Reduction of cytotoxicity of benzalkonium chloride and octenidine by Brilliant Blue G

The irritative effects of preservatives found in ophthalmologic solution, or of antiseptics used for skin disinfection is a consistent problem for the patients. The reduction of the toxic effects of these compounds is desired. Brilliant Blue G (BBG) has shown to meet the expected effect in presence of benzalkonium chloride (BAK), a well known preservative in ophthalmic solutions, and octenidine dihydrochloride (Oct), used as antiseptic in skin and wound disinfection. BBG shows a significant protective effect on human corneal epithelial (HCE) cells against BAK and Oct toxicity, increasing the cell survival up to 51 % at the highest BAK or Oct concentration tested, which is 0.01 %, both at 30 min incubation. Although BBG is described as a P2x7 receptor antagonist, other selective P2x7 receptor antagonists, OxATP (adenosine 5’-triphosphate-2’,3’-dialdehyde) and DPPH (N’-(3,5-dichloropyridin-4-yl)-3-phenylpropanehydrazide), did not reduce the cytotoxicity of neither BAK nor Oct. Therefore we assume that the protective effect of BBG is not due to its action on the P2x7 receptor. Brilliant Blue R (BBR), a dye similar to BBG, was also tested for protective effect on BAK and Oct toxicity. In presence of BAK no significant protective effect was observed. Instead, with Oct a comparable protective effect was seen with that of BBG. To assure that the bacteriostatic effect is not affected by the combinations of BAK/BBG, Oct/BBG and Oct/BBR, bacterial growth inhibition was analyzed on different Gram-negative and Gram-positive bacteria. All combinations of BAK or Oct with BBG hinder growth of Gram-positive bacteria. The combinations of 0.001 % Oct and BBR above 0.025 % do not hinder the growth of B. subtilis. For Gram-negative bacteria, BBG and BBR reduce, but do not abolish, the antimicrobial effect of BAK nor of Oct. In conclusion, the addition of BBG at bacterial inhibitory concentrations is suggested in the ready-to-use ophthalmic preparations and antiseptic solutions.

[1]  A. Pitkänen,et al.  Effect of lacosamide on structural damage and functional recovery after traumatic brain injury in rats , 2014, Epilepsy Research.

[2]  D. Gabel,et al.  Brilliant Blue G as protective agent against trypan blue toxicity in human retinal pigment epithelial cells in vitro , 2013, Graefe's Archive for Clinical and Experimental Ophthalmology.

[3]  A. Salminen,et al.  The preservative polyquaternium-1 increases cytoxicity and NF-kappaB linked inflammation in human corneal epithelial cells , 2012, Molecular vision.

[4]  K. Kang,et al.  Structure-activity relationships and optimization of 3,5-dichloropyridine derivatives as novel P2X(7) receptor antagonists. , 2012, Journal of medicinal chemistry.

[5]  C. Baudouin,et al.  Reduced in vivo Ocular Surface Toxicity with Polyquad-Preserved Travoprost versus Benzalkonium-Preserved Travoprost or Latanoprost Ophthalmic Solutions , 2012, Ophthalmic Research.

[6]  S. Sidhu,et al.  A dye binding method for measurement of total protein in microalgae. , 2012, Analytical biochemistry.

[7]  H. Bramlett,et al.  A reassessment of P2X7 receptor inhibition as a neuroprotective strategy in rat models of contusion injury , 2012, Experimental Neurology.

[8]  B. Bean,et al.  Inhibition of Neuronal Voltage-Gated Sodium Channels by Brilliant Blue G , 2011, Molecular Pharmacology.

[9]  M. Kahook,et al.  Effects of benzalkonium chloride- or polyquad-preserved fixed combination glaucoma medications on human trabecular meshwork cells , 2011, Molecular vision.

[10]  A. Kramer,et al.  Standardized comparison of antiseptic efficacy of triclosan, PVP-iodine, octenidine dihydrochloride, polyhexanide and chlorhexidine digluconate. , 2010, The Journal of antimicrobial chemotherapy.

[11]  A. Kramer,et al.  Octenidine Dihydrochloride, a Modern Antiseptic for Skin, Mucous Membranes and Wounds , 2010, Skin Pharmacology and Physiology.

[12]  T. Silhavy,et al.  The bacterial cell envelope. , 2010, Cold Spring Harbor perspectives in biology.

[13]  T. Takano,et al.  Systemic administration of an antagonist of the ATP-sensitive receptor P2X7 improves recovery after spinal cord injury , 2009, Proceedings of the National Academy of Sciences.

[14]  K. Kümmerer,et al.  The toxicity of the quaternary ammonium compound benzalkonium chloride alone and in mixtures with other anionic compounds to bacteria in test systems with Vibrio fischeri and Pseudomonas putida. , 2008, Ecotoxicology and environmental safety.

[15]  A. Kramer,et al.  Biocompatibility index of antiseptic agents by parallel assessment of antimicrobial activity and cellular cytotoxicity. , 2008, The Journal of antimicrobial chemotherapy.

[16]  Y. Goto,et al.  BRILLIANT BLUE G SELECTIVELY STAINS THE INTERNAL LIMITING MEMBRANE/BRILLIANT BLUE G–ASSISTED MEMBRANE PEELING , 2006, Retina.

[17]  M. Dutot,et al.  Fluoroquinolone eye drop-induced cytotoxicity: role of preservative in P2X7 cell death receptor activation and apoptosis. , 2006, Investigative ophthalmology & visual science.

[18]  S. Goldman,et al.  P2X7 receptor inhibition improves recovery after spinal cord injury , 2004, Nature Medicine.

[19]  A. Michel,et al.  Complexities of measuring antagonist potency at P2X(7) receptor orthologs. , 2001, The Journal of pharmacology and experimental therapeutics.

[20]  S. Cha,et al.  ATP‐induced [Ca2+]i changes and depolarization in GH3 cells , 2000, British journal of pharmacology.

[21]  R. North,et al.  Brilliant blue G selectively blocks ATP-gated rat P2X(7) receptors. , 2000, Molecular pharmacology.

[22]  A. D. Russell,et al.  Antiseptics and Disinfectants: Activity, Action, and Resistance , 2001, Clinical Microbiology Reviews.

[23]  S. Rubio,et al.  Study of the formation of dye-induced premicellar aggregates and its application to the determination of quaternary ammonium surfactants. , 1997, Talanta.

[24]  Henry D. Isenberg,et al.  Manual of Clinical Microbiology , 1991 .

[25]  D. M. Bailey,et al.  Microbicidal activity of octenidine hydrochloride, a new alkanediylbis[pyridine] germicidal agent , 1985, Antimicrobial Agents and Chemotherapy.

[26]  M Tal,et al.  Why does Coomassie Brilliant Blue R interact differently with different proteins? A partial answer. , 1985, The Journal of biological chemistry.

[27]  Ellen Jo Baron,et al.  Manual of clinical microbiology , 1975 .

[28]  Xiaodong Ma,et al.  Two Spectrophotometric Methods for the Assay of Benzalkonium Chloride in Bandage Samples , 2014 .

[29]  S. Chamran,et al.  The Disinfectant Effects of Benzalkonium Chloride on Some Important Foodborne Pathogens , 2012 .

[30]  S. Paik,et al.  Self-oligomerization and protein aggregation of alpha-synuclein in the presence of Coomassie Brilliant Blue. , 2001, European journal of biochemistry.