Krebs Joseph, Stealey Samuel, Brown Alyssa, Krohn Austin, Zustiak Silviya Petrova, Case Natasha
Department of Biomedical Engineering, Saint Louis University, Saint Louis, MO 63103, USA.
Department of Physiology and Pharmacology, School of Medicine, Saint Louis University, Saint Louis, MO 63104, USA.
Gels. 2023 Sep 1;9(9):705. doi: 10.3390/gels9090705.
Connective tissue models grown from cell monolayers can be instrumental in a variety of biomedical fields such as drug screening, wound healing, and regenerative engineering. However, while connective tissues contain abundant fibrillar collagen, achieving a sufficient assembly and retention of fibrillar collagen in vitro is challenging. Unlike the dilute cell culture environment, the body's environment is characterized by a high density of soluble macromolecules (crowding) and macromolecular networks (confinement), which contribute to extracellular matrix (ECM) assembly in vivo. Consequently, macromolecular crowding (MMC) has been successfully used to enhance the processing of type I procollagen, leading to significant increases in fibrillar collagen assembly and accumulation during in vitro culture of a variety of cell types. In this study, we developed a combination approach using a carrageenan hydrogel, which released soluble macromolecules and served as a confinement barrier. We first evaluated the local carrageenan release and then confirmed the effectiveness of this combination approach on collagen accumulation by the human MG-63 bone cell line. Additionally, computational modeling of oxygen and glucose transport within the culture system showed no negative effects of the hydrogel and its releasates on cell viability.
从细胞单层生长而来的结缔组织模型在药物筛选、伤口愈合和再生工程等多种生物医学领域中可能发挥重要作用。然而，尽管结缔组织含有丰富的纤维状胶原蛋白，但在体外实现纤维状胶原蛋白的充分组装和保留具有挑战性。与稀释的细胞培养环境不同，体内环境的特点是可溶性大分子密度高（拥挤）和大分子网络（限制），这有助于体内细胞外基质（ECM）的组装。因此，大分子拥挤（MMC）已成功用于增强I型前胶原的加工，导致在多种细胞类型的体外培养过程中纤维状胶原蛋白的组装和积累显著增加。在本研究中，我们开发了一种联合方法，使用卡拉胶水凝胶，其可释放可溶性大分子并作为限制屏障。我们首先评估了局部卡拉胶的释放，然后通过人MG-63骨细胞系证实了这种联合方法对胶原蛋白积累的有效性。此外，培养系统内氧气和葡萄糖运输的计算模型表明水凝胶及其释放物对细胞活力没有负面影响。
