Chemical Engineering Faculty, Sahand University of Technology, Tabriz, Iran.
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
J Cell Biochem. 2019 Sep;120(9):15069-15082. doi: 10.1002/jcb.28768. Epub 2019 Apr 24.
Modular bone tissue engineering is touted as an alternative approach to replace the damaged bone tissue. Hydrogel microcapsules could promote therapeutic properties by providing 3D condition and an increased cell-to-cell interaction. We investigated the osteogenic properties of alginate-nano-silica hydrogels enriched with collagen and gelatin on human osteoblast-like MG-63 cells. For encapsulation, cells were divided into three groups; control (alginate+ nano-silica), collagen (alginate + collagen + nano-silica), and gelatin (alginate + gelatin + nano-silica) and expanded for 28 days. Cell survival was determined by trypan blue staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. To confirm the osteogenic potential, we measured the alkaline phosphatase activity. Alizarin red S staining was used to reveal the existence of hydroxyapatite and transcription BMP-2, osteocalcin and osteonectin evaluated by the real-time polymerase chain reaction. Collagen substrate caused a reduced swelling ratio compared with the control and gelatin groups (P < 0.05). Compared with other groups, collagen had potential to improve mechanical strength and generate porous membrane structure. The addition of collagen (4-fold) and gelatin (1.5-fold) increased cell proliferation rate compared with the control (P < 0.05). Biochemical analysis and Alizarin red S staining showed that collagen-induced osteogenesis by induction of alkaline phosphatase and matrix mineralization. The expression of osteocalcin and BMP-2 was increased in cells from the collagen group. As a result, the combination of natural polymers collagen and gelatin with alginate + nano-silica can increase the osteogenic potential of human osteoblasts.
模块化骨组织工程被吹捧为一种替代方法，可替代受损的骨组织。水凝胶微胶囊可通过提供 3D 条件和增加细胞间相互作用来促进治疗特性。我们研究了富含胶原蛋白和明胶的藻酸盐-纳米二氧化硅水凝胶对人成骨样 MG-63 细胞的成骨特性。为了封装，将细胞分为三组;对照组（藻酸盐+纳米二氧化硅）、胶原蛋白组（藻酸盐+胶原蛋白+纳米二氧化硅）和明胶组（藻酸盐+明胶+纳米二氧化硅），并扩展培养 28 天。通过台盼蓝染色和 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法确定细胞存活率。为了确认成骨潜能，我们测量了碱性磷酸酶活性。茜素红 S 染色用于揭示羟基磷灰石的存在，并用实时聚合酶链反应评估 BMP-2、骨钙素和骨粘连蛋白的转录。与对照组和明胶组相比，胶原蛋白基质的溶胀率降低（P<0.05）。与其他组相比，胶原蛋白具有改善机械强度和产生多孔膜结构的潜力。与对照组相比，添加胶原蛋白（4 倍）和明胶（1.5 倍）可提高细胞增殖率（P<0.05）。生化分析和茜素红 S 染色表明，胶原蛋白通过诱导碱性磷酸酶和基质矿化来诱导成骨。胶原蛋白组细胞中骨钙素和 BMP-2 的表达增加。结果表明，天然聚合物胶原蛋白和明胶与藻酸盐+纳米二氧化硅的结合可提高人成骨细胞的成骨潜能。
