Valiani Ali, Hashemibeni Batool, Esfandiary Ebrahim, Ansar Malek Masoud, Kazemi Mohammad, Esmaeili Nafiseh
Department of Anatomical Sciences and Molecular Biology, Medical School, Isfahan University of Medical Sciences, Isfahan, I.R., Iran.
Department of Anatomical Sciences, Medical School, Guilan University of Medical Sciences, Guilan, I.R, Iran.
Int J Prev Med. 2014 Jul;5(7):825-34.
Osteoarthritis is one of the most common diseases in middle-aged populations in the World and could become the fourth principal cause of disability by the year 2020. One of the critical properties for cartilage tissue engineering (TE) is the ability of scaffolds to closely mimic the extracellular matrix and bond to the host tissue. Therefore, TE has been presented as a technique to introduce the best combination of cells and biomaterial scaffold and to stimulate growth factors to produce a cartilage tissue resembling natural articular cartilage. The aim of study is to improve differentiation of adipose derived stem cells (ADSCs) into chondrocytes in order to provide a safe and modern treatment for patients suffering from cartilage damages.
After functionalization, dispersions and sterilizing carbon nano-tubes (CNTs), a new type of nanocomposite gel was prepared from water-soluble CNTs and alginate. ADSCs seeded in 1.5% alginate scaffold and cultured in chondrogenic media with and without transforming growth factor-β1 (TGF-β1) for 7 and 14 days. The genes expression of sex determining region Y-box 9 (SOX9), types II and X collagens was assessed by real-time polymerase chain reaction and the amount of aggrecan (AGC) and type I collagen was measured by ELISA.
Our findings showed that the expression of essential cartilage markers, SOX9, type II collagen and AGC, in differentiated ADSCs at the concentration of 1 μg/ml CNTs in the presence of TGF-β1 were significantly increased in comparison with the control group (P < 0.001). Meanwhile, type X collagen expression and also type I collagen production were significantly decreased (P < 0.001).
The results showed that utilized three-dimensional scaffold had a brilliant effect in promoting gene expression of chondrogenesis.
骨关节炎是世界中年人群中最常见的疾病之一，到2020年可能成为第四大致残主要原因。软骨组织工程（TE）的关键特性之一是支架能够紧密模拟细胞外基质并与宿主组织结合。因此，TE已被视为一种引入细胞与生物材料支架最佳组合并刺激生长因子以产生类似天然关节软骨的软骨组织的技术。本研究的目的是改善脂肪来源干细胞（ADSCs）向软骨细胞的分化，以便为软骨损伤患者提供一种安全且现代的治疗方法。
在对碳纳米管（CNTs）进行功能化、分散和灭菌后，由水溶性CNTs和藻酸盐制备了一种新型纳米复合凝胶。将ADSCs接种到1.5%藻酸盐支架中，并在添加和不添加转化生长因子-β1（TGF-β1）的软骨形成培养基中培养7天和14天。通过实时聚合酶链反应评估性别决定区Y盒9（SOX9）、II型和X型胶原蛋白的基因表达，并通过酶联免疫吸附测定法测量聚集蛋白聚糖（AGC）和I型胶原蛋白的含量。
我们的研究结果表明，在TGF-β1存在的情况下，浓度为1μg/ml CNTs时，分化的ADSCs中关键软骨标志物SOX9、II型胶原蛋白和AGC的表达与对照组相比显著增加（P < 0.001）。同时，X型胶原蛋白表达以及I型胶原蛋白产生显著降低（P < 0.001）。
结果表明，所使用的三维支架在促进软骨形成的基因表达方面具有显著效果。
