In vitro viability, mitogenicity and clonogenic capacity of periodontal ligament cells after storage in six different media

Abstract

The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objectives of this study were to evaluate the effectiveness of six different media: culture medium, alpha minimal essential medium (alpha-MEM), milk, Hank's balanced salt solution (HBSS), ViaSpan and conditioned medium (CM) to preserve cultured periodontal ligament fibroblasts (PDLF). Periodontal ligament fibroblasts were obtained from explants of human healthy extracted teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at 4 degrees C. A control group was incubated with culture medium at 37 degrees C. After incubation, cell viability was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Storage of PDLF up to 24 h decreased their vitality by only 2%-14%. Vitality of the PDLF after 2, 8 and 24 h was highest when stored in milk or HBSS (91%-97%) and lowest when stored in ViaSpan or CM (82%-93%). PDLF stored for 2-8 h in various media had a mitogenic capacity comparable to the control. However, increasing the storage period to 24 h decreased the mitogenicity of the cells by 3%-39%. The highest mitogenicity was found in PDLF stored in milk or HBSS and the lowest in CM or ViaSpan. The clonogenic capacity of the cells dropped by 38%-71% after 24 h and was the best indicator of the deteriorating effect of long storage. Milk and HBSS were the most effective in preserving the clonogenic capacity. Nevertheless, reduction in the viability, mitogenicity or clonogenic capacity was statistically significant in nearly all the tested media only after 24 h of incubation. In conclusion, HBSS and milk were the most effective media for preserving the viability, mitogenicity and clonogenic capacity after storage for up to 24 h at 4 degrees C.

Medios de almacenamiento y transporte para dientes avulsionados

“…3,12 Respecto a la temperatura de transporte, los estudios no son concordantes; pues unos aconsejan conservar en leche a temperatura ambiente (20°C) y otros proponen leche fría (4°C) para mantener l a c a p a c i d a d c l o n o g é n i c a celular. 3,19,21,22,27 Leche evaporada entera 6,1 313 1 25…”

Egg White-Milk Mixture as a Novel Biological Storage Media for Avulsed teeth Periodontal Ligament Cells: An In-Vitro Study

“…They were promising concerning the organization of collagen fibres and the number of viable cells. Some suggest that they can be perfect media for storing avulsed teeth but they need additional studies [4,6,[18][19][20][21]. Some considered milk as a gold standard for transporting avulsed teeth.…”

“…Also, it contains many Minerals (Ca mg 33, Fe mg 0.39, Mg mg 6, P mg 74, K mg 114, Na mg 78 & Zn mg 1.18) and Vitamins (Thiamine mg 0.012 & Riboflavin mg 0.088). The pH of milk is about (6.5-7) and its osmolality is about (290-375 mOsm/kg) [4,18,30].…”

Effect of Milk Renewal on Cell Viability In Vitro at Different Time Frames

“…While some demonstrated effectiveness for only up to 3 h (15,19) or 6 h (4), other showed effectiveness for 24 h (6,17,(20)(21)(22) or for up to 48 h (7,8). When culture plates were maintained at 20 °C and 37 °C, milk was the best storage medium for up to 48 and 24 h, respectively (7).…”

“…Although there has been some controversy (6,8), it has been suggested that milk with a lower fat content might be more appropriate to maintain PDLF viability than milk with a higher fat content (14). Previous studies indicate that cold milk is more effective than milk at room temperature (3,10,(15)(16)(17). Low temperatures have the advantage of reducing cellular metabolism (18) and limiting bacterial growth (3), which may affect the prognosis of tooth replantation.…”