Porous hydrogel of wool hair keratin as a substrate for cell culture
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1
Osaka Prefecture Univertisy, Department of Biological Science, Graduate School of Science,, Japan
Introduction: Keratin is contained rich in animal epidermal tissues including hair, nail, horn, and also common proteins such as wool fibers. Keratins have not been so much utilized as Type I collagen as biomaterials although much amount is available as a bio-resource. In this study, we show a novel simple method to prepare a sponge-like porous keratin hydrogel. Wool keratin was extracted for 18 h at 60 oC in a solution containing 8 M guanidine hydrochloride and 2-mercaptoethanol, followed by dialysis against water. Specific water content (%) was calculated as an index for swelling of the gel according to the following equation.
Specific water content = (Ww-Wd)/Wd×100 (%)
Wd: dry weight of the gel after lyophilization; Ww: wet weight of the gel after swelling in either PBS(-) or DMEM. We tried cell culture on the gel and evaluated the cell growth by WST assay. All animal experiments were conducted in accordance with institutional guidelines and national standards with approval from the Animal Experiment Committee of Osaka Prefecture University.
Results and Discussion: Extracted keratin was aggregated and recrosslinked to form a porous hydrogel like a sponge. Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) proved that Type I and Type II keratin subunits were extracted by that procedure without degradation by hydrolysis. The porous keratin gel had a highly porous structure and a fast-swelling property in rehydration after freeze-drying. The gel could be autoclaved without significant change of mechanical strength and shapes. It had also high mechanical strength both in the tensile test and the measurement of dynamic viscoelasticity. The gel is flexible but mechanically durable especially for tensile strain. Those property was different from that of the chemically crosslinked type I collagen gel. Three types of animal cells, PC12 cells, HOS cells and murine embryonic fibroblasts, well attached and grew on the surface of the porous hydrogel. We observed those cells by fluorescent microscopy after live/dead staining and also by a scanning electron microscope. The keratin hydrogel has properties suitable for scaffold in tissue engineering. We tried some preliminary studies on application of the keratin hydrogel for bone tissue engineering by culturing osteoblasts.
Reference: Ozaki, Y; Takagi, Y; Mori, H; Hara, M. Mater. Sci. Eng. C 2014, 42, 146–154.
Keywords:
Tissue Engineering,
stem cell,
3D scaffold,
Biodegradable material
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Regenerative medicine: biomaterials for control of tissue induction
Citation:
Yuki
O,
Takagi
Y,
Mori
H and
Hara
M
(2016). Porous hydrogel of wool hair keratin as a substrate for cell culture.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.01232
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.
*
Correspondence:
Dr. Ozaki Yuki, Osaka Prefecture Univertisy, Department of Biological Science, Graduate School of Science,, Sakai-city, Osaka Prefecture, Japan, yuki.ozaki@cira.kyoto-u.ac.jp
Dr. Yusuke Takagi, Osaka Prefecture Univertisy, Department of Biological Science, Graduate School of Science,, Sakai-city, Osaka Prefecture, Japan, st304016@edu.osakafu-u.ac.jp
Dr. Hideki Mori, Osaka Prefecture Univertisy, Department of Biological Science, Graduate School of Science,, Sakai-city, Osaka Prefecture, Japan, morihide@b.s.osakafu-u.ac.jp
Dr. Masayuki Hara, Osaka Prefecture Univertisy, Department of Biological Science, Graduate School of Science,, Sakai-city, Osaka Prefecture, Japan, hara@b.s.osakafu-u.ac.jp