Formation of silk fibroin matrices with different texture and its cellular response to normal human keratinocytes.
Three forms of silk fibroin (SF) matrices, woven (microfiber), non-woven (nanofiber), and film form, were used to perform a conformational analysis and cell culture using normal human oral keratinocytes (NHOK). To obtain the SF microfiber (SF-M) matrix, natural grey silk was degummed, while the SF film (SF-F) and nanofiber (SF-N) matrices were prepared by casting and electrospinning the formic acid solutions of the regenerated SF, respectively. For insolubilization, as-prepared SF-F and SF-N matrices were chemically treated with an aqueous methanol solution of 50%. The conformational structures of as-prepared and chemically treated SF matrices were investigated using attenuated total reflectance infrared spectroscopy (ATR-IR) and solid-state 13C CP/MAS nuclear magnetic resonance (NMR) spectroscopy. The as-cast SF-F matrix formed a mainly beta-sheet structure that was similar to the SF-M matrix, whereas the as-spun SF-N matrix had a random coil conformation as the predominant secondary structure. Conformational transitions from random coil to beta-sheet of the as-spun SF-N occurred rapidly within 10 min following aqueous methanol treatment, and were confirmed by solid-state 13C NMR analysis. To assess the cytocompatibility and cells behavior on the different textures of SF, we examined the cell attachment and spreading of NHOK that was seeded onto the SF matrices, as well as the interaction between the cells and SF matrices. Our results indicate that the SF nanofiber matrix may be more preferable than SF film and SF microfiber matrices for biomedical applications, such as wound dressings and scaffolds for tissue engineering.
Min
Byung-Moo
BM
Department of Oral Biochemistry and Dental Research Institute, IBEC and BK21 HLS, College of Dentistry, Seoul National University, Seoul 110-749, Republic of Korea.
Jeong
Lim
L
Nam
Young Sik
YS
Kim
Jin-Man
JM
Kim
Jin Young
JY
Park
Won Ho
WH
D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D024042:Collagen Type I / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D005349:Fibroins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D007797:Laminin / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D011487:Protein Conformation; D013055:Spectrophotometry, Infrared / Q000379:methods
D024042:Collagen Type I; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D007797:Laminin; D005349:Fibroins