In vitro viability, mitogenicity and clonogenic capacities of periodontal ligament fibroblasts after storage in four media supplemented with growth factors

The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objective of this stu dy was to evaluate the effectiveness of growth factors (IGF-1 and PDGF-BB) when added to storage media in preserving the functional abilities of cultu red periodontal ligament fibroblasts (PDLF). The evaluated storage media we re: ViaS-pan, Hanks' balanced salt solution (HBSS), alpha minimal essential medium (alpha MEM), and alpha MEM supplemented with FCS and antibiotic (al pha MEM-S). PDLF were obtained from explants of human healthy extracted tee th. Plates with confluent PDLF were soaked in the various media supplemente d with IGF-1 (10 ng/ml) and PDGF-BB (4 ng/ml) for 2, 8 and 24 h at room tem perature (24 degreesC). The control group was incubated with the examined s torage media without growth factors at 24 degreesC. An additional control g roup was incubated with culture medium at 37 degreesC without growth factor s. After incubation, the viability of the cells was determined by Trypan bl ue exclusion test. Viable cells were then analyzed for mitogenic (with thym idine) and clonogenic (by culturing one cell/well) capacities. Storage of P DLF with growth factors (GE) for 2, 8 and 24 h decreased their vitality by only 3% (not statistically significant). The mitogenicity of PDLF stored fo r 2, 8 and 24 h in various media with GF was statistically comparable to th at of the control group. Generally, the highest mitogenic capacity of PDLF stored with or without GF was found after 8 h of storage. Increasing the st orage period to 24 h decreased the mitogenic capacity of the cells stored w ith GF by only 10-40% compared to the control group. In contrast, the clono genic capacity of PDLF stored with GF increased with increasing storage per iods by 100-300%, and the highest clonogenic capacity was found in most sto rage media after 24 h of storage with GF. The highest clonogenic and mitoge nic capacities were found in cells stored in HBSS followed by alpha MEM-S. The mitogenic and clonogenic capacities of PDLF stored in various media sup plemented with GF for 2-8 h were generally lower than without GF supplement ation. The mitogenic and clonogenic effects of GF-supplementation was obser ved only after 24 h of storage. After 24 h of storage with GF, the clonogen ic capacity increased by 8-224% and the mitogenicity by 20-37%, except in c ells stored in alpha MEM (-1%). However, these differences were generally n ot statistically significant. In conclusion, the mitogenic and clonogenic e ffects of GF were observed only after 24 h of storage at room temperature. HBSS and alpha MEM-S supplemented with GF were the most effective media for preserving the viability, mitogenicity and clonogenic capacity of PDLF sto red for 24 h at room temperature. For short periods of storage (2 and 8 h), HBSS and alpha MEM-S without GF were preferable.