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.
The choice of storage medium for preserving traumatically avulsed teeth is important for the success of future replantation. The objective of this study was to compare the effectiveness of four recommended storage media (Hank's balanced salt solution [HBSS], culture medium, alpha minimal essential medium [alpha-MEM], and ViaSpan) to preserve cultured periodontal ligament fibroblasts (PDLF) at room temperature (22 degrees C). PDLF were obtained from explants of extracted healthy human teeth. Plates with confluent PDLF were soaked in the various media for 2, 8 and 24 h at room temperature. A control group was incubated with culture medium at 37 degrees C. After incubation, viability of the cells was determined by trypan blue exclusion test. Viable cells were then analyzed for mitogenic (with thymidine) and clonogenic capacity (by culturing one cell/well). Viability of PDLF stored up to 24 h was comparable in all tested media, and the differences were limited to 1%-3%. PDLF stored for up to 24 h in various media had statistically comparable mitogenicity to the control group. After 8 h of storage, the differences were limited to 2%-9%, except for the alpha-MEM group which had 23%-29% lower mitogenic capacity compared to the control group. Increasing the storage time up to 24 h further decreased the mitogenicity of the cells by 22%-47%. The highest mitogenicity after 24 h of storage was found in PDLF stored in culture medium or HBSS, and the lowest in alpha-MEM. PDLF stored for 2-8 h in various media had a comparable clonogenic capacity to the control group. However, after 24 h, the cells' clonogenic ability dropped by 14%-66%. A similar trend of reduction was noted in the mitogenic and clonogenic capacity, although it was statistically significant only in the clonogenic capacity. Culture medium and ViaSpan, followed by HBSS, were the most effective in preserving the clonogenic capacity of PDLF after 24 h of storage. The lowest clonogenic capacity after 24 h of storage was in the alpha-MEM group (66%, P < 0.0025). In conclusion, culture medium, followed by HBSS and ViaSpan, was the most effective media for preserving the viability, mitogenicity and clonogenic capacity of PDLF stored for up to 24 h at room temperature. The lowest functional abilities were found in PDLF stored in alpha-MEM.
Aim . To evaluate the incidence and severity of postendodontic treatment pain (PEP) subsequent to root canal treatment (RCT) in vital and necrotic pulps and after retreatment. Methodology . A prospective study. Participants were all patients ( n = 274) who underwent RCT in teeth with vital pulp, necrotic pulp, or vital pulp that had been treated for symptomatic irreversible pulpitis or who received root canal retreatment, by one clinician, during an eight-month period. Exclusion criteria were swelling, purulence, and antibiotic use during initial treatment. A structured questionnaire accessed age, gender, tooth location, and pulpal diagnosis. Within 24 h of treatment, patients were asked to grade their pain at 6 and 18 hours posttreatment, using a 1–5 point scale. Results . RCT of teeth with vital pulp induced a significantly higher incidence and severity of PEP (63.8%; 2.46 ± 1.4, resp.) than RCT of teeth with necrotic pulp (38.5%; 1.78 ± 1.2, resp.) or of retreated teeth (48.8%; 1.89 ± 1.1, resp.). No statistical relation was found between type of pain (spontaneous or stimulated) and pulp condition. Conclusion . RCT of teeth with vital pulp induced a significantly higher incidence and intensity of PEP compared to teeth with necrotic pulp or retreated teeth.
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