Gharravi Anneh Mohammad, Orazizadeh Mahmoud, Ansari-Asl Karim, Banoni Salem, Izadi Sina, Hashemitabar Mahmoud
Cellular and Molecular Research Center (CMRC), Faculty of Medicine, Ahvaz Jundishapour University of Medical Sciences (AJUMS), Ahvaz, Iran.
Avicenna J Med Biotechnol. 2012 Apr;4(2):65-74.
The aim of the present study was to develop a tissue-engineering approach through alginate gel molding to mimic cartilage tissue in a three-dimensional culture system. The perfusion biomimetic bioreactor was designed to mimic natural joint. The shear stresses exerting on the bioreactor chamber were calculated by Computational Fluid Dynamic (CFD). Several alginate/bovine chondrocyte constructs were prepared, and were cultured in the bioreactor. Histochemical and immunohistochemical staining methods for the presence of glycosaminoglycan(GAG), overall matrix production and type II collagen protein were performed, respectively. The dynamic mechanical device applied a linear mechanical displacement of 2 mm to 10 mm. The CFD modeling indicated peak velocity and maximum wall shear stress were 1.706×10(-3)m/s and 0.02407 dyne/cm(2), respectively. Histochemical and immunohistochemical analysis revealed evidence of cartilage-like tissue with lacunas similar to those of natural cartilage and the production of sulfated GAG of matrix by the chondrons, metachromatic territorial matrix-surrounded cells and accumulation of type II collagen around the cells. The present study indicated that when chondrocytes were seeded in alginate hydrogel and cultured in biomimetic cell culture system, cells survived well and secreted newly synthesized matrix led to improvement of chondrogenesis.
本研究的目的是通过藻酸盐凝胶成型开发一种组织工程方法，以在三维培养系统中模拟软骨组织。灌注仿生生物反应器旨在模拟天然关节。通过计算流体动力学（CFD）计算施加在生物反应器腔室上的剪切应力。制备了几种藻酸盐/牛软骨细胞构建体，并在生物反应器中培养。分别采用组织化学和免疫组织化学染色方法检测糖胺聚糖（GAG）的存在、整体基质产生和II型胶原蛋白。动态机械装置施加2毫米至10毫米的线性机械位移。CFD建模表明，峰值速度和最大壁面剪应力分别为1.706×10（-3）米/秒和0.02407达因/厘米（2）。组织化学和免疫组织化学分析显示，存在类似软骨样组织，具有与天然软骨类似的腔隙，软骨细胞产生基质硫酸化GAG，异染性区域基质包围细胞，细胞周围有II型胶原蛋白积累。本研究表明，当软骨细胞接种到藻酸盐水凝胶中并在仿生细胞培养系统中培养时，细胞存活良好，分泌新合成的基质，从而促进软骨生成。
