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 Table of Contents  
ORIGINAL RESEARCH
Year : 2021  |  Volume : 13  |  Issue : 5  |  Page : 499-507

Antimicrobial activity of Salvia officinalis against Streptococcus mutans causing dental implant failure: An in vitro study


1 Department of Life Science, Central University of Technology, Free State, Bloemfontein, South Africa
2 Department of Mechanical Engineering, P/Bag, Central University of Technology, Free State, Bloemfontein, South Africa

Date of Submission02-Feb-2021
Date of Decision04-Mar-2021
Date of Acceptance05-Apr-2021
Date of Web Publication11-Oct-2021

Correspondence Address:
Ms. Sinazo S Ntondini
Department of Life Science, P/Bag X20539, Central University of Technology, Free State, Bloemfontein.
South Africa
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/JIOH.JIOH_26_21

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  Abstract 

Aim: The current study aimed at examining the antimicrobial activity and efficacy of Salvia officinalis against Streptococcus mutans that affect titanium dental implant (Ti6Al4V). Materials and Methods: An in vitro study was conducted in which Salvia officinalis essential oil was investigated against antibiotic-resistant S. mutans. Salvia officinalis chemical components were analyzed using the gas chromatography–mass spectrometry (GC–MS) method. Six samples of titanium dental implant (Ti6Al4V) were manufactured using direct metal laser sintering (DMLS) manufacturing technology. The bioassay method was conducted to confirm the inhibitory effect of Salvia officinalis essential oil against S. mutans; microdilution assay was also performed on six samples to examine the minimum inhibition concentration (MIC). Two samples that showed MIC were selected to undergo scanning electron microscopy (SEM) to investigate or observe bacterial structural changes upon treatment with Salvia officinalis essential oil. Results: Salvia officinalis essential oil exhibited a great inhibition diameter of 40 mm against S. mutans, and it showed an MIC value of 12.5 μg/mL on the titanium implant material surface and an MIC of 5 μg/mL. The SEM results have shown drastic structural changes on S. mutans on treatment with Salvia officinalis essential oil at its MIC value. Conclusion: Salvia officinalis essential oil has shown potential to be used as an antimicrobial agent; therefore, essential oils in future can be considered possible antimicrobials because of the structural changes that they induce on bacterial cells. Salvia officinalis essential oil, which was investigated in the current study, succeeded in penetrating the bacterial cell wall and caused disruption; therefore, the essential oil stands a chance of being used to inhibit the growth of S. mutans bacteria.

Keywords: Antimicrobial Activity, Essential Oils, Implant Failures


How to cite this article:
Ntondini SS, Lenetha G, Dzogbewu TC. Antimicrobial activity of Salvia officinalis against Streptococcus mutans causing dental implant failure: An in vitro study. J Int Oral Health 2021;13:499-507

How to cite this URL:
Ntondini SS, Lenetha G, Dzogbewu TC. Antimicrobial activity of Salvia officinalis against Streptococcus mutans causing dental implant failure: An in vitro study. J Int Oral Health [serial online] 2021 [cited 2021 Dec 6];13:499-507. Available from: https://www.jioh.org/text.asp?2021/13/5/499/327865


  Introduction Top


Implant failures can take place at an initial stage or a later stage, therefore early implant failure transpires before osseointegration whereas late failure occurs years or decades after insertion.[1] Implant surfaces are prone to being colonized by pathogenic bacteria that cause peri-implant tissue destruction and implant failures.[2] Osseointegrated dental implants withhold a longstanding 90% success degree, although pathogenic bacteria colonization threatens them.[3] Peri-implant colonization with pathogenic microorganisms is the source of infection.[4] Implants during surgery are more prone to bacterial infection on the mucous membranes and the skin and that results in biofilm forming around the implant’s surface.[5] The biofilm formed around the implant’s surface triggers dental inflammation.[4],[5],[6] This biofilm is known as collective microorganisms that grow on surfaces. Biofilm infestation causes inflammation on the tooth surface and may result in an inflammatory condition such as peri-implant mucositis or gingivitis that occurs on the gums.[4],[6] Peri-implantitis, which is additionally caused by bacterial colonization, is an irreversible inflammation of the soft and hard peri-implant tissues.[1] Moreover, gingivitis is the slightest type of periodontal diseases that is triggered by bacteria biofilm that gathers on the teeth and gums.[1],[4],[7] Biomaterials used for implants manufacturing are not themselves antibacterial agents; hence their surfaces are coated with antibiotics to prevent bacterial colonization. However, microorganisms have developed diverse mechanisms against antibiotics; therefore, essential oils, which are natural products, show potential to be antimicrobial agents against oral bacteria. The current study concentrated on using a natural product such as Salvia officinalis essential oil against Streptococcus mutans, which is known to colonize dental implants and later trigger implant failure. Up to date, numerous articles have been concerned with the effectiveness of essential oils in inhibiting oral microbiota and also microorganisms that are particularly found in food. The chief benefit of natural products is their non-susceptibility to antibiotic resistance as compared with the long-term use of synthetic antibiotics.[8],[9] According to Kavanaugh and Ribbeck,[10] essential oils that are extracted from plants have been used for many years as natural medicines to fight numerous microorganisms such as fungi, bacteria, and viruses. Therefore, recently there is an increased consideration toward the utilization of essential oils because they possess antifungal, antibacterial, and antioxidant properties.[11],[12] In addition, essential oils have been reported to show antiviral, antimycotic, antioxygenic, antiparasitic, and insecticidal properties.[13]Salvia officinalis (Sage) essential oil, which has been investigated in the current study, is extracted from the leaves of the Salvia officinalis herb, also known as common sage, blue mountain sage, true sage, garden sage, and Dalmatian sage, by the steam distillation method.[14] Sage oil can inhibit fungal infections, provide help against fungal infections such as dysentery, and further protect against bacterial infections in wounds.[1] Hamidpour et al.[15] believe that Salvia officinalis prohibits the growth of microorganisms that are known as causative agents of dental caries, such as Lactobacillus rhamnosus, S. mutans, and Actinomyces viscosus. S. mutans is reported as the main pathogenic bacteria in dental caries. This S. mutans colonizes the surface of the teeth and gathers as plaque by liquefying the constructions of the tooth during the existence of fermentable carbohydrates, such as sucrose, glucose, and fructose, by means of glucosyl transferases.[16],[17],[18] Bacterial colonization around the mouth can cause dental caries that trigger tooth loss and one of the rehabilitation options includes the replacement of a lost tooth with biomaterials such as titanium alloy (Ti6Al4V).

Using of titanium alloys for dental implants

Titanium alloys are the most utilized dental implants that are used to replace a missing tooth and they have been utilized ever since 1981.[19],[20],[21] The highly valuable alloys are pure titanium (cpTi) and Ti6Al4V, and both exhibit success of up to 99% at intervals of 10 years. The alloys can attach to the gingival tissues and the bones.[20],[21] Titanium alloys are widely utilized in medical applications because they exhibit excellent biocompatibility, as well as excellent mechanical properties such as less elastic modulus as compared with stainless steel or CoCr alloys.[19],[20],[22] Abdulgader[20] explained that although titanium alloys are good biomaterials, they can fail due to microbial colonization on the surface of the implant, triggering infections; therefore, their surface should be modified to prevent implant infections.

The current study aimed at examining the antimicrobial activity and efficacy of Salvia officinalis against S. mutans that affect the titanium dental implant (Ti6Al4V). Therefore, the titanium implant material (Ti6Al4V) surface was coated with Salvia officinalis to prevent S. mutans bacterial growth or bacterial colonization of the implant surface. The inhibitory effect of Salvia officinalis against S. mutans was investigated using the bioassay and microdilution assay method.


  Materials and Methods Top


Materials

Salvia officinalis oil, which is steam distilled from the leaves of the Salvia officinalis herb, was purchased from a local supplier in Bloemfontein, which is situated in the Free State province, South Africa.

Microorganism

S. mutans (ATCC 25175) bacterial strain was purchased from Laboratory Specialities (Pty) Ltd Company, which is situated in South Africa.

Methods

This in vitro study was conducted at the Central University of Technology laboratory. One essential oil, namely Salvia officinalis, was analyzed using the GC–MS method. Six samples of Ti6Al4V dental samples were manufactured to be used in the current study. All the samples were incubated at 37°C for 24h with the bioassay method and microdilution method (a total of six samples were used). For SEM, two samples from the microdilution method were investigated for bacterial and structural changes induced by the essential oil.

Gas chromatography–mass spectrometry method

In order to analyze the chemical components of the essential oil, GC–MS was carried out. Briefly, Salvia officinalis was analyzed on an Agilent 7890B GCMS system coupled directly to a 5977A mass spectrometer. A volume of 0.2 μL sample (clear liquid) was diluted with n-hexane (2% v/v) and inserted by a split ratio (100:1) with an auto sampler at 24.79 psi and an inlet temperature of 220°C. The GC–MS system was armed with an Agilent 19091S 433 UI: HP5-MS UI (30 m × 250 μm × 0.25 μm) column. The temperature of the oven ranged from 60°C to 280°C at various rate increments. Helium was used as carrier gas at the same flow of 30 cm/s. Spectra were attained by an electron impact at 70eV, scanning from 40.5 to 415 m/z. The peak areas of the selected GC elements were each expressed as percentages of the overall of all the Total Ion Count (TIC) peak areas as determined by mass spectrometry detection (MSD, transfer line 240°C; source 230°C; MS Quad 150°C) without utilizing correction factors. The compounds were identified using the NIST11 and Agilent Flavor2 mass spectra libraries, and by a comparison of peak retention times and mass spectra with data in the Adams’ database.[23],[24]

Bioassay method

A quantitative bioassay was applied to examine the antimicrobial activity of Salvia officinalis against the growth of S. mutans bacteria. S. mutans regrown on petri dishes was incubated for 24h. Later, the immersion of S. mutans in sterile distilled water (dH20) and 0.2 mL smeared on PCA (0.5% mv 1 agar) resulted in the formation of a homogenous lawn across the surface of the agar.[24],[25] Then, a 0.5 cm diameter of a hole was created in the center of a petri dish and Salvia officinalis (46 μg) was poured along with ethanol. Ethanol (96%) served as a control. The petri dishes were then incubated for 24h at 37°C; thereafter, growth inhibition zones were detected, and diameters (mm) were measured.

Microdilution assay

Salvia officinalis essential oil was further tested for MIC. The inoculate of S. mutans was prepared in Mueller-Hinton Broth (MHB) for a period of 24h. Thereafter, the S. mutans bacteria was distributed into the 96-well sterile microtiter plate.[24],[26]Salvia officinalis was poured into the first row of wells and serial dilutions were done in order to attain concentrations of 80, 40, 20, 10, 5, and 2.5 μg/mL. The microtiter plate was enclosed with a sterilized plate sealer and incubated overnight at 37°C. To indicate growth after the incubation period, the experiment was done twice in duplicate for each concentration. After incubation, p-iodonitrotetrazolium violet (INT) of 20 µL was added to each well. The microtiter plate was then incubated for 20 min at 37°C. Bacterial growth was indicated by color changes ranging from pink to violet.[24],[27]

Scanning electron microscopy (SEM)

The SEM evaluated structural variations caused by Salvia officinalis essential oils on S. mutans bacteria cell structure. Treated and untreated S. mutans was fixed in sodium phosphate and then buffered in glutaraldehyde solution at pH 7.0 for 3 h. The samples were centrifuged and buffered in osmium tetroxide solution for 1 h. Afterward, the sample was dehydrated in a series of ethanol solution of 30%, 50%, 70%, 90%, and 100% for 20 min for each solution and the 100% dehydration was conducted twice during 1h. Later, the samples with ethanol were critically dehydrated, placed on a filter paper, and shielded with uranium for the samples to be electrically conductive. The samples were then imaged using SEM. Images were taken to witness the structural variations induced by the Salvia officinalis on the S. mutans.[24],[28],[29]

Manufacturing of the experimental Ti6Al4V dental samples

The samples were additively manufactured (3D printing) using EOS M280 DMLS manufacturing technology, which is a subset set of the laser powder bed fusion (LPBF).[30],[31],[32] The machine mainly comprises a process chamber that consists of a computer controlled elevator system, a recoating system, a platform heating module, an optical system equipped with a 400-watt fiber laser, a process gas management system, and a process computer with control software. [33],[34]

Spherical argon atomized Ti6Al4V (ELI) powder procured from TLS Technik was used [Figure 1]. The elemental composition of the powder in weight percent (wt%) is presented in [Table 1]. The powder materials and substrate were similar in terms of chemical constituents. Argon was utilized as a protecting atmosphere; the oxygen level inside the built chamber was 0.07%–0.1%. The titanium dental implant samples were manufactured with optimum process parameters: laser power of 170 W, scanning speed of 1.25 m/s, hatch distance of 80 µm, and powder layer thickness of 30 µm with a zigzag scanning strategy.[35],[36],[37] The samples where stress relieve in Ar atmosphere at a temperature of 650°C for 3h and were cut from the based plates using Electrical Discharge Machining. The samples were cleaned in an ultrasonic bath to remove the “loose attached” powder residues.
Figure 1: SEM micrograph of the employed Ti6Al4V powder

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Table 1: Elemental composition of the Ti6Al4V

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Microdilution assay on titanium implant materials

Salvia officinalis essential oil was further tested for MIC. The inoculate of S. mutans was prepared in MHB for a period of 24h; then, S. mutans was prepared according to 0.5 McFarland turbidity standard (approximately 1.5 × 108 cfu/mL). S. mutans was then distributed into glass tubes with the implant material inside a 96-well sterile microtiter plate.[24],[26]Salvia officinalis was poured into the first row of glass tubes and serial dilutions were done to attain final concentrations of 100, 50, 25, 12.5, 6.25, and 3.125 μg/mL. Thereafter, the glass tubes were incubated in a shaker incubator at 37°C overnight. To indicate growth after 24h, the experiment was run twice in duplicate for each concentration. Next after incubation, p-iodonitrotetrazolium violet (INT) of 20 µL was added to each glass tube. The glass tube was then incubated for 20 min at 37°C. Bacterial growth was indicated by color changes ranging from pink to violet.[24],[27]

Scanning electron microscopy on titanium implant samples

The scanning electron microscopy (SEM) evaluated structural variations caused by Salvia officinalis essential oils on the S. mutans bacterial cell structure on the surface of a titanium dental implant sample. Treated and untreated S. mutans was fixed in sodium phosphate and then buffered in glutaraldehyde solution at pH 7.0 for 3 h. The samples were centrifuged and buffered in osmium tetroxide solution for 1 h. Thereafter, the titanium implant sample was dried out in a graded series of ethanol solution (30%, 50%, 70%, 90%, and 100% for 20 min per solution and the 100% drying was conducted twice for 1h). Later, the dried titanium implant sample with ethanol was heated at 40°C and then sputter coated with uranium for 30 min to make it electrically conductive. The implant sample coated with uranium was then imaged by SEM. Images were taken to observe the structural variations caused by the Salvia officinalis on the S. mutans plated on the surface of a titanium dental implant sample.[24],[28],[29]


  Results Top


The GC–MS was carried out to recognize the chemical constituents of Salvia officinalis that might be responsible for inhibiting S. mutans bacterial growth. The analyzed Salvia officinalis essential oil consisted of major constituents, namely, 13.62% of β-Myrcene, 9.62% of δ-3-Carene, and 6.79% of 2-Bornanone (Camphor), as shown in [Table 4].
Table 4: The chemical components of Salvia officinalis

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The bioassay method was performed to investigate whether Salvia officinalis inhibits the growth of Streptococcus mutants and subsequently microdilution was performed to discover at what minimum concentration this occurred. From the bioassay method, Salvia officinalis showed 40 mm inhibition diameter on S. mutans bacteria [Figure 2]. The microdilution results have shown that Salvia officinalis has an MIC value of 5 µg/mL against S. mutans [Table 2].
Figure 2: Bioassay of Salvia officinalis against Staphylococcus aureus cell (A), control (B), inhibition zone (J)

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Table 2: The MIC of Salvia officinalis against S. mutans

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Thereafter, a microdilution assay was performed on the titanium implant sample and an MIC value of 12.5 µg/mL was obtained, as shown in [Table 3]. S. mutans treated with Salvia officinalis at its respective MIC values in [Table 2] and [Table 3] was further tested for structural changes induced by the essential oil on bacteria using SEM. The images from SEM are depicted in [Figure 3] and [Figure 4], and they have shown remarkable alterations on the structure of the S. mutans bacterial cell when treated with Salvia officinalis essential oil.
Table 3: The MIC of Salvia officinalis against S. mutans on the surface of titanium dental implant

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Figure 3: Image of SEM of S. mutans cells control (A), S. mutans cells treated with Salvia officinalis (B) displaying loss in bacteria’s cell content (LCC), division of the cell division (CD), cell wall damage (DCW)

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Figure 4: Image of SEM of S. mutans bacterial cells plated on the surface of the titanium implant sample. Control (A), S. mutans cells treated with Salvia officinalis (B), displaying loss in cell content (LCC), division of the cell (CD), disrupted bacteria cell wall (DCW), elongated (L), and roughage (R)

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These SEM images revealed that Salvia officinalis possesses antibacterial activity. Salvia officinalis essential oil can be confirmed to be effective in inhibiting the growth of S. mutans, which is regarded as one of the primary pathogenic bacteria responsible for dental caries.[38]


  Discussion Top


Antimicrobial activity of Salvia officinalis against S. mutans

Chemical agents appear to control peri-implant infections; however this area requires further research. The usage of antimicrobial mouth rinses as a form of chemical methods has become common during the past decade to control biofilm formation. Mouth rinses have active ingredients that are used for chemical biofilm control and these ingredients include bis-biguanide, essential oils, and 0.12% chlorhexidine gluconate. However, chlorhexidine gluconate has several undesirable side effects such as irritation and staining of the teeth and it needs to be prescribed with caution.[39],[40] The most common therapeutic agents that mouth rinses consist of are components that are found in essential oils such as thymol, eucalyptol, menthol, and methyl salicylate. The other chemical components are chlorhexidine gluconate, cetylpyridinium chloride, hexetidine, benzalkonium chloride, hydrogen peroxide, and occasionally domiphen bromide, fluoride, and xylitol, which are known to be responsible for inhibiting bacterial growth.[39] Further research on use of essential oils to control bacteria causing too loss or decay can be useful in oral health or dentistry. In the present study, Salvia officinalis has revealed antimicrobial activity against S. mutans bacteria.

Further, the ever-increasing resistance of pathogens to conventional chemicals and drugs is a severe and evident universal problem that has driven research into identifying new biocides that possess far-reaching activity.[40],[41] Antibiotics fail to inhibit bacterial growth, because the bacteria changes or limits the number of openings in the cell wall. The antibiotic drug molecules usually gain access to the cell by diffusion through the porins found in the gram-positive bacteria’s outer membrane; however, reduction of the quantity of porins channels leads to the reduced entrance of antibiotics inside the cell.[41],[42],[43] The cytoplasmic membrane of the bacteria has pumps that take antibiotics out of the cell and these are called efflux mechanisms pumps; for example, Tetracyclines, Macrolides, Lincosamides, Streptogramins, Oxazolidinones, Phenicols, Cationic peptides, Lipopeptides, Quinolones, Pyrimidines, Rifamycins, and Sulfonamides groups of antibiotics are pumped out.[42],[44] The pumps use the same speed that the antibiotics use to gain access in the cells by pumping the antibiotic out before it reaches the target. The outer membrane of the gram-positive bacteria keeps the antibiotics from gaining access to the bacteria’s cell.[44] Therefore, microorganisms are becoming more antibiotic resistant as they can survive and increase in the presence of antibiotics; they also tend to prevent the antimicrobial agents from meeting their target by limiting their capability to infiltrate the bacteria, whereas essential oils consume excessive potential in the field of biomedicine because they successfully inhibit the growth of numerous bacterial, fungal, and viral pathogens. Essential oils inhibit the growth of pathogens by aiming at the membrane and the cytoplasm and they can totally change the structure of the cells; the current study shows the mechanism of essential oil against oral bacteria.[4],[41],[45]

Increased antibiotic resistance is a serious public health challenge and it is due to the increasing number of dental implants inserted. The number of peri-implant infections is likewise increasing, and bacteria are becoming more resistant to antibiotics, for example, Staphylococcus aureus is known to be resistant against penicillin and methicillin.[42],[43] This study focused on using natural-based products, such as essential oils, to control the pathogens affecting dental implants.

The bioassay method was performed to investigate whether Salvia officinalis inhibits the growth of Streptococcus mutants and subsequently microdilution was performed to discover at what minimum concentration this occurred. The results have shown the antimicrobial activity of Salvia officinalis with an MIC value of 5 µg/mL against S. mutans [Table 2]. Thereafter, microdilution assay was performed on the titanium implant sample and an MIC value of 12.5 µg/mL was obtained, as shown in [Table 3]. In the bioassay method, Salvia officinalis showed significant inhibitory influence against S. mutans with an inhibition diameter of 40 mm [Figure 2]. This demonstrates that Salvia officinalis possesses remarkable antibacterial activity against the S. mutans bacterial cell, and it is one of the primary pathogenic bacteria causing dental caries.[38] Previous studies were done on the antimicrobial activity of essential oils; for example, Salvia officinalis exhibited the lowest antimicrobial potential against candida, a causative agent of several human oral disorders, as compared with other essential oils such as H. officinalis, R. officinalis.[46] However, in this study, Salvia officinalis showed remarkable antimicrobial potential against S. mutans. A similar study about the antimicrobial activity of Sage oil was conducted and it was found that the potential of sage as a therapeutic agent is remarkable as it has been shown to control the development of opportunistic pathogens such as S. aureus, S. epidermidis, S. mutans, C. albicans, C. tropicalis, and glabrata.[47] Adams[48] consider that the essential oils of Salvia comprise monoterpenoid major constituents that are known to be effective antibacterial agents; therefore, similar results were observed in the current study. Salvia officianalis, which inhibited the growth of S. mutans, has been shown to contain monoterpenoids such as 13.62% of β-Myrcene, 6.79% of Bornanone (Camphor), 6.09% of α-Pinene, 1.06% of Borneol, and 1.91% of 1, 8-cineole (eucalyptol), as shown in [Table 4]. A similar study was conducted in which the antimicrobial effect of Salvia officinalis showed the presence of numerous bioactive compounds such 34.7 of α-thujone, 23.5% of camphor, 11.5% of 1, 8-cineole, and 7.4% of carvacrol.[49] In addition, in literature, essential oil known as Sage or Salvia officinalis consists of α-thujone, camphor, and 1, 8-cineole that serve as major chemical components and these are known to have antimicrobial activity against human bacterial pathogens such as Staphylococcus aureus and Providencia stuarti. The existence of camphor and camphene in Salvia officinalis provides it with antifungal and antibacterial properties.[1]

Essential oils are well known to target the cell membrane’s lipid bilayer, making the bilayer extra penetrable and resulting in leakage or loss of vital cell contents. Therefore, Salvia officinalis has been shown to inhibit the growth of the S. mutans cell by destroying the cell wall and further leading to a loss of cellular content.[48] The permeability and loss of cellular content characteristics is the cause of cell death, as shown in [Figures 3] and [4].

Structural changes induced by Salvia officinalis on S. mutans

Essential oils, which are regarded as natural products, are deemed as strong antimicrobial agents due to their mechanism of action, such as corruption of the cell wall, destruction of the cytoplasmic membrane, and cytoplasm coagulation when they meet a bacterial cell.[50],[51] In the current study, Salvia officinalis partitioned into the cell membrane’s lipid bilayer of S. mutans bacteria and caused leakage in cell contents.

S. mutans was treated with Salvia officinalis, MICs are shown in [Tables 2] and [3], and the images from SEM are shown in [Figures 3] and [4]. The images show notable changes on the bacterial structure when treated with Salvia officinalis. In the literature, Tardugno et al.[24],[52] stated that natural products such as essential oils are capable of effortlessly entering bacterial cell membranes and similar results were obtained in the study [Figure 3B]. The structure of the S. mutans shows loss of cell impairment, loss of cellular content, division of the cell wall, and roughage when equated to the untreated. S. mutans. A few S. mutans cells looked distorted, producing leakage of the intracellular contents [Figure 2B]. [Figure 4] presents cell wall damage, division of the bacterial cell, loss of cell content, elongated cells, and roughage of the bacterial cell plated on the surface of a titanium implant material. These remarks indicate that Salvia officinalis with major compounds β-Myrcene (13.62%), δ-3-Carene (9.62%), and 2-Bornanone (Camphor, 6.79%) are shown in [Table 4]; these could be connected to the disruption of the S. mutans bacteria’s structure or cell wall of the bacteria. In the literature, Salvia officinalis consists of α-thujone, camphor, and 1, 8-cineole that serve as the main chemical components and these are known to possess antimicrobial activity against human bacterial pathogens such as S. aureus and Providencia stuarti. Therefore, similar results were also observed as the essential oil consists of camphor and inhibits the bacterial growth of S. mutans.[14] Sage essential oil or Salvia officinalis contains a variety of healing properties plus antibacterial, antiviral, antifungal, and antioxidant effects.[48] It would be of importance to define whether Salvia officinalis can be valuable in dentistry.


  Conclusion Top


Bacteria have become antibiotic resistant; however, Salvia officinalis essential oils have proven their antimicrobial activity against drug-resistant S. mutans, the inhibition of which by conventional antibiotics has proved to be problematic.[24],[43]Salvia officinalis essential oil that was investigated in the current study succeeded in permitting the S. mutans bacteria’s cell wall, causing disruption; therefore, Salvia officinalis has shown capability to be considered as an antimicrobial agent against S. mutans bacteria. The images from the SEM have also shown radical structural variations; such S. mutans cells became pleomorphic, unequal in size and some ruptured and divided when treated with Salvia officinalis. The bioassay results also revealed that Salvia officinalis essential oil can perform as a strong antimicrobial agent against S. mutans. Therefore, the current study established that Salvia officinalis is effective in reducing microbial colonization on the surface of the dental implant.

For future studies, essential oils require to be considered as potential future antimicrobials due to their mechanism of action on contact with a bacterial cell. The current research has discovered the following potential future research: exploring new preventative methods such as use essential oils for inhibiting oral pathogen growth affecting dental implants; expanding the use of essential oils in dentistry for biomedical applications; coating titanium implant materials with essential oils to prevent implant failures; investigating how long the essential oil adheres to the surface of a titanium dental implant; and possibly using Salvia officinalis in mouth rinses against Staphylococcus aureus, S. mutans.

Acknowledgment

The authors acknowledge the Centre for Rapid Prototyping and Manufacturing (CRPM) for producing the experimental dental samples.

Financial support and sponsorship

The research project was sustained by the National Research Fund (NRF Scholarship) and the Collaborative Program in Additive Manufacturing (Contract No. CSIR-NLC-CPAM-18-MOA-CUT-01).

Conflicts of interest

There are no conflicts of interest.

Authors’ contributions

SSN conducted the laboratory work and prepared the article, TCD and GL were involved in article preparation.

Ethical policy and institutional review board statement

No ethical clearance was needed, as the research did not involve any patients or individuals in data collection.

Patient declaration of consent

No patients were involved.

Data availability statement

Data are obtainable on reasonable request with the corresponding author’s contact details.

 
  References Top

1.
Resnik R. Misch’s Contemporary Implant Dentistry. e-book. Ottawa: Elsevier Health Sciences; 2020.  Back to cited text no. 1
    
2.
Ramanauskaite A, Becker K, Schwarz F. Clinical characteristics of peri-implant mucositis and peri-implantitis. Clin Oral Implants Res 2018;29:551-6.  Back to cited text no. 2
    
3.
Khammissa RAG, Feller L, Meyerov R, Lemmer J. Peri-implant mucositis and peri-implantitis: Bacterial infection. S Afr Dent J2012;67:70-4.  Back to cited text no. 3
    
4.
Pedrazzi V, Escobar EC, Cortelli JR, Haas AN, Andrade AKPD, Pannuti CM. Antimicrobial mouthrinse use as an adjunct method in peri-implant biofilm control. Braz Oral Res2014;28.  Back to cited text no. 4
    
5.
Chouirfa H, Bouloussa H, Migonney V, Falentin-Daudré C. Review of titanium surface modification techniques and coatings for antibacterial applications. Acta Biomater 2019;83:37-54.  Back to cited text no. 5
    
6.
Costerton JW. Overview of microbial biofilms. J Ind Microbiol 1995;15:137-40.  Back to cited text no. 6
    
7.
Pihlstrom BL, Michalowicz B, Atkinson J, Kingman A. Clinical trials involving oral diseases. Pharm Sci Ency: DDDM2010;15:1-26.  Back to cited text no. 7
    
8.
Högberg LD, Heddini A, Cars O. The global need for effective antibiotics: Challenges and recent advances. Trends Pharmacol Sci 2010;31:509-15.  Back to cited text no. 8
    
9.
Fournomiti M, Kimbaris A, Mantzourani I, Plessas S, Theodoridou I, Papaemmanouil V, et al. Antimicrobial activity of essential oils of cultivated oregano (Origanum vulgare), sage (Salvia officinalis), and thyme (Thymus vulgaris) against clinical isolates of Escherichia coli, Klebsiella oxytoca, and Klebsiella pneumoniae. Microb Ecol Health Dis 2015;26:23289.  Back to cited text no. 9
    
10.
Kavanaugh NL, Ribbeck K. Selected antimicrobial essential oils eradicate Pseudomonas spp. and Staphylococcus aureus biofilms. Appl Environ Microbiol 2012;78:4057-61.  Back to cited text no. 10
    
11.
Dagli N, Dagli R. Possible use of essential oils in dentistry. J Int Oral Health 2014;6:i-ii.  Back to cited text no. 11
    
12.
Pandey AK, Kumar P, Singh P, Tripathi NN, Bajpai VK. Essential oils: Sources of antimicrobials and food preservatives. Front Microbiol 2016;7:2161.  Back to cited text no. 12
    
13.
Predoi D, Iconaru SL, Buton N, Badea ML, Marutescu L. Antimicrobial activity of new materials based on lavender and basil essential oils and hydroxyapatite. Nanomaterials 2018;8:291.  Back to cited text no. 13
    
14.
Swamy MK, Akhtar MS, Sinniah UR. Antimicrobial properties of plant essential oils against human pathogens and their mode of action: An updated review. Evid Based Complement Alternat Med 2016;2016:3012462.  Back to cited text no. 14
    
15.
Hamidpour M, Hamidpour R, Hamidpour S, Shahlari M. Chemistry, pharmacology, and medicinal property of sage (salvia) to prevent and cure illnesses such as obesity, diabetes, depression, dementia, lupus, autism, heart disease, and cancer. J Tradit Complement Med 2014;4:82-8.  Back to cited text no. 15
[PUBMED]  [Full text]  
16.
Ishihara K, Miyakawa H, Hasegawa A, Takazoe I, Kawai Y. Growth inhibition of Streptococcus mutans by cellular extracts of human intestinal lactic acid bacteria. Infect Immun 1985;49:692-4.  Back to cited text no. 16
    
17.
Chen L, Jia L, Zhang Q, Zhou X, Liu Z, Li B, et al. A novel antimicrobial peptide against dental-caries-associated bacteria. Anaerobe 2017;47:165-72.  Back to cited text no. 17
    
18.
Matsumoto-Nakano M. Role of Streptococcus mutans surface proteins for biofilm formation. Jpn Dent Sci Rev 2018;54:22-9.  Back to cited text no. 18
    
19.
Moztarzadeh A. Biocompatibility of implantable materials focused on titanium dental implants. 2017. Available from: https://dspace.cuni.cz/handle/20.500.11956/93643. [Last accessed Sep 17, 2021].  Back to cited text no. 19
    
20.
Abdulgader RS. Examination of the biocompatibility and anti‑ microbial activity of coated dental implants. Masters dissertation, University of the Western Cape. Available from: http://hdl.handle.net/11394/6136.  Back to cited text no. 20
    
21.
Nicholson JW. Titanium alloys for dental implants: A review. Prosthesis 2020;2:100-16.  Back to cited text no. 21
    
22.
Baltatu MS, Tugui CA, Perju MC, Benchea M, Spataru MC, Sandu AV, et al. Biocompatible titanium alloys used in medical applications. Rev Chim 2019;70:4.  Back to cited text no. 22
    
23.
Desam NR, Al-Rajab AJ, Sharma M, Mylabathula MM, Gowkanapalli RR, Albratty M. Chemical constituents, in vitro antibacterial and antifungal activity of Mentha × Piperita L.(peppermint) essential oils. J King Saud Univ Sci 2019;31:528-33.  Back to cited text no. 23
    
24.
Ntondini SS, Lenatha GG, Dzogbewu TC. Inhibitory effect of essentials oil against oral pathogens. Int J Dent Oral Sci 2021;8:1018-23.  Back to cited text no. 24
    
25.
Kock JL, Sebolai OM, Pohl CH, van Wyk PW, Lodolo EJ. Oxylipin studies expose aspirin as antifungal. FEMS Yeast Res 2007;7:1207-17.  Back to cited text no. 25
    
26.
Zainal-Abidin Z, Mohd-Said S, Adibah F, Majid A, Mustapha WA, Jantan I. Anti-bacterial activity of cinnamon oil on oral pathogens. Open Conf Proc J2013;4:12-16.  Back to cited text no. 26
    
27.
Bouyahya A, Abrini J, Dakka N, Bakri Y. Essential oils of origanum compactum increase membrane permeability, disturb cell membrane integrity, and suppress quorum-sensing phenotype in bacteria. J Pharm Anal 2019;9:301-11.  Back to cited text no. 27
    
28.
Al-Bayaty F, Taiyeb-Ali T, Abdulla MA, Hashim F. Antibacterial effect of chlorine dioxide and hyaluronate on dental biofilm. Afr J Microbiol Res 2010;4:1525-31.  Back to cited text no. 28
    
29.
Al-Bayaty FH, Taiyeb-Ali TB, Abdulla MA, Mahmud ZB. Antibacterial effects of Oradex, Gengigel and Salviathymol-n mouthwash on dental biofilm bacteria. Afr J Microbiol Res 2011;5:636-42.  Back to cited text no. 29
    
30.
Chua CK, Leong KF, Lim CS. Rapid Prototyping: Principles and Applications (with Companion CD-ROM). Singapore: World Scientific Publishing Company; 2010.  Back to cited text no. 30
    
31.
Tao W, Leu MC. Design of lattice structure for additive manufacturing. International Symposium on Flexible Automation (ISFA) 2016;1:325-32.  Back to cited text no. 31
    
32.
Dzogbewu TC. Additive manufacturing of porous Ti-based alloys for biomedical applications—A review. J New Gener Sci 2017;15:278-94.  Back to cited text no. 32
    
33.
Yadroitsev I, Shishkovsky I, Bertrand P, Smurov I. Manufacturing of fine-structured 3D porous filter elements by selective laser melting. Appl Surf Sci 2009;255:5523-7.  Back to cited text no. 33
    
34.
Dzogbewu T, Yadroitsev I, Krakhmalev P, Yadroitsava I, Du Plessis A. Optimal process parameters for in-situ alloyed Ti15Mo structures by direct metal laser sintering. A paper presented at SSF 2017-The 28th Annual International Solid Freeform Fabrication Symposium. Austin: University of Texas; 2017. p. 75-96.  Back to cited text no. 34
    
35.
Popa M, Măruțescu L, Oprea E, Bleotu C, Kamerzan C, Chifiriuc MC, et al. In vitro evaluation of the antimicrobial and immunomodulatory activity of culinary herb essential oils as potential perioceutics. Antibiotics 2020;9:428.  Back to cited text no. 35
    
36.
Niinomi M. Mechanical properties of biomedical titanium alloys. Mater Sci Eng 1998;243:231-6.  Back to cited text no. 36
    
37.
Dzogbewu TC, Monaheng L, Els J, van Zyl I, Du Preez WB, Yadroitsava I, et al. Evaluation of the compressive mechanical properties of cellular DMLS structures for biomedical applications. In 17th Annual Conference of the Rapid Product Development Association of South Africa, Vaal University of Technology, Johannesburg, November 2-6; 2016.  Back to cited text no. 37
    
38.
Serra E, Hidalgo-Bastida LA, Verran J, Williams D, Malic S. Antifungal activity of commercial essential oils and biocides against Candida albicans. Pathogens 2018;7:15.  Back to cited text no. 38
    
39.
Rokaya D, Srimaneepong V, Wisitrasameewon W, Humagain M, Thunyakitpisal P. Peri-implantitis update: Risk indicators, diagnosis, and treatment. Eur J Dent 2020;14:672-82.  Back to cited text no. 39
    
40.
Figuero E, Graziani F, Sanz I, Herrera D, Sanz M. Management of peri-implant mucositis and peri-implantitis. Periodontology 2014;66:255-73.  Back to cited text no. 40
    
41.
Hamilton WL, Wenlock R. Antimicrobial resistance: A major threat to public health. Cambridge Med J2016;1-10.  Back to cited text no. 41
    
42.
Lambert PA. Bacterial resistance to antibiotics: Modified target sites. Adv Drug Deliv Rev 2005;57:1471-85.  Back to cited text no. 42
    
43.
Smith RA, M’ikanatha NM, Read AF. Antibiotic resistance: A primer and call to action. Health Commun 2015;30: 309-14.  Back to cited text no. 43
    
44.
Warnke PH, Becker ST, Podschun R, Sivananthan S, Springer IN, Russo PA, et al. The battle against multi-resistant strains: Renaissance of antimicrobial essential oils as a promising force to fight hospital-acquired infections. J Craniomaxillofac Surg 2009;37: 392-7.  Back to cited text no. 44
    
45.
Chouhan S, Sharma K, Guleria S. Antimicrobial activity of some essential oils—Present status and future perspectives. Medicines 2017;4:58.  Back to cited text no. 45
    
46.
Imane NI, Fouzia H, Azzahra LF, Ahmed E, Ismail G, Idrissa D, et al. Chemical composition, antibacterial and antioxidant activities of some essential oils against multidrug resistant bacteria. Eur J Integr Med 2020;35:101074.  Back to cited text no. 46
    
47.
Sridhar S, Wilson TG Jr, Palmer KL, Valderrama P, Mathew MT, Prasad S, et al. In vitro investigation of the effect of oral bacteria in the surface oxidation of dental implants. Clin Implant Dent Relat Res 2015;17(Suppl 2):e562-75.  Back to cited text no. 47
    
48.
Adams RP. Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry. Carol Stream, IL: Allured Publishing Corporation; 2007.  Back to cited text no. 48
    
49.
Beheshti-Rouy M, Azarsina M, Rezaie-Soufi L, Alikhani MY, Roshanaie G, Komaki S. The antibacterial effect of sage extract (Salvia officinalis) mouthwash against Streptococcus mutans in dental plaque: A randomized clinical trial. Iran J Microbiol 2015;7:173-7.  Back to cited text no. 49
    
50.
Zhang Y, Liu X, Wang Y, Jiang P, Quek S. Antibacterial activity and mechanism of cinnamon essential oil against Escherichia coli and Staphylococcus aureus. Food Control 2016;59:282-9.  Back to cited text no. 50
    
51.
Tariq S, Wani S, Rasool W, Shafi K, Bhat MA, Prabhakar A, et al. A comprehensive review of the antibacterial, antifungal and antiviral potential of essential oils and their chemical constituents against drug-resistant microbial pathogens. Microb Pathog 2019;134:103580.  Back to cited text no. 51
    
52.
Tardugno R, Pellati F, Iseppi R, Bondi M, Bruzzesi G, Benvenuti S. Phytochemical composition and in vitro screening of the antimicrobial activity of essential oils on oral pathogenic bacteria. Nat Prod Res2018;32:544-51.  Back to cited text no. 52
    


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