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The initiation of the effect of commercially-prepared dried apple on dental caries in albino rats
0003-9969/83/040369-02$O3.00/0 Copyrighr 0 1983 Pergamon Press Ltd
Archs orul Biol. Vol. 28, No. 4, pp. 369-370, 1983 Printed in Great Britain. All rights reserved
SHORT
COMMUNICATION
THE INITIATION OF THE EFFECT OF COMMERCIALLY-PREPARED DRIED APPLE DENTAL CARIES IN ALBINO RATS
ON
L. 2. G. TOUYZ Department
of General
Physiology, School of Dentistry, University Johannesburg, South Africa
of the Witwatersrand,
Summary-Group I of Sprague-Dawley rats was fed diet MIT 200 with 25 parts/lo6 F’ in the drinking water. Group II received MIT 200, as did Group IV in which the sucrose was replaced by sugars in the same proportions as in dried apple. Group III received MIT 200 with minced dried apple replacing the sucrose. The diet containing commercially-prepared dried apple induced more dental caries than did similar diets containing equivalent amounts of sugars (p < 0.01).
The destructive or zariogenic effects on teeth of apple cider (Brunel et a/., 1966) and apple juice (Wynn and Haldi, 1948; Touyz, Glassman and Naidu, 1981) have been reported, but dried-apple preparations have not been studied. SpragueeDawley rats were bred and infected with cariogenic bacteria according to guidelines outlined by Navia (1977). The pups were weaned at 18 days and lived together for 3 days after which they were re-inoculated orally with fresh faecai cariogenic inoculum. They were fed arl lihitlrnl diet MIT 200 (Navia, Lopez and Harris, 1969) and ordinary tap-water, each supplied from a common source. Four groups (8 rats each) were fed as follows: Group I, MIT 200 with 25 parts/lo6 F in the drinking water; Group II, MIT 200; Group III, MIT 200 with its 67 per cent sucrose replaced by adding freshly-minced dried apple; and Group IV, MIT Z!OOwith its sucrose replaced with sugars proportional to those found in dried apple viz. sucrose 18 per cent, fructose 38 per cent and glucose I I per cent. Groups II-IV received distilled water udlihiturn. All the rats were housed individually and fed simultaneously, nocturnally, with 18 equal feeds in a programmed feeding cage (designed after Kanig, Schmid and Schmid, 1968:~.Rats in Groups I, 11 and IV were fed 4g of the dry diet each night; the rats in Group III were given I g extra food per night to compensate for the water cortent of the commercially-prepared dried apple. The rats were kept on the experimental diets for I5 nights. and then killed in a carbon dioxide gas chamber at 36 days of age. Carious lesions were stained with AgNO, (Navia, 1977). The jaws were dissected out, cleaned using bromelain proteolytic enzymes and then the teeth were scored for caries, using a x 30 magnification, initially whole, and again after embedding in plastic, from mesiodistal sections, as described by Keyes (1958b). A second operator unaware of the design of the experiment checked scores. The rats were weighed before and after the experiment. 369
Commercially-prepared dried apple is preserved by adding sulphur (150&2000 parts/106) and drying to below 29 per cent water content (mass/mass) (Mrak and McKinney, 1951). The mean carbohydrate content, excluding cellulose, of dried apple (water content 24 _t 2 per cent) was regarded as 67 + 3 per cent, based on information from Widdowson and McCance (1935). Strachan et al. (1951). Watt and Merrill (1963) and Whiting (1978). The health and diet of the rats appeared to be adequate, all rats gaining weight during the experiment (Table 1). Post-mortem examination of visceral contents by two examiners revealed nothing abnormal. Comparison of weight gains to normal growth cannot be made because the rats were kept hungry, though not starving, ensuring they ate everything presented (Kiinig er al., 1968). The trend to gain weight confirmed the healthy state of the rats, the increase in Group III being attributed to extra apple nutriment. Maxillae and mandibles, one each from every rat in all groups, were analysed. Two molars in each jaw were examined, a total of 16 jaws and 32 teeth analysed in each group. Buccal sulcal and proximal lesions were scored as E (involving enamel only) and D (penetrating to dentine) (Table 2). Data for carious lesions and enamel caries scores were compared by Student’s t-tests.
Table 1. The masses (means i SD) of the pups (n = 8) in grams at the commencement (before) and at termination (after) of the experiment Before Means f SD Group I Group II Group III Group IV
37.1 36.8 37.1 37.1
f k * f
1.4 1.6 0.5 1.2
After Means i SD 39.2 38.9 40.1 39.3
_t f * *
1.5 1.4 1.2 0.5
370
L. Z. G. Touyz
Table 2. The caries scores of enamel (E) and dentine involvement proximal surfaces Caries scores Buccal Group I II III IV
Sulcal
E
D
E
D
0.44 0.88 0.38
0.19 0.69 0.25
0.25 3.81 6.13 3.75
0.19 1.56 5.00 1.81
Total No. of carious lesions
Proximal E D 0.12 0.63 :.38 0.75
0.63 0.38
1 0.44
(D) for buccal, sulcal and
0.38 3.46 4.00 3.50
+ & + +
0.63 1.55 0.89 1.62
Total No. of E scores 0.43 4.87 8.37 4.87
* & & f
0.72 1.28 1.76 1.62
Means and standard deviations are shown for the number of carious lesions and the total E scores. Scores according to the method of Keyes (1958b). Student’s t-tests showed the following: all groups versus Group I p < 0.001 for carious lesions and enamel caries scores; Groups II and IV versus Group III p < 0.001 for enamel caries scores.
The physical form of the diets was identical except for the dried apple in Group III. The extra driedapple fibre did not appear to reduce tooth decay. The diets produced caries in Groups II-IV, more caries being observed in Group III than in the other groups. The lower caries rate in Group I was attributed to the water fluoride. The caries experience in Groups II and IV was as expected from previous observations (Navia, 1977). Group III shows that in vulnerable mouths, with a cariogenic organism, dried apple produces tooth decay. It is not cariostatic like fluoride under these conditions (p < O.OOl), but seems to promote caries. Comparison of Groups II-IV, demonstrates that dried apple produces more caries than the equivalent amount of sucrose or of sugars similar to those in dried apple. This may be due to the malic acid (Eheart and Mason. 1967) and other fresh apple constituents which may enhance the cariogenicity of fermentable apple sugars (Touyz, 1980). It should, however, be remembered that experiments like these, in which the diets of animals are varied, allow only a qualified assessment and ranking of foods with regard to their cariogenic potential in man.
REFERENCES
Briinel A. L., Causeret J., Houssiaux C. and Hugot D. (1966) Etude de I’effet cariogene du cidre chez le rat. Bull. Grpmi. int. Rech. scient. Stomat. 9, 157-169.
Eheart J. E. and Mason B. S. (1967) Sugar and acid in the edible portion of fruits. J. Am. diet. Ass. 50, 13@132. Keyes P. H. (1958b) Dental caries in the molar teeth of rats
II. A method
for diagnosing and scoring types of lesions simultaneously. J. dent. Res. 37, 1088-1099. KGnig K. G., Schmid P. and Schmid R. (1968) An apparatus for frequency controlled feeding of small rodents and its use in dental caries experiments. Archs oral Biol. 13, 13-26. Mrak E. M. and MacKinney G. (1951) The dehydration of fruits. In: The Chemistry and Technology of Foods (Edited by Jacobs M. B.), Vol. III, pp. 1773-1821. Interscience, New York. Navia J. M., Lopez H. and Harris R. S. (1969) Purified diet for dental caries research with rats. J. Nutr. 97, 133-140. Navia J. M. (1977) Experimental dental caries. In: Animal Models in Dental Research, Chap. 13, pp. 257-297. Alabama Press, Alabama. Strachan C. C., Moyls A. W., Atkinson F. E. and Britton J. E. (1951) The sugar content of apples. Dept. Agr.. Ottawa, Canada, Publ., No. 862. Touyz L. Z. G. (1980) Apples, acids and teeth. S. Afr. J. Sci. 76, 200-201. Touyz L. Z. G., Glassman R. M. and Naidu S. (1981) Erosive action of apple and orange juice on rat’s molars in vivo. S. Afr. J. Sci. 77, 423424. Watt B. K. and Merrill A. L. (1963) In: Composition of Foods, p. 6. Agr. Hand Book No. 8. Consumer and Food Economics Res. Div. Agricultural Research Science. U.S. Department of Agriculture. U.S. Government Printing Office, Washington, U.S.A. Whiting G. C. (1978) Sugars. In: Biochemistry of Fruits and their Products, 3rd edn (Edited by Hulme A. C.), Vol. I, Chap. I. pp. I-31. Academic Press, London. Widdowson E. M. and McCance R. A. (1935) The sugar contents of the edible portion of fruits. Biochem. J. 29, 151Ll56. Wynn W. and Haldi J. (1948) The erosive action of various fruit juices on the lower molar teeth of the albino rat. J. Nutr. 35. 488491.
Archs orul Biol. Vol. 28, No. 4, pp. 369-370, 1983 Printed in Great Britain. All rights reserved
SHORT
COMMUNICATION
THE INITIATION OF THE EFFECT OF COMMERCIALLY-PREPARED DRIED APPLE DENTAL CARIES IN ALBINO RATS
ON
L. 2. G. TOUYZ Department
of General
Physiology, School of Dentistry, University Johannesburg, South Africa
of the Witwatersrand,
Summary-Group I of Sprague-Dawley rats was fed diet MIT 200 with 25 parts/lo6 F’ in the drinking water. Group II received MIT 200, as did Group IV in which the sucrose was replaced by sugars in the same proportions as in dried apple. Group III received MIT 200 with minced dried apple replacing the sucrose. The diet containing commercially-prepared dried apple induced more dental caries than did similar diets containing equivalent amounts of sugars (p < 0.01).
The destructive or zariogenic effects on teeth of apple cider (Brunel et a/., 1966) and apple juice (Wynn and Haldi, 1948; Touyz, Glassman and Naidu, 1981) have been reported, but dried-apple preparations have not been studied. SpragueeDawley rats were bred and infected with cariogenic bacteria according to guidelines outlined by Navia (1977). The pups were weaned at 18 days and lived together for 3 days after which they were re-inoculated orally with fresh faecai cariogenic inoculum. They were fed arl lihitlrnl diet MIT 200 (Navia, Lopez and Harris, 1969) and ordinary tap-water, each supplied from a common source. Four groups (8 rats each) were fed as follows: Group I, MIT 200 with 25 parts/lo6 F in the drinking water; Group II, MIT 200; Group III, MIT 200 with its 67 per cent sucrose replaced by adding freshly-minced dried apple; and Group IV, MIT Z!OOwith its sucrose replaced with sugars proportional to those found in dried apple viz. sucrose 18 per cent, fructose 38 per cent and glucose I I per cent. Groups II-IV received distilled water udlihiturn. All the rats were housed individually and fed simultaneously, nocturnally, with 18 equal feeds in a programmed feeding cage (designed after Kanig, Schmid and Schmid, 1968:~.Rats in Groups I, 11 and IV were fed 4g of the dry diet each night; the rats in Group III were given I g extra food per night to compensate for the water cortent of the commercially-prepared dried apple. The rats were kept on the experimental diets for I5 nights. and then killed in a carbon dioxide gas chamber at 36 days of age. Carious lesions were stained with AgNO, (Navia, 1977). The jaws were dissected out, cleaned using bromelain proteolytic enzymes and then the teeth were scored for caries, using a x 30 magnification, initially whole, and again after embedding in plastic, from mesiodistal sections, as described by Keyes (1958b). A second operator unaware of the design of the experiment checked scores. The rats were weighed before and after the experiment. 369
Commercially-prepared dried apple is preserved by adding sulphur (150&2000 parts/106) and drying to below 29 per cent water content (mass/mass) (Mrak and McKinney, 1951). The mean carbohydrate content, excluding cellulose, of dried apple (water content 24 _t 2 per cent) was regarded as 67 + 3 per cent, based on information from Widdowson and McCance (1935). Strachan et al. (1951). Watt and Merrill (1963) and Whiting (1978). The health and diet of the rats appeared to be adequate, all rats gaining weight during the experiment (Table 1). Post-mortem examination of visceral contents by two examiners revealed nothing abnormal. Comparison of weight gains to normal growth cannot be made because the rats were kept hungry, though not starving, ensuring they ate everything presented (Kiinig er al., 1968). The trend to gain weight confirmed the healthy state of the rats, the increase in Group III being attributed to extra apple nutriment. Maxillae and mandibles, one each from every rat in all groups, were analysed. Two molars in each jaw were examined, a total of 16 jaws and 32 teeth analysed in each group. Buccal sulcal and proximal lesions were scored as E (involving enamel only) and D (penetrating to dentine) (Table 2). Data for carious lesions and enamel caries scores were compared by Student’s t-tests.
Table 1. The masses (means i SD) of the pups (n = 8) in grams at the commencement (before) and at termination (after) of the experiment Before Means f SD Group I Group II Group III Group IV
37.1 36.8 37.1 37.1
f k * f
1.4 1.6 0.5 1.2
After Means i SD 39.2 38.9 40.1 39.3
_t f * *
1.5 1.4 1.2 0.5
370
L. Z. G. Touyz
Table 2. The caries scores of enamel (E) and dentine involvement proximal surfaces Caries scores Buccal Group I II III IV
Sulcal
E
D
E
D
0.44 0.88 0.38
0.19 0.69 0.25
0.25 3.81 6.13 3.75
0.19 1.56 5.00 1.81
Total No. of carious lesions
Proximal E D 0.12 0.63 :.38 0.75
0.63 0.38
1 0.44
(D) for buccal, sulcal and
0.38 3.46 4.00 3.50
+ & + +
0.63 1.55 0.89 1.62
Total No. of E scores 0.43 4.87 8.37 4.87
* & & f
0.72 1.28 1.76 1.62
Means and standard deviations are shown for the number of carious lesions and the total E scores. Scores according to the method of Keyes (1958b). Student’s t-tests showed the following: all groups versus Group I p < 0.001 for carious lesions and enamel caries scores; Groups II and IV versus Group III p < 0.001 for enamel caries scores.
The physical form of the diets was identical except for the dried apple in Group III. The extra driedapple fibre did not appear to reduce tooth decay. The diets produced caries in Groups II-IV, more caries being observed in Group III than in the other groups. The lower caries rate in Group I was attributed to the water fluoride. The caries experience in Groups II and IV was as expected from previous observations (Navia, 1977). Group III shows that in vulnerable mouths, with a cariogenic organism, dried apple produces tooth decay. It is not cariostatic like fluoride under these conditions (p < O.OOl), but seems to promote caries. Comparison of Groups II-IV, demonstrates that dried apple produces more caries than the equivalent amount of sucrose or of sugars similar to those in dried apple. This may be due to the malic acid (Eheart and Mason. 1967) and other fresh apple constituents which may enhance the cariogenicity of fermentable apple sugars (Touyz, 1980). It should, however, be remembered that experiments like these, in which the diets of animals are varied, allow only a qualified assessment and ranking of foods with regard to their cariogenic potential in man.
REFERENCES
Briinel A. L., Causeret J., Houssiaux C. and Hugot D. (1966) Etude de I’effet cariogene du cidre chez le rat. Bull. Grpmi. int. Rech. scient. Stomat. 9, 157-169.
Eheart J. E. and Mason B. S. (1967) Sugar and acid in the edible portion of fruits. J. Am. diet. Ass. 50, 13@132. Keyes P. H. (1958b) Dental caries in the molar teeth of rats
II. A method
for diagnosing and scoring types of lesions simultaneously. J. dent. Res. 37, 1088-1099. KGnig K. G., Schmid P. and Schmid R. (1968) An apparatus for frequency controlled feeding of small rodents and its use in dental caries experiments. Archs oral Biol. 13, 13-26. Mrak E. M. and MacKinney G. (1951) The dehydration of fruits. In: The Chemistry and Technology of Foods (Edited by Jacobs M. B.), Vol. III, pp. 1773-1821. Interscience, New York. Navia J. M., Lopez H. and Harris R. S. (1969) Purified diet for dental caries research with rats. J. Nutr. 97, 133-140. Navia J. M. (1977) Experimental dental caries. In: Animal Models in Dental Research, Chap. 13, pp. 257-297. Alabama Press, Alabama. Strachan C. C., Moyls A. W., Atkinson F. E. and Britton J. E. (1951) The sugar content of apples. Dept. Agr.. Ottawa, Canada, Publ., No. 862. Touyz L. Z. G. (1980) Apples, acids and teeth. S. Afr. J. Sci. 76, 200-201. Touyz L. Z. G., Glassman R. M. and Naidu S. (1981) Erosive action of apple and orange juice on rat’s molars in vivo. S. Afr. J. Sci. 77, 423424. Watt B. K. and Merrill A. L. (1963) In: Composition of Foods, p. 6. Agr. Hand Book No. 8. Consumer and Food Economics Res. Div. Agricultural Research Science. U.S. Department of Agriculture. U.S. Government Printing Office, Washington, U.S.A. Whiting G. C. (1978) Sugars. In: Biochemistry of Fruits and their Products, 3rd edn (Edited by Hulme A. C.), Vol. I, Chap. I. pp. I-31. Academic Press, London. Widdowson E. M. and McCance R. A. (1935) The sugar contents of the edible portion of fruits. Biochem. J. 29, 151Ll56. Wynn W. and Haldi J. (1948) The erosive action of various fruit juices on the lower molar teeth of the albino rat. J. Nutr. 35. 488491.