Department of BIN Convergence Technology, Department of PolymerNano Science & Technology and Polymer Materials Fusion Research Center, Chonbuk National University, 567 Baekje-daero, Deokjin-gu, Jeonju-si, Jeollabuk-do 54896, Republic of Korea.
Department of Orthopaedic Surgery, Medical School and Research Institute of Clinical Medicine, Chonbuk National University, Jeonju, Jeonbuk 561-756, Republic of Korea.
Mater Sci Eng C Mater Biol Appl. 2019 Apr;97:347-355. doi: 10.1016/j.msec.2018.12.001. Epub 2018 Dec 3.
Tissue engineered scaffolds, made of natural derived materials, have the potential to be used in bone regeneration fields due to the biocompatible and biodegradable features. In this study, we propose duck's feet-derived collagen (DC) sponges blended with hydroxyapatite (HAp), incorporated with different concentrations of silymarin (Smn), for improved bone regeneration. The morphological and structural properties of DC/HAp and DC/HAp loaded with 25, 50 and 100 μM of Smn sponges were analyzed using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). In vitro evaluations were carried out on rabbit bone marrow stem cells (rBMSCs) using MTT assay for cell proliferation, ALP assay for osteogenic differentiation and reverse transcription-polymerase chain reaction (RT-PCR) for expression of mRNAs. For the evaluation of new bone formation in vivo, histological analysis and micro computed tomography (μCT) were used. Preliminary results, on Smn/DC/HAp morphology and mechanical properties, showed an interconnected porosity suitable for cells ingrowth and a higher compressive strength with the presence of Smn. Similarly, the cells proliferation and ALP activity modulation were positively influenced by the Smn content. Especially, the 100 μM Smn/DC/HAp sponge efficiently enhances the rBMSCs adhesion, growth and gene expression of osteogenic markers. The enhanced osteoinductive effects of sponges blended with Smn were confirmed using μ-CT and histological evaluations. In conclusion, results suggest that collagen sponges represent an excellent environment for cells growth and proliferation, while Smn plays an important role to improve materials osteogenic properties.
基于其生物相容性和可降解性，由天然衍生材料制成的组织工程支架具有应用于骨再生领域的潜力。在这项研究中，我们提出了鸭脚来源的胶原蛋白（DC）海绵与羟基磷灰石（HAp）混合，并掺入不同浓度的水飞蓟素（Smn），以提高骨再生能力。使用扫描电子显微镜（SEM）和傅里叶变换红外光谱（FTIR）分析了 DC/HAp 和负载 25、50 和 100µM Smn 的 DC/HAp 海绵的形态和结构特性。通过 MTT 测定法进行细胞增殖、ALP 测定法进行成骨分化和逆转录-聚合酶链反应（RT-PCR）进行 mRNAs 表达的体外评价，在兔骨髓间充质干细胞（rBMSCs）上进行了评估。为了评估体内新骨形成，进行了组织学分析和微计算机断层扫描（μCT）。Smn/DC/HAp 形态和力学性能的初步结果表明，具有适合细胞向内生长的互连孔隙率和更高的抗压强度，且具有 Smn。同样，细胞增殖和 ALP 活性的调节也受到 Smn 含量的积极影响。特别是，100µM Smn/DC/HAp 海绵有效地增强了 rBMSCs 的黏附、生长和成骨标志物的基因表达。使用 μCT 和组织学评估证实了与 Smn 混合的海绵具有增强的成骨作用。总之，结果表明胶原蛋白海绵为细胞生长和增殖提供了极好的环境，而 Smn 对提高材料的成骨性能起着重要作用。
