Mapping human serum-induced gene networks as a basis for the creation of biomimetic periosteum for bone repair
Mapping human serum-induced gene networks as a basis for the creation of biomimetic periosteum for bone repair
复制标题
DOI：
10.1016/j.jcyt.2020.03.434
复制
发表时间：
2020-08-01
期刊：
影响因子：
4.5
通讯作者：
Roberts, Scott J.
中科院分区：
文献类型：
--
作者：
Al Hosni, Rawiya;Shah, Mittal;Roberts, Scott J.
背景：骨膜是一种高度血管化、富含胶原的组织，在引导骨修复中起着至关重要的作用。这主要是由其驻留的祖细胞数量来安排的。事实上，骨膜完整性的保存对骨愈合至关重要。从骨膜中提取的细胞保留了它们的骨软骨形成特性，因此是修复骨缺损的组织工程策略的有希望的基础。然而，培养扩增条件和细胞被重新引入缺陷部位的方式是成功翻译的关键方面。目的：本研究旨在开发一种在仿生骨膜样环境中扩增人骨膜衍生细胞(HPDC)的方案。方法：基于对人血清中培养的细胞的转录转录分析，通过研究与增强hPDC增殖和多潜能特性有关的生物活性线索，确定了扩增条件。含雌二醇、FGF2、肿瘤坏死因子α、转化生长因子β、胰岛素样生长因子-1和血小板衍生生长因子-BB)。在PD-GFC中扩增的hPDCs在细胞形态、增殖能力和软骨分化方面表现出血清拟态。当将hPDCs结合到三维I型胶原基质中并在PD-GFC中培养时，hPDCs迁移到代表骨膜形成层基质形态的表面。此外，基因表达分析显示WNT和转化生长因子β的表达下调，CREB的表达上调，这可能表明hPDCs正在重建其前体细胞的签名。结论：本研究强调了仿生骨膜的第一阶段，这可能在骨修复中有应用。(C)2020年国际细胞和基因治疗学会。爱思唯尔公司出版，版权所有。
Background: The periosteum is a highly vascularized, collagen-rich tissue that plays a crucial role in directing bone repair. This is orchestrated primarily by its resident progenitor cell population. Indeed, preservation of periosteum integrity is critical for bone healing. Cells extracted from the periosteum retain their osteochondrogenic properties and as such are a promising basis for tissue engineering strategies for the repair of bone defects. However, the culture expansion conditions and the way in which the cells are reintroduced to the defect site are critical aspects of successful translation. Indeed, expansion in human serum and implantation on biomimetic materials has previously been shown to improve in vivo bone formation.Aim: This study aimed to develop a protocol to allow for the expansion of human periosteum derived cells (hPDCs) in a biomimetic periosteal-like environment.Methods: The expansion conditions were defined through the investigation of the bioactive cues involved in augmenting hPDC proliferative and multipotency characteristics, based on transcriptomic analysis of cells cultured in human serum.Results: Master regulators of transcriptional networks were identified, and an optimized periosteum-derived growth factor cocktail (PD-GFC; containing beta-estradiol, FGF2, TNF alpha, TGF beta, IGF-1 and PDGF-BB) was generated. Expansion of hPDCs in PD-GFC resulted in serum mimicry with regard to the cell morphology, proliferative capacity and chondrogenic differentiation. When incorporated into a three-dimensional collagen type 1 matrix and cultured in PD-GFC, the hPDCs migrated to the surface that represented the matrix topography of the periosteum cambium layer. Furthermore, gene expression analysis revealed a down-regulated WNT and TGF beta signature and an up-regulation of CREB, which may indicate the hPDCs are recreating their progenitor cell signature.Conclusion: This study highlights the first stage in the development of a biomimetic periosteum, which may have applications in bone repair. (C) 2020 International Society for Cell and Gene Therapy. Published by Elsevier Inc. All rights reserved.