Comparative Evaluation of Viability and Function of Albino Rabbit Periodontal Ligament Fibroblasts in Various Media (In Vitro Study)

Abstract

Background: Tooth avulsion is one of traumatic injuries that cause damage to periodontal ligament (PDL) tissues. Avulsed tooth should be stored in a suitable storage medium to preserve the vitality of the PDL fibroblasts (PDLFs). Aim: To assess and compare the effect of various media on albino rabbit’s PDLFs viability and basic fibroblast growth factor (b-FGF) protein and gene expressions at different time intervals. Material and Methods: Periodontal ligament stem cells (PDLSCs) were isolated from molars of albino rabbit then differentiated into PDLFs. The PDLFs were cultured in different media (contact lens solution, skimmed milk, Dentosafe and alcohol free mouthwash) for 1h, 12h and 24h. Phosphate buffered saline (PBS) was used as control medium. Cell viability was assessed by MTT assay. b-FGF protein and gene expression levels were detected by immunofluorescence (IF) staining and Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) respectively. Data from MTT assay and H-score of IF staining were statistically analyzed using ANOVA test and multiple two groups comparisons pos hoc test. Results: Examination of isolated PDLSCs at different passages showed various shapes of cells from rounded to spindle shaped. The differentiated PDLFs showed positive IF reaction. Statistical analysis of tested media at different time intervals represented that; PBS had the highest mean of fibroblastic cell viability% and H-score, followed by Dentosafe, skimmed milk, contact lens solution and mouthwash respectively. Gene expression of b-FGF was up-regulated in Dentosafe and skimmed milk while it was down-regulated in mouthwash. Conclusions: Dentosafe is more effective in maintenance of PDLFs viability and function than the other tested media. Mouthwash is not recommended as a storage media for PDLFs.