Cell–Material Interactions in Covalent Adaptable Thioester Hydrogels

IF 5.4 2区 医学 Q2 MATERIALS SCIENCE, BIOMATERIALS
Shivani Desai, Benjamin Carberry, Kristi S. Anseth and Kelly M. Schultz*, 
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引用次数: 0

Abstract

Covalent adaptable networks (CANs) are polymeric networks with cross-links that can break and reform in response to external stimuli, including pH, shear, and temperature, making them potential materials for use as injectable cell delivery vehicles. In the native niche, cells rearrange the extracellular matrix (ECM) to undergo basic functions including migration, spreading, and proliferation. Bond rearrangement enables these hydrogels to mimic viscoelastic properties of the native ECM which promote migration and delivery from the material to the native tissue. In this work, we characterize thioester CANs to inform their design as effective cell delivery vehicles. Using bulk rheology, we characterize the rearrangement of these networks when they are subjected to strain, which mimics the strain applied by a syringe, and using multiple particle tracking microrheology (MPT) we measure cell-mediated remodeling of the pericellular region. Thioester networks are formed by photopolymerizing 8-arm poly(ethylene glycol) (PEG)-thiol and PEG-thioester norbornene. Bulk rheology measures scaffold properties during low and high strain and demonstrates that thioester scaffolds can recover rheological properties after high strain is applied. We then 3D encapsulated human mesenchymal stem cells (hMSCs) in thioester scaffolds. Using MPT, we characterize degradation in the pericellular region. Encapsulated hMSCs degrade these scaffolds within ≈4 days post-encapsulation. We hypothesize that this degradation is mainly due to cytoskeletal tension that cells apply to the matrix, causing adaptable thioester bonds to rearrange, leading to degradation. To verify this, we inhibited cytoskeletal tension using blebbistatin, a myosin-II inhibitor. Blebbistatin-treated cells can degrade these networks only by secreting enzymes including esterases. Esterases hydrolyze thioester bonds, which generate free thiols, leading to bond exchange. Around treated cells, we measure a decrease in the extent of pericellular degradation. We also compare cell area, eccentricity, and speed of untreated and treated cells. Inhibiting cytoskeletal tension results in significantly smaller cell area, more rounded cells, and lower cell speeds when compared to untreated cells. Overall, this work shows that cytoskeletal tension plays a major role in hMSC-mediated degradation of thioester networks. Cytoskeletal tension is also important for the spreading and motility of hMSCs in these networks. This work informs the design of thioester scaffolds for tissue regeneration and cell delivery.

Abstract Image

共价可适应硫酯水凝胶中的细胞-材料相互作用
共价可适应网络(CANs)是一种具有交联的聚合物网络,可在 pH 值、剪切力和温度等外部刺激下断裂或重组,因此有可能用作注射细胞输送载体。在原生龛位中,细胞重新排列细胞外基质(ECM),以实现迁移、扩散和增殖等基本功能。粘结重排使这些水凝胶能够模拟原生 ECM 的粘弹性,从而促进材料向原生组织的迁移和输送。在这项研究中,我们对硫酯 CAN 进行了表征,以便为设计有效的细胞输送载体提供依据。利用体流变学,我们描述了这些网络在受到应变(模拟注射器施加的应变)时的重新排列;利用多粒子跟踪微流变学(MPT),我们测量了细胞介导的细胞周围区域重塑。硫酯网络由 8 臂聚乙二醇(PEG)-硫醇和 PEG-硫酯降冰片烯光聚合而成。大体积流变学测量了支架在低应变和高应变时的特性,并证明硫酯支架在施加高应变后可恢复流变特性。然后,我们将人间质干细胞(hMSCs)三维包裹在硫代酯类支架中。我们使用 MPT 分析了细胞周围区域的降解特性。被包裹的间充质干细胞会在包裹后≈4天内降解这些支架。我们假设这种降解主要是由于细胞对基质施加了细胞骨架张力,导致可适应的硫酯键重新排列,从而导致降解。为了验证这一点,我们使用肌球蛋白II抑制剂blebbistatin抑制细胞骨架张力。经blebbistatin处理的细胞只能通过分泌包括酯酶在内的酶来降解这些网络。酯酶水解硫酯键,产生游离硫醇,导致键交换。在经处理的细胞周围,我们测量到细胞周围降解的程度有所下降。我们还比较了未处理细胞和处理细胞的细胞面积、偏心率和速度。与未经处理的细胞相比,抑制细胞骨架张力可使细胞面积明显缩小,细胞更圆,细胞速度更低。总之,这项研究表明,细胞骨架张力在 hMSC 介导的硫酯网络降解中起着重要作用。细胞骨架张力对 hMSC 在这些网络中的扩散和运动也很重要。这项研究为硫酯支架的组织再生和细胞输送设计提供了参考。
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来源期刊
ACS Biomaterials Science & Engineering
ACS Biomaterials Science & Engineering Materials Science-Biomaterials
CiteScore
10.30
自引率
3.40%
发文量
413
期刊介绍: ACS Biomaterials Science & Engineering is the leading journal in the field of biomaterials, serving as an international forum for publishing cutting-edge research and innovative ideas on a broad range of topics: Applications and Health – implantable tissues and devices, prosthesis, health risks, toxicology Bio-interactions and Bio-compatibility – material-biology interactions, chemical/morphological/structural communication, mechanobiology, signaling and biological responses, immuno-engineering, calcification, coatings, corrosion and degradation of biomaterials and devices, biophysical regulation of cell functions Characterization, Synthesis, and Modification – new biomaterials, bioinspired and biomimetic approaches to biomaterials, exploiting structural hierarchy and architectural control, combinatorial strategies for biomaterials discovery, genetic biomaterials design, synthetic biology, new composite systems, bionics, polymer synthesis Controlled Release and Delivery Systems – biomaterial-based drug and gene delivery, bio-responsive delivery of regulatory molecules, pharmaceutical engineering Healthcare Advances – clinical translation, regulatory issues, patient safety, emerging trends Imaging and Diagnostics – imaging agents and probes, theranostics, biosensors, monitoring Manufacturing and Technology – 3D printing, inks, organ-on-a-chip, bioreactor/perfusion systems, microdevices, BioMEMS, optics and electronics interfaces with biomaterials, systems integration Modeling and Informatics Tools – scaling methods to guide biomaterial design, predictive algorithms for structure-function, biomechanics, integrating bioinformatics with biomaterials discovery, metabolomics in the context of biomaterials Tissue Engineering and Regenerative Medicine – basic and applied studies, cell therapies, scaffolds, vascularization, bioartificial organs, transplantation and functionality, cellular agriculture
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