A COMPARATIVE STUDY OF PRESERVING ABILITY OF HUMAN PERIODONTAL LIGAMENT CELLS STORED IN DIFFERENT STORAGE MEDIA

수종의 저장용액에서 치주인대세포의 생존율 비교

Preservation of the remaining periodontal ligament cells on an avulsed tooth is very important to the successful outcome of replantation. HBSS is recommended as the most suitable storage medium for the avulsed tooth that cannot be replanted immediately. But their availability near the site of an accident is doubtful. The purpose of this in vitro study was to compare periodontal ligament cells stored in different storage media obtained easily on the spot. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ culture medium containing 20% FBS, at $37^{\circ}C$ 100% humidity, in a 5% $CO_2$ incubator. Cells were cultured in 96 well culture plate, $5{\times}10^4$ cells per well with ${\alpha}-MEM$ and incubated for 24 hours. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS, pasteurized milk, sterilized saline, unstimulated saliva and bench-dried state at $25^{\circ}C$ room temperature for 30, 60, 90, 120, 180 minutes respectively. And then each group was measured using MTT assay. The results were as follows. 1. Between the group of each time, there was statistically significant difference. Periodontal ligament cells viability was highest in pasteurized milk and was reduced stepwisely in sterilized saline, unstimulated saliva and bench-dried state(p<0.05). 2. between the time of each group, there was statistically significant difference(p<0.05) but was no statistically significant difference at 90-120 minutes in pasteurized milk and at 60-90 minutes and 120-180 minutes in sterilized saline(p>0.05). In conclusion, HBSS as storage medium of an avulsed tooth is not practical on the spot. Insteadily pasteurized milk can be recommended to maintain the periodontal ligament cells viability.

치아탈구 발생시 사고 현장에서 쉽게 구할 수 있는 저장용액을 이용하여 치주인대세포의 생존율을 비교해보고자 치주인대세포를 10% FBS함유 ${\alpha}-MEM$에서 배양한 후 ${\alpha}-MEM$에서 저장한 군을 대조군으로 하고 동일한 조건하에서 저온살균된 우유군, 멸균된 생리적 식염수군, 비자극성 타액군, 건조된 상태로 방치한 군을 실험군으로 하여 $25^{\circ}C$ 실온에서 세포 배양 30, 60, 90, 120, 180분 후에 각 군에 대해 MTT assay를 시행하여 다음과 같은 결론을 얻었다. 1. 각 조사시점에서 실험군간의 비교에서는 모두 유의성있는 차이를 보였으며 저온살균된 우유군에서 치주인대세포의 생존율이 가장 높았고, 멸균된 생리적 식염수군, 비자극성 타액군, 건조된 상태로 방치한 군 순으로 점차 감소하였다(P<0.05). 2. 각 실험군내에서 조사시점간의 비교에서는 치주인대세포의 생존율이 시간이 경과함에 따라 유의성있게 감소하였으나(p<0.05), 저온살균된 우유군에서 90-120분 경과 사이, 멸균된 생리적 식염수군에서 60-90분과 120-180분 경과 사이에서는 유의성있는 차이는 없었다(P>0.05). 이상의 결론을 종합해볼 때 외상으로 인한 치아탈구 발생시 저장용액으로 HBSS가 추천되고 있으나 사고 현장에서 쉽게 구할 수 있고 치주인대세포의 생활력 보존에도 유리한 저온살균된 우유에 탈구된 치아를 담가오도록 추천할 수 있겠다.