1 Shiley Center for Orthopaedic Research and Education at Scripps Clinic , La Jolla, California.
2 Department of Material Science and Engineering, University of California , San Diego, La Jolla, California.
Tissue Eng Part A. 2018 Jan;24(1-2):81-93. doi: 10.1089/ten.TEA.2016.0205. Epub 2017 Jun 13.
Hydrogel and electrospun scaffold materials support cell attachment and neotissue development and can be tuned to structurally and mechanically resemble native extracellular matrix by altering either electrospun fiber or hydrogel properties. In this study, we examined meniscus tissue generation from different human cell sources including meniscus cells derived from vascular and avascular regions, human bone marrow-derived mesenchymal stem cells, synovial cells, and cells from the infrapatellar fat pad (IPFP). All cells were seeded onto aligned electrospun collagen type I scaffolds and were optionally encapsulated in a tricomponent hydrogel. Single or multilayered constructs were generated and cultivated in defined medium with selected growth factors for 2 weeks. Cell viability, cell morphology, and gene-expression profiles were monitored using confocal microscopy, scanning electron microscopy, and quantitative polymerase chain reaction (qPCR), respectively. Multilayered constructs were examined with histology, immunohistochemistry, qPCR, and for tensile mechanical properties. For all cell types, TGFβ1 and TGFβ3 treatment increased COL1A1, COMP, Tenascin C (TNC), and Scleraxis (SCX) gene expression and deposition of collagen type I protein. IPFP cells generated meniscus-like tissues with higher meniscogenic gene expression, mechanical properties, and better cell distribution compared to other cell types studied. We show proof of concept that electrospun collagen scaffolds support neotissue formation and IPFP cells have potential for use in cell-based meniscus regeneration strategies.
水凝胶和静电纺丝支架材料支持细胞附着和新生组织的发育,并可以通过改变静电纺丝纤维或水凝胶的性质来调整结构和机械性能,使其类似于天然细胞外基质。在这项研究中,我们研究了不同的人类细胞来源(包括源自血管和非血管区域的半月板细胞、人骨髓间充质干细胞、滑膜细胞和髌下脂肪垫(IPFP)细胞)的半月板组织生成。所有细胞均接种到排列的胶原 I 型静电纺丝支架上,并可选择封装在三组分水凝胶中。生成单层或多层结构,并在含有选定生长因子的定义培养基中培养 2 周。使用共聚焦显微镜、扫描电子显微镜和定量聚合酶链反应(qPCR)分别监测细胞活力、细胞形态和基因表达谱。使用组织学、免疫组织化学、qPCR 和拉伸力学性能检查多层结构。对于所有细胞类型,TGFβ1 和 TGFβ3 处理均增加了 COL1A1、COMP、Tenascin C(TNC)和 Scleraxis(SCX)基因的表达以及 I 型胶原蛋白的沉积。与其他研究的细胞类型相比,IPFP 细胞生成的半月板样组织具有更高的半月板生成基因表达、机械性能和更好的细胞分布。我们证明了概念,即静电纺丝胶原支架支持新生组织的形成,并且 IPFP 细胞具有用于基于细胞的半月板再生策略的潜力。