Data for: Plasmid BMP-2–Embedded Gelatin Sponge As A Gene-Activated Matrix for Preosteoblast Differentiation

Supplementary Figure 1 To allow endocytosis of DNA complexes into the cells, 5 × 104 cells and TransIT-2020/pEGFP-C1 complexes were mixed for 5, 15, 30, or 60 min (n = 3). Flow cytometry analysis was performed to evaluate GFP expression at these time points using FACS Calibur flow cytometer (Becton Dickinson, Heidelberg, Germany). Cells were gated at 1 × 104 cells/sample in FL1 channel setting, and BMP-2 transfection samples under the same condition at each time point were used as controls(Figure S1A). Transfection efficiencies of these four mixing time points were compared using Becton Dickinson Cell Quest software(Figure S1B). We found that duration of 30 min was the optimal mixing time, as the average GFP expression (9.92%) at 30 min was the highest compared to that at the other three time points. Live and dead assay : The scaffolds were stained using a Live/Dead Cytotoxicity/Viability kit (Invitrogen, Carlsbad, CA)from day 7 to day 28 .The live cells ,dead cells and merge images were all included. Figure 5 ALP supplementary data : all ALP activity corresponding to Figure 5 Figure 6 ARS supplementary data : all ARS activity corresponding to Figure 6 Figure 7 BMP-2 supplementary data: all BMP-2 expression corresponding to Figure 7补充图1 为使DNA复合物可被细胞内吞，将5×10⁴个细胞与TransIT-2020/pEGFP-C1复合物分别孵育5、15、30或60分钟（n=3）。采用FACSCalibur流式细胞仪（Becton Dickinson，德国海德堡）在各时间点通过流式细胞术分析以评估绿色荧光蛋白（Green Fluorescent Protein, GFP）的表达水平。每个样本以1×10⁴个细胞设门于FL1通道，并以相同条件下各时间点的骨形态发生蛋白2（Bone Morphogenetic Protein 2, BMP-2）转染样本作为对照（补充图S1A）。采用碧迪（Becton Dickinson）Cell Quest软件比较该四个孵育时间点的转染效率（补充图S1B）。结果显示，30分钟为最优孵育时长，相较于其余三个时间点，30分钟时的平均GFP表达率（9.92%）最高。 活死细胞检测： 分别于第7天至第28天，采用活/死细胞毒性/活力检测试剂盒（Invitrogen，美国加利福尼亚州卡尔斯巴德）对支架进行染色，结果包含活细胞、死细胞及合并荧光的图像。 图5碱性磷酸酶（Alkaline Phosphatase, ALP）补充数据：对应图5的全部碱性磷酸酶活性数据 图6茜素红S（Alizarin Red S, ARS）补充数据：对应图6的全部茜素红S活性数据 图7骨形态发生蛋白2（Bone Morphogenetic Protein 2, BMP-2）补充数据：对应图7的全部骨形态发生蛋白2表达数据