Department of Chemical Engineering, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
Acta Biomater. 2010 Apr;6(4):1446-52. doi: 10.1016/j.actbio.2009.10.004. Epub 2009 Oct 8.
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 H(2)O(2) as an electron acceptor. This study evaluates the effect of H(2)O(2) and HRP concentrations on cellular adhesiveness and proliferation on the resultant enzymatically crosslinked Alg-Ph gels. After 4h of seeding, 81.1% of L929 fibroblast cells adhere to an Alg-Ph hydrogel prepared with 1 U ml(-1) HRP and 1mM H(2)O(2). Increasing the concentration of H(2)O(2) to 15 mM decreases the percentage of adhering cells to 28.4%. The cellular adhesion at this H(2)O(2) 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.
具有酚羟基（Alg-Ph）基团的海藻酸钠水溶液可通过辣根过氧化物酶（HRP）催化的 Ph 基团之间的氧化交联反应凝胶化，消耗 H（2）O（2）作为电子受体。本研究评估了 H（2）O（2）和 HRP 浓度对细胞黏附性和增殖的影响，研究对象是由此酶交联的 Alg-Ph 凝胶。接种后 4 小时，1 U ml（-1）HRP 和 1mM H（2）O（2）制备的 Alg-Ph 水凝胶中 81.1%的 L929 成纤维细胞黏附。将 H（2）O（2）浓度增加到 15 mM 会将黏附细胞的百分比降低到 28.4%。将 HRP 浓度增加到 10 U ml（-1）可将该 H（2）O（2）浓度下的细胞黏附率提高到 82.6%。在培养 168 小时期间，黏附在 Alg-Ph 水凝胶上的细胞会建立一个细胞单层。然后可以在含有 1.0 mg ml（-1）的海藻酸钠酶的培养基中浸泡 5 分钟，收获细胞片。收获的细胞片重新黏附，并在转移到另一个细胞培养皿后，细胞片中的细胞增殖。
