The Application of the Starfish Hatching Enzyme for the Improvement of Scar and Keloid Based on the Fibroblast-Populated Collagen Lattice
Various bioactivities of the starfish hatching enzyme (HE) including collagen gel contraction, MMPs activity, hydroxyproline release, and gene regulation based on the fibroblast-populated collagen lattice (FPCL) in three-dimensional medium were investigated for the improvement of scar and keloid. The starfish HE significantly inhibited the collagen gel contraction over 2 days of culture. MMP-2 and MMP-9 activities were also identified by gelatin zymography and RT-PCR products with both HE and collagenase treatments, which resulted in the high amount of hydroxyproline release. The HE treatment on the FPCL significantly inhibited the fibroblast proliferation at 3 days of culture. The LPS-induced NO level and iNOS mRNA expression at low concentrations of HE presented a certain ability to inflammatory response. The COX-2 mRNA from the FPCL indicated no significant inflammation-mediated activity at 5 μg/mL of HE, whereas the cytokines of TNF-α and IL-1β were significantly higher than those of the control. Hence, the starfish hatching enzyme can regulate the fibroblast-populated collagen gel conditions by the contraction, MMP production, inflammatory gene expression, etc. Therefore, the starfish HE could be a potential cosmeceutical to heal the scar and keloid tissue.
KeywordsHatching enzyme Collagen gel contraction MMP Gene regulation
Tissue inhibitors metalloproteases
Fibroblast-populated collagen lattice
Inducible nitric oxide synthase
Tumor necrosis factor-alpha
Transfer growth factor
This research was supported by the Korea Sea Grant Program (GangWon Sea Grant) funded by the Ministry of Oceans and Fisheries in Korea.
- 3.Abergel, R. P., Pizzurro, D., Meeker, C. A., Lask, G., Matsuoka, L. Y., Minor, R. R., et al. (1985). Biochemical composition of the connective tissue in keloids and analysis of collagen metabolism in keloid fibroblast cultures. Journal of Investigative Dermatology, 84, 384–390.CrossRefGoogle Scholar
- 6.Stricklin, G. P., Li, L., Jancic, V., Wenczak, B. A., & Nanney, L. B. (1993). Localization of mRNAs representing collagenase and TIMP in sections of healing human burn wounds. American Journal of Pathology, 143, 1657–1666.Google Scholar
- 8.Oriente, A., Fedarko, N. S., Pacocha, S. E., Huang, S. K., Lichtenstein, L. M., & Essayan, D. M. (2000). Interleukin-13 modulates collagen homeostasis in human skin and keloid fibroblasts. Journal of Pharmacology and Experimental Therapeutics, 292, 988–994.Google Scholar
- 10.Eickelberg, O., Köhler, E., Reichenberger, F., Bertschin, S., Woodtli, T., Erne, P., et al. (1999). American Journal of Physiology, 276, 814–824.Google Scholar
- 12.Norway. Aqua Bio Technology, Norway, December. 2014. http://www.aquabiotechnology.com/index.php?id=5.
- 13.Li, Z. J. & Kim, S. M. (2014). Structural identification and proteolytic effects of the hatching enzyme from starfish Asterias amurensis. Protein and Peptide Letter. doi: 10.2174/0929866521666140221153026.
- 15.Fang, Q., Schulte, N. A., Kim, H., Kobayashi, T., Wang, X., Miller-Larsson, A., et al. (2013). Effect of budesonide on fibroblast-mediated collagen gel contraction and degradation. Journal of Inflammation Research, 6, 25–33.Google Scholar
- 18.Creemers, L. B., Jansen, D. C., van Veen-Reurings, A., van den Bos, T., & Everts, V. (1997). Microassay for the assessment of low levels of hydroxyproline. Biotechniques, 22, 656–658.Google Scholar
- 25.Wang, C., Rong, Y., Ning, F., & Zhang, G. (2011). The content and ratio of type I and III collagen in skin differ with age and injury. African Journal of Biotechnology, 10, 2524–2529.Google Scholar
- 31.Kauh, Y. C., Rouda, S., Mondragon, G., Tokarek, R., diLeonardo, M., Tuan, R. S., et al. (1997). Major suppression of pro-alpha1(I) type I collagen gene expression in the dermis after keloid excision and immediate intrawound injection of triamcinolone acetonide. Journal of the American Academy of Dermatology, 37, 586–589.CrossRefGoogle Scholar
- 34.Lavnikova, N., & Laskin, D. L. (1995). Unique patterns of regulation of nitric oxide production in fibroblasts. Journal of Leukocyte Biology, 58, 451–458.Google Scholar
- 36.Tayler, B. S., & Geller, D. A. (2001). Regulation of the inducible nitric oxide synthase (iNOS) gene. In D. Salvemini, T. R. Billiar, Y. Vodovotz (Eds.), Nitric oxide and inflammation (pp. 1–27). Birkhauser Verlag: Springer.Google Scholar
- 37.Cuzzocre, S. (2011). Role of nitric oxide and reactive oxygen species in arthritis. In D. Salvemini, T. R. Billiar, Y. Vodovotz (Eds.), Nitric oxide and inflammation (pp. 145–160). Birkhauser Verlag: Springer.Google Scholar
- 38.Kang, K. W., Wagley, Y., Kim, H. W., Pokharel, Y. R., Chung, Y. Y., Chang, I. Y., et al. (2007). Novel role of IL-8/SIL-6R signaling in the expression of inducible nitric oxide synthase (iNOS) in murine B16, metastatic melanoma clone F10.9 cells. Free Radical Biology and Medicine, 42, 215–227.CrossRefGoogle Scholar