羧乙基壳聚糖-纳米羟基磷灰石复合材料细胞相容性和生物力学性能的实验研究  

Cellular biocompatibflity and biomechanical properties of N-carboxyethyl chitosan / nano-hydroxyapatite scaffolds

作　　者：史宏灿 王万鹏 陆妍[2] 朱爱萍[2] 陈林松 李海甲 

机构地区：[1]扬州大学临床医学院胸心外科,江苏省扬州225001 [2]扬州大学化学化工学院

出　　处：《中华生物医学工程杂志》2011年第3期230-233,共4页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金（30672080、30972968）;江苏省“科教兴卫工程”医学重点人才基金（200728031）;江苏省“青蓝工程”学术带头人基金（20081215）;江苏省“六大人才高峰”高层次人才基金.

摘　　要：目的制备羧乙基壳聚糖一纳米羟基磷灰石（NCECS／nHA）复合材料，研究其生物力学性能以及与气管软骨细胞的生物相容性。方法气管软骨片段取自8周龄大耳白兔，Ⅱ型胶原酶消化，将所获得软骨细胞传代培养。将体外制备的NCECS／nHA复合材料分别进行干态标本和湿态标本的生物力学检测。将第3代软骨细胞种植到NCECS／nHA复合材料，分别计算材料表面软骨细胞在2h、6h、12h细胞贴壁率，并用噻唑蓝（MTT）法测定细胞增殖活性。结果NCECS／nHA复合材料具有良好的生物力学性能。兔气管软骨细胞在NCECS／nHA复合材料表面上12h的贴壁率达（88．4＋2．1）％，与其他组差异无统计学意义（P〉0．05）。同时MTF显示气管软骨细胞在NCECS／nHA复合材料表面生长状态良好。扫描电镜结果显示软骨细胞在NCECS／nHA薄膜上增殖和分化良好。结论NCECS／nHA复合材料具备良好的细胞相容性和适宜的生物力学强度，作为一种具有开发潜力的生物材料，可用于组织工程气管的体外构建。Objective To prepare N- carboxyethyl chitosan/nano-hydroxyapatite （NCECS/nHA） scaffolds, and to investigate its cellular biocompatibility and biomechanical properties. Methods Samples of tracheal cartilage were obtained from an 8-week-oid Japanese white rabbit. The cartilage fragments were digested with 0.2% collagenase type Ⅱ to obtain chondrocytes which were then passage cultured. Biomechanical test was performed for the dried and wet films of NCECS/nHA complex. Then, the thirdpassage chondrocytes were seeded onto the NCECS/nHA films. Cell adhesion rate was measured at 2, 6, and 12 hours after seeding, and the proliferation index of the cells on NCECS/nHA was quantified using MTF assay. Results The unseeded composite film showed excellent biomechanical properties. The cell adhesion rate of the third-passage chondrocytes seeded on NCECS/nHA films after 12 hours was （88.4±2.1）%, which was not significantly different compared to the other groups. MTT assay indicated good proliferation of chondroeytes on NCECS/nHA films. Satisfactory growth and differentiation of chondrocytes on NCECS/nHA films were also revealed by SEM. Conclusion NCECS/nHA composite film shows favorable biomechanical properties and excellent biocompatibility with rabbit tracheal chondrocytes, and therefore can be used as a scaffold material for tissue-engineered trachea.

关 键 词：组织工程 生物力学 生物相容性 羧乙基壳聚糖 纳米羟基磷灰石 

分 类 号：R318.08[医药卫生—生物医学工程]