人源和鼠源成骨前体细胞在三维组织工程支架中表现出不同的成骨模式。
Human and mouse osteoprogenitor cells exhibit distinct patterns of osteogenesis in three-dimensional tissue engineering scaffolds.
机构信息
Redondo Beach and Los Angeles, Calif. From the Division of Plastic Surgery, Department of Surgery, University of California, Los Angeles; the Department of Surgery, Harbor UCLA Medical Center; and the Plastic Surgery Laboratory and the Research Service, Veterans Affairs Greater Los Angeles Healthcare System.
出版信息
Plast Reconstr Surg. 2009 Dec;124(6):1869-1879. doi: 10.1097/PRS.0b013e3181bf81ab.
BACKGROUND
Understanding interspecies variation between animal models and humans is essential to develop tissue-engineered bone. The authors studied osteogenic and angiogenic marker expression in human and murine osteoblasts and mesenchymal stem cells.
METHODS
Three human cells (human mesenchymal stem cells, multilineage progenitor cells, and normal human osteoblasts) and three murine cells (MC3T3-E1, C3H10T1/2, and M2-10B4) were used. Cells were seeded onto poly-lactide-glycolic acid-coated tissue culture plates or three-dimensional poly-lactide-glycolic acid scaffolds, incubated in osteogenic medium, and harvested at 1, 4, and 7 days. mRNA expression was analyzed using quantitative real-time reverse-transcriptase polymerase chain reaction for osteogenic markers, including alkaline phosphatase, osteocalcin, bone sialoprotein, and core-binding factor alpha-1, and angiogenic markers, including vascular endothelial growth factor and interleukin-8. Data were analyzed using analysis of variance.
RESULTS
All human cells had significantly increased expression of osteogenic markers in three dimensions compared with two dimensions (alkaline phosphatase by 220 percent, osteocalcin by 323 percent, bone sialoprotein by 534 percent, and core-binding factor alpha-1 by 357 percent). However, all murine cells exhibited significant decreases in the expression of osteogenic markers in three-dimensional compared with two-dimensional cultures (alkaline phosphatase by 89 percent, osteocalcin by 64 percent, bone sialoprotein by 76 percent, and core-binding factor alpha-1 by 73 percent). In contrast, all human and murine cells showed markedly elevated expression of angiogenic factors interleukin-8 and vascular endothelial growth factor in three-dimensional compared with two-dimensional cultures. Measurement of alkaline phosphatase activity confirmed this pattern of osteogenic differentiation.
CONCLUSIONS
In three-dimensional versus two-dimensional cultures, osteogenesis increased significantly in human cells but decreased in murine cells; angiogenesis increased regardless of species. Since three-dimensional cultures represent in vivo conditions more closely, this species variation has important translational implications to tissue-engineered bone research.
背景
了解动物模型和人类之间的种间差异对于开发组织工程骨至关重要。作者研究了人源成骨细胞和间充质干细胞的成骨和成血管标记物的表达。
方法
使用了三种人源细胞（人骨髓间充质干细胞、多能祖细胞和成骨细胞）和三种鼠源细胞（MC3T3-E1、C3H10T1/2 和 M2-10B4）。将细胞接种到聚乳酸-羟基乙酸涂层的组织培养板或三维聚乳酸-羟基乙酸支架上，在成骨培养基中孵育，并在第 1、4 和 7 天收获。使用定量实时逆转录聚合酶链反应分析成骨标记物（碱性磷酸酶、骨钙素、骨涎蛋白和核心结合因子α-1）和血管生成标记物（血管内皮生长因子和白细胞介素-8）的 mRNA 表达。使用方差分析对数据进行分析。
结果
与二维相比，所有的人源细胞在三维中均表现出成骨标记物的表达显著增加（碱性磷酸酶增加 220%，骨钙素增加 323%，骨涎蛋白增加 534%，核心结合因子α-1 增加 357%）。然而，所有的鼠源细胞在三维中均表现出成骨标记物表达的显著降低（碱性磷酸酶降低 89%，骨钙素降低 64%，骨涎蛋白降低 76%，核心结合因子α-1 降低 73%）。相比之下，所有的人源和鼠源细胞在三维中均表现出血管生成因子白细胞介素-8 和血管内皮生长因子的显著升高。碱性磷酸酶活性的测量证实了这种成骨分化模式。
结论
与二维相比，三维培养中人类细胞的成骨作用显著增加，而鼠源细胞的成骨作用则减少；无论物种如何，血管生成均显著增加。由于三维培养更接近体内条件，这种种间差异对组织工程骨研究具有重要的转化意义。
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