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Control of cellular adhesiveness in an alginate-based hydrogel by varying peroxidase and H2O2 concentrations during gelation
Authors:S Sakai  K Hirose  K Moriyama  K Kawakami
Institution:1. Chemical Engineering Faculty, Sahand University of Technology, Tabriz 51335-1996, Iran;2. Division of Chemical Engineering, Department of Materials Science and Engineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka 560-8531, Japan;1. Department of Applied Science, National Hsinchu University of Education, Hsinchu 30014, Taiwan, ROC;2. Department of Cardiac Surgery, Boston Children’s Hospital, Harvard Medical School, Boston, MA 02115, USA;3. Material and Chemical Research Laboratories, Industrial Technology Research Institute, Hsinchu 31040, Taiwan, ROC;1. State Key Laboratory of Chemical Engineering, East China University of Science and Technology, Shanghai, 200237, China;2. Shanghai Key Laboratory of Multiphase Materials Chemical Engineering, East China University of Science and Technology, Shanghai, 200237, China;3. Engineering Research Center of Materials Chemical Engineering of Xinjiang Bingtuan, Key Laboratory of Materials Chemical Engineering of Xinjiang Uygur Autonomous Region, Shihezi University, Shihezi, 832000, China
Abstract:An aqueous solution of alginate possessing phenolic hydroxyl (Alg-Ph) groups is gellable via a horseradish peroxidase (HRP)-catalyzed oxidative crosslinking reaction between Ph groups, consuming H2O2 as an electron acceptor. This study evaluates the effect of H2O2 and HRP concentrations on cellular adhesiveness and proliferation on the resultant enzymatically crosslinked Alg-Ph gels. After 4 h of seeding, 81.1% of L929 fibroblast cells adhere to an Alg-Ph hydrogel prepared with 1 U ml?1 HRP and 1 mM H2O2. Increasing the concentration of H2O2 to 15 mM decreases the percentage of adhering cells to 28.4%. The cellular adhesion at this H2O2 concentration is increased to 82.6% by increasing the HRP concentration to 10 U ml?1. The cells adhering to the Alg-Ph hydrogels with higher cellular adhesiveness establish a confluent monolayer during 168 h of culture. A cell sheet can then be harvested within 5 min of immersion in a medium containing alginate lyase at 1.0 mg ml?1. The harvested cell sheet re-adhere, and the cells contained in the sheet proliferate after being transferred to another cell culture dish.
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