载红细胞海藻酸钠-聚赖氨酸-海藻酸钠微囊的制备及质量评价

Preparation and quality evaluation of alginate/poly-L-lysine/alginate microencapsules loaded with red blood cells

目的:制备载红细胞海藻酸钠-聚赖氨酸-海藻酸钠微囊(APA微囊)并进行质量评价。方法:采用高压静电成囊技术制备海藻酸钙微胶珠,与聚左赖氨酸反应后再包覆海藻酸钠形成三层膜结构,柠檬酸钠液化囊心后得APA微囊;光学显微镜下检测微囊的粒径,振摇法考察微囊的机械强度,不同酸碱度溶液测定微囊的化学强度,同时紫外分光光度法检测透过微囊膜血红蛋白的吸光度来评价囊膜的通透性,小鼠腹腔移植观察微囊生物相容性。结果:在成囊电压4kV,推进速度50mL·h-1,针头直径0.7mm的条件下制得的微囊表面圆整光滑,95%以上微囊的粒径集中分布于1.5～2.1mm;72h内机械振摇破损率约为5%,在pH4.2～9.4的磷酸缓冲液中稳定;小鼠腹腔内移植于2周后回收的微囊外形圆整、光滑,无破损及明显结缔组织包绕。结论:本方法制备得到的APA微囊具有良好的机械、化学强度及生物相容性,可以作为微囊化细胞移植的载体。

Objective：To prepare alginate/poly-L-lysine/alginate（APA） microencapsules loaded with red blood cells and evaluate it＇s quality.Methods：The alginate calcium microbeeds were prepare useing high static voltage sponsor,then reacted with poly-L-lysine and alginate sodium,and citrate sodium was used to wash away the alginate in the central.Particle diameter was measured under microscope,mechanical strength of membrane was evaluated by continuous shaking,and chemical intensity was investigated by adding different pH buffers into microcapsules solution.The absorbance of hemoglobin was determined with ultraviolet spectrophotometry to evaluate the permeability of the membrane.The biocompatibility of microcapsules was evaluated by transplanting them into mice＇ abdominal cavity.Results：Using the optimal coefficient：under the conditions of static voltage 4kv,push speed 50 ml·h-1,needle diameter 0.7 mm,and the height between needle and liquid level 1.5 cm,the shape of microcapsules was round,smooth and uniform,and 95% of its diameter ranged from 1.6 to 2.1 mm.The damage ratio of mechanical shaking was about 5% and no microcapsules were destroyed in different phosphate buffer in 72 h.The biocompatibility of microcapsules was well after 2 weeks in the abdominal cavity of mouse.Conclusion：APA microencapsules shows its good mechanical strength,chemical intensity and biocompatibility in vivo,which provide a potential carrier for cell transplantation.