Tissue Engineering, Biomaterials and Nanobiotechnology Laboratory, Ankara University Faculty of Science, Ankara University Stem Cell Institute, Ankara, Turkey.
Tissue Engineering, Biomaterials and Nanobiotechnology Laboratory, Ankara University Faculty of Science, Ankara University Stem Cell Institute, Ankara, Turkey; Biovalda Health Technologies, Inc., Ankara, Turkey.
Mater Sci Eng C Mater Biol Appl. 2020 May;110:110703. doi: 10.1016/j.msec.2020.110703. Epub 2020 Jan 28.
In this study, three-dimensional macroporous cryogels were developed from platelet lysate (PL) and different concentrations of oxidized dextran (OD; 0.5, 1, 2, 4%). Subsequently, PL/OD scaffolds were characterized for potential use in tissue engineering applications. The pore size and morphology of the resulting cryogels were visualized using scanning electron microscopy (SEM). The pore size distributions were determined using mercury intrusion porosimetry (MIP). In vitro hydrolytic degradation, water uptake, mechanical properties and hemocompatibility were investigated. Extraction test was carried out to evaluate potential in vitro toxic effects of the PL/OD. The in vitro adhesion, proliferation, chondrogenic differentiation, and extracellular matrix production of human adipose stem cells (hASCs) on PL/OD cryogels were evaluated. In vivo biodegradation of the cryogels was investigated at the subcutaneous dorsal site of rats. SEM and MIP results indicated that PL/OD had a macroporous pore structure with pore sizes ranging between 10 and 200 μm. The cryogels completely degraded within 90-240 days post-implantation, depending on OD concentration. Histochemical analysis revealed high levels of cell and tissue infiltration into the pores of PL/OD. In vitro cytotoxicity findings indicated that the extracts of PL/OD, PL/OD and PL/OD showed no cytotoxic effect, whereas that of PL/OD exhibited a moderate cytotoxic effect on cell cultures. hASCs seeded on PL/OD retained their viability and showed extensive spreading and filopodia formation after 7 days. PL/OD also supported the chondrogenesis of hASCs in the presence of chondro-inductive factors. Given all the results, PL/OD could have potential as a scaffold for tissue engineering applications.
在这项研究中，从血小板裂解液（PL）和不同浓度的氧化葡聚糖（OD；0.5、1、2、4%）中开发出了三维大孔冷冻凝胶。随后，对 PL/OD 支架进行了表征，以评估其在组织工程应用中的潜力。使用扫描电子显微镜（SEM）观察了所得冷冻凝胶的孔径和形态。使用压汞法（MIP）测定了孔径分布。研究了体外水解降解、吸水率、力学性能和血液相容性。进行了提取试验以评估 PL/OD 的潜在体外毒性作用。评估了人脂肪干细胞（hASC）在 PL/OD 冷冻凝胶上的体外黏附、增殖、软骨分化和细胞外基质产生。研究了冷冻凝胶在大鼠皮下背部的体内生物降解。SEM 和 MIP 结果表明，PL/OD 具有 10-200 μm 孔径的大孔多孔结构。根据 OD 浓度的不同，冷冻凝胶在植入后 90-240 天内完全降解。组织化学分析显示细胞和组织大量渗透到 PL/OD 的孔中。体外细胞毒性研究结果表明，PL/OD、PL/OD 和 PL/OD 的提取物没有细胞毒性作用，而 PL/OD 提取物对细胞培养物表现出中度细胞毒性作用。PL/OD 上接种的 hASC 在 7 天后仍保持活力，并表现出广泛的扩散和丝状伪足形成。PL/OD 还支持软骨诱导因子存在下 hASC 的软骨形成。考虑到所有结果，PL/OD 可能具有作为组织工程应用支架的潜力。
