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Research Article
 

Anticaries Effects of Baphia nittida Extract in Albino Rats



E.S. Amadi, C.A. Oyeka, I. Okoli, J.I. Ihedioha and I.R. Iroha
 
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ABSTRACT

The possible curative and preventive effect of Baphia nittida extract on insipient caries and caries-free rats were investigated. About 200 mg mL-1 of Baphia nittida extract were intra-orally administered to the teeth surfaces of different rat groups. Evidence of development of caries was determined by direct observation, probing, microbial count and radiography. The result of the study showed that there was no evidence of dental caries among the non-insipient caries rat groups fed simultaneously with 70% sucrose-in-diet and extract of Baphia nittida. Similarly, the rat groups having insipient caries did not exhibit signs of dental caries after the administration of Baphia nittida extract. There was a significant difference (p<0.05) between the pre and post microbial count of the insipient caries rat groups given Baphia nittida extract. Also, there was a significant difference (p<0.05) between the pre and post microbial count of the caries-free rat groups given Baphia nittida extract twice daily. The result generally suggested that Baphia nittida extract has in vivo preventive and curative potential on dental caries in rats simultaneously fed with 70% sucrose-in-diet.

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  How to cite this article:

E.S. Amadi, C.A. Oyeka, I. Okoli, J.I. Ihedioha and I.R. Iroha, 2009. Anticaries Effects of Baphia nittida Extract in Albino Rats. European Journal of Dentistry and Medicine, 1: 1-7.

DOI: 10.3923/ejdm.2009.1.7

URL: https://scialert.net/abstract/?doi=ejdm.2009.1.7
 

INTRODUCTION

Dental caries is an infectious and communicable diseases which results in the gradual destruction of enamel, dentine and cementum by acid forming bacteria found in dental plaque, in the presence of sugar (Johnson and Beighton, 1992; Ingraham et al., 2001). It is one of the most common oral diseases showing marked geographical variation and severity of distribution all over the world (Ettinger, 1999; Talaro and Talaro, 2002). The prevalence is reported to be on the increase as a result of excessive sugar consumption especially in the developing countries (Adegbembo et al., 1995; El-Nadeef et al., 1998; Patro et al., 2008; Declerk et al., 2008; Umesi-Koleoso et al., 2007). Evidence on the association between amount and frequency of free sugar intake and dental caries has been previously highlighted (Moynihan and Peterson, 2004; Stecksen-Blicks et al., 2007; Kolker et al., 2007). Sugar free chewing gums has been reported to significantly slow the progression of caries (Morgan et al., 2008).

Dental caries is often associated with severe pain and discomfort especially when the pulp chamber is affected. It considerably impacts on self-esteem and quality of life and are expensive to treat. Dental caries ultimately results to loss of tooth minerals, which begins with the outer surface of the tooth and can progress through the dentine, to the pulp, thus culminating to tooth-loss (Johnson and Beighton, 1992; Moynihan and Peterson, 2004).

Synthetic drugs and antimicrobial chemicals has been reported to be useful in the reduction of dental caries. They probably inhibits the development and maturation of biofilms as well as affect bacterial metabolism (Baehni and Takeuchi, 2003). Most of these drugs, especially the very effective ones, are either not available or are expensive, beyond the reach of the common people in the developing nations. Furthermore, as with the case with most drugs in recent times, these agents and chemicals are becoming increasingly prone to microbial resistance.

The usefulness of traditional medicinal plants for prophylaxis and treatment of ailments and their important role in the developmental progress of the pharmaceutical industry has long been recognized. In Nigeria, many a plant material has been used since ancient days in the treatment of diseases including oral problems. Although the antimicrobial potentials of some of these local plants has been enumerated (Prashant et al., 2007; Amadi et al., 2007), most are yet to be scientifically studied in the laboratory. The present study investigates the in vivo effect of Baphia nittida, a local plant used widely in southeastern Nigeria as chewing stick, on dental caries in albino rats.

MATERIALS AND METHODS

Collection of Plant Materials
The leaves and tender stems of Baphia nittida (Abosi) were collected from Abakpa Nike, Enugu, Enugu State, Nigeria. The plant was authenticated by a taxonomist of the Department of Applied Biology, Ebonyi State University, Ebonyi State, Nigeria. This study was conducted in 2003.

Extraction
The washed leaves and tender stems of Baphia nittida were dried at room temperature. They were then macerated using mechanical grinder (Fondicion, Corona). Forty gram of the ground plant material were introduced into 95% ethanol and allowed to stand for 24 h with constant agitation. They were subsequently filtered using Whatman’s No.1 filter paper and evaporated to dryness in a water bath at 40°C (Ibrahim et al., 1997).

Experimental Rats
Twenty seven male albino rats were selected from the animal house of the Faculty of Veterinary Medicine, University of Nigeria, Nsukka. Only rats showing no signs of dental caries and whose anterior teeth plaque samples contain Streptococcus rattus were included in the selection. The plaque samples were cultured in crystal violet blood agar and identified using standard methods (Chessbrough, 2002; Collins et al., 1995).

Curative Effect of Baphia nittida on Albino Rats
A total of (12) 8-10 week old male albino rats weighing 62-120 g harboring Streptococcus rattus on their teeth surface, without signs of dental caries were used. They were divided into four groups (A-D). To establish base-line data, the upper and lower anterior teeth of the rats were visually observed and their texture determined using dental probe. Plaque samples from the upper and lower anterior teeth surfaces were also obtained using sterile swab sticks and probe and subsequently cultured in crystal violet blood agar to establish pre-microbial count.

Seventy percent sucrose-in-diet were fed to rat group A and B while group C and D rats were fed with normal diet without sucrose. Rat groups A and B were observed daily for development of white spot lesion. On determination of early signs of dental caries among the two rat groups, administration of Baphia nittida extract started. Two hundred milligram per milliliter of Baphia nittida extract was intra-orally administered on the anterior teeth of rat group A. However, while rat group A were given normal drinking water (containing 0.1 mg L-1 of fluoride), group B were given water containing 0.7 mg L-1 of fluoride . At this juncture, rat group D started receiving 70% sucrose-in-diet along with rat groups A and B, while group C rat continued with normal diet without sucrose. Rat group D was observed for the development of early caries signs. On confirmation of early caries on rat group D, the teeth surface of all the rat groups were observed visually and with dental probe. Also, plaque samples were collected from all the rats in all the groups for post microbial count. The radiograph of the lower interior teeth of all the rats were also done.

Preventive Effect of Baphia nittida Extract on Albino Rats
A total of (15) 8-10 week old male albino rat weighing 62-120 g harboring Streptococcus rattus on their teeth surface and without signs of dental caries were used. They were divided into groups A-E. To obtain baseline data, the upper and lower anterior teeth of the rats were observed visually and their texture determined using dental probe. Plaque samples from the upper and lower anterior teeth surface were also obtained to establish pre-microbial count.

Rat groups A, B, D and E were all fed with 70% sucrose-in-diet while group C were fed with normal diet. While group A rats were given 200 mg mL-1 Baphia nittida extract intra-orally once daily, group B were given same quantity of extract twice daily. However, whereas all the rat groups were given normal drinking water (containing 0.1 mg L-1 of fluoride), group D were given water containing 0.7 mg L-1 of fluoride . Group E rats served as control without plant extract or fluoride. The rat groups were fed and observed on daily basis until white spot lesions were observed in the anterior teeth surface of the group E rats.

Upon observation of the early caries sign on the teeth surface of the group E rats, the anterior teeth of all the rats in all the groups were observed visually and with dental probe. Plaque samples were also collected from all the rats to determine the post treatment microbial count. The radiograph of the lower anterior teeth of all the rats were also taken.

Determination of Insipient Caries
The early signal of insipient dental caries were determined by direct observation and probing. The plate count technique was used to determine the microbial count in the dental plaque sample (Ingraham et al., 2001). The radiographic examination of the lower anterior teeth of the rats were determined using an x-ray machine (ST-300) with a capacity of 0-150 kV and 50-300 mA at the Radiography Department of Ebonyi State University Teaching Hospital, Abakaliki, Ebonyi State, Nigeria.

Statistical Analysis
The t-distribution was used to analyze the microbial counts obtained in this study.

RESULTS

Curative Effects of Baphia nittida Extract
There was no observable difference between the appearance of the anterior teeth surface of the rat groups showing early signs of dental caries and simultaneously given Baphia nittida extract and 70% sucrose-in-diet and the control group given normal diet, on visual observation. The result thus indicated that insipient caries signs became visually imperceptible with the administration of Baphia nittida extract.

Also, the texture and translucency of the upper and lower anterior teeth surface of the rats were similar to those of the control group given normal diet. However, the anterior teeth surface of the rat group given 0.7 mg L-1 of fluoride appeared grayish white and were non-sticky on probing.

There was a significant difference (p<0.05) between the pre and post microbial count of the insipient caries rat group simultaneously given 70% sucrose-in-diet and Baphia nittida extract. Thus, the microbial count of the rat showing early signs of dental caries drastically reduced upon simultaneous administration of Baphia nittida extract and 70% sucrose-in-diet. However, there was no significant difference (p<0.05) between the pre and post microbial count in the control groups given normal diet (Table 1).

The radiographic study indicated no signs of radiolucency among the rat groups given Baphia nittida extract as well as those of the control groups given normal diet. There was however some evidence of radiolucency among the rat group given 70% sucrose-in-diet without plant extract.

Preventive Effects of Baphia nittida Extract
The visual appearance of the upper and lower anterior teeth surface of caries-free rats given simultaneously 70% sucrose-in-diet and Baphia nittida extracts were similar to those of the control group given normal diet without sucrose. Similarly, there was no difference in the texture and translucency of the same rat groups given 70% sucrose-in-diet and Baphia nittida extract and those of the control group given normal diet. However, the anterior teeth surface of the rats given only 70% sucrose-in-diet had milky-white spots and were sticky on probing after a period of about six weeks. Those of the rat group given 0.7 mg L-1 fluoride were however grayish white in color and non sticky on probing.

There was a significant difference between the pre and post microbial count (p<0.05) of the caries-free rats simultaneously given 70% sucrose-in-diet and Baphia nittida extract twice daily. No significant difference (p<0.05) however were observed for the rat groups given the same plant extract once daily as well as the control groups given normal diet (Table 2).

No signs of radiolucency was observed among the rat groups simultaneously given 70% sucrose-in-diet and Baphia nittida extract once and twice daily. The same was the case for the control groups given normal diet. There was however, evidence of radiolucency among the rat group given 70% sucrose-in-diet without plant extract or fluoride.

Table 1: Effect of Baphia nittida extract and fluoride on the microbial count of albino rats showing early signs of dental caries
Image for - Anticaries Effects of Baphia nittida Extract in Albino Rats
abDifferent alphabetical superscript in a row indicate that the means are significantly different at the probability level p<0.05, but similar superscripts in a row indicate a lack of significant difference between the means (p>0.05)

Table 2: Effect of Baphia nittida extract and fluoride on the microbial count of caries free albino rats
Image for - Anticaries Effects of Baphia nittida Extract in Albino Rats
abDifferent alphabetical superscript in a row indicate that the means are significantly different at the probability level p<0.05, but similar superscripts in a row indicate a lack of significant difference between the means (p>0.05)

DISCUSSION

The result of this study revealed that there was no evidence of early signs of dental caries among the insipient caries rat groups fed simultaneously with 70% sucrose-in-diet and Baphia nittida extract. This is suggestive of probable curative potential of the plant extract. This was indicated by the lack of white spot lesion, rough texture and bright light reflection of the anterior teeth surfaces of the rat groups given the plant extract. Further corroboration was obtained by the significant reduction (p<0.05) in the microbial count of the rat groups given the plant extract. Reduction in the number of cariogenic bacteria on the teeth surface has been reported to result to decline in the incidence and prevalence of dental caries in a community (Adetunji et al., 1996). Previous studies has shown that Streptococcus mutans usually increase at the sites that later becomes carious especially in the presence of sucrose diet (Linossier et al., 2003).

However, that the microbial count in the rat group given fluoride water (0.7 mg L-1) did not reduce significantly is somewhat surprising, since the anti-caries properties of fluoride has long been established (Marinho et al., 2003; Reelick et al., 2003; Addai et al., 2002; Moynihan and Peterson, 2004). This was quite obvious among the insipient caries rat groups were the mean microbial counts before and after fluoride treatment were drastically reduced and yet statistically insignificant. It is most probable, that the simultaneous administration of 70% sucrose-in-diet and fluoride may have affected the anti-caries activity of the fluoride. The caries inhibitory activity of fluoride containing dentifrices in rats has been previously highlighted (Guggenheim et al., 1997).

The insipient-caries curative property of Baphia nittida extract was further highlighted by the fact that the radiograph of the rat group simultaneously given 70% sucrose-in-diet and Baphia nittida extract were not radiolucent compared with those of the group given only 70% sucrose-in-diet that had evidence of radiolucency.

In the same vein, that no early sign of dental caries were observed among the caries-free rat groups simultaneously given 70% sucrose-in-diet and Baphia nittida extract once and twice daily tends to suggest that the plant extract probably posses preventive activity. Unlike the rat group given only 70% sucrose-in-diet where white spot lesion, rough texture and dull light reflection were recorded, the anterior teeth surface of the rats given plant extract were the same with those of the control group given normal diet.

The result of the study further showed that significant difference (p<0.05) between the pre and post microbial count was only obtained for caries-free rat group given Baphia nittida extract twice daily. Although one may not absolutely assert that twice daily administration of plant extract is essential for the preventive activity of the plant extract, since other criteria including visual observation, probing and radiography did not indicate same, it is not unlikely however, that twice daily administration may have some degree of advantage.

Previous study had indicated the anticaries property of polyphenol compounds from Camellia sinensis in Streptococcus mutans infected rats (Saito, 1990). Linke and LeGeros (2003) also reported the in vivo anticaries properties of black tea extract in hamsters. Further, the inhibitory activity of mango and neem extracts against Streptococcus mutans, S. mitis, S. salivarius and S. sanguis has also been highlighted (Prashant et al., 2007). It is important to note, that the in vitro inhibitory activity of the plant material used in this study against S. mutans has been reported (Amadi et al., 2007). Further studies to identify and characterize the active ingredients in the medicinal plant used in this study is recommended.

CONCLUSION

It is therefore most probable that the extract of Baphia nittida posses some degree of preventive and curative potential against dental caries in albino rat. Further investigation of this plant material is hereby suggested.

ACKNOWLEDGMENTS

The authors are grateful to Dr. Okeke Clement, of the Department of Botany, Nnamdi Azikiwe University, Awka and The Association for Scientific, Identification, Conservation and Utilization of Medicinal Plants of Nigeria (ASICUMPON) Enugu, Nigeria, for their inestimable assistance in the accomplishment of this study.

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