Volkov Vadim, Vasconcelos Andreia, Sárria Marisa P, Gomes Andreia C, Cavaco-Paulo Artur
Centro de Engenharia Biológica, Universidade do Minho, Campus de Gualtar, Braga, Portugal.
Biotechnol J. 2014 Oct;9(10):1267-78. doi: 10.1002/biot.201400302. Epub 2014 Aug 29.
Silk fibroin demonstrates great biocompatibility and is suitable for many biomedical applications, including tissue engineering and regenerative medicine. Current research focuses on manipulating the physico-chemical properties of fibroin, and examining the effect of this manipulation on firobin's biocompatibility. Regenerated silk fibroin was modified by in vitro enzymatic phosphorylation and cast into films. Films were produced by blending, at several ratios, the phosphorylated and un-phosphorylated fibroin solutions. Fourier transform infra-red spectroscopy was used to determine the specific P-OH vibration peak, confirming the phosphorylation of the regenerated silk fibroin solution. Differential scanning calorimetry showed that phosphorylation altered the intra- and inter-molecular interactions. Further experiments demonstrated that phosphorylation can be used to tailor the hydrophylicity/hydrophobicity ratio as well as the crystalinity of silk fibroin films. Release profiling of a model drug was highly dependent on silk modification level. Cytotoxicity assays showed that exposure to lixiviates of phosphorylated films only slightly affected cellular metabolism and proliferation, although direct contact resulted in a strong direct correlation between phosphorylation level and cell proliferation. This new method for tuning silk biomaterials to obtain specific structural and biochemical features can be adapted for a wide range of applications. Phosphorylation of silk fibroins may be applied to improve the cytocompatibility of any silk-based device that is considered to be in contact with live animals or human tissues.
丝素蛋白具有良好的生物相容性,适用于许多生物医学应用,包括组织工程和再生医学。目前的研究重点是控制丝素蛋白的物理化学性质,并研究这种控制对丝素蛋白生物相容性的影响。通过体外酶促磷酸化对再生丝素蛋白进行修饰,并制成薄膜。通过将磷酸化和未磷酸化的丝素蛋白溶液按几种比例混合来制备薄膜。利用傅里叶变换红外光谱法确定特定的P-OH振动峰,证实再生丝素蛋白溶液发生了磷酸化。差示扫描量热法表明磷酸化改变了分子内和分子间的相互作用。进一步的实验表明,磷酸化可用于调整丝素蛋白薄膜的亲水性/疏水性比例以及结晶度。模型药物的释放曲线高度依赖于丝素蛋白的修饰水平。细胞毒性试验表明,尽管直接接触导致磷酸化水平与细胞增殖之间存在很强的直接相关性,但暴露于磷酸化薄膜的浸出液仅对细胞代谢和增殖有轻微影响。这种调整丝素生物材料以获得特定结构和生化特性的新方法可适用于广泛的应用。丝素蛋白的磷酸化可用于改善任何被认为与活体动物或人体组织接触的丝基装置的细胞相容性。