Journal of the Society of Cosmetic Scientists of Korea (대한화장품학회지)

Society of Cosmetic Scientists of Korea (대한화장품학회)
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Analysis of Procollagen Biosynthesis of Functional Peptides Utilizing Stiffness Controlled Artificial Skin Dermis

강도가 제어된 인공피부 진피를 활용한 기능성 펩타이드의 프로콜라겐 생합성 분석

  • Byun, Jina (Dept. of Chemical Engineering, Soongsil University) ;
  • Shin, Sung Gyu (Dept. of Chemical Engineering, Soongsil University) ;
  • Han, Sa Ra (Dept. of Chemical Engineering, Soongsil University) ;
  • Cho, Sung Woo (Dept. of Chemical Engineering, Soongsil University) ;
  • Lim, Jun Woo (Dept. of Chemical Engineering, Soongsil University) ;
  • Jeong, Jae Hyun (Dept. of Chemical Engineering, Soongsil University)
  • 변진아 (숭실대학교 화학공학과) ;
  • 신성규 (숭실대학교 화학공학과) ;
  • 한사라 (숭실대학교 화학공학과) ;
  • 조성우 (숭실대학교 화학공학과) ;
  • 임준우 (숭실대학교 화학공학과) ;
  • 정재현 (숭실대학교 화학공학과)
  • Received : 2018.10.01
  • Accepted : 2018.11.20
  • Published : 2018.12.30
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Abstract

In this study, cross-linked collagen gels were successfully prepared with varying of elastic modulus from 0.7 to 17.7 kPa using a chemical cross-linker. Then, human dermal fibroblasts were encapsulated into the porous pores introduced into the gels, and cell growth and behavior were examined by gel's mechanical properties. Specifically, increasing elastic modulus of the gel led to decreases in procollagen synthesis from 47 to 32 ng. In addition, there could be optimum elastic modulus for procollagen production, when the gels were treated with adenosine. However, interestingly, this study discovered that the procollagen production level was not influenced by the elastic modulus of the gel for functional peptide. In conclusion, these results would be highly useful for designing reconstructed skins with varying of elastic modulus to examine functional materials in cosmetics.

본 연구에서는 가교 분자를 사용하여 0.7 kPa에서 17.7 kPa까지 다양한 강도를 갖는 콜라겐 겔을 성공적으로 제조하였다. 가교된 콜라겐 겔에 다공성 기공을 도입하고 진피세포를 내부에 담지하여, 겔 강도에 따른 세포 성장 및 거동을 확인하였다. 상대적으로 강도가 높은 겔에서 진피세포의 프로콜라겐 생합성이 47 ng에서 32 ng까지 감소하는 것을 확인하였다. 이렇게 제조된 인공피부 진피에 아데노신을 처리하였을 때, 특정 강도를 갖는 콜라겐 겔에서 프로콜라겐 생합성이 감소하는 것을 확인하였다. 반면에 기능성 펩타이드를 처리하였을 때는 프로콜라겐 생합성이 콜라겐 겔의 강도에 크게 영향을 받지 않는 것을 확인할 수 있었다. 이러한 결과는 강도가 제어된 인공피부 제조 및 응용, 나아가 다양한 조직공학 분야의 기반 기술로 활용 가능하리라 기대된다.

Keywords

HJPHBN_2018_v44n4_419_f0001.png 이미지

Figure 1. Preparation and structural characterization of a cross-linked collagen gel (A-D). the cross-linked collagen gel is standardized with 8 mm of diameter and 1 mm of height. HDF cells were encapsulated into the freeze dried collagen gel and incubated (C-D). the prepared collagen gel encapsulated HDF cells were used as a reconstructed dermis for pro-collagen synthesis (E-F).

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Figure 3. Scanning electron microscopic (SEM) images of cross-linked collagen gel with glutaraldehyde of 0.10% (w/v) shown in (A) 1.0 KX and (B) 3.0 KX. (scale bars = 10 μm).

HJPHBN_2018_v44n4_419_f0003.png 이미지

Figure 5. (A) Ratios of synthesized pro-collagen in the gels with varying of GHK-Cu peptide concentrations. The pro-collagen was normalized to the pro-collagen characterized in pure medium as a control. (B) The type I pro-collagen levels measured on pure media was assessed by elastic modulus of gels. The statistical analysis of the data was derived from T-test (*p < 0.05).

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Figure 2. (A) Elastic modulus (E) of cross-linked collagen gels with varying of glutaraldehyde concentration. (B) Compressive strain and stress curves.

HJPHBN_2018_v44n4_419_f0005.png 이미지

Figure 4. (A) The image was captured after 7 days of cell culture in a cross-linked collagen gel with 0.7 kPa of elastic modulus. (B) Phase-contrast images of HDFs positively stained by MTT reagents. (C) Ratios in the number of HDFs remained metabolically active in the gels with 0.7 kPa of elastic modulus. The cell viability measured at each gel was normalized to the viability characterized in pure medium.

HJPHBN_2018_v44n4_419_f0006.png 이미지

Figure 6. The pro-collagen measured at each gel was normalized to the pro-collagen characterized in pure medium by each modulus of gel. (A) Ratios are decreased by increasing elastic modulus of gels, when the adenosine was treated. (B) Ratios are maintained with elastic modulus of gels, when the GHK-Cu peptide was treated. The statistical analysis of the data was derived from T-test (*p < 0.05).

References

  1. Y. Ahn, C. Y. Lee, S. Baek, T. Kim, P. Kim, S. Lee, D. Min, H. Lee, J. Kim, and W. G. Jung, Quantitative monitoring of laser-treated engineering skin using optical coherence tomography, Biomed. Opt. Express., 7(3), 1030 (2016). https://doi.org/10.1364/BOE.7.001030
  2. E. Ruoslahti and M. D. Pierschbacher, New perspectives in cell adhesion: RGD and integrins, Science, 238, 491 (1987). https://doi.org/10.1126/science.2821619
  3. M. Jung, S. G. Shin, J. W. Lim, S. R. Han, H. Kim, and J. H. Jeong, Tuning the stiffness of dermal fibroblast-encapsulating collagen gel by sequential cross-linking, J. Soc. Cosmet. Sci. Korea, 44(1), 23 (2018). https://doi.org/10.15230/SCSK.2018.44.1.23
  4. M. W. Tibbitt and K. S. Anseth, Hydrogels as extracellular matrix mimics for 3D cell culture, Biotechnol. Bioeng., 103, 665 (2009).
  5. E. Proksch, M. Schunck, V. Zague, D. Segger, J. Degwert, and S. Oesser, Oral intake of specific bioactive collagen peptides reduces skin wrinkles and increases dermal matrix synthesis, Skin Pharmacol. Physiol., 27(3), 113 (2014). https://doi.org/10.1159/000355523
  6. J. M. Ruijgrok, J. R. Wijn, and M. E. Boon, Glutaraldehyde crosslinking of collagen: effects of time, temperature, concentration and presoaking as measured by shrinkage temperature, Clin. Mater., 17, 23 (1994). https://doi.org/10.1016/0267-6605(94)90044-2
  7. J. H. Jeong, Y. Liang, M. Jang, C. Cha, C. Chu, H. Lee, W. Jung, J. W. Kim, S. A. Boppart, and H. Kong, Stiffness-modulated water retention and neovascularization of dermal fibroblast-encapsulating collagen gel, J. Tissue Eng. Part A, 19(11), 1275 (2013). https://doi.org/10.1089/ten.tea.2012.0230
  8. P. Zorlutuna, J. H. Jeong, H. Kong, and R. Bashir, Stereolithography-based hydrogel microenvironments to examine cellular interactions, Adv. Funct. Mater., 21(19), 3642 (2011). https://doi.org/10.1002/adfm.201101023
  9. S. J. Oh, H. Kim, S. W. Cho, S. R. Ham, S. G. Shim, B. H. Han, and J. H. Jeong, Tuning the toughness of an alginate micro-gel for cell-instructed delivery, Polym. Kor., 42(1), 140 (2018). https://doi.org/10.7317/pk.2018.42.1.140
  10. H. Kim, Y. N. Cho, S. W. Cho, Y. Kim, J. H. Jeong, and H. W. Ryu, Tuning the hydrophobicity of agar hydrogel with substituent effect, Polym. Kor., 40(2), 1 (2016). https://doi.org/10.7317/pk.2016.40.1.1
  11. S. Jose, M. L. Hughbanks, B. Y. Binder, G. C. Ingavle, and J. K. Leach, Enhanced trophic factor secretion by mesenchymal stem/stromal cells with glycine-histidine-lysine (GHK)-modified alginate hydrogels, Acta Biomater., 10(5), 1955 (2014). https://doi.org/10.1016/j.actbio.2014.01.020
  12. T. Kwon, C. T. Oh, E. J. Choi, S. R. Kim, Y. Jang, E. J. Ko, K. H. Yoo, and B. J. Kim, Conditioned medium from human bone marrow-derived mesenchymal stem cells promotes skin moisturization and effacement of wrinkles in UVB-irradiated SKH-1 hairless mice, Photodermatol. Photoimmunol. Photomed., 32(3), 120 (2016). https://doi.org/10.1111/phpp.12224
  13. J. H. Jeong, J. J. Schmidt, C. Cha, and H. J. Kong, Tuning responsiveness and structural integrity of a pH responsive hydrogel using a poly (ethylene glycol) cross-linker, Soft Matter, 6, 3930 (2010). https://doi.org/10.1039/c0sm00094a
  14. T. L. Tuan, L. C. Keller, D. Sun, M. E. Nimni, D. Cheung, Dermal fibroblasts activate keratinocyte outgrowth on collagen gels, J. Cell. Sci., 107, 2285 (1994).
  15. M. El-Domyati, S. Attia, F. Saleh, D. Brown, D. E. Birk, F. Gasparro, H, Ahmad, and J. Uitto, Intrinsic aging vs. photo-aging: a comparative histopathological, immunohistochemical and ultrastructural study of skin, Exp. Dermatol., 11(5), 398 (2002). https://doi.org/10.1034/j.1600-0625.2002.110502.x