플라즈마 메디신; 저온 상압 플라즈마는 어떻게 의학분야에 적용될 수 있는가?

박상례(Sang Rye Park),홍진우(Jin Woo Hong),이해준(Hae June Lee),김규천(Gyoo Cheon Kim) 한국생명과학회 2013 생명과학회지 Vol.23 No.6

플라즈마는 물질의 제4의 상태로서 이온화된 가스로 불리며 다양한 활성종 및 수산화기, 하전입자, 이온, 자유전자, 산소등을 발생시키는 특징을 가지고 있다. 20세기 후반에 플라즈마는 산업현장에서 활발히 사용되고 있으며, 최근 저온 상압 플라즈마 장치가 개발되면서 의생명 분야에 적용되기 시작했다. 저온 상압 플라즈마는 인체조직에 열 손상을 입히지 않을 뿐만 아니라, 암 치료, 살균, 치아미백, 지혈, 상처치유 등에서 높은 효율성을 보이고 있다. 이때 발생되는 활성종은 포유세포나 병원균에 영향을 미치는 것으로 알려져 있다. 또한, 다학제간의 연구를 통해 저온 상압 플라즈마의 활용범위는 다양한 영역으로 넓어지고 있으며, 새로운 첨단 의료기술로서의 가치가 높아지고 있다. 저온 상압 플라즈마가 포유세포와 미생물에 적용된 이후, 지난 10여 년간 급속한 발전을 통해 최근 플라즈마 메디신이란 학문영역으로 성장했다. 본 논문은 저온 플라즈마가 적용되고 있는 분야에 대해 소개하고, 플라즈마 메디신에 대한 이해를 돕고자 한다. As a forth state of material, plasma is ionized gas, which generates characteristically various reactive species. After late of 20th century, plasma has been widely used in industry. After nonthermal atmospheric plasma was developed, it has been applied to biomedical fields. Nonthermal atmospheric plasma does not give thermal damages to human tissues, and it shows the high efficiency in cancer treatment, sterilization, tooth bleaching, coagulation, and wound healing. Because the application of plasma to biomedicine has been expanded through interdisciplinary studies, its value of high medical technology is increasing now. Since nonthermal atmospheric plasma was first applied to the mammalian cells and microorganisms, many valuable studies has been performed for about last 10 years, so that now the new research area called ‘plasma medicine’ has been formed. This article introduces the recent data resulted from plasma medicine and helps to understand the plasma medicine.

Hairless 마우스 조직에서의 저온 상압 플라즈마의 안정성 평가

박상례 ( Sang Rye Park ),김규천 ( Gyoo Cheon Kim ),최별보라 ( Byul Bora Choi ),김지영 ( Ji Young Kim ) 대한예방치과·구강보건학회 2014 大韓口腔保健學會誌 Vol.38 No.3

Objectives: The aim of the present study was to evaluate the stability of non-thermal atmosphericpressure plasma on Candida albicans in hairless mouse-2 (HRM-2) tissues. Methods: HRM-2 mice were subjected to non-thermal atmospheric-pressure plasma jet treatment using an optical fiber probe and monitored using a thermometer. The skin of HRM-2 mice was treated with plasma jet for 0, 60, 180, and 300 s per day for 5 days. After plasma treatment, morphological changes in Candida albicans on the skin of these mice were examined using a scanning electron microscope. Biopsy of the plasma-treated skin was performed and the tissues were histologically analyzed using hematoxylin and eosin (H&E) and Masson’s trichrome stains. Results: The scanning electron microscopic images revealed the morphological changes in the membrane structure of the plasma-treated Candida albicans. Histological analysis showed that non-thermal plasma treatment did not cause epidermal damage or tissue inflammation and did not significantly modify the collagen layers of the mouse skin. Conclusions: The results of this study suggest that non-thermal atmospheric-pressure plasma might be safe and effective for clinical applications in the field of dentistry.

바로잡음 : Hairless 마우스 조직에서의 저온 상압 플라즈마의 안전성 평가

박상례 ( Sang Rye Park ),김규천 ( Gyoo Cheon Kim ),최별보라 ( Byul Bora Choi ),김지영 ( Ji Young Kim ) 대한구강보건학회 2014 大韓口腔保健學會誌 Vol.38 No.4

구강세치제에 함유된 SLS(Sodium lauryl Sulfate)가 HaCaT 세포와 NIH-3T3 세포에 미치는 독성 효과

박상례 ( Sang Rye Park ),김영민 ( Young Min Kim ),최별보라 ( Byul Bora Choi ),김지영 ( Ji Young Kim ) 한국치위생학회 2015 한국치위생학회지 Vol.15 No.4

Objectives: The purpose of this study was to determine the toxic effects of sodium lauryl sulfate(SLS) in human keratinocyte HaCaT cells and mouse fibroblast NIH-3T3 cells. Methods: The effect of sodium lauryl sulfate(SLS) cell viability and proliferation were determined by WST-1 assay and changes shape of nucleus were evaluated by Hoechst staining under fluorescence microscopy. Additionally, observation of cell morphological changes under light microscopy. Results: SLS induced cytotoxicity and a marked apoptosis in both HaCaT and NIH-3T3 cell lines. With the result of the WST-1 assay, SLS induced the cytotoxicity of 0.005% and 0.0075%, 0.01% SLS for 24 h after HaCaT and NIH-3T3 cells in time and dose-dependent manner(p<0.005). SLS inhibited cell growth and caused apoptosis as evidenced by nuclear fragmentation and condensation. Thus, determination of the morphological changes to define apoptosis was visualized using inverted phase contrast microscopy. Conclusions: SLS had toxicity of the human keratinocyte cells and mouse fibroblast cells and this study will provide the basic data for the development of proper SLS concentration in dentifrice.

저온 상압플라즈마에 의한 Hairless Mouse-2 마우스 조직의 Candida albicans 사멸 효과

박상례 ( Sang Rye Park ),김규천 ( Gyoo Cheon Kim ) 한국치위생과학회 2014 치위생과학회지 Vol.14 No.1

본 연구는 저온 상압 플라즈마 장치를 이용하여 구강점막 질환을 일으키는 C. albicans 균을 효과적으로 사멸하기 위해 시행하였다. 조직에 적합하게 처리될 수 있도록 저온 상압 플라즈마 장치를 고안하고, 먼저 agar plate에 C. albicans 균을 처리하여 플라즈마를 조사한 결과 agar plate 표면에 C. albicans 균을 처리 후 저온 상압 플라즈마 장치를 적용한 결과 60초 처리시 1.2 cm, 180초 처리시 1.4 cm, 300초 처리시 1.7 cm의 박테리아 생장 억제 구간이 나타나는 것을 확인하였다. 또한, 조직에서의 구강병원균 사멸 효과를 확인하기 위해 HRM-2 마우스 조직에 C. albicans 균을 처리하여 저온 상압 플라즈마를 조사 시 마우스 조직 표면에 C. albicans 균을 오염시켜 저온 상압 플라즈마 처리후 CFU 방법으로 측정한 결과 300초간 1회 처리시 2 log CFU/ml, 300초간 2회 처리시 3 log CFU/ml, 300초간 3 회처리시 6 log CFU/ml의 균 수 감소 효과가 나타나는 것을 확인하였다(p<0.05). 따라서, 저온의 저온 상압 플라즈마 장치는 효과적으로 구강 병원균을 사멸시킬 수 있으며, 구강점막질환 치료 장비로서 사용될 수 있을 것으로 생각된다. The purpose of this study was to investigate the killing effect of Candida albicans on hairless mouse-2 (HRM-2) mouse tissues. We tested theeffectiveness of a non-thermal atmospheric pressure plasma in killing C. albicans strains. The viability of C. albicans was determined by countingthe colony forming units (CFU), after non-thermal atmospheric pressure plasma treatment. When non-thermal atmospheric pressure plasma wasrepeatedly treated on mouse skin which inoculated with C. albicans. The C. albicans cells were planted on skin tissue, and then the infected mousetissue was exposed to non-thermal atmospheric pressure plasma for 0 sec, 60 sec, 180 sec and 300 sec. The death rate of C. albicans wasincreased in dependent with treatment times. The three times of non-thermal atmospheric pressure plasma at the interval of 10 minutes significantlyshowed the 6 log CFU/ml reduction of death rate on HRM-2 mouse tissues. Thus, non-thermal atmospheric pressure plasma could be used forthe disinfection of C. albicans on oral surface.

임상에서 쓰이는 저 출력 초음파(Low Intensity Pulsed Ultrasound)가 조골세포에 미치는 영향

박상례 ( Sang Rye Park ),김지영 ( Ji Young Kim ),최별보라 ( Byul Bo Ra Cho ) 한국치위생과학회 2014 치위생과학회지 Vol.14 No.2

In this study, we investigated the proliferative and adhesional effect of human osteoblast like MG-63 cell treated with low intensity pulsed ultrasound (LIPUS). We tested the effectiveness of LIPUS in human osteoblast like MG-63 cells. Cell proliferation was measured using a water soluble tetrazolium salts-1 assay. The mRNA expression of alkaline phosphate, vascular endothelial growth factor (VEGF), integrin alpha 2, colla 1A1 were performed by reverse transcription-polymerase chain reaction. LIPUS was no cytotoxicity in human osteoblast like MG-63 cells. In addition, the data show that treatment with 1 MHz and 3 MHz LIPUS on increased proliferation 7 days after. There were significant increased in mRNA expression of alkaline phosphatase, osteocalcin, VEGF, integrin alpha 2 and colla 1A1 (p<0.05). Therefore, the LIPUS significantly increased differential expression of mRNA levels in osteoblast like MG-63 cell and new possibilities in dental clinical practice.

승마추출물이 MDPC-23세포의 분화에 미치는 영향

최별보라 ( Byul-bora Choi ),김지영 ( Ji-young Kim ),박상례 ( Sang-rye Park ) 한국치위생학회 2017 한국치위생학회지 Vol.17 No.3

Objectives: The purpose of this study was to examine the cell proliferation and expression of alkaline phosphatase (ALP) during the differentiation of murine odontoblast-like cells (MDPC-23) by Cimicifuga rhizoma extract. Cimicifuga rhizoma extract was prepared using 70% ethanol. Then, the cells were treated with 25, 50, 100, 150, and 200 μg of Cimicifuga rhizoma extract. Methods: We determined the Cimicifuga rhizoma effects of MDPC-23 using WST-1 (water soluble tetrazolium salt-1) assay, ALP activity assay and histochemical staining. Results: 25-200 μg of Cimicifuga rhizoma extract did not inhibit the growth of MDPC-23 cells; 100±0, 100±3.29, 99±4.86, 98±3.80, 98±1.73, 99±5.05% (p<0.794). 50 μg of Cimicifuga rhizoma extract stimulated ALP activity on MDPC-23; 5.1±0.20 units/μℓ (p<0.001). Conclusions: It was proven that Cimicifuga rhizoma promoted differentiation of MDPC- 23 cells.

박하추출물의 구강미생물에 대한 항균효과

최별보라 ( Byul Bo Ra Choi ),윤세은 ( Se Eun Yun ),박상례 ( Sang Rye Park ),김규천 ( Gyoo Cheon Kim ) 대한예방치과·구강보건학회 2020 大韓口腔保健學會誌 Vol.44 No.2

Objectives: Dental caries and periodontal disease are infectious and chronic diseases. The aim of the study was to investigate the antimicrobial effect of mentha extracts against Streptococcus mutans (S. mutans ) and Porphyromonas gingivalis (P. gingivalis ). Methods: This activity of mentha extracts were confirmed by the disk diffusion test and minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and colony forming unit (CFU) assays. Results: S. mutans and P. gingivalis showed the highest antimicrobial activity within the inhibition zones. The antimicrobial activity was interrupted as the MIC and MBC of the herbal extracts against the two bacteria were 1 mg/ml and 10 mg/ml, respectively. The antimicrobial effect was determined by the CFU assay. Conclusions: Mentha herb extract demonstrated potential antimicrobial activity against S. mutans and P. gingivalis that cause dental caries and periodontal disease.

체외에서 배양된 구강 내 정상세포에 불화나트륨이 미치는 영향

최별보라 ( Byul Bora Choi ),김다혜 ( Da Hye Kim ),김지영 ( Ji Young Kim ),박상례 ( Sang Rye Park ) 한국치위생학회 2016 한국치위생학회지 Vol.16 No.3

Objectives: Fluoride is widely used in the prevention and control of dental caries. The purpose of this study is to examine the biological effects of Sodium fluoride on the proliferation of oral normal cell in vitro(MDPC-23, HaCaT, HGF-1 cells). Methods: The proliferation of normal cells and the cyto-skeletal change of normal cells were assessed by WST-1 assay and F-actin stain assay. The statistical significances of the resulting data were analyzed using SPSS(Window 12.0). Results: The sodium fluoride(0-12 mM) treatment decreased the cell viability in a dose and time dependent manner: HaCaT(6 h): 100±0, 98±0.39, 82±2.68, 75±0.83, 69±1, 67±1.42%(p<0.005); HaCaT(24 h): 100±0, 98±1.85, 54±0.64, 43±0.4, 38±0.32, 36±0.13%(p<0.006), MDPC-23(6 h): 100±0, 93±1.48, 85±0.28, 82±1.58, 79±1.48, 76±1.93%(p<0.009); MDPC-23(24 h): 100±0, 91±1.26, 58±0.65, 49±1, 44±0.74, 2±0.05%(p<0.005), HGF-1(6 h): 100±0, 97±2.93, 89±5, 71±5.42, 58±4.82, 43±3.47%(p<0.009); HGF-1(24 h): 100±0, 97±2.05, 73±1.73, 22±1.61, 14±1.73, 7±0.85%(p<0.005). Thus, changes in cell morphology and disruption of filamentous(F)-actin organization were observed in higher concentration. Conclusions: These results suggest that higher concentrations of fluoride lead to a reduce the number of cells and morphology change of normal cell.

승마추출액을 처리한 G361세포에서의 apoptosis 효과에 대한 연구

최별보라 ( Byul Bo Ra Choi ),김규천 ( Gyoo Cheon Kim ),홍진우 ( Jin Woo Hong ),박상례 ( Sang Rye Park ) 대한예방치과·구강보건학회 2019 大韓口腔保健學會誌 Vol.43 No.2

Objectives: To investigate whether the cytotoxic effect of Cimicifuga rhizoma extract is associated with cell death in the human keratinocyte (HaCaT) and human melanoma cell lines (G361). Methods: Apoptosis induced by Cimicifuga rhizoma extract was confirmed by water-soluble tetrazolium salts-1 (WST-1) assay, immunocytochemistry, and western blot. Additionally, the release of cytochrome c and apoptosis-inducing factor (AIF) was visualized by confocal laser scanning microscopy. Results: The results showed that Cimicifuga rhizoma extract significantly reduced the viability of G361 cells with half-maximal inhibitory concentration (IC 50) of 200 mg/ml, and the apoptotic process was found to occur via the activation of caspase-3 and caspase-9 pathways. Besides, the release of cytochrome c and AIF was also detected. Conclusions: This study suggests that Cimicifuga rhizoma extract causes apoptosis of human melanoma cells through the intrinsic apoptotic pathway.