|Year : 2019 | Volume
| Issue : 2 | Page : 57-60
Antibacterial effect of Cissus quadrangularis against cariogenic microorganisms: An in vitro study
B Kesavaraj1, Sharath Asokan1, PR GeethaPriya1, V Vijayasankari1, S Rubavathi2
1 Department of Pedodontics and Preventive Dentistry, KSR Institute of Dental Science and Research, Tiruchengode, Tamil Nadu, India
2 Department of Biotechnology, K S Rangasamy College of Technology, Tiruchengode, Tamil Nadu, India
|Date of Web Publication||9-Jul-2019|
Dr. Sharath Asokan
Department of Pedodontics and Preventive Dentistry, KSR Institute of Dental Science and Research, Tiruchengode - 637 215, Tamil Nadu
Aim: The aim is to evaluate the antibacterial activity of Cissus quadrangularis (CQ) extract and calcium hydroxide against Streptococcus mutans and Lactobacillus acidophilus. Materials and Methods: The ethanolic extract of CQ was obtained by Soxhlet method. The antibacterial activity of CQ extract and calcium hydroxide powder in three different solvents (saline, glycerol, and olive oil) was evaluated by agar well-diffusion method. The zones of inhibition formed against S. mutans and L. acidophilus were measured. Results: CQ in saline showed zone of inhibition against both S. mutans and L. acidophilus, whereas in olive oil, zone of inhibition was evident only against L. acidophilus. Calcium hydroxide exhibited zone of inhibition against S. mutans when dissolved in saline and glycerol. On the other hand, zone of inhibition against L. acidophilus was evident only in glycerol. Calcium hydroxide showed maximum zone of inhibition against both the microorganisms compared to CQ. Conclusion: CQ showed antibacterial activity against both S. mutans and L. acidophilus, but it was not comparable to calcium hydroxide.
Keywords: Antibacterial activity, calcium hydroxide, Cissus quadrangularis
|How to cite this article:|
Kesavaraj B, Asokan S, GeethaPriya P R, Vijayasankari V, Rubavathi S. Antibacterial effect of Cissus quadrangularis against cariogenic microorganisms: An in vitro study. SRM J Res Dent Sci 2019;10:57-60
|How to cite this URL:|
Kesavaraj B, Asokan S, GeethaPriya P R, Vijayasankari V, Rubavathi S. Antibacterial effect of Cissus quadrangularis against cariogenic microorganisms: An in vitro study. SRM J Res Dent Sci [serial online] 2019 [cited 2019 Jul 21];10:57-60. Available from: http://www.srmjrds.in/text.asp?2019/10/2/57/262378
| Introduction|| |
Dental caries is a biofilm (plaque)-induced acid demineralization of enamel or dentin, mediated by saliva. The biofilm comprises diverse microbial community, in which Streptococcus mutans and Lactobacillus acidophilus play a key role in the initiation and progression of caries. Thus, reducing the bacterial load of oral cavity is one of the primitive steps in preventing dental caries. Despite several chemical agents being commercially available, these might alter the oral microbiota and have adverse side effects. Hence, in the search for alternative product, natural phytochemical isolated from the plants are considered as promising alternative to synthetic chemicals.
According to WHO, medicinal plants would be the best source to obtain a variety of drugs. The natural products from plant may offer new agents for antimicrobial use. This might be due to the presence of secondary metabolites such as tannins, terpenoids, alkaloids, and flavonoids. In the current research, studies based on extraction of biologically active compounds from plant species used for therapeutic purposes are intensively increased. One such plant with numerous beneficial effects is Cissus quadrangularis (CQ) L. It is an indigenous medicinal plant of India belonging to the family Vitaceae. The whole plant helps in oral rehydration, while the root extracts, leaves, and stem are supportive in the treatment of various ailments. It has various other unique properties such as antimicrobial, antiulcer, antioxidative, antiosteoporotic, gastro protective, and cholinergenic activity.
According to the studies done by Selvamaleeswaran et al. and Kashikar and George, CQ exhibited antibacterial activity against various microorganisms such as Bacillus subtilis, Pseudomonas aeruginosa, Salmonella More Details typhi, Escherichia More Details coli, Proteus mirabilis, Staphylococcus aureus, and Streptococcus pyogenes. There is no evidence of the antibacterial effect of CQ against S. mutans and L. acidophilus. Therefore, the current study aimed to evaluate the antibacterial activity of CQ against these microorganisms in different solvents.
| Materials and Methods|| |
Preparation of Cissus quadrangularis extract
The stems of CQ were collected from areas around Tiruchengode, Namakkal, Tamil Nadu, India. The steps involved in the extraction process are shown in [Figure 1]. The ethanolic extract of CQ was obtained by Soxhlet method. The fleshy stems of about 3 kg were washed, cut into small pieces, shade dried, and crushed into powder. Two hundred and fifty grams of powdered stem was Soxhlet extracted with 95% ethanol. The extract was evaporated in vacuum under reduced pressure, and the final extract was stored in glass container at room temperature. The control group was calcium hydroxide powder (Dento Kem, Ballabgarh, India).
|Figure 1: Method of ethanolic extraction of Cissus quadrangularis. (a) collected stem, (b) shade dried stem, (c) crushed stem powder, (d) soxhlet extraction, (e) ethanolic extract of stem, and (f) final extract|
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The antibacterial activity was evaluated by agar well-diffusion method (Kirby–Bauer method). The bacterial strains used in the study were S. mutans (MTCC 10449) and L. acidophilus (MTCC 10307). The cultures were obtained from the Institute of Medical Technology, Chandigarh, India. A sum of 12 agar plates was used in the study. The bacterial strain cultures were swabbed on the surface of the nutrient agar medium using sterile cotton swabs. In each of these plates, three wells were created at equidistance to each other using 4 mm sterile borer. The extract powder and calcium hydroxide were mixed with the following solvents: saline, glycerol, and olive oil. The powders were diluted in the solvent at a concentration of 1 g/100 μl. The allocation of agar plates for extract/powder, bacterial strains, and solvents was described in [Figure 2]. All the plates were then incubated for 48–72 h at 37°C. The zone of inhibition formed around the wells was observed, and diameter of the zones was measured. The data were tabulated and analyzed.
| Results|| |
[Table 1] depicts the mean values of zone of inhibition against S. mutans. CQ in saline showed zone of inhibition, whereas no zones were formed when it mixed with glycerol and olive oil. Calcium hydroxide in saline and glycerol showed zone of inhibitions, but no zone was observed in olive oil.
|Table 1: Antibacterial activity of Cissus quadrangularis and calcium hydroxide against Streptococcus mutans|
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[Table 2] depicts the mean values of zone of inhibition against L. acidophilus. CQ in saline and olive oil revealed zone of inhibitions, but no zones were formed in glycerol. Calcium hydroxide in glycerol showed zone of inhibition, while no zones were formed by calcium hydroxide in saline and olive oil.
|Table 2: Antibacterial activity of Cissus quadrangularis and calcium hydroxide against Lactobacillus acidophilus|
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Calcium hydroxide showed a maximum zone of inhibition than CQ against S. mutans and L. acidophilus.
[Figure 3] describes the zone of inhibition against cariogenic microorganisms.
|Figure 3: Zone of inhibition. (a) Calcium hydroxide, (b) Cissus quadrangularis|
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| Discussion|| |
Phytochemicals derived from plants have been receiving much attention in the recent decades. It serves as a prototype to develop less toxic and more effective medicine in controlling the growth of organisms. The ancient system of medicine, Ayurveda, uses CQ to treat various diseases and disorders. Its osteogenic potential stemmed the notion to investigate its potential as an indirect pulp-capping agent. It was found to have variable antibacterial activity against different microorganisms,, and the existing literature did not show any studies on its antibacterial activity against S. mutans and L. acidophilus. This study was planned as the preliminary step toward its use as a potential pulp-capping agent. In dentistry, calcium hydroxide has been widely used as the gold standard pulp-capping agent. Its strong alkaline nature renders high antibacterial activity against cariogenic bacteria. Hence, in this study, it was planned to compare the antibacterial effect of CQ with calcium hydroxide as the control.
The diffusibility and solubility of powder may vary according to the nature of solvents used which in turn affect the antibacterial activity., Thus, saline, glycerol, and olive oil (aqueous, viscous, and oil) were used in this study. The results of the present study revealed that calcium hydroxide in all the solvents exhibited antibacterial effects. Contrary to our results, a study done by Leonardo et al. and Siqueira and de Uzeda concluded that calcium hydroxide in saline/glycerol/distilled water had little or no antibacterial effects. This might be due to the concentration ratio of the powder/solvent used.
In the present study, we have chosen ethanolic extract of CQ to evaluate the antibacterial activity, and it showed better results. Selvamaleeswaran et al. evaluated the antibacterial activity of various extracts of CQ such as ethanol, ethyl acetate, isopropyl alcohol, and petroleum ether. They reported that ethanolic extract had reasonable antibacterial effect and it was most commonly used. In contrary, Kashikar and George reported that no antibacterial effect was shown by ethanolic extract of CQ.
The antibacterial effect of CQ varies according to the location and season where they were cultivated. The reason might be the plants growing in different geographical localities were not alike in their phytochemical content. Their environmental factor affects the secondary metabolite of the plants. Basker and Mohammed showed that CQ collected from different provinces such as Tamil Nadu, Kerala, and Andhra Pradesh exhibited variable antibacterial activity. In our study, CQ collected around areas of Tiruchengode, Tamil Nadu, India, also showed antibacterial activity against cariogenic microorganisms.
In the present study, calcium hydroxide in glycerol showed maximum zone of inhibition. This is in contrary to the results stated by Siqueira and de Uzeda, who evaluated the antibacterial activity of calcium hydroxide mixed with different solvents such as saline, camphorated monochlorophenol, and glycerol and concluded that calcium hydroxide in glycerol had no antibacterial activity.
- The minimum inhibitory concentration of powder/extract was not standardized
- The powder/solvent ratio used in this study was restricted to a single concentration.
| Conclusion|| |
The results of the present study confirmed the antibacterial potential of CQ against cariogenic microorganisms. The antibacterial effect was not better than calcium hydroxide. The therapeutic application of this plant has to be considered only after further detailed research of their active components.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3]
[Table 1], [Table 2]