2023年 新着論文 46 蛋白質発現分野から論文が発表されました

β-defensin 2 synthesized by a cell-free protein synthesis system and encapsulated in liposomes inhibits adhesion of Porphyromonas gingivalis to oral epithelial cells

Odontology. 2023 Oct;111(4):830-838. doi: 10.1007/s10266-023-00789-x. Epub 2023 Feb 6.

Authors

Yuka Hiroshima  1 Jun-Ichi Kido  2 Rie Kido  2 Kaya Yoshida  3 Mika Bando  2 Kazuaki Kajimoto  4 Hiromichi Yumoto  2 Yasuo Shinohara  5

Affiliations

  • 1 Department of Oral Microbiology, Tokushima University, 3-18-15, Kuramoto, Tokushima, 770-8504, Japan. yuka.hiroshima@tokushima-u.ac.jp.
  • 2 Department of Periodontology and Endodontology, Tokushima University, Tokushima, Japan.
  • 3 Department of Oral Healthcare Education, Graduate School of Biomedical Sciences, Tokushima University, Tokushima, Japan.
  • 4 Health and Medical Research Institute, National Institute of Advanced Industrial Science and Technology, Tokyo, Japan.
  • 5 Institute of Advanced Medical Sciences, Tokushima University, Tokushima, Japan.

Abstract

β-defensin 2 (BD-2), an antimicrobial peptide (AMP), is expressed by oral epithelial cells and plays an important role in innate immunity of the oral cavity. Cell-free protein synthesis (CFPS) systems have been studied for the synthesis of various proteins, however, the synthesis of BD-2 by a CFPS system has not been extensively explored. Liposomes have been developed as tools for drug delivery. A delivery of liposome-encapsulated AMP to oral epithelium may be useful to prevent oral infectious diseases. In the present study, we investigated the antimicrobial activity of the BD-2 protein, artificially synthesized using a CFPS system and encapsulated in liposomes. BD-2 protein was artificially synthesized using template DNA and a reconstituted CFPS system and was identified by western blotting. Bilayer liposomes were prepared using 1,2-dioleoyl-sn-glycero-3-phospho-choline and 3-sn-phosphatidylcholine from egg yolk. The artificially synthesized BD-2 was encapsulated in liposomes, collected by ultrafiltration, and detected by western blotting. Human oral epithelial cells were cultured with the liposome-encapsulated BD-2 and the concentration of BD-2 in the cell lysate of the culture with the synthesized BD-2 was higher than that of the control cultures. The antimicrobial activity of the synthesized BD-2 was investigated by an adhesion assay of Porphyromonas gingivalis to oral epithelial cells. The artificially synthesized BD-2 and its liposome significantly inhibited adhesion of P. gingivalis to oral epithelial cells. These results suggest that artificially synthesized BD-2 and liposome-encapsulated BD-2 show antimicrobial activity and can potentially play a role in oral healthcare for periodontal diseases.

Keywords: Cell-free protein synthesis system; Drug delivery system; Liposome; Porphyromonas gingivalis; β-defensin 2.

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