用于ADSCs增殖分化的海藻酸钙/骨粉微胶珠的制备与检测  

Preparation and detection of calcium alginate/bone powder microbeads for expansion and differentiation of ADSCs

作　　者：宋克东[1] 闫新宇[1] 宋飞[2] 杨延飞[1] 李士晓[1] 朱艳霞[3] 刘天庆[1] 

机构地区：[1]大连理工大学精细化工国家重点实验室干细胞与组织工程研究室,辽宁大连116024 [2]大连医科大学第二附属医院神经外科,辽宁大连116027 [3]深圳大学医学部深圳抗衰老与再生医学重点实验室,广东深圳518060

出　　处：《功能材料》2015年第20期20117-20122,共6页Journal of Functional Materials

基　　金：国家自然科学基金资助项目(31370991/81301597/31170945);教育部留学回国科研启动基金资助项目(42P);精细化工国家重点实验室开放基金资助项目(KF1111);中央高校基本科研业务费资助项目(DUT14YQ016);深圳市战略性新兴产业发展专项资金资助项目(JCYJ20120821110533438/JCYJ20130326110154156);霍英东基金资助项目(132027)

摘　　要：海藻酸钙微胶珠具有良好的生物相容性、可降解、机械强度高、韧性好等优点,已被广泛应用于组织工程研究。骨粉中含有大量的羟基磷灰石及BMP等生长因子,具有良好的骨诱导活性。结合上述两种材料的优点,制备了一种复合海藻酸钙/骨粉微胶珠,并系统研究了人源脂肪干细胞(adipose-derived stem cells,ADSCs)在其中的最佳包埋密度。通过正交实验综合考虑海藻酸钠溶液浓度、氯化钙溶液浓度和骨粉密度对海藻酸钙/骨粉微胶珠机械强度及传质性能的影响,并采用液滴法制备了最优化的用于ADSCs体外增殖分化的微胶珠。结果表明,海藻酸钠溶液的浓度对微胶珠传质能力和机械强度的影响最大,CaCl2溶液浓度其次,骨粉密度的影响最小。当海藻酸钠溶液浓度、CaCl2溶液浓度和骨粉密度分别为2.5%,4.5%和5.0mg/mL时,微胶珠的性能最好。对细胞扩增倍数的比较和死活染色等检测表明,当ADSCs的初始包埋密度为5×106 cells/mL时,更适于ADSCs的扩增与分化。Calcium alginate/bone-powder microbeads have been widely used in tissue engineering research due to their excellent biocompatibility,biodegradability,enhanced mechanical strength and toughness.Bone-powder containing abundant hydroxylapatite and growth factors such as BMP,possess an excellent osteoinductive activity.Herein,a hybrid calcium alginate/bone-powder microbeads combining both advantages of the abovementioned materials was prepared,afterwards the optimal inoculating density of the Adipose-derived stem cells(ADSCs)embedded in hybrid microbeads was explored systematically.The optimized microbeads suitable to the in vitro expansion and differentiation of ADSCs,were prepared using the droplet method under overall considering the concentration of calcium alginate and calcium chloride,the density of bone powder,also by the orthogonal experiment.It demonstrates that the concentration of the sodium alginate has the strongest influence on the mass transfer and mechanical strength of the microbeads,secondly the calcium chloride,then the density of bone powder.The microbeads perform best when the concentrations of the sodium alginate and calcium chloride are 2.5%and 4.5%respectively,as well as the density of bone powder was 5.0mg/mL.Additionally,the comparison about cell expanding fold and live/dead staining are carried out,which shows the ADSCs expand and differentiate well with the initial embedding density of 5×106 cells/mL.

关 键 词：海藻酸钙微胶珠 脂肪干细胞 骨粉 骨组织工程 细胞载体 

分 类 号：R318.08[医药卫生—生物医学工程]