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ORIGINAL ARTICLE
Year : 2017  |  Volume : 7  |  Issue : 1  |  Page : 36-39

Efficacy of garlic extract and sodium hypochlorite on dental pulp dissolution: An in vitro study


1 Department of Conservative Dentistry and Endodontics, Mamata Dental College, Khammam, Telangana, India
2 Department of Microbiology, Mamata Medical College, Khammam, Telangana, India

Date of Web Publication10-Jan-2017

Correspondence Address:
Lakkireddy Sunitha
Department of Endodontics, Mamata Dental College, Khammam - 507 002, Telangana
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1658-5984.197986

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  Abstract 

Aim: The present study was aimed to assess and compare the human pulp dissolution capacity of sodium hypochlorite (NaOCl) (2.5%) and garlic extract.
Materials and Methods: Several pulp tissues are obtained from both endodontically treated teeth and also by orthodontically extracted premolar teeth and were sectioned with a surgical blade into seventy-five pieces of same size and weight. Pulp tissues were then divided into Allium sativum extract (ASE) group, NaOCl group, and NaCl group and tissues were immersed in the test solutions for 30, 60, and 90 min, respectively. Further tissues were weighed using a precision balance at tested intervals, and the percentage weight loss was calculated and statistically analyzed.
Results: In this study, pulp tissue treated with 2.5% NaOCl solution at all tested time intervals showed significant (P < 0.001) pulp tissue dissolution capacity and a maximum of dissolution was observed at 90 min. Pulp tissue treated with ASE showed little dissolution ability at all tested time periods. Whereas, 0.9% NaCl showed no ability to dissolve human pulp tissue.
Conclusion: It can be concluded that 2.5% NaOCl had the maximum tissue dissolving capacity when compared to different concentrations of ASE. Saline had no effect on the human pulp dissolution.

Keywords: Allium sativum extract, dental pulp, sodium hypochlorite, tissue dissolution capacity


How to cite this article:
Rao S A, Sunitha L, Rao B N, Naik J P, Shekar V C. Efficacy of garlic extract and sodium hypochlorite on dental pulp dissolution: An in vitro study. Saudi Endod J 2017;7:36-9

How to cite this URL:
Rao S A, Sunitha L, Rao B N, Naik J P, Shekar V C. Efficacy of garlic extract and sodium hypochlorite on dental pulp dissolution: An in vitro study. Saudi Endod J [serial online] 2017 [cited 2023 Feb 5];7:36-9. Available from: https://www.saudiendodj.com/text.asp?2017/7/1/36/197986


  Introduction Top


The practice of endodontics has been revolutionized with the advancements in techniques and materials; this resulted in an increase in the success rate of endodontic treatment over the past few decades. Several studies have shown that mechanical instrumentation of the root canal produces a smear layer which consists of residual pulp tissue, infected dentin or bacteria in the root canal which may result in endodontic treatment failure.[1] Disinfecting and cleaning by chemo-mechanical procedures are the prerequisites for successful root canal treatment.[2] With advanced instrumentation technology, though root canal shaping can be efficiently attained, effective cleaning of the entire root canal system, still, remains a challenge.[3]

It is an endodontic tenet that the chance of successful root canal treatment directly depends on an irrigant that aids in removal of organic and inorganic debris from the root canal system. A wide range of synthetic solutions has been used over the years as endodontic irrigants. Different auxiliary chemical agents for tissue dissolution have been proposed (sodium hypochlorite [NaOCl], chlorine dioxide, calcium hypochlorite, peracetic acid).[4],[5] NaOCl most commonly used in endodontics has been proved as an effective agent against a broad spectrum of bacteria and has the capability to dissolve vital as well as necrotic tissue.[6] However, NaOCl with alkaline pH of approximately 11–12 shows toxic effects on organic tissues, ranging from skin ulceration to rupture or destruction of the red blood cells leading to hemolysis, eventually leading to cell death or necrosis.[7] Because of the constant increase in antimicrobial resistance and side effects caused by synthetic agents, there is a need for alternative agents. The natural products made from the medicinal plants have shown immense potential as an alternate source of combating infections in human beings.[8]

Herbs or natural products in dentistry have become more popular due to their easy availability, cost effectiveness, low toxicity, and lack of microbial resistance and increased shelf life.[9] Garlic (Allium sativum) is a bulbous perennial medicinal plant which belongs to the family Liliaceae and widely used in medicine for its antibacterial, antifungal, antiviral, antimutagenic, antioxidative, anticancerous, antiageing, immunomodulatory effects.[10] Previous investigations on garlic have reported its main active compound is allicin which is a sulfurous compound responsible for all the properties of garlic.[11] To date, there are no studies reported on the human pulp tissue dissolving effect of garlic. Hence, the purpose of this present study was to evaluate the tissue dissolution effect of garlic and compare it with NaOCl on human pulp tissue.


  Materials and Methods Top


Preparation of extract

The dried seeds of AS commonly known as garlic were commercially procured. The seeds of AS were cleaned, shade-dried, and powdered and subjected to sequential extraction using Soxhlet apparatus. AS was extensively extracted in distilled water at 60°C and dried at 50°C. The aqueous extract thus obtained was then cooled and concentrated by evaporation and this stock of extract was stored at –20°C until use. The pH value of the fresh AS was <7.

Preparation of pulp tissue and dissolution test

The present study was carried out with the prior approval from the Institutional Ethics Committee (Ref No. MDC_R_088116). The pulp tissues used in this study were obtained from endodontically treated teeth and also from freshly obtained noninfected premolar teeth collected from individuals who underwent various orthodontic procedures. After the extraction, periodontal tissues were removed with a brush and these teeth were placed in distilled water containing 0.2% sodium azide. Each tooth was then split into two using micromotor in buccolingual direction. The tooth pulps were removed carefully by an anatomic forceps and washed in distilled water to remove the debris and blood and stored in distilled water at 4°C until use. Thereafter, immediately before the dissolution tests, the entire tissue was blotted dry and sectioned into seventy-five pieces with a surgical blade of similar size, shape, with an average weight of 20 mg [12] which was determined using precision balance in an airtight container (Sartorius Practum 124-1S analytical).

Pulp tissues were then divided randomly into five groups and with each group containing 15 pulp tissues.

  • Allium sativum extract (ASE) Group: Pulp specimens (n = 45) were placed in 5 ml of 10, 20, and 30 mg/ml ASE solution
  • NaOCl group: Pulp specimens (n = 15) were placed in 5 ml of 2.5% NaOCl solution (Merck Pvt Ltd., India)
  • NaCl group: Pulp specimens (n = 15) were placed in 5 ml of 0.9% NaCl solution (Merck Pvt Ltd., India).


Each group had three subgroups having five pulp tissues each (n = 5) according to different time periods for which the sample is immersed in the irrigant, i.e., 30, 60, and 90 min. The pulp specimens in each group were immersed in test solution and are placed in a vibrator to agitate the test solution. The pulp tissues placed in all the three experimental groups were weighed after the passage time of 30, 60, and 90 min. All the tissue pieces were washed with distilled water to remove any precipitate of the test solution, blotted on a filter paper, dried, and then weighed. All the samples were weighed by a single investigator who was unaware of the test solutions. The difference in weight of the tissue sample before and after exposure to the test solution was divided by the original weight and multiplied by 100 to obtain the percentage of tissue weight loss or gain.

Statistical analysis

The experimental data were expressed as mean ± standard error of mean from the data of a minimum of three separate experiments. The statistical analysis was performed using Statistical Package for Social Sciences software version 16.0 (Chicago, SPSS Inc). Analysis of variance was used to compare the differences between treatments groups (P < 0.05 considered as a significant difference).


  Results Top


Human pulp tissue treated with 0.9% NaCl showed no ability of dissolution. Whereas, pulp tissue treated with 2.5% NaOCl solution at all tested time intervals showed significantly (P < 0.001) pulp tissue dissolution capacity when compared with other groups. ASE-treated pulp tissue at various time periods showed little dissolution ability [Table 1]. There was no statistically significant (P > 0.05) difference noted between different concentrations of ASE solution at all the time intervals tested.
Table 1: Intercomparison of the pulp tissue dissolving potential of the test solutions at different time intervals

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  Discussion Top


Chemomechanical preparation forms the key requisite for the success of root canal instrumentation.[1] Callahan and Grossman group has demonstrated the importance of the solvent ability of endodontic irrigant and emphasized the elimination of pulp tissue from the root canal.[11],[13] As a sign of continued concern in this field encouraged to attempt on a known dietary antioxidant garlic for its ability of pulp dissolution.

The dissolving capacity of any irrigant depends on the amount of irrigant, type and contact time with the tissues, area, and mass of the organic tissue.[1] Therefore, in the present study, these factors were rigorously standardized using the same volume of solutions and human pulp tissue of approximately the same shape and weight. In the present study, a precision scale is used to evaluate the weight of the pulp tissue. The pulp tissue is weighed before and after its contact with the irrigating solutions. This method is more accurate because the exact dissolution of the tissue can be calculated by subtracting the weight of residual pulp from the weight of the pulp before contact with the irrigating solution.

The amount of pulp dissolution can be assessed using various methods like analyzing the dissolved material for hydroxyproline content,[14] measuring the amount of total phosphate extracted from the dissolved material, loss of tissue collagen,[15] and dissolution by agitation method.[16] None of the above methods proved to be the dissolution of whole tissue, as these methods assess the amount of tissue dissolution based on one content which is the major drawback. Therefore, in this study, a simple, yet reliable, dry weight method was used to quantify specimen dissolution. Earlier several studies have demonstrated the efficacy of various solutions on tissue dissolving capacity using different types of organic tissues such as bovine pulp,[17] human umbilical cord,[18] pig palatal mucosa,[19] rat dermal connective tissue,[20] and rabbit liver. However, none of these tissues are relevant to the human pulp tissue.

In the present study, human pulp tissue was selected to assess the tissue dissolution in response to various irrigants. Present study results showed that 2.5% NaOCl solution at all tested time intervals showed significantly more pulp tissue dissolution. Our observations are in accordance with various other studies that have reported NaOCl to be an effective pulp tissue dissolving agent.[4],[5],[6] The tissue dissolution property of NaOCl is due to the presence of their free available chlorine in the solution. However, the successful use of NaOCl in medical practice is not appreciated much due to its toxic side effects at their optimal dose that limit their use in clinics.[21] Therefore, the quest for effective, nontoxic compound with its optimal tissue dissolution capability is of immediate need which could be more clinically useful for the patients undergoing endodontic procedures. In view of this, the interest was shifted more on the naturally occurring dietary antioxidants. Various natural plant extracts have been reported for antimicrobial effect suggesting their ability to be used as an endodontic irrigant.[22],[23]

Garlic has been reported to possess various properties such as antibacterial, antioxidant, anti-inflammatory, and immune modulatory actions.[10] ASE is also one of the strongest disinfectants with antimicrobial property, but a comprehensive quantitative study assessing human pulp tissue dissolution was missing. In the present study, an attempt has been made to screen the tissue dissolution potential of garlic extract. Studies have shown that factors such as concentration, duration of exposure, area of application, freshness of garlic and individual reactivity contribute to the development of burn wounds.[24],[25],[26] Hence, based on the earlier observations, in the present study lower concentrations of ASE was used. Treatment of pulp tissue with ASE showed the pulp dissolving capacity to some extent at 30 mg/ml ASE at 90 min when compared with 20 and 10 mg/ml, but there was no significant difference in between tested ASE concentrations. ASE showed lower pulp dissolving capacity at 30 min. Studies have suggested that pH, concentration and incubation time might have an effect on tissue dissolution.[11] Several earlier studies have shown that higher pH levels of a solution resulted in greater tissue dissolution. In addition, they also suggested that as more pH is lowered, more time is needed for a solution to dissolve the tissue.[27] Since the pH of ASE is very low (<7) this might be the reason for less effectiveness of ASE in dissolution of pulp tissue at 30 min than at 60 and 90 min. With increase in contact time, from 30 to 90 min, there is increase in the tissue dissolution, thus indicating that high concentrations should be used if the contact time is less. The present study also shows that 0.9% NaCl exhibited no ability to dissolve human pulp tissue.

Tissue dissolution of ASE is attributed to its characteristic organosulfur compounds such as allicin which is the principle active substance of fresh garlic extract may be one of the factors responsible for its tissue dissolution activity.[28] The enzyme alliinase present in garlic converts alliin to allicin, immediately after it is crushed. Allicin participates in the metabolism of cysteine in proteins, thus showing the potential to disrupt the epidermal junction and cause coagulative necrosis of the tissues.[29] Based on the results, of this study 2.5% NaOCl is the most effective pulp tissue solvent whose action increases with time. Further clinical studies are required to substantiate the findings of this study.


  Conclusion Top


To conclude, 2.5% NaOCl had the maximum tissue dissolving capacity at 90 min when compared to different concentrations of ASE. ASE of 30 mg/ml at 90 min dissolved the human pulp tissue more than 10 mg/ml and 20 mg/ml of ASE. Saline had no effect on the human pulp tissue dissolution.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
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