The Journal of Korean Academy of Prosthodontics (대한치과보철학회지)

The Korean Academy of Prosthodonitics (대한치과보철학회)
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Effect and mechanism of chitosan-based nano-controlled release system on the promotion of cell cycle progression gene expression

키토산 기반 나노방출제어시스템의 세포주기진행 유전자 발현 증진 효과 및 기전

  • Lee, Won Joong (Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, School of Dentistry, Kyung Hee University) ;
  • Park, Kwang Man (Department of Dentistry, Graduate School, Kyung Hee University) ;
  • Lee, sungbok Richard (Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, School of Dentistry, Kyung Hee University) ;
  • Hwang, Yu Jeong (Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, School of Dentistry, Kyung Hee University) ;
  • Lee, Suk Won (Department of Biomaterials & Prosthodontics, Kyung Hee University Hospital at Gangdong, School of Dentistry, Kyung Hee University)
  • 이원중 (강동경희대학교 치과병원 치과(생체재료)보철과, 경희대학교 치과대학 치과보철학교실) ;
  • 박광만 (경희대학교 대학원 치의학과) ;
  • 이성복 (강동경희대학교 치과병원 치과(생체재료)보철과, 경희대학교 치과대학 치과보철학교실) ;
  • 황유정 (강동경희대학교 치과병원 치과(생체재료)보철과, 경희대학교 치과대학 치과보철학교실) ;
  • 이석원 (강동경희대학교 치과병원 치과(생체재료)보철과, 경희대학교 치과대학 치과보철학교실)
  • Received : 2021.05.20
  • Accepted : 2021.08.23
  • Published : 2021.10.31
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Abstract

Purpose. In our previous studies, application of trichloroacetic acid (TCA) to gingival fibroblasts or to canine palatal soft tissue was verified to alter the expression of several genes responsible for cell cycle progression. In order to confirm this effect in a system allowing sequential release of TCA and epidermal growth factor (EGF), expression of various cell cycle genes following the application of the agents, using hydrophobically modified glycol chitosan (HGC)-based nano-controlled release system, was explored in this study. Materials and methods. HGC-based nano-controlled release system was developed followed by loading TCA and EGF. The groups were defined as the control (CON); TCA-loaded nano-controlled release system (EXP1); TCA- and EGF- individually loaded nano-controlled release system (EXP2). At 24- and 48 hr culture, expression of 37 cell cycle genes was analyzed in human gingival fibroblasts. Correlations and the influential genes were also analyzed. Results. Numerous genes such as cyclins (CCNDs), cell division cycles (CDCs), cyclin-dependent kinases (CDKs), E2F transcription factors (E2Fs), extracellular signal-regulated kinases (ERKs) and other cell cycle genes were significantly up-regulated in EXP1 and EXP2. Also, cell cycle arrest genes of E2F4, E2F5, and GADD45G were up-regulated but another cell cycle arrest gene SMAD4 was down-regulated. From the multiple regression analysis, CCNA2, CDK4, and ANAPC4 were determined as the most influential factors on the expression of ERK genes. Conclusion. Application of TCA and EGF, using the HGC-based nano-controlled sequential release system significantly up-regulated various cell cycle progression genes, leading to the possibility of regenerating oral soft tissue via application of the proposed system.

목적: 이전 연구에서 치은섬유아세포 혹은 성견 구개 연조직에 trichloroacetic acid (TCA)를 적용하는 것이 세포주기진행 유전자 발현의 변화를 일으키는 것으로 밝혀졌다. 이에 따라 본 연구에서는, hydrophobically modified glycol chitosan (HGC)기반의 나노방출제어시스템을 이용한 TCA 및 상피세포성장인자(EGF)의 순차적 방출시스템에서 이 효과를 검증하기 위하여 다양한 세포주기진행 유전자들의 발현을 규명하였다. 재료 및 방법: TCA와 EGF를 담지하는 HGC기반 나노방출제어시스템을 제작하였다. 실험군은 대조군(CON); TCA-담지형 나노방출제어시스템 투여군(EXP1); TCA- 및 EGF-담지형 나노방출제어시스템 투여군(EXP2)으로 정의되었다. 24시간 및 48시간 배양 시 37개 세포주기 유전자들의 발현을 분석하였다. 영향인자로서의 유전자 및 상관관계에 대해서도 분석하였다. 결과: Cyclins (CCNDs), cell division cycles (CDCs), cyclin-dependent kinases (CDKs), E2F transcription factors (E2Fs), extracellular signal-regulated kinases (ERKs)와 같은 다수의 유전자들과 기타 다른 세포주기 유전자들의 발현이 EXP1과 EXP2에서 상향조절되었다. E2F4, E2F5, GADD45G와 같은 세포주기차단 유전자들의 발현도 상향조절되었으나, 또다른 세포주기차단 유전자인 SMAD4의 발현은 하향조절되었다. 다중회귀분석에서 CCNA2, CDK4 그리고 ANAPC4가 ERK 유전자 발현에 가장 영향력 있는 유전자로 선정되었다. 결론: HGC기반 순차적 나노방출제어시스템을 이용한 TCA 및 EGF의 적용은 다양한 세포주기진행 유전자들의 발현을 상향조절함이 밝혀졌고, 이를 토대로 한 구강연조직증대시스템 개발의 가능성이 확보되었다.

Keywords

Acknowledgement

This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant number: HI16C1838).

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