Brief Resume of the intended work :
6.1 Need for the study :
According to the World Health Organization classification for traumatized teeth, avulsion or exarticulation is the complete displacement of a tooth from its alveolar socket due to traumatic injury.1 The viability of the periodontal ligament fibroblast is critical for successful healing of replanted avulsed teeth.2 When an injury occurs, the avulsed tooth should be replanted immediately to prevent further injury to periodontal ligament cells.. 1
Two factors that have a profound effect on the prognosis of replanted teeth are extra oral time of the avulsed tooth and the medium used to preserve the tooth before replantation.2
Storage media such as milk, saline, water, saliva, culture media and Hanks Balanced Salt Solution (HBSS) have been suggested to preserve the viability of PDL cells.1
The American Association of Endodontists recommends the use of HBSS as the storage medium of choice for the treatment of avulsed teeth. However, HBSS is not available in most places where these traumatic events usually occur.
The aim of the present study is to evaluate the effectiveness of Aloevera as a storage media for avulsed tooth in maintaining the periodontal cell viability along with Rice water & Egg white.
6.2 Review of literature :
Trauma to permanent and deciduous dentition is a common problem faced by dental practitioners. Injuries to the PDL can result in tooth avulsion. If replantation is done immediately, the tooth will remain in the dental arch without resorption and with normal function. The highest percentage of success can be achieved by immediate replantation or replantation within 30 minutes of avulsion . If the tooth cannot be immediately replanted , the viability PDL cells on the surface of the root can be maintained in a suitable transport medium which will lead to a decreased incidence of root resorption .1,2
Blomlof et al considered milk as a gold standard for transporting avulsed tooth to the laboratory. It has the physiologic osmolality similar to that of serum and contains fewer bacteria as compared to the saliva.3
Collagenase and dispase enzymes disrupt the extracellular matrix and cause the release of cells without disruption and destruction of their own membrane. Further this method is more representative of the actual clinical situation; because the cells are not subjected to long processing times to determine their viability status.4
Aleo vera is remarkable because it is one of the most durable plants known to mankind. The plant has a miraculous ability to self seal and heal, and contains all the nutrients within its leaf in order to survive. In recent studies human kidney cells were utilized to determine the effectiveness of aleo vera on cellular longevity. The cellular death rate was found to be reduced when cultured with the Aloe gel.5
Rice is an extremely healthy food for a number of reasons. Rice has low sodium content and contains useful quantities of potassium, Vitamin B, thiamin and niacin. In the Medicinal Book of Malayan Medicine, it is prescribed that boiled rice "greens" can be used as an eye lotion and for use with acute inflammation of the inner body tissues. Rice water is prescribed by the Pharmacopoeia of India as an ointment to counteract inflamed surface.6
Egg white can be a suitable storage medium for an avulsed tooth because it has a ph of 8.6 and 258 osmolality and is more likely to be available at the site of a traumatic event .1
Several techniques have been used to determine the viability of the periodontal cells following avulsion. However, most of the experiments use techniques in which cell are cultured and /or trypsinized for longer periods of time. Because the extra cellular matrix has a high content of collagen and other proteins, it seems reasonable that the use of enzymatic degradation would provide a greater number of cells within a shorter time frame.
Thus the present study aims at evaluating aloevera, rice water and egg white as storage media for avulsed tooth using collagenase assay and trypan blue exclusion test.
6.3 Objectives of the study :
To evaluate the efficacy of Aloe vera, Rice water and Egg white, as storage media for transporting an avulsed tooth.
To compare the efficacy of Aloe vera, Rice water and Egg white , as storage media for transporting an avulsed tooth.
Material and Methods:
Source of Data:
Aloe vera, Rice water, Egg white
Forty non-carious premolar teeth, extracted for orthodontic therapeutic purposes will be selected for the study. Immediately after the extraction, teeth will be left in mud for 15 minutes to simulate an avulsion injury. Then the teeth will be collected and randomly assigned to four groups of 10 teeth each:
Group I : Teeth specimens stored in Milk (Control )
Group II : Teeth specimens stored in Aloevera
Group III: Teeth specimens stored in rice water
Group IV: Teeth specimens stored in egg white
All the experimental and control group specimens will be stored for a time period of 30 minutes. Following 30 minute of storage in media, the teeth will be rinsed in saline and the root surfaces will be scraped with the scalpel leaving the cervical third of root surface intact. These periodontal ligament scrapings will be collected in falcon tubes having collagenase in 2.5 ml of phosphate buffer saline and this will be incubated for 30 minutes and centrifuged for 5minutes at 800 rpm. Then the PDL cells will be stained with Trypan Blue and will be observed under optical microscope.
The cell count will be done using the following formulae
: Total cells - Stained cells x 100
All the results will be tabulated and statistically analyzed by Kruskal-Wallis test for inter-group comparison and Mann-Whitney Test for group wise comparison.
7.3 Does the study require any investigation or intervention to be conducted on patients or other humans or animals? If so please describe briefly:
Yes, this study requires therapeutically extracted human permanent premolars.
7.4 Has ethical clearance been obtained from your institution in of “ yes” for
Yes, the recommendations of ethical committee are being attached.
Khademi AA, Saeed SA. New Storage Medium for an Avulsed Tooth. The Journal of Contemporary Dental Practice 2008;9(6):25-32
Saad Al-Nazhan, Afaf Al-Nasser. Viability of Human Periodontal Ligament Fibroblasts in Tissue Culture After Exposure to Different Contact Lens Solutions. The Journal of Contemporary Dental Practice 2006;7(4):1-9.
Blomlof H, Lindskog S, Andersson L, Hedstrom KG, Hammarsstrom L. Storage of experimentally avulsed teeth in milk prior to replantation. J Dent Res 1983;62(8):912-6.
Pillegi R, Dumsha TC, Nor JE. Assessment of post traumatic PDL cell viability by a novel collagenase assay. Dent Traumatol 2002;18:186-9.
Wolfe B. Aloe Vera: An ancient plant for modern dentistry.[Online] [Cited 2008 Nov 6]. Available from: URL: http://www. dr wolfe. Com
Nutritional facts about rice[Online] [Cited 2008 Nov 6]. Available from : URL:http://www.rice-trade.com/rice-nutritional-facts.html.