Nanotechnology and Nanomedicine Division, Hacettepe University, Graduate School of Science and Engineering, Beytepe, Ankara, Turkey.
Bioengineering Division, Hacettepe University, Graduate School of Science and Engineering, Beytepe, Ankara, Turkey.
Mater Sci Eng C Mater Biol Appl. 2021 Dec;131:112528. doi: 10.1016/j.msec.2021.112528. Epub 2021 Oct 31.
In skeletal muscle tissue engineering, success has not been achieved yet, since the properties of the tissue cannot be fully mimicked. The aim of this study is to investigate the potential use of poly-3-hydroxybutyrate (P3HB)/poly-β-alanine (PBA) fibrous tissue scaffolds with piezoelectric properties for skeletal muscle regeneration. Random and aligned P3HB/PBA (5:1) fibrous matrices were prepared by electrospinning with average diameters of 951 ± 153 nm and 891 ± 247 nm, respectively. X-ray diffraction (XRD) analysis showed that PBA reinforcement and aligned orientation of fibers reduced the crystallinity and brittleness of P3HB matrix. While tensile strength and elastic modulus of random fibrous matrices were determined as 3.9 ± 1.0 MPa and 86.2 ± 10.6 MPa, respectively, in the case of aligned fibers they increased to 8.5 ± 1.8 MPa and 378.2 ± 4.2 MPa, respectively. Aligned matrices exhibited a soft and an elastic behaviour with ~70% elongation in similar to the natural tissue. For the first time, d piezoelectric modulus of P3HB/PBA matrices were measured as 5 pC/N and 5.3 pC/N, for random and aligned matrices, respectively. Cell culture studies were performed with C2C12 myoblastic cell line. Both of random and aligned P3HB/PBA fibrous matrices supported attachment and proliferation of myoblasts, but cells cultured on aligned fibers formed regular and thick myofibril structures similar to the native muscle tissue. Reverse transcription polymerase chain reaction (RT-qPCR) analysis indicated that MyoD gene was expressed in the cells cultured on both fiber orientation, however, on the aligned fibers significant increase was determined in Myogenin and Myosin Heavy Chain (MHC) gene expressions, which indicate functional tubular structures. The results of RT-qPCR analysis were also supported with immunohistochemistry for myogenic markers. These in vitro studies have shown that piezoelectric P3HB/PBA aligned fibrous scaffolds can successfully mimic skeletal muscle tissue with its superior chemical, morphological, mechanical, and electroactive properties.
在骨骼肌肉组织工程中，尚未取得成功，因为组织的性能无法完全模拟。本研究旨在探讨具有压电特性的聚 3-羟基丁酸酯（P3HB）/聚-β-丙氨酸（PBA）纤维组织支架在骨骼肌再生中的潜在用途。通过静电纺丝制备了随机和定向排列的 P3HB/PBA（5:1）纤维基质，平均直径分别为 951±153nm 和 891±247nm。X 射线衍射（XRD）分析表明，PBA 增强和纤维的定向排列降低了 P3HB 基质的结晶度和脆性。而随机纤维基质的拉伸强度和弹性模量分别为 3.9±1.0MPa 和 86.2±10.6MPa，定向纤维的拉伸强度和弹性模量分别增加到 8.5±1.8MPa 和 378.2±4.2MPa。定向基质表现出柔软和弹性的行为，伸长率约为 70%，类似于天然组织。这是首次测量 P3HB/PBA 基质的压电模量，随机和定向基质分别为 5pC/N 和 5.3pC/N。使用 C2C12 成肌细胞系进行细胞培养研究。随机和定向的 P3HB/PBA 纤维基质均支持成肌细胞的附着和增殖，但在定向纤维上培养的细胞形成了类似于天然肌肉组织的规则和厚实的肌原纤维结构。逆转录聚合酶链反应（RT-qPCR）分析表明，MyoD 基因在两种纤维取向的细胞中均有表达，但在定向纤维上，Myogenin 和肌球蛋白重链（MHC）基因的表达显著增加，表明存在功能性管状结构。RT-qPCR 分析结果也得到了肌生成标志物免疫组织化学的支持。这些体外研究表明，具有压电特性的 P3HB/PBA 定向纤维支架可以成功模拟具有优异的化学、形态、机械和电活性特性的骨骼肌组织。
