首页> 外文会议>NATO Advanced Study Institute on Polymer Based Systems on Tissue Engineering, Replacement and Regeneration Oct 15-25, 2001 Alvor, Algarve, Portugal >TISSUE ENGINEERING OF ELASTIC CARTILAGE BY USING SCAFFOLD/CELL CONSTRUCTS WITH DIFFERENT PHYSICAL AND CHEMICAL PROPERTIES
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TISSUE ENGINEERING OF ELASTIC CARTILAGE BY USING SCAFFOLD/CELL CONSTRUCTS WITH DIFFERENT PHYSICAL AND CHEMICAL PROPERTIES

机译:通过使用具有不同理化特性的脚手架/细胞结构进行弹性软骨的组织工程

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In this study, we compared three scaffold/cell constructs with different physical and chemical properties, in their potential for tissue engineering elastic cartilage. Group Ⅰ consisted of PCL scaffolds Group Ⅱ custom-made non-wovt n made of PGA fibers in combination with agarose beads; collagen sponges were used in-group Ⅲ. Chondrocytes were isolated via enzyme digestion from an ear cartilage biopsy of 2 year old male piglets. 250,000 cells in 30 ul were seeded into three different scaffold types. The specimens were then cultured for 1 week. The scaffold/cell constructs and controls were placed subcutaneously on the paravertebral fascia for 4 1/2 month. Explanted scaffolds were cut in cubical blocks and mechanically tested in phosphate buffered saline solution at 37℃ in accordance with ASTM F451-99a Standard Specification for Acrylic Bone Cement. After explantatior the non-seeded PCL specimens (control) had compressive stiffness and 1 % offset yield strength in plane of 5.6 +- 0.4 MPa and out of plane 4.9 +- 0.5 MPa, respectively. In comparison, the scaffolds with a seeded specimens had compressive stiffness of 4.9 +- 0.5 MPa and 3.9+- 0.3 MPa, respectively. The polymer molecular weight distribution was determined by gel permeation chromatography to study the in vivo degradation of the polymer matrix. Sections of explants were prepared using standard cryo-histochemical techniques and stained with anti-type Ⅱ Ⅲ, Ⅸ collagen antibodies, F-actin and anti-integrin beta, and Ⅰ. There was considerably more organized extracellular matrix formation in group Ⅰ, whereas group Ⅱ and Ⅲ exhibiting a loosely scattered patterns. Results of the toluidine blue, trichrome Goldner's staining and collagen Ⅱ expression showed cartilage-like tissue formation in the PCL scaffolds whereas all the other matrices showed fibrous tissue in combination with calcification.
机译:在这项研究中,我们比较了三种具有不同物理和化学特性的支架/细胞构建体在组织工程弹性软骨中的潜力。第一组由PCL支架组成,第二组由PGA纤维与琼脂糖珠结合制成的定制非织造布。 Ⅲ组使用胶原蛋白海绵。通过酶消化从2岁雄性仔猪的耳软骨活检中分离软骨细胞。将30ul中的250,000个细胞接种到三种不同的支架类型中。然后将样品培养1周。将支架/细胞构建体和对照皮下放置在椎旁筋膜上4 1/2个月。按照ASTM F451-99a丙烯酸骨水泥标准规范,将外植的支架切成立方体块,并在37℃的磷酸盐缓冲盐溶液中进行机械测试。种植后,非种子PCL样品(对照)在平面内分别具有5.6±0.4 MPa的抗压刚度和4.9±0.5 MPa的平面外的1%偏移屈服强度。相比之下,具有种子样本的支架的抗压刚度分别为4.9±0.5 MPa和3.9±0.3 MPa。通过凝胶渗透色谱法确定聚合物分子量分布,以研究聚合物基质的体内降解。用标准的冷冻组织化学技术制备外植体切片,并用抗Ⅱ型Ⅲ,Ⅰ型胶原抗体,F-肌动蛋白和抗整合素β以及Ⅰ染色。 Ⅰ组细胞外基质的形成更为有组织,而Ⅱ组和Ⅲ组则表现出松散的分布。甲苯胺蓝,三色戈德纳氏染色和Ⅱ型胶原的表达结果表明,PCL支架中软骨样组织形成,而其他所有基质均显示出纤维组织并伴有钙化。

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