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Conlaining Lidocaine Or Benzocaine Mark E Anesth Prog 35:9-13 1988 Antimicrobial Properties of Topical Anestletc Liquids Conlaining Lidocaine or Benzocaine Mark E. Morrow, DDS, MSD,* and Charles W. Berry, PhDt *Department of Pediatric Dentistry, tDepartment of Microbiology, Baylor College of Dentistry, Dallas, Texas Six species of microorganisms commonly found ganisms have been isolated from the oral cavity,3 it is within the oral cavity were exposed for either one logical that an antiseptic agent should be wiped on the minute or two hours to 5% lidocaine liquid mucosa before the administration of an injectable local topical anesthetic and benzocaine liquid topical anesthetic in the dental setting. However, few dentists anesthetic. Mixtures of microorganisms and currently follow this regimen.4 Instead, the use of a anesthetics were diluted and plated onto a brain topical anesthetic just before administration of a local heart infusion medium. Reduction in cell viability anesthetic to block the pain of the needle puncture is very was 73-100% after exposure to the anesthetic common.5 agents when compared with the saline/buffer The possible antimicrobial properties of local anesthe- controls. A significant reduction (p < .005) in cell tic agents were originally described by Jonnesco.6 He growth by Streptococcus mutans, S. sanguis, S. indicated that there was no need to sterilize local anesthe- mitis, S. salivarius, Actinomyces viscosus, and tics used for spinal anesthesia because of their antiseptic Candida albicans was associated with a one- characteristics. The earliest work documenting the in vitro minute and two-hour exposure to lidocaine, antimicrobial activity of anesthetic agents was reported by benzocaine, 5% lidocaine, and the benzocaine Schlegal and Swan,7 who described the inhibition of vehicle control. Five percent lidocaine reduced growth of Micrococcus and Pseudomonas with 0.2% be- growth of the test orgainisms more than noxinate. Likewise, the effect of anesthetic agents on benzocaine in one-minute exposures to S. microorganisms found in clinical specimens was recog- mutans, A. viscosus and S. salivarius and with a nized.8-1 Each of these studies concluded that local two-hour exposure to S. salivarius. anesthetics reduced microbial growth. Sculley and Dun- Five percent lidocaine was bacteriocidal or ley12 were the first to report the antibacterial efficacy of fungicidal to all microorganisms for both time several anesthetic preparations used in dentistry. They periods whereas, benzocaine liquid topical examined the antimicrobial activity of two percent lido- anesthetic was predominately bacteriostatic or caine with and without epinephrine against 10 different fungistatic after one-minute exposures and species of nonoral microorganisms. Lidocaine with and bacteriocidal or fungicidal after two hours. without epinephrine demonstrated antimicrobial prop- The results indicated that two dental liquid erfies. topical anesthetics containing lidocaine or Because the application of topical anesthetic agents is benzocaine possessed considerable antimicrobial the only usual pre-injection preparation of the oral mu- activity to selected oral microorganisms. The cosa, the purpose of this study is to report the in vitro exclusive use of a topical liquid anesthetic may be antimicrobial properties of two commercially prepared an adequate means to render the oral mucosa topical anesthetic agents against microorganisms com- aseptic before injection of a local anesthetic. monly found in the oral cavity. The effect of the length of exposure of the test organisms to the topical anesthetic is also described. B efore the needle penetration of a local anesthetic injection, the tissue at the site of injection should METHODS be as aseptic as possible.",2 Because numerous microor- Two of the commercially available liquid topical anesthe- Received July 13, 1987; accepted for publication October 2, 1987. tics that frequently are used in the dental setting were Address correspondence to Dr. Charles W. Berry, 3302 Gaston obtained from the manufacturers. Five percent lidocaine Avenue, Dallas, TX 75246. (Xylocaine liquid, Astra Pharmaceutical Products Inc., C) 1988 by the American Dental Society of Anesthesiology ISSN 0003-3006/88/$3.50 9 10 Antimicrobial Properties of Topical Anesthetics Anesth Prog 35:9-13 1988 Worchester, Massachusetts), an amide-type local anes- the C. albicans plates and tubes were incubated for 48 thetic, contains a mixture of 5% lidocaine, propylene hours aerobically, all at 37°C. Subsequently, the colonies glycol, glycerine, and flavoring agents. Benzocaine (Hur- on each plate were counted using a Quebec Colony ricane liquid, Beutlich Inc., Niles, Illinois), an ester-type Counter (American Optical Co., Buffalo, New York) and local anesthetic, contains 20% benzocaine with flavoring a hand-held counter. The broth tubes were observed for and coloring agents in a polyethylene glycol base. positive or negative growth. Six species of oral microorganisms (Streptococcus Bacteriostatic or bacteriocidal properties were deter- mutans, Streptococcus sanguis, Streptococcus mitis, mined by the ratio of microorganisms killed or altered by Streptococcus salivarius, Actinomyces viscosus, and Can- the test material when compared to a sterile control dida albicans) were chosen for study because of their medium. By definition, if a test material killed 99.9% of presence on the oral mucosal surfaces, or in the saliva or the microorganisms, the substance was described as gingival crevice exudate that coats the mucosal surfaces bacteriocidal; if less than 99.9%, the agent was bacterio- onto which topical anesthetics are applied before injec- static. 13 tion of local anesthetics. Subcultures of these microor- Two negative controls of sterile 0.85% saline and ganisms were acquired from the Baylor College of Den- sterile 0.1 M phosphate buffer was used in place of the tistry, Department of Microbiology, stock culture two anesthetic preparations. Also, Hurricaine vehicle collection and maintained on brain infusion agar (Difco without benzocaine and a 5%, by weight, lidocaine/ Inc., Chicago, Illinois). polyethylene glycol mixture served as vehicle controls. The cell populations for each assay were propagated Triplicate analyses were carried out with each exper- by inoculation of brain heart infusion broth. The strepto- iment. coccal organisms were incubated in an atmosphere en- The Peritz' F test14 was utilized to determine the level riched with carbon dioxide, A. viscosus was grown of statistical significance between the antimicrobial action anaerobically and C. albicans was propagated aerobically of the anesthetic agents and their control solutions. at 37°C for 24 hours. Cell pellets were obtained by refrigerated centrifugation and were rinsed with either sterile 0.85% saline (Streptococcus) or 0.1 M buffered RESULTS phosphate solution (Actinomyces and Candida). A Coul- ter Counter (Coulter Electronics, Hialeah, Florida) was Six species of oral microorganisms were exposed for one used to confirm cell inoculum concentrations, that were minute and two hours to both 5% lidocaine liquid topical found to be between 2.3 and 7.4 x 107 cells/mL. anesthetic or benzocaine liquid topical anesthetic. The A procedure described by Sculley and Dunley was mixture of the microorganisms and anesthetics were modified and used for this experiment.12 Specifically, a diluted and subsequently plated to brain heart infusion 0.01 mL portion of each of the six cultures, harvested in agar and broth medium to observe for organism via- logarithmic growth phase, was placed into 1.99 mL of bility. each undiluted anesthetic (as received from the manufac- The growth of all six species of microorganisms was turer) and was kept at room temperature for either one significantly reduced (p < .005) by exposure to either minute or two hours. After the appropriate reaction time, 5% lidocaine liquid topical anesthetic or benzocaine a 0.01 mL aliquot of the reactants was removed with a liquid topical anesthetic for one minute or two hours fixed-volume pipette and transferred into 9.99 mL of when compared with sterile saline/buffer controls (Table 0.85% sterile saline for streptococcal species and 0.1 M 1). There was no significant difference (p < .05) in the buffered phosphate solution for A. viscosus and C. killing ability of either topical anesthetic on S. sanguis, S. albicans. The tubes containing cells, anesthetic and dilu- mitis, or C. albicans for both periods. There was also little ent were tightly capped and thoroughly agitated on a difference (p < .05) between lidocaine or benzocaine for vortex mixer (Scientific American Co., McGaw Park, two hour exposures to A. viscosus, S. mutans, and S. Illinois) for 15 seconds to promote cell dispersion. To salivarius. However, the antibacterial effect of 5% lido- determine the bacteriostatic properties of each anesthetic, caine liquid topical anesthetic was significantly greater (p brain heart infusion pour plates were made with 0.1 mL < .005) than benzocaine liquid topical anesthetic in one of each diluted reaction solution. Another 0.1 mL aliquot minute exposures to A. viscosus, S. mutans, and S. of reaction mixture was placed into 8.0 mL of brain heart salivarius. The 5% lidocaine/propylene glycol and ben- infusion broth. The plates and broth tubes containing zocaine vehicle also significantly reduced (p < .005) the streptococcal organism were incubated for 24 hours in a viability of all six microorganisms for both exposure carbon dioxide enriched environment. A. viscosus plates
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