Influence of Hexetidine Upon Plaque and Gingivitis in the Beagle Dog JAMES L. MCDONALD, JR., BRUCE R. SCHEMEHORN, and GEORGE K. STOOKEY
Oral Health Research Institute, Indiana University School of Dentistry, 410 Beauty Avenue, Indianapolis, Indiana 46202, USA Two studies were performed in beagle dogs periodontal disease'0 the beagle dog is to determine the effect of 0.2% hexetidine widely used,5-7 and generally accepted, as upon plaque and gingizvtzs. In both in- one of the preferred models for such invesstances, hexetidine exerted no signzficant tigations. The similarities between sponeffect while 0.2% chlorhexidine resulted in taneous periodontal disease in dogs and reductions in both plaque and gingivitis of that observed in human beings has been reabout 45 and 50%, respectzvely. ported on several occasions."' 5 The objectives of this study in beagle dogs were twoJ Dent Res 57(2): 345-348, February 1978. fold: (1) to test two antimicrobial agents for their influence on plaque and gingivitis In view of the intimate role of oral micro- in the beagle dog; and (2) to determine the organisms in the diseases of both the denti- reproducibility of our animal periodontal tion and the periodontium, the use of oral disease model by conducting the test in antimicrobial agents for disease control has duplicate. been vigorously pursued for the past decade.' Oral rinses with chlorhexidine gluMethods and Materials conate have repeatedly been shown effecMale beagle dogs approximately 13 to 15 tive in both human beings2-4 and experimental animals5-7 in reducing dental months of age were utilized; 15 and 20 aniplaque and gingivitis to the extent that this mals were used in Studies 1 and 2, respecagent is now a yardstick to which other po- tively. One week prior to the initiation of the study, all dogs received a thorough tential agents can be compared. dental prophylaxis (scaling and polishing In addition to chlorhexidine, a wide variety of antimicrobial agents have been in- with a pumice slurry). Again, 24 hours vestigated for their potential ability to pre- prior to the study initiation, baseline gingivent plaque formation and the develop- vitis scores were obtained, and any plaque ment of periodontal disease. One such accumulations removed with a rubber proagent is hexetidine and the findings with phylactic cup and pumice. The dogs were this material have been equivocal. Dargent then stratified into comparable groups usand co-workers8 reported that the clinical ing the baseline gingivitis scores, and treatuse of a dentifrice containing hexetidine ments begun. Throughout the conduct of the studies, resulted in the virtual elimination of dental the dogs were daily provided approxiplaque and gingival inflammation. However, the results of a clinical study of Ber- mately 200 gm of a commercial ration genholtz and Hanstroim9 failed to confirm (Purina Dog Chow) and tap water (1.0 these effects and indicated that this agent ppm F). The dietary regimen was prewas decidedly inferior to chlorhexidine in moistened with tap water and provided at a regular time each afternoon, with the the prevention of dental plaque. While numerous animal models have food bowls being removed after approxibeen suggested for preclinical studies of mately one hour. The 3 treatment regimens were as folReceived for publication May 19, 1977. lows: Group A, deionized water; Group B, Accepted for publicationJuly 13, 1977. 345 Downloaded from jdr.sagepub.com at CARLETON UNIV on May 4, 2015 For personal use only. No other uses without permission.
McDONALD, SCHEMEHORN & STOOKEY
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1978
TABLE 1
EFFECTI OF CHLORHEXIDINE AND HEXETIDINE ON PLAQUE AND GINGIVITIS IN DOGS (STUDY 1) Plaque
Number of Dogs
Treatment
Control (Water) Hexetidine Chlorhexidine
5 5 5
Mean Final Plaque Score
% Difference
2.38±0.11*1t 2.37±0.09 1.32+0.22
0 45
Gingivitis Mean Gingivitis Difference Increment
0.46+0.11 0.55 + 0.07] 0.23 ± 0.07
+ -
20 50
*Standar error of the mean. WValues within brackets are not significantly different (P < 0.05).
surfaces were then stained with a 0.075% basic fuchsin solution and assigned scores according to the following criteria: 0, no plaque observable; 1, scattered plaque covering less than 10% of the total tooth surfaces; 2, plaque covering between 10 and 33% of the total tooth surfaces; 3, plaque covering between 33 and 66% of the total tooth surfaces; and, 4, plaque covering more than 66% of the total tooth surfaces. The individual tooth scores were then totaled and the sum divided by the total number of teeth scored to give a mean plaque score for each dog. The mean gingivitis and plaque scores were then analyzed for homogeneity of variance using the Bartlett chi square test (at a = 0.10). Assuming homogeneity of variance occurred, a one-way analysis of variance was then utilized. In cases lacking homogeneity of variance, the Welch test modified the procedure. Whenever a significant "F" value was found, the Newman-Keul's procedure was used to test for significant differences between individual group means.
0.2% chlorhexidine solution; and, Group C, 0.2% hexetidine solution. Treatments were provided twice daily, 5 days a week over a 4-week experimental period. The treatments consisted of spraying a stream of the appropriate experimental solution upon the buccal gingival segments and tooth surfaces in the areas to be treated (I3, Cl, Pi, P3, P4, and MI, bilaterally). Each quadrant received about a 5-second exposure to the treatment. At the conclusion of the study, the dogs were clinically evaluated for gingivitis and plaque. The 22 different gingival areas mentioned previously were evaluated clinically for gingival health status using the following criteria: 0, no apparent inflammation; 0. 5, mild inflammation not extending completely around the tooth; 1, mild or moderate inflammation extending completely around the tooth; 2, moderate inflammation with hemorrhaging on gentle probing; and, 3, severe inflammation with spontaneous hemorrhaging. The scores for each gingival area were totaled and the sum divided by the total number of surfaces scored to give an average gingivitis score for each animal. To determine the amount of plaque accumulation, the 22 different buccal tooth
Results
The results of the two studies are presented in Tables 1 and 2, respectively. In
TABLE 2 EFFECT OF CHLORHEXIDINE AND HEXETIDINE ON PLAQUE AND GINGIVITIS IN DOGS (STUDY 2) Gingivitis
Plaque Number of Dogs
Treatment
Control (Water) Hexetidine Chlorhexidine
7 7 6
Mean Final Plaque Score
2.58 + 0.13*11 2.60+0.11J 1.45 ± 0.16
N Difference
Mean Gingivitis Increment
0 44
0.55 + 0.12 0.67 + 0.09] 0.26±0.06
% Difference
*Standard error of the mean. tValues within brackets are not significantly different.
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+ 22 -
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PLAQUE & GINGIVITIS PREVENTION IN THE DOG
the first study (Table 1) the chlorhexidine treatments resulted in a 45% reduction in the amount of plaque formed during the 4week experimental period while the hexetidine treatments resulted in plaque scores which were essentially identical to the control regimen. Correspondingly, chlorhexidine resulted in a 50% decrease in gingivitis while hexetidine was without any sig-
nificant effect. The results of the second study (Table 2) were quite comparable to the first study. In this instance, the chlorhexidine resulted in reductions of plaque and gingivitis of 44 and 53%, respectively, while hexetidine was again without any significant effect. Discussion
Under these experimental conditions daily applications of a 0.2% hexetidine solution failed to exert any significant effect upon either plaque formation or the development of gingivitis in the dog. These results thus support the clinical findings of Bergenholtz and H-instrom9 and are in contrast to those of Dargent and coworkers.'0 Interestingly, the 0.2% hexetidine solution resulted in numerical increases in gingivitis of 20 and 22% in the two studies. Bergenholtz and Hainstrom9 reported that a 0.2% hexetidine solution resulted in appreciable oral irritation and they utilized concentrations of 0.1 and 0.14% in their clinical investigation both of which were significantly less effective than 0.2% chlorhexidine. It is possible that the numerical increase in gingivitis observed in the present studies thus reflects oral irritation caused by the agent directly rather than any effect mediated through the plaque. The results from these two studies are quite reproducible. The plaque and gingivitis scores observed after 4 weeks in the control animals were comparable in the two studies even though different animals were utilized. Further, the plaque and gingivitis data with both chlorhexidine and hexetidine were essentially identical for the two studies. Conclusions
The results of these two studies indicate
347
that under these experimental conditions, hexetidine exerted no significant effect upon either plaque accumulation or gingivitis in the beagle dog. Conversely, the anticipated beneficial influence of chlorhexidine upon these parameters was observed. Further, the findings in the two studies demonstrate the reproducibility of this model for the preclinical evaluation of agents suspected to influence plaque accumulation and gingivitis formation. The authors wish to acknowledge the technical assistance of Mr. F. M. Swearingen in the conduct of these studies.
References 1. PARSONS, J.C.: Chemotherapy of Dental Plaque - A Review, J Perio 45:177-186, 1974. 2. LOE, H., and RINDOM-SCHIOTT, C.: The
Effect of Mouthrinses and Topical Application of Chlorhexidine on the Development of Dental Plaque and Gingivitis in Man, J Periodont Res 5:79-83, 1970. 3. GJERMo, P., and ERIKSEN, H.M.: Unchanged Plaque Inhibiting Effect of Chlorhexidine in Human Subjects After 2 Years of Continuous Use, Arch Oral Biol 18:317319, 1974. 4. LOE, H.; RINDOM-SCHIOTT, C.; GLAVIND, L.; and KARRING, T.: Two Years Use of Chlorhexidine Against Human Dental Diseases. I. General Design and Clinical Effects,JPeriodontRes 11:135-144, 1976. 5. LINDHE, J.; HAMP, S.E.; LOE, H.; and RINDOM-SCHIbTT, C.: Influence of Topical Applications of Chlorhexidine on Chronic
Gingivitis and Gingival Wound Healing in the Dog, Scand J Dent Res 78:471-478, 1970. 6. HAMP, S.E.; LINDHE, J.; and LOE, H.: Long Term Effect of Chlorhexidine on Developing Gingivitis in the Beagle Dog, J Periodont Res 8:63-70, 1973. 7. BRINER, W.W., and WUNDER, J.A.: Sensitivity of Dog Plaque Microorganisms to
Chlorhexidine During Longitudinal Studies,JPeriodont Res 12:135-139, 1977. 8. DARGENT, P.; TREVOUS, M.; and GAILLARD, M.: Dental Plaque: Inhibiting Effect of Hexetidine, Chir Dent France 43:63-68, 1973. 9. BERGENHOLTZ, A., and HANSTROM, L.: The Plaque-Inhibiting Effect of Hexetidine
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348
McDONALD, SCHEMEHORN & STOOKE
(Oraldene) - Mouthwash Compared to that of Chlorhexidine, Community Dent Oral Epidemiol 2:70-74, 1974. 10. SHAFER, W.G.: Use of Laboratory Animals for Periodontal Studies, Ann NY Acad Sci 153:223-229, 1968. 11. LINDE, H.; HAMP, S.E.; and LOE, H.: Experimental Periodontitis in the Beagle Dog, JPeriodont Res 8:1-10, 1973. 12. SCHROEDER, H.E.; LINDHE, J.; HUGOSON, A.; and MUNZEL-PEDRAZOLE, S.: Structural Constituents of Clinically Normal and Slightly Inflamed Dog Gingiva. A. Mor-
J Dent Res February 1978 phometric Study, Helv Odont Acta 17:7083, 1973. 13. ERICSSON, I.; LINDHE, J.; RYLANDER, H.; and OKAMOTO, H.: Experimental Periodontal Breakdown in the Dog, ScandJ Dent Res 83:189-192, 1975. 14. LINDHE, J., and RYLANDER, H.: Experimental Gingivitis in Young Dogs, ScandJ Dent Res 83:314-326, 1975. 15. LINDHE, J.; HAMP, S.E.; and LOE, H.: Plaque Induced Periodontal Disease in Beagle Dogs, J Periodont Res 10:243-255, 1975.
Downloaded from jdr.sagepub.com at CARLETON UNIV on May 4, 2015 For personal use only. No other uses without permission.
Oral Health Research Institute, Indiana University School of Dentistry, 410 Beauty Avenue, Indianapolis, Indiana 46202, USA Two studies were performed in beagle dogs periodontal disease'0 the beagle dog is to determine the effect of 0.2% hexetidine widely used,5-7 and generally accepted, as upon plaque and gingizvtzs. In both in- one of the preferred models for such invesstances, hexetidine exerted no signzficant tigations. The similarities between sponeffect while 0.2% chlorhexidine resulted in taneous periodontal disease in dogs and reductions in both plaque and gingivitis of that observed in human beings has been reabout 45 and 50%, respectzvely. ported on several occasions."' 5 The objectives of this study in beagle dogs were twoJ Dent Res 57(2): 345-348, February 1978. fold: (1) to test two antimicrobial agents for their influence on plaque and gingivitis In view of the intimate role of oral micro- in the beagle dog; and (2) to determine the organisms in the diseases of both the denti- reproducibility of our animal periodontal tion and the periodontium, the use of oral disease model by conducting the test in antimicrobial agents for disease control has duplicate. been vigorously pursued for the past decade.' Oral rinses with chlorhexidine gluMethods and Materials conate have repeatedly been shown effecMale beagle dogs approximately 13 to 15 tive in both human beings2-4 and experimental animals5-7 in reducing dental months of age were utilized; 15 and 20 aniplaque and gingivitis to the extent that this mals were used in Studies 1 and 2, respecagent is now a yardstick to which other po- tively. One week prior to the initiation of the study, all dogs received a thorough tential agents can be compared. dental prophylaxis (scaling and polishing In addition to chlorhexidine, a wide variety of antimicrobial agents have been in- with a pumice slurry). Again, 24 hours vestigated for their potential ability to pre- prior to the study initiation, baseline gingivent plaque formation and the develop- vitis scores were obtained, and any plaque ment of periodontal disease. One such accumulations removed with a rubber proagent is hexetidine and the findings with phylactic cup and pumice. The dogs were this material have been equivocal. Dargent then stratified into comparable groups usand co-workers8 reported that the clinical ing the baseline gingivitis scores, and treatuse of a dentifrice containing hexetidine ments begun. Throughout the conduct of the studies, resulted in the virtual elimination of dental the dogs were daily provided approxiplaque and gingival inflammation. However, the results of a clinical study of Ber- mately 200 gm of a commercial ration genholtz and Hanstroim9 failed to confirm (Purina Dog Chow) and tap water (1.0 these effects and indicated that this agent ppm F). The dietary regimen was prewas decidedly inferior to chlorhexidine in moistened with tap water and provided at a regular time each afternoon, with the the prevention of dental plaque. While numerous animal models have food bowls being removed after approxibeen suggested for preclinical studies of mately one hour. The 3 treatment regimens were as folReceived for publication May 19, 1977. lows: Group A, deionized water; Group B, Accepted for publicationJuly 13, 1977. 345 Downloaded from jdr.sagepub.com at CARLETON UNIV on May 4, 2015 For personal use only. No other uses without permission.
McDONALD, SCHEMEHORN & STOOKEY
346
j Dent
Res
February
1978
TABLE 1
EFFECTI OF CHLORHEXIDINE AND HEXETIDINE ON PLAQUE AND GINGIVITIS IN DOGS (STUDY 1) Plaque
Number of Dogs
Treatment
Control (Water) Hexetidine Chlorhexidine
5 5 5
Mean Final Plaque Score
% Difference
2.38±0.11*1t 2.37±0.09 1.32+0.22
0 45
Gingivitis Mean Gingivitis Difference Increment
0.46+0.11 0.55 + 0.07] 0.23 ± 0.07
+ -
20 50
*Standar error of the mean. WValues within brackets are not significantly different (P < 0.05).
surfaces were then stained with a 0.075% basic fuchsin solution and assigned scores according to the following criteria: 0, no plaque observable; 1, scattered plaque covering less than 10% of the total tooth surfaces; 2, plaque covering between 10 and 33% of the total tooth surfaces; 3, plaque covering between 33 and 66% of the total tooth surfaces; and, 4, plaque covering more than 66% of the total tooth surfaces. The individual tooth scores were then totaled and the sum divided by the total number of teeth scored to give a mean plaque score for each dog. The mean gingivitis and plaque scores were then analyzed for homogeneity of variance using the Bartlett chi square test (at a = 0.10). Assuming homogeneity of variance occurred, a one-way analysis of variance was then utilized. In cases lacking homogeneity of variance, the Welch test modified the procedure. Whenever a significant "F" value was found, the Newman-Keul's procedure was used to test for significant differences between individual group means.
0.2% chlorhexidine solution; and, Group C, 0.2% hexetidine solution. Treatments were provided twice daily, 5 days a week over a 4-week experimental period. The treatments consisted of spraying a stream of the appropriate experimental solution upon the buccal gingival segments and tooth surfaces in the areas to be treated (I3, Cl, Pi, P3, P4, and MI, bilaterally). Each quadrant received about a 5-second exposure to the treatment. At the conclusion of the study, the dogs were clinically evaluated for gingivitis and plaque. The 22 different gingival areas mentioned previously were evaluated clinically for gingival health status using the following criteria: 0, no apparent inflammation; 0. 5, mild inflammation not extending completely around the tooth; 1, mild or moderate inflammation extending completely around the tooth; 2, moderate inflammation with hemorrhaging on gentle probing; and, 3, severe inflammation with spontaneous hemorrhaging. The scores for each gingival area were totaled and the sum divided by the total number of surfaces scored to give an average gingivitis score for each animal. To determine the amount of plaque accumulation, the 22 different buccal tooth
Results
The results of the two studies are presented in Tables 1 and 2, respectively. In
TABLE 2 EFFECT OF CHLORHEXIDINE AND HEXETIDINE ON PLAQUE AND GINGIVITIS IN DOGS (STUDY 2) Gingivitis
Plaque Number of Dogs
Treatment
Control (Water) Hexetidine Chlorhexidine
7 7 6
Mean Final Plaque Score
2.58 + 0.13*11 2.60+0.11J 1.45 ± 0.16
N Difference
Mean Gingivitis Increment
0 44
0.55 + 0.12 0.67 + 0.09] 0.26±0.06
% Difference
*Standard error of the mean. tValues within brackets are not significantly different.
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+ 22 -
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Vol. 5 7 No. 2
PLAQUE & GINGIVITIS PREVENTION IN THE DOG
the first study (Table 1) the chlorhexidine treatments resulted in a 45% reduction in the amount of plaque formed during the 4week experimental period while the hexetidine treatments resulted in plaque scores which were essentially identical to the control regimen. Correspondingly, chlorhexidine resulted in a 50% decrease in gingivitis while hexetidine was without any sig-
nificant effect. The results of the second study (Table 2) were quite comparable to the first study. In this instance, the chlorhexidine resulted in reductions of plaque and gingivitis of 44 and 53%, respectively, while hexetidine was again without any significant effect. Discussion
Under these experimental conditions daily applications of a 0.2% hexetidine solution failed to exert any significant effect upon either plaque formation or the development of gingivitis in the dog. These results thus support the clinical findings of Bergenholtz and H-instrom9 and are in contrast to those of Dargent and coworkers.'0 Interestingly, the 0.2% hexetidine solution resulted in numerical increases in gingivitis of 20 and 22% in the two studies. Bergenholtz and Hainstrom9 reported that a 0.2% hexetidine solution resulted in appreciable oral irritation and they utilized concentrations of 0.1 and 0.14% in their clinical investigation both of which were significantly less effective than 0.2% chlorhexidine. It is possible that the numerical increase in gingivitis observed in the present studies thus reflects oral irritation caused by the agent directly rather than any effect mediated through the plaque. The results from these two studies are quite reproducible. The plaque and gingivitis scores observed after 4 weeks in the control animals were comparable in the two studies even though different animals were utilized. Further, the plaque and gingivitis data with both chlorhexidine and hexetidine were essentially identical for the two studies. Conclusions
The results of these two studies indicate
347
that under these experimental conditions, hexetidine exerted no significant effect upon either plaque accumulation or gingivitis in the beagle dog. Conversely, the anticipated beneficial influence of chlorhexidine upon these parameters was observed. Further, the findings in the two studies demonstrate the reproducibility of this model for the preclinical evaluation of agents suspected to influence plaque accumulation and gingivitis formation. The authors wish to acknowledge the technical assistance of Mr. F. M. Swearingen in the conduct of these studies.
References 1. PARSONS, J.C.: Chemotherapy of Dental Plaque - A Review, J Perio 45:177-186, 1974. 2. LOE, H., and RINDOM-SCHIOTT, C.: The
Effect of Mouthrinses and Topical Application of Chlorhexidine on the Development of Dental Plaque and Gingivitis in Man, J Periodont Res 5:79-83, 1970. 3. GJERMo, P., and ERIKSEN, H.M.: Unchanged Plaque Inhibiting Effect of Chlorhexidine in Human Subjects After 2 Years of Continuous Use, Arch Oral Biol 18:317319, 1974. 4. LOE, H.; RINDOM-SCHIOTT, C.; GLAVIND, L.; and KARRING, T.: Two Years Use of Chlorhexidine Against Human Dental Diseases. I. General Design and Clinical Effects,JPeriodontRes 11:135-144, 1976. 5. LINDHE, J.; HAMP, S.E.; LOE, H.; and RINDOM-SCHIbTT, C.: Influence of Topical Applications of Chlorhexidine on Chronic
Gingivitis and Gingival Wound Healing in the Dog, Scand J Dent Res 78:471-478, 1970. 6. HAMP, S.E.; LINDHE, J.; and LOE, H.: Long Term Effect of Chlorhexidine on Developing Gingivitis in the Beagle Dog, J Periodont Res 8:63-70, 1973. 7. BRINER, W.W., and WUNDER, J.A.: Sensitivity of Dog Plaque Microorganisms to
Chlorhexidine During Longitudinal Studies,JPeriodont Res 12:135-139, 1977. 8. DARGENT, P.; TREVOUS, M.; and GAILLARD, M.: Dental Plaque: Inhibiting Effect of Hexetidine, Chir Dent France 43:63-68, 1973. 9. BERGENHOLTZ, A., and HANSTROM, L.: The Plaque-Inhibiting Effect of Hexetidine
Downloaded from jdr.sagepub.com at CARLETON UNIV on May 4, 2015 For personal use only. No other uses without permission.
348
McDONALD, SCHEMEHORN & STOOKE
(Oraldene) - Mouthwash Compared to that of Chlorhexidine, Community Dent Oral Epidemiol 2:70-74, 1974. 10. SHAFER, W.G.: Use of Laboratory Animals for Periodontal Studies, Ann NY Acad Sci 153:223-229, 1968. 11. LINDE, H.; HAMP, S.E.; and LOE, H.: Experimental Periodontitis in the Beagle Dog, JPeriodont Res 8:1-10, 1973. 12. SCHROEDER, H.E.; LINDHE, J.; HUGOSON, A.; and MUNZEL-PEDRAZOLE, S.: Structural Constituents of Clinically Normal and Slightly Inflamed Dog Gingiva. A. Mor-
J Dent Res February 1978 phometric Study, Helv Odont Acta 17:7083, 1973. 13. ERICSSON, I.; LINDHE, J.; RYLANDER, H.; and OKAMOTO, H.: Experimental Periodontal Breakdown in the Dog, ScandJ Dent Res 83:189-192, 1975. 14. LINDHE, J., and RYLANDER, H.: Experimental Gingivitis in Young Dogs, ScandJ Dent Res 83:314-326, 1975. 15. LINDHE, J.; HAMP, S.E.; and LOE, H.: Plaque Induced Periodontal Disease in Beagle Dogs, J Periodont Res 10:243-255, 1975.
Downloaded from jdr.sagepub.com at CARLETON UNIV on May 4, 2015 For personal use only. No other uses without permission.
