Toxic Chemical Cross-linker-free Cryosponges Made from Chitosan-Gluconic Acid Conjugate for Chondrocyte Culture
Toxic Chemical Cross-linker-free Cryosponges Made from Chitosan-Gluconic Acid Conjugate for Chondrocyte Culture
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DOI：
10.1252/jcej.16we145
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发表时间：
2017-02-01
影响因子：
0.8
通讯作者：
Yoshida, Masahiro
中科院分区：
文献类型：
--
作者：
Takei, Takayuki;Yoshitomi, Hiroki;Yoshida, Masahiro
壳聚糖已被广泛用作软骨再生的多孔支架材料。在壳聚糖支架的制备过程中，有毒的化学交联剂如戊二醛被用来交联壳聚糖分子。我们以前开发的化学无交联剂壳聚糖海绵只由壳聚糖-葡萄糖酸结合物(CG)的水溶液冻融形成。在本研究中，我们将CG冻融材料作为软骨组织工程的支架材料。低葡萄糖酸含量的CG冻球在添加胎牛血清的细胞培养液中表现出明确的孔道结构和高稳定性，而胎牛血清最初含有低水平的溶菌酶，这是一种壳聚糖的水解酶。通过在-30℃下冷冻CG溶液，可以获得有利于软骨再生的平均孔径(100-300 Im)的冻球。与用戊二醛交联的冻球相比，在普通的CG冻球中牛软骨细胞的增殖和糖胺聚糖的产生更容易。这些结果表明，CG冻融体是一种很有前途的软骨组织工程支架材料。
Chitosan has been widely utilized as a porous scaffold material for cartilage regeneration. During the preparation process of chitosan scaffolds, toxic chemical cross-linkers, such as glutaraldehyde, have been used for cross-linking of chitosan molecules. We previously developed chemical cross-linker-free chitosan sponges formed only by freeze-thawing an aqueous solution of chitosan-gluconic acid conjugate (CG). In the present study, we applied the CG cryosponges as scaffolds for cartilage tissue engineering. CG cryosponges with low gluconic acid content showed a defined porous structure and high stability in cell culture medium supplemented with fetal bovine serum that originally contained a low level of lysozyme, a hydrolysis enzyme for chitosan. A favorable mean pore size of the cryosponges for cartilage regeneration (100-300 Im) was accomplished by freezing the CG solution at -30 degrees C. Proliferation and glycosaminoglycan production of bovine chondrocytes were facilitated in plain CG cryosponges compared with those cross-linked with glutaraldehyde. These results demonstrated that the CG cryosponges were promising scaffolds for cartilage tissue engineering.