Comparative Evaluation of Efficacy of Three Different Storage Media in Maintaining the Viability of Periodontal Ligament Cells: An In Vitro Study

  • Venu Verma
  • Sundeep Hegde K
  • Sham S Bhat
Keywords: Casein phosphopeptide amorphous calcium phosphate, Hank’s balanced salt solution, Oral rehydration solution, Storage Medium


Introduction: Dental trauma is the most common injury which occurs in oro-facial region. Traumatic dental injuries are often seen among injuries to the face. Among them, tooth avulsion (0.5%-16%) is a complex traumatic injury characterized by the rupture of the neurovascular bundle and periodontal ligament (PDL) exposing the tooth to the outer environment. It occurs most often in the age group of 7-10 years, when the alveolar bone is resilient and offers minimal resistance to extrusive forces. Avulsion is a potential threat to the vitality of Periodontal ligament cells which are essential for the healing of replanted avulsed teeth. Hence management protocols should include management of the pulp and the periodontal ligament cells in the long-term survival and prognosis of avulsed teeth.2 The types of healing that takes place after the avulsion injury are as follows: 1.Favorable healing: a. Healing with a normal periodontal ligament (without root resorption) b. Healing with surface resorption (repair-related resorption) 2.Unfavorable healing: a. Healing with ankylosis (replacement). b. Healing with inflammatory resorption (infection related resorption).3 Two of the most critical factors affecting the prognosis of an avulsed tooth after replantation are extra oral dry time and the storage medium in which the tooth is placed.4
As replantation of avulsed teeth occurs more frequently between 1 and 4 hours after avulsion, degeneration of cemental periodontal ligament fibers is a common event and the presence of necrotic Periodontal ligament remnants on root surface stimulates the occurrence of inflammatory root resorption, which is the major cause of loss of replanted teeth.5
Secondly, storage or transport medium to support cell viability is more important than the extra oral Introduction: An ideal storage medium should be one that is capable of preserving the viability, mitogenicity and clonogenic capacity of the damaged Periodontal ligament cells to facilitate proliferation of these cells over the denuded root surface, thereby preventing further root resorption.
Aim: The aim of this study is to evaluate the effectiveness of Casein phosphopeptide amorphous calcium phosphate as a storage media for avulsed tooth in maintaining periodontal ligament cell viability in comparison with Hank’s Balanced Salt Solution and Oral Rehydration Solution.
Materials and Methods: Forty freshly extracted human premolar teeth with normal periodontium and closed apices were taken. Forty teeth were randomly assigned into five experimental groups. It was then incubated for 30 minutes in falcon tubes with 2.5 ml solution of 0.2 mg/ml of collagenase II and 2.4 mg/ml solution of dispase grade II in phosphate buffered saline. After incubation, 50 μl of fetal bovine serum was added to each tube with the help of micropipette. Cells were labelled with 0.4% trypan blue for determination of viability. The number of viable cells in a grid of Neubauer’s chamber were counted under a light microscope at 40X magnification.
Results: Results were analysed using Kruskul-Wallis test and Mann-Whitney U test.
Conclusion: GC Tooth Mousse, Hanks balanced salt solution and Oral rehydration solution can be used as storage medium. GC Tooth Mousse is better than Hanks balanced salt solution and Oral rehydration solution as a storage medium.


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Venu Verma, Sundeep Hegde K, & Sham S Bhat. (2017). Comparative Evaluation of Efficacy of Three Different Storage Media in Maintaining the Viability of Periodontal Ligament Cells: An In Vitro Study. International Healthcare Research Journal, 1(5), 25-34. Retrieved from
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