Normal flora: living vehicles for non-invasive protein drug delivery

• Published in: International journal of pharmaceutics 2004-11, Vol.286 (1), p.117-124
• Main Authors: Shao, Jun, Kaushal, Gagan
• Format: Article
• Language: English
• Subjects: Administration, Oral; Area Under Curve; Bacterial Proteins - administration & dosage; Bacterial Proteins - chemistry; Bacterial Proteins - pharmacokinetics; beta-Lactamases - chemistry; beta-Lactamases - pharmacology; beta-Lactamases - secretion; Biological and medical sciences; Caco-2; Caco-2 Cells; Cell integrity; Cell Survival - drug effects; Cell Survival - physiology; Coculture Techniques - methods; Culture Media - chemistry; Culture Media - classification; Culture Media - pharmacology; Drug Delivery Systems - methods; Electric Impedance; Epithelial Cells - drug effects; Epithelial Cells - physiology; Gastrointestinal Tract - microbiology; Gastrointestinal Tract - physiology; General pharmacology; Humans; Lactococcus lactis; Lactococcus lactis - genetics; Lactococcus lactis - metabolism; Mannitol - metabolism; Mannitol - pharmacology; Medical sciences; Medicine, Traditional; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Plasmids - genetics; Probiotic bacteria; Propranolol - metabolism; Propranolol - pharmacology; Protein delivery; TEM β-lactamase; Tritium; Trypan Blue
• ISSN: 0378-5173; 1873-3476
• DOI: 10.1016/j.ijpharm.2004.08.004

Feasibility to use probiotic bacteria as a living protein delivery system through oral route was assessed in vitro. Lactococcus lactis transformed with a plasmid to express and secret β-lactamase was used to deliver β-lactamase through Caco-2 monolayer, an intestine epithelium. Transport of β-lactamase through Caco-2 monolayer was carried out in the transwells. The viability and integrity of the cell monolayers co-cultured with L. lactis was examined by trypan blue exclusion method and by measuring the transport of mannitol and propranolol as well as the transepithelial electrical resistance (TEER). Results show that it is feasible to use cell culture technique to evaluate the drug delivery by normal flora. The transport rate of β-lactamase when delivered by L. lactis was 2.0 ± 0.1 × 10 −2 h −1 ( n = 9) and through free solution form was 1.0 ± 0.1 × 10 −2 h −1. When co-cultured with L. lactis, Caco-2 cell viability decreased to 98, 96, and 94% at 6, 8, and 10 h, respectively. Transport of mannitol through Caco-2 cell monolayer was significantly increased and the transport of propranolol through Caco-2 cell monolayer was significantly decreased in the presence of L. lactis. Increase in the amount of protein delivered is probably due to the concentrate of the protein by L. lactis on the monolayer (absorption surface) and the opening of the tight junction of Caco-2 monolayer by L. lactis.