Biosynthesis of Cissus rotundifolia Stem-Mediated Titanium Dioxide Nanoparticles and Their Anticariogenic Activity against Streptococcus mutans and Lactobacillus sp.

ABSTRACT

Introduction:

The green synthesis of metal oxide nanoparticles using plant extracts has emerged as an eco-friendly method. Titanium dioxide nanoparticles (TiO2NPs) were synthesized using Cissus rotundifolia in this study. Titanium dioxide nanoparticles were utilized in restorative medicine for enhanced medicinal properties and in dental composites for their antimicrobial activities. Cissus rotundifolia is recognized as a medicinal plant due to its diverse properties, including mild laxatives, anti-inflammatory, and hyperglycemic activities.

Materials and Methods:

The antimicrobial activity of the prepared nanoparticles against Lactobacillus Sp. and Streptococcus mutans was evaluated using agar well diffusion method. The bactericidal and bacteriostatic activity of the prepared TiO2NPs was examined using time-kill kinetic analysis.

Results:

The prepared nanoparticles exhibited potential antimicrobial activity against Lactobacillus sp. (12 mm) at the highest concentration of 100 μg/mL. The prepared nanoparticles also exhibited excellent bactericidal activity against Lactobacillus Sp. and mild bacteriostatic activity against Staphylococcus mutans at the highest concentration of 100 μg/mL.

Conclusion:

The synthesized TiO2NPs showed significant antimicrobial activity against dental pathogens. The observed anticariogenic activity shows the potential of nanoparticles for dental applications. Hence, the prepared nanoparticles can be used in the field of dentistry as an antimicrobial agent instead of synthetic drugs causing more side effects.

INTRODUCTION

Materials created with a size between 1 and 100 nm are referred to as nanoparticles. Because of their physicochemical characteristics, they have fantastic potential to act as anticariogenic agents. The study of materials has been greatly impacted by nanoparticles,[1] which have drawn a lot of attention. The technological significance of nanoparticles and the implementation of ongoing research in this area will ensure their dominance in the coming years. In most applications, metal oxide nanoparticles are getting more important attention. In comparison with chemical approaches, the green production of titanium dioxide nanoparticles (TiO2 NPs) offers greater benefits.[2] In this study, it is revealed how herbal plants can biosynthesize TiO2 NPs. Due to its bottom-up nature, the reduction or oxidation process is the main reaction in the production of nanoparticles.[3]

Titanium dioxide is a cheap, non-toxic, and inert substance. TiO2 NPs are commonly employed to give items, such as toothpaste and food colorant brightness and opacity. Natural materials have been used to create TiO2 NPs.[4] The origin of the plant extract is known to affect the properties of the nanoparticles. Plant extracts have the potential to function as both reducing and stabilizing agents in the creation of nanoparticles.[5] This is because different extracts have various combinations and quantities of organic reducing agents.[6,7]

An environmentally beneficial method was the green production of metal oxide utilizing Cissus rotundifolia plant extract. This plant is mostly regarded as a medicinal plant because of its wide range of qualities, including gentle laxatives for kids and pregnant women used to treat chronic disorders and anti-inflammatory and hypoglycemic properties.[8] We need more efficient techniques for doing every task because we live in a world that is constantly evolving and changing. This holds true for nanoparticles and drug delivery methods. The manufacturing of nanoparticles also uses plant sources.[9,10,11] The anticariogenic activity and time–kill curve assay were used to achieve the study’s objectives. TiO2 NPs protect against cariogenic infections through the stem of Cissus rotundifolia. Because of its technological significance and the implementation of ongoing research in the field, nanoparticle supremacy may persist in future years.

MATERIALS AND METHODS

Preparation of plant extract

5 gm of Cissus rotundifolia stem was weighed and washed with water to remove the dust particles. After that, the stem was crushed and added to 100 ml of distilled water [Figure 1]. After that, Cissus rotundifolia stem extract was boiled at 50–60°C for 20 minutes using a heating mantle. After boiling, the plant extract was filtered using Muslim cloth. The filtration of the boiling extract was used for further research purposes.

Figure 1.

Preparation of green synthesis of Cissus rotundifolia stem-mediated titanium dioxide nanoparticles

Preparation of nanoparticles

0.395 g of titanium oxide was mixed with 50 mL of distilled water. After that, 50 mL of Cissus rotundifolia stem-mediated solution was added to the titanium oxide solution [Figure 2]. The titanium oxide solution was used to convert the TiO2 NPs. For the preparation of nanoparticles, a solution of nanoparticles is kept in a rotating orbital shaker. The nanoparticles were then visually synthesized using an ultraviolet (UV)–visible spectroscopy reading.

Figure 2.

Preparation of Cissus rotundifolia stem-mediated titanium dioxide nanoparticles

Anticariogenic activity

The anticariogenic activity was followed by the previous research article (Rajeshkumar et al., 2021).[2]

Time-kill kinetics analysis

A 1 mL aliquot of the bacterial and fungal suspension (Streptococcus (S.) mutans and Lactobacillus sp.) was added to 9 mL of Mueller–Hinton broth containing the TiO2 NPs at concentrations of 25 μg, 50 μg, and 100 μg. The final microbial concentration was approximately 106 colony-forming unit (CFU)/mL. The mixture was then incubated at room temperature with shaking at 200 rpm for varied time durations (0, 4, 6, 8, 10, 12, and 24 hr). Then, the dead cell percentage was calculated at 600 nm at the time interval.

RESULTS

The graph represents the antimicrobial activity and the results obtained in the presence of Cissus rotundifolia stem-mediated TiO2 NPs [Figure 3]. The error bars indicate the zone of inhibition. The statistical analysis revealed that the observed differences in the zone of inhibition between the standard group and the treatment group were found to be statistically significant (P < 0.05).

Figure 3.

Graphical representation of antimicrobial activity of Cissus rotundifolia stem-mediated TiO2 NPs

The graph represents the antimicrobial activity and results obtained in the presence of Cissus rotundifolia stem-mediated TiO2 NPs against S. mutans [Figure 4]. The error bars indicate the zone of inhibition. The statistical analysis revealed that the observed differences in the zone of inhibition between the standard group and the treatment group were found to be statistically significant (P < 0.05).

Figure 4.

Graphical representation of the antimicrobial activity of Cissus rotundifolia stem-mediated TiO2 NPs against S. mutans

The graph represents the antimicrobial activity and results obtained in the presence of Cissus rotundifolia stem-mediated TiO2 NPs against Lactobacillus sp [Figure 5]. The error bars indicate the zone of inhibition. The statistical analysis revealed that the observed differences in the zone of inhibition between the standard group and the treatment group were found to be statistically significant (P < 0.05).

Figure 5.

Graphical representation of the antimicrobial activity of Cissus rotundifolia stem-mediated TiO2 NPs against Lactobacillus sp.

Antimicrobial activity

In this study, the agar well-diffusion method was employed to check for antibacterial activity. To compare the inhibitory effects on Lactobacillus species and Staphylococcus mutans, two agar plates were used. There were four wells on each plate, each containing a different concentration of nanoparticles: 25 μg/ml, 50 μg/ml, 100 μg/ml, and a standard. At 50 μg/ml and 100 μg/ml, the nanoparticles’ zones of inhibition against S. mutans and Lactobacillus species have been measured to be 15 mm and 24 mm, respectively. At concentrations of 25 μg/ml, 50 μg/ml, and 100 μg/ml, the diameter of the nanoparticle-inhibiting zone in S. mutans was measured to be 9 mm, 10 mm, and 11 mm, respectively.

DISCUSSION

This was also seen in research that compared Cissus rotundifolia efficacy against a range of cariogenic infections. Additionally, they discovered that the zone of inhibition surrounding S. mutans had been quantified.[12] When compared to the research, we can see that the delivery strategy is much more effective when using nanoparticles because the inhibition zone has a bigger width. This work also advances our understanding of how effective nanoparticles are at treating a range of disorders.[13] We previously knew that it was efficient against cancer and that it had cytotoxic activity. In addition, we can also state that it was effective against germs.[14,15]

Zn-TiO2 nanocomposite (NC) showed effective antibacterial activity at 150 μL concentration against S. mitis (28 ± 1.0 mm), followed by S. mutans (25 ± 0.6 mm), and these were more or less equal to bactericidal activity at 150 μL concentration of amoxyrite (34 ± 1.4 mm). This result suggests that the green synthesized Zn-TiO2 NC effectively acts against oral pathogens such as S. mitis and S. mutans.[16]

After incubation at room temperature under anaerobic conditions, the antimicrobial activity of clove and ginger-mediated TiO2 NP-based dental varnish was observed, producing the least amount of bacterial growth inhibition at the different concentrations on the culture plate. On the culture plate, the freshly made Mueller–Hinton broth culture was applied. The average minimum bactericidal concentration (MBC) values were 8, 4, and 1 μg/mL for the concentrations of 25, 50, and 100 μL, respectively. Significant growth inhibition was seen at 100 μL as compared to 25 and 50 μL in the statistical analysis for MBC performed on all test samples of all concentrations against S. mutans and Lactobacilli sp.[17]

CONCLUSION

TiO2 nanoparticles were effectively synthesized utilizing a Cissus rotundifolia stem extract. Analytical instruments, such as the UV–visible spectrophotometer, were used. The microscope aided in understanding the TiO2 nanoparticles. TiO2 nanoparticles were synthesized in a spherical shape. This could lead to a slew of new uses. Thus, we can conclude that the stem extract of Cissus rotundifolia can be effective against Lactobacillus sp. and S. mutans. The Cissus rotundifolia mediated TiO2 NPs using anticariogenic activity. They showed good results for anticariogenic activity. They produce a higher zone of inhibition for Lactobacillus sp. In future studies, they will develop oral medicine.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.