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骨涎蛋白涂层三维打印磷酸钙支架对原代人成骨细胞的影响。

Effect of bone sialoprotein coated three-dimensional printed calcium phosphate scaffolds on primary human osteoblasts.

机构信息

Department of Orthopaedics and Traumatology, Biomatics Group, University Medical Center, Johannes Gutenberg University, Mainz, Germany.

Platform for Biomaterial Research, Biomatics Group, University Medical Center, Johannes Gutenberg University, Mainz, Germany.

出版信息

J Biomed Mater Res B Appl Biomater. 2018 Oct;106(7):2565-2575. doi: 10.1002/jbm.b.34073. Epub 2018 Jan 9.

Abstract

The combination of the two techniques of rapid prototyping 3D-plotting and bioactive surface functionalization is presented, with emphasis on the in vitro effect of Bone Sialoprotein (BSP) on primary human osteoblasts (hOBs). Our primary objective was to demonstrate the BSP influence on the expression of distinctive osteoblast markers in hOBs. Secondary objectives included examinations of the scaffolds' surface and the stability of BSP-coating as well as investigations of cell viability and proliferation. 3D-plotted calcium phosphate cement (CPC) scaffolds were coated with BSP via physisorption. hOBs were seeded on the coated scaffolds, followed by cell viability measurements, gene expression analysis and visualization. Physisorption is an effective method for BSP-coating. Coating with higher BSP concentrations leads to enhanced BSP release. Two BSP concentrations (50 and 200 μg/mL) were examined in this study. The lower BSP concentration (50 µg/mL) decreased ALP and SPARC expression, whereas the higher BSP concentration (200 μg/mL) did not change gene marker expression. Enhanced cell viability was observed on BSP-coated scaffolds on day 3. hOBs developed a polygonal shape and connected in an intercellular network under BSP influence. Quantitative cell morphology analyses demonstrated for BSP-coated CPCs an enhanced cell area and reduced circularity. The strength of the above-mentioned effects of BSP-coated scaffolds in vivo is unknown, and future work is focusing on bone ingrowth and vascularization in vivo. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2565-2575, 2018.

摘要

呈现了快速原型 3D 绘图和生物活性表面功能化这两种技术的组合,重点是骨唾液蛋白(BSP)对原代人成骨细胞(hOB)的体外影响。我们的主要目标是证明 BSP 对 hOB 中独特成骨细胞标志物表达的影响。次要目标包括检查支架的表面和 BSP 涂层的稳定性以及研究细胞活力和增殖。通过物理吸附将 BSP 涂覆到 3D 绘制的磷酸钙水泥(CPC)支架上。将 hOB 接种到涂有涂层的支架上,然后进行细胞活力测量、基因表达分析和可视化。物理吸附是 BSP 涂层的有效方法。更高的 BSP 浓度会导致 BSP 释放增强。本研究检查了两个 BSP 浓度(50 和 200 μg/mL)。较低的 BSP 浓度(50 µg/mL)降低了 ALP 和 SPARC 的表达,而较高的 BSP 浓度(200 µg/mL)没有改变基因标记的表达。在 BSP 涂覆的支架上,在第 3 天观察到增强的细胞活力。在 BSP 的影响下,hOB 呈多边形形状并在细胞间网络中连接。定量细胞形态分析表明,BSP 涂覆的 CPC 的细胞面积增加,圆形度降低。BSP 涂覆的支架在体内的上述效果的强度尚不清楚,未来的工作重点是体内的骨向内生长和血管化。© 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2565-2575, 2018.

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