Ionically and Enzymatically Dual Cross-Linked Oxidized Alginate Gelatin Hydrogels with Tunable Stiffness and Degradation Behavior for Tissue Engineering,ACS Biomaterials Science & Engineering

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Ionically and Enzymatically Dual Cross-Linked Oxidized Alginate Gelatin Hydrogels with Tunable Stiffness and Degradation Behavior for Tissue Engineering
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2020-05-20 , DOI: 10.1021/acsbiomaterials.0c00677
Thomas Distler ₁ , Kilian McDonald ₁ , Susanne Heid ₁ , Emine Karakaya ₁ , Rainer Detsch ₁ , Aldo R. Boccaccini ₁

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Hydrogels that allow for the successful long-term in vitro culture of cell–biomaterial systems to enable the maturation of tissue engineering constructs are highly relevant in regenerative medicine. Naturally derived polysaccharide-based hydrogels promise to be one material group with enough versatility and chemical functionalization capability to tackle the challenges associated with long-term cell culture. We report a marine derived oxidized alginate, alginate dialdehyde (ADA), and gelatin (GEL) system (ADA-GEL), which is cross-linked via ionic (Ca²⁺) and enzymatic (microbial transglutaminase, mTG) interaction to form dually cross-linked hydrogels. The cross-linking approach allowed us to tailor the stiffness of the hydrogels in a wide range (from <5 to 120 kPa), without altering the initial ADA and GEL hydrogel chemistry. It was possible to control the degradation behavior of the hydrogels to be stable for up to 30 days of incubation. Increasing concentrations of mTG cross-linker solutions allowed us to tune the degradation behavior of the ADA-GEL hydrogels from fast (<7 days) to moderate (14 days) and slow (>30 days) degradation kinetics. The cytocompatibility of mTG cross-linked ADA-GEL was assessed using NIH-3T3 fibroblasts and ATDC-5 mouse teratocarcinoma cells. Both cell types showed highly increased cellular attachment on mTG cross-linked ADA-GEL in comparison to Ca²⁺ cross-linked hydrogels. In addition, ATDC-5 cells showed a higher proliferation on mTG cross-linked ADA-GEL hydrogels in comparison to tissue culture polystyrene control substrates. Further, the attachment of human umbilical vein endothelial cells (HUVEC) on ADA-GEL (+) mTG was confirmed, proving the suitability of mTG+Ca²⁺ cross-linked ADA-GEL for several cell types. Summarizing, a promising platform to control the properties of ADA-GEL hydrogels is presented, with the potential to be applied in long-term cell culture investigations such as cartilage, bone, and blood-vessel engineering, as well as for biofabrication.

中文翻译：

具有可调刚度和降解行为的离子和酶双交联氧化藻酸明胶水凝胶，用于组织工程

在再生医学中，能够成功进行细胞生物材料系统的长期体外培养以使组织工程构造成熟的水凝胶与再生医学高度相关。天然衍生的基于多糖的水凝胶有望成为具有足够的多功能性和化学功能化能力的一类材料，以应对与长期细胞培养相关的挑战。我们报告了海洋衍生的氧化藻酸盐，藻酸盐二醛（ADA）和明胶（GEL）系统（ADA-GEL），其通过离子（Ca ²⁺）和酶（微生物转谷氨酰胺酶，mTG）相互作用形成双重交联的水凝胶。交联方法使我们能够在宽范围（<5至120 kPa）范围内调整水凝胶的硬度，而无需改变初始ADA和GEL水凝胶的化学性质。可以控制水凝胶的降解行为，使其在长达30天的孵育时间内保持稳定。mTG交联剂溶液浓度的增加使我们能够将ADA-GEL水凝胶的降解行为从快速（<7天）调整为中度（14天）和慢速（> 30天）降解动力学。使用NIH-3T3成纤维细胞和ATDC-5小鼠畸胎癌细胞评估mTG交联的ADA-GEL的细胞相容性。与Ca相比，这两种细胞类型均在mTG交联的ADA-GEL上显示出高度附着的细胞²⁺交联水凝胶。另外，与组织培养聚苯乙烯对照基质相比，ATDC-5细胞在mTG交联的ADA-GEL水凝胶上显示出更高的增殖。此外，证实了人脐静脉内皮细胞（HUVEC）在ADA-GEL（+）mTG上的附着，证明了mTG + Ca ²⁺交联ADA-GEL对几种细胞类型的适用性。总而言之，提出了一种有前途的平台来控制ADA-GEL水凝胶的性能，其潜力可用于长期的细胞培养研究，例如软骨，骨骼和血管工程以及生物制造。

更新日期：2020-07-13

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