ABSTRACT
Objectives:
This research was done to assess the periodontal ligament cells viability using propolis, coconut water, aloe vera, and soy milk storage media.
Materials and Methods:
Periodontal ligament cells were taken from freshly extracted teeth and stored into four groups of storage media: Group A-propolis, Group B-coconut water, Group C-aloe vera, and Group D-soy milk. Later, the cell viability was assessed at 1, 12, and 24 hours.
Results:
The periodontal ligament cell viability was significantly greater in propolis, followed by coconut water, aloe vera, and soy milk.
Conclusion:
We found that propolis, coconut water, aloe vera, and soy milk can be used as an effective storage media. These are naturally occurring and easily available storage medium.
KEYWORDS: Aloe vera, avulsion, coconut water, propolis, soy milk, storage media
INTRODUCTION
Tooth avulsion accounts for 1%–16% of all dental injuries that affect the permanent teeth. The age range of 7–10 years old is most affected by this injury as a result of the participation of kids in contact sports. Immediate tooth replacement improves PDL repair and significantly lowers root resorption occurrence.[1]
In case of delayed reimplantation condition, avulsed teeth can be saved in suitable tooth storage media. The extra oral dry time and tooth storage media are the two most important factors in the success of replanted avulsed teeth.[2] There have been investigations into a variety of wet storage media for avulsed teeth.[1,2]
Hank’s balanced salt solution (HBSS) has been developed for preservation of cell, and it helps in preservation of tissues for longer duration. Since it is expensive and not easily available, alternative storage media were tried, such as Viaspan, culture media—Minimum Essential Medium (MEM), saline, bovine milk, water, green tea, saliva, egg white, propolis, and coconut water; rehydrating solutions, like Gatorade solutions.[1]
Propolis is a biological substance that is non-toxic and has properties that are antibacterial, anti-inflammatory, antioxidant, antifungal, and tissue regenerative.[3] Iron and zinc are also present in propolis, both of which are crucial for collagen synthesis.[1] The viability of PDL cells was found to be most effectively maintained by coconut water when compared to other media like HBSS, propolis, and milk. Coconut water contains many nutrients.[3] A. vera (Barbadensis) gel has an antibacterial, anti-inflammatory, antioxidant, immune-stimulating, and hypoglycemic properties.[2] Soy milk is advised as a storage medium because recent studies have shown that it promoted good cell viability when in contact with periodontal ligament cells, comparable to HBSS and milk.[3]
There are limited reported researches on natural storage medias; hence, the current research was planned to assess the periodontal ligament cell viability using propolis, coconut water, aloe vera, and soy milk storage media.
MATERIALS AND METHODS
In this current in vitro research, freshly extracted premolar teeth for orthodontic purpose with intact PDL fibers were used for this research. These teeth were stored in each group of storage media: Group A-propolis, Group B-coconut water, Group C-aloe vera, and Group D-soy milk.
Using a mortar and pestle, solid propolis was ground into tiny particles. Propolis (50%) storage media was made by mixing 250 ml of a 0.4% ethanol solution with 50 mg of ground propolis.[2] Coconut water is obtained from fresh coconut and stored for later usage. Soybeans are soaked, ground, and then boiled to create soy milk, which is then filtered to remove any remaining impurities. Aloe vera medium was prepared according to Babaji et al.[2] study.
Immediately after extraction, the coronal third of the PDL was scraped with a curette and damaged cells were removed by holding only crown portion. Then the teeth were kept in four different kinds of storage for 1, 12, and 24 hours. Later, scraped PDL cells were incubated in the Falcon tubes and centrifuged for five minutes at 800 rpm.
The centrifuged residue was collected, and the supernatant was discarded. This was thoroughly mixed with an equal volume of 0.4% Trypan blue stain. Non-viable cells are stained blue by Trypan blue, while healthy cells are either colorless or pink. After staining, the cells were examined under an optical microscope using a hemocytometer.
Using the SPSS package (22.0 version, Inc.; Chicago, USA) and one-way ANOVA, post-hoc test, and Mann–Whitney test, the obtained data was statistically evaluated with P > 0.05.
RESULTS
The mean periodontal cells after 24 hours of storage were 138.6 cumm in Group A, 132.3 cumm in Group B, 122.7 cumm in Group C, and 110.2 cumm in Group D, and the difference was statistically significant [Table 1] (p<0.001). The average absorbance value recorded at different time intervals in various groups was considerable [Table 2]. The periodontal ligament cell viability was significantly higher in propolis, followed by coconut water, aloe vera, and soy milk.
Table 1.
Comparison of mean viable and non-viable periodontal cells among various groups
| Group | Mean viable cells (cumm) | Mean non-viable cells (cumm) | SD | SE | P |
|---|---|---|---|---|---|
| Group A propolis | 138.6 | 24.9 | 3.12 | 1.40673 | 0.001 |
| Group B coconut water | 132.3 | 26.7 | 4.43 | 1.20456 | |
| Group C aloe vera | 122.7 | 31.8.5 | 4.08 | 0.87452 | |
| Group D soy milk | 110.2 | 36.3 | 3.12 | 0.98563 |
P<0.05, SD=standard deviation, SE=standard error, one=way analysis of variance, Tukey’s test. cumm=cubic millimeter
Table 2.
Average absorbance values at various time periods
| Time | Group A | Group B | Group C | Group D | P |
|---|---|---|---|---|---|
| 1 hour | 140.2 | 136.7 | 127.4 | 118.3 | 0.05 |
| 12 hours | 139.4 | 134.4 | 124.5 | 115.8 | |
| 24 hours | 138.6 | 132.3 | 122.7 | 110.2 |
Test used=Mann–Whitney test, P<0.05
DISCUSSION
The current research was done to assess the periodontal ligament cell viability using propolis, coconut water, aloe vera, and soy milk storage media. Aloe vera extract can be suggested as a appropriate storage medium for avulsed teeth, according to Babaji et al.[2]
Propolis had the highest level of periodontal cell viability, followed by Viaspan solution, aloe vera, and Gatorade solution, according to Misurya et al.[4] Bijlani et al.[5] concluded that propolis was the most efficient medium for maintaining PDL cell viability compared to 30% aloe vera extract and ice apple water.
According to Thomas et al.,[6] coconut water may be a better substitute to milk or HBSS in terms of preserving viability of cells and to store avulsed teeth. Gopikrishna et al.[7] concluded that coconut water kept considerably more PDL cells viable compared with HBSS, propolis, and milk.
Soy milk and coconut water demonstrated hopeful results as storage media for avulsed teeth, and maintaining the viability for up to 24 hours, according to Moura et al.[8]
The limitation of the current study is an in vitro study. Further studies are needed to validate the results.
CONCLUSION
We found higher viable periodontal ligamental cells with propolis, followed by coconut water, aloe vera, morus rubra, and soy milk. These natural media are easily available and economical for management of avulsed teeth.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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