0.4, 2, and 4 mg/kg body weight) revealed no mortality, and no gross morphological abnormalities were noticed. There were no dose-dependent toxic effects of free folic acid (FA) and FA-Chi-NPs in haematological, histological, plasma biochemical indices, oxidative stress enzymes, or other parameters when compared to the control group(s). The plasma and tissue levels of FA from FA-Chi-NPs were significantly higher. Results demonstrated enhanced bioavailability of FA from FA-Chi-NPs with no toxicity. Practical applicationsIn the current study, nanoencapsulation is used as a potential approach to enhance folic acid bioavailability. The results showed that encapsulating folic acid in chitosan improved bioavailability with no apparent adverse effects, and thus chitosan could be a promising polymer for safe folic acid delivery.

Explore related subjects

pharmacology and toxicology Preprint

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Fathima, E., Kumar, S., Patil, M. M., Khanum, F., Anand, T.. 2022-12-25. Folic acid-loaded chitosan nanoparticles: Cytotoxicity, acute and sub-acute oral toxicity and bioavailability in Wistar rats. https://doi.org/10.1101/2022.12.24.521870

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Lipid-ASO therapeutics exhibit differential tissue targeted delivery upon systemic or local CNS administration

Antisense oligonucleotides (ASOs) are a powerful therapeutic modality, but their full potential is hindered by pharmacokinetic properties that affect tissue and cellular delivery. Lipid conjugation is increasingly used to modulate ASO's biodistribution and promote extrahepatic activity, yet lipid dependent effects on in vivo functional delivery, particularly in the central nervous system (CNS), remain less explored. Here, we performed a side by side in vivo comparison of cholesterol, palmitic acid (C16:0), docosanoic acid (C22:0), and eicosapentaenoic acid (C20:5) conjugated to a fully phosphorothioated 3 10 3 LNA gapmer ASO targeting the Malat1 long non coding RNA. Lipid-ASO conjugates were administered systemically or locally in the brain of mice and evaluated for tissue level and cellular level distribution by imaging, qPCR and single-cell RNA sequencing, simultaneously annotating cell origin and global transcriptional changes within the cell. Following systemic administration in mice, lipid conjugation improved overall multi organ efficacy compared to unconjugated ASO, but with pronounced tissue specific differences. Single cell sequencing of liver and heart transcriptomes revealed lipid dependent cellular uptake patterns and transcriptional responses distinct from administration of unconjugated ASO. After intracerebroventricular administration, selected fatty acid conjugates enhanced silencing in deep brain regions such as the striatum, whereas cholesterol conjugation impaired functional delivery despite increased CNS retention. Light-sheet microscopy showed restricted parenchymal penetration of cholesterol ASOs compared with broader but heterogeneous distribution of palmitic acid conjugate. Together, these findings demonstrate that lipid identity critically determines ASO efficacy, productive cellular uptake, and regional CNS engagement, emphasizing the need for context specific lipid design in ASO therapeutic development.

pharmacology and toxicology↗

bioRxiv2026

Novel Dissymmetric Ionizable Lipid-Assembled Lipid Nanoparticles for Delivery of Ferroptosis-Related siRNA in Diabetic Treatment

Small interfering RNA (siRNA) enables precise post-transcriptional gene silencing for refractory diseases, yet its clinical translation remains limited by the lack of safe and efficient delivery vectors. Inspired by the dissymmetric alkyl chain architecture of natural membrane phospholipids, we designed and synthesized 34 novel ionizable lipids with dissymmetric hydrophobic tails and formulated them into lipid nanoparticles (LNPs). Through systematic physicochemical and biological assessments, we established clear structure-activity relationships and identified two lead LNPs (O14-LNP, H18a-LNP) with superior endosomal escape capacity, enhanced in vivo gene silencing potency, and favorable biosafety relative to the clinical benchmark MC3-LNP. In both streptozotocin-induced and spontaneous db/db type 2 diabetes (T2D) mouse models, lead LNPs delivering ferroptosis-related siRNAs effectively ameliorated glucose and lipid metabolic disorders, restored islet function, and alleviated hepatic steatosis. This study not only lays a theoretical foundation for the rational design of novel ionizable lipids, but also validates the therapeutic potential of siRNA therapy targeting ferroptosis, providing a versatile delivery platform and targeted therapeutic strategy for the treatment of T2D.

pharmacology and toxicology↗

bioRxiv2024

Blockade of TREM2 ameliorates pulmonary inflammation and fibrosis by modulating sphingolipid metabolism

Pulmonary fibrosis is a chronic interstitial lung disease involving systemic inflammation and abnormal collagen deposition. Dysregulations in lipid metabolism, such as macrophage-dependent lipid catabolism, have been recognized as critical factors for the development of pulmonary fibrosis. However, little is known about the signaling pathways involved and the key regulators. Here we found that triggering receptor expressed on myeloid cells 2 (TREM2) plays a pivotal role in regulating the lipid handling capacities of pulmonary macrophages and triggering fibrosis. By integrating analysis of single-cell and bulk RNA sequencing data from patients and mice with pulmonary fibrosis, we revealed that pulmonary macrophages consist of heterogeneous populations with distinct pro-fibrotic properties, and found that both sphingolipid metabolism and the expression of chemotaxis-related genes are elevated in fibrotic lungs. TREM2, a sensor recognizing multiple lipid species, is specifically upregulated in a subset of monocyte- derived macrophages. Blockade of TREM2 by gene knock-out or soluble TREM2 administration can both attenuate bleomycin-induced pulmonary fibrosis. By utilizing scRNA Seq and lipidomics, we found that Trem2 deficiency downregulates the synthesis of various sphingomyelins, and inhibits the expression of chemokines such as Ccl2. Together, our findings not only reveal the alterations in lipidomic profiles and the atlas of pulmonary macrophages during pulmonary fibrosis, but also suggest that targeting TREM2, the crucial regulator affecting both pulmonary sphingolipid metabolism and the chemokines secretion, can benefit pulmonary fibrosis patients in the future.

pharmacology and toxicology↗