Impact of increasingly complex cell culture conditions on the proteome of human periodontal ligament stem cells.

IF 2.4 4区 医学 Q4 CELL & TISSUE ENGINEERING
Asier Fullaondo, Mar Zalduendo, Nerea Osinalde, Mohammad H Alkhraisat, Eduardo Anitua, Ana M Zubiaga
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引用次数: 0

Abstract

Aims: Human periodontal ligament stem cells (hPDLSCs) exhibit an enormous potential to regenerate periodontal tissue. However, their translatability to the clinical setting is constrained by technical difficulties in standardizing culture conditions. The aim was to assess complex culture conditions using a proteomic-based protocol to standardize multi-layer hPDLSC cultivation methodology.

Materials and methods: hPDLSC-derived constructs were created with varying biological complexity. The simplest constructs were monolayer sheets of hPDLSCs cultured with fetal bovine serum (FBS) or Plasma Rich in Growth Factors supernatant (PRGFsn). The most complex constructs were triple-layered cell structures cultured with PRGFsn, with or without PRGF fibrin membrane (mPRGF). Ultrastructure and proteomic analyses were performed on these constructs.

Results: PRGF supernatant improved protein expression related to extracellular matrix, adhesion, proliferation, and migration in hPDLSCs. PRGF fibrin scaffold upregulates proteins for cell activation, respiration, and electron transport. hPDLSCs on fibrin membrane show robust osteogenic potential through differential protein expression (ossification, tissue remodeling, morphogenesis, or cell migration) and overall homeostasis relative to less complex structures.

Conclusion: Our data reveal the far-reaching potential of 3-dimensional constructs in combination with PRGF technology in periodontal regenerative applications.

日益复杂的细胞培养条件对人牙周韧带干细胞蛋白质组的影响。
目的:人牙周韧带干细胞(hPDLSCs)具有再生牙周组织的巨大潜力。然而,它们在临床环境中的可翻译性受到标准化培养条件的技术困难的限制。目的是使用基于蛋白质组学的协议来评估复杂的培养条件,以标准化多层hPDLSC培养方法。材料和方法:hpdlsc衍生的构建物具有不同的生物复杂性。最简单的构建是用胎牛血清(FBS)或富生长因子血浆上清(PRGFsn)培养单层hPDLSCs。最复杂的结构是用PRGFsn培养的三层细胞结构,有或没有PRGF纤维蛋白膜(mPRGF)。对这些构建体进行了超微结构和蛋白质组学分析。结果:PRGF上清液改善了hPDLSCs中细胞外基质、粘附、增殖和迁移相关蛋白的表达。PRGF纤维蛋白支架上调细胞活化、呼吸和电子传递的蛋白。纤维蛋白膜上的hPDLSCs通过不同的蛋白表达(骨化、组织重塑、形态发生或细胞迁移)和相对于较不复杂的结构的整体稳态显示出强大的成骨潜力。结论:我们的数据揭示了三维构建体结合PRGF技术在牙周再生应用中的深远潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Regenerative medicine
Regenerative medicine 医学-工程:生物医学
CiteScore
4.20
自引率
3.70%
发文量
82
审稿时长
6-12 weeks
期刊介绍: Regenerative medicine replaces or regenerates human cells, tissue or organs, to restore or establish normal function*. Since 2006, Regenerative Medicine has been at the forefront of publishing the very best papers and reviews covering the entire regenerative medicine sector. The journal focusses on the entire spectrum of approaches to regenerative medicine, including small molecule drugs, biologics, biomaterials and tissue engineering, and cell and gene therapies – it’s all about regeneration and not a specific platform technology. The journal’s scope encompasses all aspects of the sector ranging from discovery research, through to clinical development, through to commercialization. Regenerative Medicine uniquely supports this important area of biomedical science and healthcare by providing a peer-reviewed journal totally committed to publishing the very best regenerative medicine research, clinical translation and commercialization. Regenerative Medicine provides a specialist forum to address the important challenges and advances in regenerative medicine, delivering this essential information in concise, clear and attractive article formats – vital to a rapidly growing, multidisciplinary and increasingly time-constrained community. Despite substantial developments in our knowledge and understanding of regeneration, the field is still in its infancy. However, progress is accelerating. The next few decades will see the discovery and development of transformative therapies for patients, and in some cases, even cures. Regenerative Medicine will continue to provide a critical overview of these advances as they progress, undergo clinical trials, and eventually become mainstream medicine.
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