Synthesis of scaffold-free, three dimensional, osteogenic constructs following culture of skeletal osteoprogenitor cells on glass surfaces.
Synthesis of scaffold-free, three dimensional, osteogenic constructs following culture of skeletal osteoprogenitor cells on glass surfaces.
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DOI：
10.1016/j.bonr.2021.101143
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发表时间：
2021-12
期刊：
影响因子：
2.5
通讯作者：
El-Serafi AT
中科院分区：
文献类型：
--
作者：
Alghfeli L;Parambath D;Manzoor S;Roach HI;Oreffo ROC;El-Serafi AT
干细胞向三维（3D）成骨构建体的有效分化仍然是一个未满足的挑战。这些结构对于由于先天性或创伤性原因导致骨缺损的患者至关重要。由于与材料的物理和化学性质的相互作用，细胞命运和功能的调节是众所周知的。目前的研究已经检查了在玻璃表面上作为单层或在玻璃管中作为颗粒培养物培养的人骨骼群体的成骨分化潜力。通过形态测定评估3D性能，并通过分子表征以及基质形成评估分化。在玻璃表面上培养后，观察到骨骼群体碱性磷酸酶表达的早期时间表达。Skeletal群体接种在玻璃管上，作为单层粘附到管基部，随后在空气-介质界面处形成3D颗粒。在玻璃上培养的颗粒显示出其在塑料管中培养的对应物的重量的4.9 ± 1.3倍和直径的2.9 ± 0.1倍，并且显示出成骨基质蛋白如胶原蛋白I和骨粘连蛋白的增加的产生。颗粒的大小和重量与表面积相关，而与接种的细胞数量相关。在玻璃上培养的小球中，整体DNA甲基化水平降低。相反，基因表达分析证实了细胞外基质蛋白和成骨相关生长因子的上调。这种在玻璃管上培养骨骼细胞的简单方法提供了一种无支架的3D构建平台，用于生成颗粒，从而能够分析和评估组织发育和多个构建体的整合，并对组织修复和再生应用产生影响。
Efficient differentiation of stem cells into three-dimensional (3D) osteogenic construct is still an unmet challenge. These constructs can be crucial for patients with bone defects due to congenital or traumatic reasons. The modulation of cell fate and function as a consequence of interaction with the physical and chemical properties of materials is well known. The current study has examined the osteogenic differentiation potential of human skeletal populations following culture on glass surfaces, as a monolayer, or in glass tubes as a pellet culture. The 3D prosperities were assessed morphometrically and the differentiation was evaluated through molecular characterization as well as matrix formation. Early temporal expression of alkaline phosphatase expression of skeletal populations was observed following culture on glass surfaces. Skeletal populations seeded on glass tubes, adhered as a monolayer to the tube base and subsequently formed 3D pellets at the air -media interface. The pellets cultured on glass displayed 4.9 ± 1.3 times the weight and 2.9 ± 0.1 the diameter of their counterpart cultured in plastic tubes and displayed enhanced production of osteogenic matrix proteins, such a collagen I and osteonectin. The size and weight of the pellets correlated with surface area in contrast to cell numbers seeded. Global DNA methylation level was decreased in pellets cultured on glass. In contrast, gene expression analysis confirmed upregulation extracellular matrix proteins and osteogenesis-related growth factors. This simple approach to the culture of skeletal cells on glass tubes provides a scaffold-free, 3D construct platform for generating pellets enabling analysis and evaluation of tissue development and integration of multiple constructs with implications for tissue repair and regenerative application on scale-up.
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