Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Autoradiographic Verification of Transdermal Penetration of Oleic Acid-conjugated Peptide Nanosomes

자가방사법에 의한 올레산이 결합된 펩타이드의 피부침투 확인

  • Lee, Kyung-Eun (School of Life Sciences and Biotechnology, Korea University) ;
  • Jung, Min-Kyo (School of Life Sciences and Biotechnology, Korea University) ;
  • Eum, Jai-Hoon (School of Life Sciences and Biotechnology, Korea University) ;
  • Jung, Se-Hui (Department of Bio-health Technology, Kangwon National University School of Medicine) ;
  • Ha, Kwon-Soo (Department of Bio-health Technology, Kangwon National University School of Medicine) ;
  • Park, Jeong-Hae (R&D Center, HwaJin Cosmetics) ;
  • Lee, Jin-Sung (R&D Center, HwaJin Cosmetics) ;
  • Han, Sung-Sik (School of Life Sciences and Biotechnology, Korea University) ;
  • Choe, Myeon (Department of Bio-health Technology, Kangwon National University School of Medicine)
  • 이경은 (고려대학교 생명과학대학 세포공학 및 생체 3차 구조 연구실) ;
  • 정민교 (고려대학교 생명과학대학 세포공학 및 생체 3차 구조 연구실) ;
  • 엄재훈 (고려대학교 생명과학대학 세포공학 및 생체 3차 구조 연구실) ;
  • 정세희 (강원대학교 의생명공학부) ;
  • 하권수 (강원대학교 의생명공학부) ;
  • 박정해 (화진화장품 R&D 센터) ;
  • 이진성 (화진화장품 R&D 센터) ;
  • 한성식 (고려대학교 생명과학대학 세포공학 및 생체 3차 구조 연구실) ;
  • 최면 (강원대학교 의생명공학부)
  • Received : 2010.08.31
  • Accepted : 2010.11.24
  • Published : 2010.12.31
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Abstract

Short peptides are potentially effective materials as cosmeceuticals, but their delivery across the skin can be problematic due to the ionic nature of peptides and the structure of the skin. For short peptide to be utilized as cosmeceuticals, its ability to penetrate the skin must be altered. In this study, we conjugated the widely used procollagen type I signal peptide, KTTKS, with oleic acid to improve the lipophilic properties of the peptide, and used the oleic acid-conjugated peptides to construct cosmeceutical nanosomes. Then we examined the penetration of cosmeceutical nanosomes prepared from isotope-labeled peptide into the skin after transdermal application using autoradiography. Because of its hydrophilic property of penta-peptide, the penta-peptide itself was not able to be penetrated through the stratum corneum of the skin. In contrast, nanosomes made from olecic acid conjugated penta-peptide were able to be penetrated through the stratum corneum effectively. Autoradiography showed the precise penetration points to dermal layer, demonstrating the appropriateness of this method for clarifying the mechanism of penetration of transdermal delivery systems.

Short 펩타이드는 약용화장품(cosmeceuticals)으로 가능성이 있지만, 펩타이드의 친수성 성질 때문에 피부를 통한 전달이 어렵다. 따라서 이러한 short 펩타이드를 약용화장품으로 이용하기 위해서는 피부를 통과할 수 있는 성질로 바꾸어야만 한다. 이 연구에서는 프로콜라겐 type I의 신호펩타이드로 작용할 수 있는 KTTKS의 5개 펩타이드에 양친매적인 특성을 부여하기 위해 올레산을 결합시켰고, 그 후 나노좀으로 제조하였다. 제조된 나노좀은 동결투과전자현미경법을 이용하여 크기가 100에서 200 nm 사이의 리포좀의 형태임이 확인되었으며, 제타전위측정장치(ELS)를 통해 24주 동안 안정적인 크기를 유지하고 있음이 확인되었다. 이렇게 제조된 나노좀을 피부에 바른 후 자가방사기록법(autoradiography)을 이용하여 피부에 침투 여부를 확인하였다. 이 5개 펩타이드의 친수성 성질로 인하여 펩타이드만 피부에 도말하였을 때는 피부에 침투되지 않았으나, 펩타이드와 올레산을 결합하여 나노좀으로 제조된 경우에는 피부의 진피층까지 잘 도달한 것을 확인하였다. 이 연구를 통해 자가방사법이 피부를 통한 효과물질의 전달을 검증할 수 있는 좋은 방법임을 보여줌과 동시에 피부에 효과를 나타내는 친수성펩타이드의 피부를 통한 전달에 올레산 결합을 이용한 나노좀 형태가 효과적인 것을 보여주었다.

Keywords

References

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