Preparation of Poly (dl-Lactide-co-Glycolide) Nanoparticles Encapsulated with Periglaucine A and Betulinic Acid for In Vitro Anti-Acanthamoeba and Cytotoxicity Activities

Abstract

Poly (dl-lactide-co-glycolide) (PLGA) microspheres were synthesized as delivery system for the natural anti-parasitic compounds, Periglaucine A (PGA) and Betulinic acid (BA). Periglaucine A and Betulinic acid were encapsulated in PLGA nanoparticles by single emulsion method with an average particle size of approximately 100–500 nm. Periglaucine A and Betulinic acid encapsulation efficiency was observed to be 90% and 35% respectively. Anti-Acanthamoeba property of Periglaucine A and Betulinic acid remained intact after encapsulation. PGA-PLGA and BA-PLGA nanoparticles demonstrated inhibition in viability of Acanthamoeba triangularis trophozoites by 74.9%, 59.9%, 49.9% and 71.2%, 52.2%, 88% respectively at concentration of 100 µg/mL, 50 µg/mL and 25 µg/mL. Cytotoxicity of PGA-PLGA and BA-PLGA nanoparticles has been evaluated against lung epithelial cell line and showed dose dependent cytotoxicity value of IC50 2 µg/mL and 20 µg/mL respectively. Futher, increased viability was observed in lung epithelial cell line in higher doses of synthesized polymeric nanoparticles. Results indicate that poly (dl-lactide-co-glycolide) (PLGA) nanoparticles could be exploratory delivery systems for natural products to improve their therapeutic efficacy.

Antiamoebic activity of plant-based natural products and their conjugated silver nanoparticles against Acanthamoeba castellanii (ATCC 50492)

“…Recently, natural products coated nanoparticles have shown potent antiacanthamoebic effects. Tannic acid, Oleic acid, Cinnamic acid, Hesperidin, Naringin, Periglaucine A, and Betulinic acid coated nanoparticles have shown the step forward in the development of nanomedicine against challenging infections caused by Acanthamoeba (Padzik et al 2018;Anwar et al 2018bAnwar et al , 2019aMahboob et al 2018). Moreover, various plant extracts coated nanoparticles have also shown promise in the drug discovery against Acanthamoeba infections.…”

PLGA nanoparticles loaded with Gallic acid- a constituent of Leea indica against Acanthamoeba triangularis

“…After removing the DCM by evaporation, the resulting NPs were collected by centrifugation for 5 minutes (10,000 rpm) and washed thrice with distilled water. The resulting nano-GA-PLGA was lyophilized in a freeze dryer, Emitech K750X (Quorum Technologies, Lewes, UK) at −53 °C overnight and stored at −20 °C for further use 33 . The control was prepared following the same method without gallic acid.…”

“…At specific time intervals, the sample was collected, and fresh medium was added to maintain the volume. The collected media were centrifuged and the amount of gallic acid released was determined by a UV-visible spectrophotometer at 230 nm 33 .…”

“…This assay is based on the binding of terminal deoxynucleotidyl transferase (TdT) and the conjugated drug and peroxidase, converting the substrate, a mixture of hydrogen peroxide and 3'3-diaminobenzidine (DAB) into a brown precipitate. Hence, apoptotic cells form a brown precipitate, detected under a light microscope 33 .…”

Antimicrobial activities of green synthesized gums-stabilized nanoparticles loaded with flavonoids

“…Previously, our research showed that Amphotericin B, Nystatin and Fluconazole conjugated AgNPs showed enhanced antiamoebic activity against brain-eating N. fowleri and A. castellanii 21,22 . Recently, natural compounds such as cinnamic acid 23 , periglaucine A, betulinic acid 24 , and tannic acid 25 coated nanomaterials have been shown to be effective against A. castellanii . Based on these reports, we synthesized HDN and NRG loaded silver and gold nanoparticles stabilized by plant gums GA and GT respectively by utilizing green synthesis protocols.…”