用于骨骼肌组织工程的基于海藻酸盐-明胶的复合支架的挤出式3D(生物)打印

Extrusion 3D (Bio)Printing of Alginate-Gelatin-Based Composite Scaffolds for Skeletal Muscle Tissue Engineering.

作者信息

Sonaye Surendrasingh Y, Ertugral Elif G, Kothapalli Chandrasekhar R, Sikder Prabaha

机构信息

Mechanical Engineering, Cleveland State University, Cleveland, OH 44115, USA.

Chemical and Biomedical Engineering, Cleveland State University, Cleveland, OH 44115, USA.

出版信息

Materials (Basel). 2022 Nov 10;15(22):7945. doi: 10.3390/ma15227945.

Abstract

Volumetric muscle loss (VML), which involves the loss of a substantial portion of muscle tissue, is one of the most serious acute skeletal muscle injuries in the military and civilian communities. The injured area in VML may be so severely affected that the body loses its innate capacity to regenerate new functional muscles. State-of-the-art biofabrication methods such as bioprinting provide the ability to develop cell-laden scaffolds that could significantly expedite tissue regeneration. Bioprinted cell-laden scaffolds can mimic the extracellular matrix and provide a bioactive environment wherein cells can spread, proliferate, and differentiate, leading to new skeletal muscle tissue regeneration at the defect site. In this study, we engineered alginate−gelatin composite inks that could be used as bioinks. Then, we used the inks in an extrusion printing method to develop design-specific scaffolds for potential VML treatment. Alginate concentration was varied between 4−12% w/v, while the gelatin concentration was maintained at 6% w/v. Rheological analysis indicated that the alginate−gelatin inks containing 12% w/v alginate and 6% w/v gelatin were most suitable for developing high-resolution scaffolds with good structural fidelity. The printing pressure and speed appeared to influence the printing accuracy of the resulting scaffolds significantly. All the hydrogel inks exhibited shear thinning properties and acceptable viscosities, though 8−12% w/v alginate inks displayed properties ideal for printing and cell proliferation. Alginate content, crosslinking concentration, and duration played significant roles (p < 0.05) in influencing the scaffolds’ stiffness. Alginate scaffolds (12% w/v) crosslinked with 300, 400, or 500 mM calcium chloride (CaCl2) for 15 min yielded stiffness values in the range of 45−50 kPa, i.e., similar to skeletal muscle. The ionic strength of the crosslinking concentration and the alginate content significantly (p < 0.05) affected the swelling and degradation behavior of the scaffolds. Higher crosslinking concentration and alginate loading enhanced the swelling capacity and decreased the degradation kinetics of the printed scaffolds. Optimal CaCl2 crosslinking concentration (500 mM) and alginate content (12% w/v) led to high swelling (70%) and low degradation rates (28%) of the scaffolds. Overall, the results indicate that 12% w/v alginate and 6% w/v gelatin hydrogel inks are suitable as bioinks, and the printed scaffolds hold good potential for treating skeletal muscle defects such as VML.

摘要

容积性肌肉损失(VML)涉及大部分肌肉组织的丧失,是军事和民用领域中最严重的急性骨骼肌损伤之一。VML中的损伤区域可能受到严重影响,以至于身体失去了再生新的功能性肌肉的固有能力。诸如生物打印等先进的生物制造方法能够开发载有细胞的支架,从而显著加速组织再生。生物打印的载有细胞的支架可以模拟细胞外基质,并提供一个生物活性环境,使细胞能够在其中扩散、增殖和分化,从而在缺损部位实现新的骨骼肌组织再生。在本研究中,我们设计了可用作生物墨水的藻酸盐-明胶复合墨水。然后,我们将这些墨水用于挤出打印方法,以开发用于潜在VML治疗的特定设计支架。藻酸盐浓度在4-12% w/v之间变化,而明胶浓度保持在6% w/v。流变学分析表明,含有12% w/v藻酸盐和6% w/v明胶的藻酸盐-明胶墨水最适合开发具有良好结构保真度的高分辨率支架。打印压力和速度似乎对所得支架的打印精度有显著影响。所有水凝胶墨水均表现出剪切变稀特性和可接受的粘度,不过8-12% w/v的藻酸盐墨水表现出适合打印和细胞增殖的理想特性。藻酸盐含量、交联浓度和持续时间在影响支架硬度方面发挥了重要作用(p < 0.05)。与300、400或500 mM氯化钙(CaCl2)交联15分钟的藻酸盐支架(12% w/v)产生的硬度值在45-50 kPa范围内,即与骨骼肌相似。交联浓度的离子强度和藻酸盐含量显著(p < 0.05)影响支架的肿胀和降解行为。较高的交联浓度和藻酸盐负载量增强了打印支架的肿胀能力并降低了其降解动力学。最佳的CaCl2交联浓度(500 mM)和藻酸盐含量(12% w/v)导致支架具有高肿胀率(70%)和低降解率(28%)。总体而言,结果表明12% w/v藻酸盐和6% w/v明胶水凝胶墨水适合作为生物墨水,并且打印的支架在治疗诸如VML等骨骼肌缺损方面具有良好的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d60e/9695625/8f161a25b524/materials-15-07945-g001.jpg

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