蚕丝支架与3T3-L1前脂肪细胞生物相容性的体外实验(英文)

Biocompatibility of silk scaffold/3T3-L1 preadipocytes compounds An in vitro experiment

背景:蚕丝是天然制品,其力学性能及生物相容性优于传统人工合成的可降解高分子材料,在医疗领域中已获得了广泛的应用而受到关注。目的:观察蚕丝对3T3-L1前脂肪细胞吸附作用及蚕丝对3T3-L1前脂肪细胞形态和功能的影响。方法:取原料蚕丝和用胰酶消化后的蚕丝任意缠绕成网状立体构型纤维条索,架空固定在自制的不锈钢支架上,支架网孔70～200μm,厚200～300μm,孔隙率为20%。消化后的蚕丝三维支架放入24孔培养板中,将浓度为6×1010L-1的3T3-L1前脂肪细胞悬液每孔滴入3滴,每孔细胞数量为1×107个。悬空孵育4h,待细胞充分吸附在支架上后加入培养液,令细胞完全浸没,隔两三天换半液,培养1～4周。结果与结论:①倒置显微镜观察3T3-L1前脂肪细胞-蚕丝复合物可见细胞伸出细长的突起沿着蚕丝不断向前迁移延伸,细胞首尾相互融合,渐渐连成一片分布于蚕丝网眼内。②扫描电镜观察3T3-L1前脂肪细胞-蚕丝复合物可见细胞与支架紧密贴附,适度伸展,并有基质分泌。提示蚕丝对3T3-L1前脂肪细胞具有良好的吸附作用,并能维持3T3-L1前脂肪细胞正常形态和功能。

BACKGROUND：Silk,a natural product,has been paid wide attention in medical field owing to a fact that silk＇s mechanical property and biocompatibility are superior to traditional artificially synthesized degradable polymers.OBJECTIVE：To observe the effects of silk on the adsorption as well as cell morphology and function of 3T3-L1 preadipocytes.METHODS：Raw silk and trypsin-digested silk were ad libitum twisted into reticular fibrous cord with three-dimensional structure：mesh size 70-200μm,thickness 200-300μm,and porosity 20%.The three-dimensional scaffolds made of digested silk were placed in the 24-well plate.3T3-L1 preadipocytes suspension（6×1010/L）was added to the culture plate,3 drops comprising 1×107 cells per well.Following 4-hour incubation in the air,when cells fully attached to the scaffold,they were thoroughly soaked with medium,which was renewed every other 2-3 days for a total of 1-4 weeks.RESULTS AND CONCLUSION：Inverted microscope results showed that silk scaffold/3T3-L1 preadipocytes compounds presented with slender prominences that stretched out and migrated ahead to gradually connect together through a head-to-end fusion fashion and enter into meshes.Scanning electron microscope results demonstrated that in the silk scaffold/3T3-L1 preadipocytes compounds,cells tightly attached to silk scaffold,appropriately spread out,and secreted matrix.Silk shows better absorption for 3T3-L1 preadipocytes and maintains the normal morphology and functions of 3T3-L1 preadipocytes.