Role of offset and gradient architectures of 3-D melt electrowritten scaffold on differentiation and mineralization of osteoblasts
Role of offset and gradient architectures of 3-D melt electrowritten scaffold on differentiation and mineralization of osteoblasts
复制标题
DOI：
10.1186/s40824-019-0180-z
复制
发表时间：
2020-01-03
影响因子：
11.3
通讯作者：
Hamlet, Stephen
中科院分区：
文献类型：
--
作者：
Abbasi, Naghmeh;Ivanovski, Saso;Hamlet, Stephen
背景基于细胞支架的疗法具有提供有效的骨再生治疗的潜力，并且PCL已被普遍用作支架，然而其有效性受到差的细胞保留特性的限制。这可以通过具有有效孔排列的多孔支架结构来改善，以增加细胞截留。为了促进这一点，熔融电写入（MEW）已被开发为能够通过可预测的纤维沉积制造具有精确微孔尺寸的细胞支撑支架的技术。然而，支架结构对细胞基因表达的影响尚未完全阐明。（250、500和750 μ m），以及两个具有不同纤维布局的偏移脚手架（30和50%）和一种具有250-500 - 750 μ m三种梯度孔径的复合支架。磷酸钙改性应用于增强PCL支架的亲水性和骨诱导性，然后接种成骨细胞，然后将其保持在培养物中长达30天。在这段时间内，成骨细胞形态，基质矿化，成骨基因表达和胶原production.ResultsThe在体外研究结果显示，梯度支架显着增加碱性磷酸酶活性的附着成骨细胞，而基质矿化较高的50%偏移支架。与其他成骨基因相比，骨钙素和骨桥蛋白基因的表达也在培养30天后上调，特别是在偏移和梯度支架结构中。免疫组化显示，在偏移和梯度支架structure.ConclusionsThis研究表明，在梯度和纤维偏移PCL支架制备MEW显着提高成骨细胞的成骨潜能，因此可以提供上级结果在骨再生应用中的异质性孔径的显着表达。
BackgroundCell-scaffold based therapies have the potential to offer an efficient osseous regenerative treatment and PCL has been commonly used as a scaffold, however its effectiveness is limited by poor cellular retention properties. This may be improved through a porous scaffold structure with efficient pore arrangement to increase cell entrapment. To facilitate this, melt electrowriting (MEW) has been developed as a technique able to fabricate cell-supporting scaffolds with precise micro pore sizes via predictable fibre deposition. The effect of the scaffold's architecture on cellular gene expression however has not been fully elucidated.MethodsThe design and fabrication of three different uniform pore structures (250, 500 and 750 mu m), as well as two offset scaffolds with different layout of fibres (30 and 50%) and one complex scaffold with three gradient pore sizes of 250-500 - 750 mu m, was performed by using MEW. Calcium phosphate modification was applied to enhance the PCL scaffold hydrophilicity and bone inductivity prior to seeding with osteoblasts which were then maintained in culture for up to 30days. Over this time, osteoblast cell morphology, matrix mineralisation, osteogenic gene expression and collagen production were assessed.ResultsThe in vitro findings revealed that the gradient scaffold significantly increased alkaline phosphatase activity in the attached osteoblasts while matrix mineralization was higher in the 50% offset scaffolds. The expression of osteocalcin and osteopontin genes were also upregulated compared to other osteogenic genes following 30days culture, particularly in offset and gradient scaffold structures. Immunostaining showed significant expression of osteocalcin in offset and gradient scaffold structures.ConclusionsThis study demonstrated that the heterogenous pore sizes in gradient and fibre offset PCL scaffolds prepared using MEW significantly improved the osteogenic potential of osteoblasts and hence may provide superior outcomes in bone regeneration applications.