Examination of Effective Buccal Absorption of Salmon Calcitonin Using Cell-Penetrating Peptide-Conjugated Liposomal Drug Delivery System

• Published in: International journal of nanomedicine 2022-01, Vol.17, p.697-710
• Main Authors: Keum, Taekwang, Noh, Gyubin, Seo, Jo-Eun, Bashyal, Santosh, Sohn, Dong Hwan, Lee, Sangkil
• Format: Article
• Language: English
• Subjects: Amino acids; Animals; Bioavailability; Blood lipids; Blood vessels; buccal drug delivery; Calcitonin; Cell-Penetrating Peptides - pharmacology; Drug delivery systems; Drug Delivery Systems - methods; Drugs; Efficiency; Enzyme-linked immunosorbent assay; Ethylenediaminetetraacetic acid; Humans; Insulin; Lipids; liposomes; Liposomes - chemistry; Membrane lipids; Microscopy; Mouth Mucosa; Oral administration; Oral Mucosal Absorption; Original Research; Patient compliance; penetratin; peptide delivery; Peptides; porcine buccal tissues; Swine; Thin films; Thiols; tr146 cells; Vehicles
• ISSN: 1178-2013; 1176-9114; 1178-2013
• DOI: 10.2147/IJN.S335774

The buccal route has been considered an attractive alternative delivery route for injectable formulations. Cell-penetrating peptides (CPPs) are gaining increased attention for their cellular uptake and tissue permeation effects. This study was aimed to evaluate the in vitro and ex vivo permeation-enhancing effect of penetratin-conjugated liposomes for salmon calcitonin (sCT) in TR146 human buccal cells and porcine buccal tissues. Penetratin was conjugated to phospholipids through a maleimide-thiol reaction. Liposomes were prepared and sCT was encapsulated using a thin-film hydration method. Physical properties such as particle size, zeta potential, encapsulation efficiency, and morphological images via transmission electron microscopy were obtained. Cellular uptake studies were conducted using flow cytometry (FACS) and confocal laser scanning microscopy (CLSM). A cell permeation study was performed using a Transwell assay, and permeation through porcine buccal tissue was evaluated. The amount of sCT permeated was quantified using an ELISA kit and was optically observed using CLSM. The particle size of penetratin-conjugated liposomes was approximately 123.0 nm, their zeta potential was +29.6 mV, and their calcitonin encapsulation efficiency was 18.0%. In the cellular uptake study using FACS and CLSM, stronger fluorescence was observed in penetratin-conjugated liposomes compared with the solution containing free sCT and control liposomes. Likewise, the amount of sCT permeated from penetratin-conjugated liposomes was higher than that from the free sCT solution and control liposomes by 5.8-fold across TR146 cells and 91.5-fold across porcine buccal tissues. Penetratin-conjugated liposomes are considered a good drug delivery strategy for sCT via the buccal route.