The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
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Effects of Chlorhexidine digluconate on Rotational Rate of n-(9-Anthroyloxy)stearic acid in Model Membranes of Total Lipids Extracted from Porphyromonas gingivalis Outer Membranes

  • Jang, Hye-Ock (Department of Dental Pharmacology and Biophysics, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Kim, Dong-Won (Department of Dental Pharmacology and Biophysics, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Kim, Byeong-Ill (Department of Dental Pharmacology and Biophysics, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Sim, Hong-Gu (Department of Dental Pharmacology and Biophysics, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Lee, Young-Ho (Department of Dental Pharmacology and Biophysics, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Lee, Jong-Hwa (Department of Dental Pharmacology and Biophysics, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Bae, Jung-Ha (Department of Skin Beauty Coordination, Yangsan College) ;
  • Bae, Moon-Kyoung (Department of Oral Physiology and Molecular Biology, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University) ;
  • Kwon, Tae-Hyuk (HANDOK PHARMACEUTICALS Co., LTD.) ;
  • Yun, Il (Department of Dental Pharmacology and Biophysics, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University)
  • Published : 2004.04.21
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Abstract

The purpose of this study was to provide a basis for studying the molecular mechanism of pharmacological action of chlorhexidine digluconate. Large unilamellar vesicles (OPGTL) were prepared with total lipids extracted from cultured Porphyromonas gingivalis outer membranes (OPG). The anthroyloxy probes were located at a graded series of depths inside a membrane, depending on its substitution position (n) in the aliphatic chain. Fluorescence polarization of n-(9-anthroyloxy)stearic acid was used to examine effects of chlorhexidine digluconate on differential rotational mobility, while changing the probes' substitution position (n) in the membrane phospholipids aliphatic chain. Magnitude of the rotational mobility of the intact six membrane components differed depending on the substitution position in the descending order of 16-(9-anthroyloxy)palmitic acid (16-AP), 12, 9, 6, 3 and 2-(9-anthroyloxy)stearic acid (12-AS, 9-AS, 6-AS, 3-AS and 2-AS). Chlorhexidine digluconate increased in a dose-dependent manner the rate of rotational mobility of hydrocarbon interior of the OPGTL prepared with total lipids extracted from cultured OPG, but decreased the mobility of membrane interface of the OPGTL. Disordering or ordering effects of chlorhexidine digluconate on membrane lipids may be responsible for some, but not all of its bacteriostatic and bactericidal actions.

Keywords

References

  1. Abrams FS, Chattopadhyay A, London E. Determination of the location of fluorescent probes attached to fatty acids using parallax analysis of fluorescence quenching: effect of carboxyl ionization state and environment on depth. Biochemistry 31: 5322-5327, 1992 https://doi.org/10.1021/bi00138a011
  2. Abrams FS, London E. Extension of parallax analysis of membrane penetration depth to the polar region of model membranes: Use of fluorescence quenching by a spin-label attached to the phospholipid polar head group. Biochemistry 32: 10826-10831, 1993 https://doi.org/10.1021/bi00091a038
  3. Audus KL, Tavakoli-Saberi MR, Zheng H, Boyce EN. Chlorhexidine effects on membrane lipid domains of human buccal epithelial cells. J Dent Res 71: 1298-1303, 1992 https://doi.org/10.1177/00220345920710060601
  4. Buck RA, Eleazer PD, Staat RH, Scheetz JP. Effectiveness of three endodontic irrigants at various tubular depths in human dentin. J Endod 27: 206-208, 2001 https://doi.org/10.1097/00004770-200103000-00017
  5. Fisher RG, Quintana RP, Boulware MA. Surface-chemical studies on chlorhexidine and related compounds. I. Effects at air-water, n-hexane-water, and hydroxyapatite-water interfaces. J Dent Res 54: 20-24, 1975 https://doi.org/10.1177/00220345750540012901
  6. Fisher RG, Quintana RP. Surface-chemical studies on chlorhexidine and related compounds. II. Interaction with monomolecular-film systems. J Dent Res 54: 25-31, 1975 https://doi.org/10.1177/00220345750540013001
  7. Folch J, Lees M, Sloane Stanley GH. A simple method for the isolation and purification of total lipides from animal tissues.J Biol Chem 226: 497-509, 1956
  8. Gabler WL, Roberts D, Harold W. The effect of chlorhexidine on blood cells. J Periodontal Res 22: 150-155, 1987 https://doi.org/10.1111/j.1600-0765.1987.tb01555.x
  9. Gerlach RW, White DJ. Removal of extrinsic using a tartar control whitening dentifrice: a randomized clinical trial. J Clin Dent 12: 42-46, 2001
  10. Hugo WB. Membrane-active antimicrobial compounds-a reappraisal of their mode of action in the light of the chemiosmotic theory. Int J Pharm 1: 127-131, 1978 https://doi.org/10.1016/0378-5173(78)90014-5
  11. Kenney EB, Saxe SR, Bosles RD. Effect of chlorhexidine on human polymorphonuclear leucocytes. Arch Oral Biol 17: 1633-1636, 1972 https://doi.org/10.1016/0003-9969(72)90051-9
  12. Knuuttila M, Söderling E. Effect of chlorhexidine on the release of lysosomal enzymes from cultured macrophages. Acta Odontol Scand 39: 285-289, 1981 https://doi.org/10.3109/00016358109162291
  13. Jang HO, Cha SK, Lee C, Choi MG, Huh SR, Shin SH, Chung IK, Yun I. Effects of chlorhexidine digluconate on rotational rate of n-(9-anthroyloxy)stearic acid in Porphyromonas gingivalis outer membranes. Korean J Physiol Pharmacol 7: 125-130, 2003
  14. Lamont RJ, Chan A, Belton CM, Izutsu KT, Vasel D, Weinbero A. Porphyromonas gingivalis invasion of gingival epithelial cells. Infect Immun 63: 3878-3885, 1995
  15. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275, 1951
  16. Madeira VMC, Antunes-Madeira MC. Lipid composition of biomembranes: a complete analysis of sarcoplasmic reticulum phospholipids. Cienc Biol (Coimbra) 2: 265-291, 1976
  17. Mason JT. Properties of phosphatidylcholine bilayers as revealed by mixed-acyl phospholipid fluorescent probes containing n- (9-anthroyloxy) fatty acids. Biochim Biophys Acta 1194: 99-108, 1994 https://doi.org/10.1016/0005-2736(94)90207-0
  18. Molitoris BA, Alfery AC, Arris RA, Simon FR. Renal apical membrane cholesterol and fluidity in regulation of phosphate transport. Am J Physiol 249: 12-19, 1985
  19. Molitoris BA, Hoilien C. Static and dynamic components of renal cortical brush border and basolateral membrane fluidity: Role of cholesterol. J Membr Biol 99: 165-172, 1987 https://doi.org/10.1007/BF01995697
  20. Schachter D. Fluidity and function of hepatocyte plasma membranes. Hepatology 4: 140-151, 1984 https://doi.org/10.1002/hep.1840040124
  21. Smalley JW, Birss AJ, Mckee AS, Marsh PD. Haemin-binding proteins of Porphyromonas gingivalis W50 grown in a chemostat under haemin-limitation. J Gen Microbiol 139: 2145-2150, 1993 https://doi.org/10.1099/00221287-139-9-2145
  22. Smalley JW, Birss AJ. Trypsin-like enzyme activity of the extracellular vesicles of Bacteroides gingivalis W50. J Gen Microbiol 133: 2883-2894, 1987
  23. Spratt DA, Pratten J, Wilson M, Gulablvala K. An in vitro evaluation of the antimicrobial efficacy of irrigants on biofilms of root canal isolates. Int Endod J 34: 300-307, 2001 https://doi.org/10.1046/j.1365-2591.2001.00392.x
  24. Stubbs CD, Rubin E. Molecular mechanism of ethanol and anesthetic actions: lipid- and protein-based theories. In: Alling C, Diamond I, Leslie SW, Sun GY, Wood WG ed, Alcohol, Cell Membranes, and Signal Transduction in Brain. Plenum Press, New York, p 1-11, 1993
  25. Thulborn KR, Tilley LM, Sawyer WH, Treloar FE. The use of n-(9-anthroyloxy) fatty acids to determine fluidity and polarity gradients in phospholipid bilayers. Biochim Biophys Acta 558: 166-178, 1979 https://doi.org/10.1016/0005-2736(79)90057-9
  26. Tilley L, Thulborn KR, Sawyer WH. As assessment of the fluidity gradient of the lipid bilayer as determined by a set of n-(9-anthroyloxy) fatty acids (n=2,6,9,12,16). J Biol Chem 254: 2592-2594, 1979
  27. Tsuchiya H. Effects of green tea catechins on membrane fluidity. Pharmacol 59: 34-44, 1999 https://doi.org/10.1159/000028303
  28. Tsutsui H, Kinouchi T, Wakano Y, Ohnishi Y. Purification and characterization of a protease from Bacteroides gingivalis 381. Infect Immun 55: 420-427, 1987
  29. Villalaín J, Prieto M. Location and interaction of N-(9-anthroyloxy)- stearic acid probes incorporated in phosphatidylcholine vesicles. Chem Phys Lipids 59: 9-16, 1991 https://doi.org/10.1016/0009-3084(91)90058-J
  30. Vincent M, De Furesta D, Gallay J, Alfsen A. Fluorescence anisotropy decays of n-(9-anthroyloxy) fatty acids in dipalmitoyl phosphatidylcholine vesicles: Localization of the effects of cholesterol addition. Biochim Biophys Res Commun 107: 914-921, 1982 https://doi.org/10.1016/0006-291X(82)90610-6
  31. Yun I, Kang J-S. The general lipid composition and aminophospholipid asymmetry of synaptosomal plasma membrane vesicles isolated from bovine cerebral cortex. Mol Cells 1: 15-20, 1990
  32. Yun I, Kang J-S. Transbilayer effects of n-alkanols on the fluidity of model membranes of total lipids extracted from synaptosomal plasma membrane vesicles. Korean J Pharmacol 28: 191-199, 1992