체외충격파치료기 카트리지의 L-929 세포를 통한 인체접촉부의 세포독성시험

Cytotoxicity test on human contact area with L-929 cells using extracorporeal shock wave therapy cartridge

본 연구는 체외충격파치료기(ESWT : Extracorporeal ShockWave Therapy) 카트리지부를 시료로 사용하여 의료기기 안전성 평가 기준인 GLP(Good Laboratory Practice)에 관한 시험 중 실험물질 인체접촉부위 용출물에 대해 L-929 세포를 이용하여 세포독성 유무를 확인하기 위하여 실시하였다. 실험물질 및 대조물질은 10% FBS가 포함된 1xMEM 배양액으로 37±1℃에서 24±2시간 용출하였다. L929 세포에 실험물질 용출액(시험군), 음성대조물질 용출액(음성대조군), 양성대조물질 용출액(양성대조군), 공시험액 용출액(용매대조군)을 적용하여 37±1℃, 5±1% Co2 배양기에서 48±2시간 배양하였다. 현미경으로 세포반응을 관찰한 결과 공시험액 용출액 및 음성대조물질 용출액을 적용한 세포는 0 등급, 양성대조물질 용출액을 적용한 세포는 4등급, 시험물질 용출액을 적용한 세포는 0 등급으로 나타났다. 세포계수를 통한 정량적 평가를 한 결과 공시험액 용출액 대비 음성대조물질 용출액을 적용한 세포는 106.28%, 양성대조물질 용출액을 적용한 세포는 0.00%, 시험물질 용출액을 적용한 세포는 99.58%의 세포생존율이 측정되었다. 따라서 음성 및 양성대조군의 결과를 확인하였을 때 시험과정은 적합하였으며, 정성적인 평가 방법에서 2등급 미만이고, 정량적인 평가 방법에서도 세포 생존율은 70% 이상이므로 세포독성을 유발하지 않는 것으로 판단된다.

This study was conducted to determine the cytotoxicity of the extracts from the human body contact area of the test substance during a test on the Good Laboratory Practice (GLP), which is the medical device safety evaluation standard, using the extracorporeal shock wave therapy (ESWT) cartridge as a sample, using L-929 cells. The test and control substances were extracted with 1xMEM culture medium containing 10% FBS at 37±1℃ for 24±2 hours. The test substance extract (test group), negative control substance extract (negative control group), positive control substance extract (positive control group), and blank test solution extract (solvent control group) were applied to L929 cells and cultured for 48±2 hours in a 37±1℃, 5±1% CO2 incubator. As a result of observing cell reactions under a microscope, the cells to which the blank test solution extract and negative control substance extract were applied were grade 0, the cells to which the positive control substance extract was applied were grade 4, and the cells to which the test substance extract was applied were grade 0. As a result of quantitative evaluation through cell counting, the cell viability rate of the cells to which the negative control substance extract was applied was 106.28% compared to the blank test solution extract, the cells to which the positive control substance extract was applied were 0.00%, and the cells to which the test substance extract was applied were 99.58%. Therefore, when the results of the negative and positive control groups were confirmed, the test process was appropriate, and it was determined that it did not cause cytotoxicity because the qualitative evaluation method was less than grade 2 and the quantitative evaluation method showed a cell viability rate of more than 70%.