Protective Effects of Membrane-Free Stem Cell Extract from H2O2-Induced Inflammation Responses in Human Periodontal Ligament Fibroblasts

무막줄기세포추출물의 H2O2에 의해 유도된 치주 세포의 염증 반응 보호 효과

  • He, Mei Tong (Department of Food Science and Nutrition, Pusan National University) ;
  • Kim, Ji Hyun (Department of Food Science and Nutrition, Pusan National University) ;
  • Kim, Young Sil (T-STEM Co., Ltd.) ;
  • Park, Hye Sook (T-STEM Co., Ltd.) ;
  • Cho, Eun Ju (Department of Food Science and Nutrition, Pusan National University)
  • Received : 2019.02.26
  • Accepted : 2019.06.07
  • Published : 2019.06.30


Periodontal inflammation, a major kind of periodontal diseases, is characterized to bleed, pain, and teeth loss, and it is resulted from oxidative stress. Membrane-free stem cell extract could avoid the immunogencity rejection by removal of cell membrane. In the present study, we investigated the protective effect of membrane-free stem cell extract from oxidative stress-induced periodontal inflammation in human periodontal ligament fibroblasts (HPLF). In the cell viability measurement, membrane-free stem cell extract showed significant increase of cell viability, compared with the $H_2O_2$-treated control group. To further investigation of molecular mechanisms, we measured inflammation and apoptosis related protein expressions. Membrane-free stem cell extract attenuated inflammation-related protein expressions such as nuclear factor kappa light chain enhancer of activated B cells, inducible nitric oxide synthase, and interleukin-6. In addition, the treatment of membrane-free stem cell extract decreased apoptotic protein expressions such as cleaved caspase-9, -3, poly (ADP-ribose) polymerase, and B-cell lymphoma 2 (Bcl-2)-associated X protein/Bcl-2 ratio in the $H_2O_2$-treated HPLF cells. In conclusion, membrane-free stem cell extract exhibited anti-oxidative stress effects by regulation of inflammation and apoptosis in HPLF, suggesting that it could be used as the treatment agents for periodontal inflammatory disease.

대표적인 치주질환인 치주염은 출혈, 통증 및 치아 손실을 초래하며, 산화적 스트레스는 치주염의 주요 원인으로 알려져 있다. 본 연구는 지방조직 유래 무막줄기세포추출물의 $H_2O_2$ 유도 산화적 손상에 대한 치주염 보호 효과를 확인하고자, 치주인대 섬유모세포(human periodontal ligament fibroblasts; HPLF)를 이용하여 세포 생존율, 염증 및 세포 사멸 관련 단백질 발현을 측정하였다. $H_2O_2$로 산화적 스트레스를 유도한 HPLF 세포에 무막줄기세포추출물 처리 시, $H_2O_2$만을 처리한 control군에 비해 유의적으로 세포 생존율이 증가함을 통해 산화적 손상에 대한 세포 보호 효과를 확인하였다. 또한, 무막줄기세포추출물은 nuclear factor kappa light chain enhancer of activated B cells, inducible nitric oxide synthase 및 interleukin-6와 같은 염증 관련 단백질 발현을 감소시켜 $H_2O_2$로 유도된 염증반응 보호 효과를 확인할 수 있었다. 뿐만 아니라, 무막줄기세포추출물 처리 군은 caspase-9, -3, poly (ADP-ribose) polymerase 단백질 발현 감소와 B-cell lymphoma 2 (Bcl-2)-associated X protein/Bcl-2 비율을 저하시켜 $H_2O_2$ 유도 산화적 손상에 대한 세포사멸 보호 효과를 보였다. 따라서 지방조직 유래 무막줄기세포추출물은 $H_2O_2$ 유도 산화적 손상에 대한 HPLF 세포의 염증반응 및 세포사멸을 저해함으로써 치주염으로부터 보호 효과가 있어, 치주질환 치료용 소재로써의 활용 가능성이 있을 것으로 기대된다.

SHGSCZ_2019_v20n6_95_f0001.png 이미지

Fig. 1. Effect of membrane-free stem cell extract on H2O2-treated HPLF cells. Values are presented as mean ± SD. The different letters (a-c) indicate significant differences (P < 0.05) by Duncan’s multiple range test.

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Fig. 2. Effect of membrane-free stem cell extract on inflammation-related protein expression in H2O2-treated HPLF cells. Values are presented as mean ± SD. The different letters (a-e) indicate significant differences (P < 0.05) by Duncan’s multiple range test.

SHGSCZ_2019_v20n6_95_f0003.png 이미지

Fig. 3. Effect of membrane-free stem cell extract on apoptosis-related protein expression such as caspase-9, -3, and PARP in H2O2-treated HPLF cells. Values are presented as mean ± SD. The different letters (a-d) indicate significant differences (P < 0.05) by Duncan’s multiple range test.

SHGSCZ_2019_v20n6_95_f0004.png 이미지

Fig. 4. Effect of membrane-free stem cell extract on apoptosis-related protein expression such as Bax and Bcl-2 in H2O2-treated HPLF cells. Values are presented as mean ± SD. The different letters (a-d) indicate significant differences (P < 0.05) by Duncan’s multiple range test.

Table 1. Cytotoxicity of membrane-free stem cell extract on HPLF cells

SHGSCZ_2019_v20n6_95_t0001.png 이미지


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