The effect of elevating extracellular CaCl2: Important considerations for tissue engineering applications

IF 2.7 4区生物学 Q1 ANATOMY & MORPHOLOGY

Tissue & cell Pub Date : 2024-11-14 DOI:10.1016/j.tice.2024.102615

Kayley Jaworska , Jessica J. Senior , Anke Brüning-Richardson , Alan M. Smith

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引用次数: 0

Abstract

Polysaccharides such as sodium alginate, pectin and gellan gum are widely used biomaterials, for their ability to easily form hydrogels in the presence of divalent metal ions, such as calcium - a process often cited as a mild crosslinking mechanism. However, when using these materials as substrates for tissue engineering, there is a lack of extensive studies that investigate the impact of elevated calcium concentrations on cell health and behaviour. In this study, we performed an in-depth exploration to understand the potential effects of raising extracellular CaCl₂ on cell viability, proliferation, morphology and migration. We used an established glioblastoma (GBM) cell line (U251), human dermal fibroblasts (HDF), and murine osteoblasts (MC3T3) to assess the consequences of using CaCl₂ in tissue engineered models to help reevaluate biomaterial suitability and enhance standardisation practices in the field of tissue engineering. Our findings revealed that the addition of CaCl₂ induced notable morphological changes in GBM cells when cultured in 3D hydrogels with excess CaCl₂ added, leading to a transition from mesenchymal to amoeboid phenotypes, even at a concentration as low as 8 mM. Furthermore, cell viability was reduced in a concentration-dependent manner across all cell types, and migration was also affected. Despite the widespread use of high CaCl₂ concentrations to facilitate scaffold gelation, our research unveils that there can be significant risks to cell viability, proliferation, morphology, and migration when such practices are not preceded by cell line-specific experimentation and thorough standardization procedures. This highlights the importance of careful consideration and optimisation of CaCl₂ concentration when used as a crosslinking agent for hydrogels intended for use in tissue engineering applications that demand accurate recapitulation of cellular responses and physiological conditions.

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提高细胞外 CaCl2 的影响：组织工程应用的重要考虑因素

海藻酸钠、果胶和结冷胶等多糖是被广泛使用的生物材料，因为它们在二价金属离子（如钙）的存在下能很容易地形成水凝胶，这一过程通常被认为是一种温和的交联机制。然而，在使用这些材料作为组织工程基底时，缺乏对钙浓度升高对细胞健康和行为影响的广泛研究。在本研究中，我们进行了深入探索，以了解细胞外 CaCl2 升高对细胞活力、增殖、形态和迁移的潜在影响。我们使用已建立的胶质母细胞瘤（GBM）细胞系（U251）、人真皮成纤维细胞（HDF）和小鼠成骨细胞（MC3T3）来评估在组织工程模型中使用 CaCl2 的后果，以帮助重新评估生物材料的适用性并加强组织工程领域的标准化实践。我们的研究结果表明，在添加过量 CaCl2 的三维水凝胶中培养 GBM 细胞时，即使浓度低至 8 mM，CaCl2 的添加也会诱导细胞发生显著的形态变化，导致细胞从间充质向变形表型转变。此外，所有细胞类型的细胞活力都以浓度依赖的方式降低，迁移也受到影响。尽管高浓度 CaCl2 被广泛用于促进支架凝胶化，但我们的研究揭示出，如果在采用这种方法之前没有进行细胞系特异性实验和彻底的标准化程序，细胞的活力、增殖、形态和迁移都可能面临重大风险。这凸显了在组织工程应用中将 CaCl2 用作交联剂时，仔细考虑和优化 CaCl2 浓度的重要性，因为组织工程应用要求准确再现细胞反应和生理条件。

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.