一种用于培养半月板形状构建体的牛共培养无支架方法的评估。

Assessment of a bovine co-culture, scaffold-free method for growing meniscus-shaped constructs.

作者信息

Aufderheide Adam C, Athanasiou Kyriacos A

机构信息

Department of Bioengineering, Rice University, Houston, Texas 77251, USA.

出版信息

Tissue Eng. 2007 Sep;13(9):2195-205. doi: 10.1089/ten.2006.0291.

Abstract

Using a self-assembly (SA), scaffoldless method, five high-density co-cultures with varied ratios of meniscal fibrochondrocytes (MFCs) and articular chondrocytes (ACs) were seeded into novel meniscus-specific, ring-shaped agarose wells. The following ratios of MFCs to ACs were used: 0% MFC, 25% MFC, 50% MFC, 75% MFC, and 100% MFC. Over 4 weeks, all ratios of cells self-assembled into three-dimensional constructs with varying mechanobiological and morphological properties. All groups stained for collagen II (Col II), and all groups except the 0% MFC group stained for collagen I (Col I). It was found that the tensile modulus was proportional to the percentage of MFCs employed. The 100% MFC group yielded the greatest mechanical stiffness with 432.2 +/- 47 kPa tensile modulus and an ultimate tensile strength of 23.7 +/- 2.4 kPa. On gross inspection, the 50% MFC constructs were the most similar to our idealized meniscus shape, our primary criterion. A second experiment was performed to examine the anisotropy of constructs as well as to directly compare the scaffoldless, SA method with a poly-glycolic acid (PGA) scaffold-based construct. When compared to PGA constructs, the SA groups were 2-4 times stiffer and stronger in tension. Further, at 8 weeks, SA groups exhibited circumferential fiber bundles similar to native tissue. When pulled in the circumferential direction, the SA group had significantly higher tensile modulus (226 +/- 76 kPa) than when pulled in the radial direction (67 +/- 32 kPa). The PGA constructs had neither a directional collagen fiber orientation nor differences in mechanical properties in the radial or circumferential direction. It is suggested that the geometric constraint imposed by the ring-shaped, nonadhesive mold guides collagen fibril directionality and, thus, alters mechanical properties. Co-culturing ACs and MFCs in this manner appears to be a promising new method for tissue engineering fibrocartilaginous tissues exhibiting a spectrum of mechanical and biomechanical properties.

摘要

采用一种自组装(SA)、无支架的方法,将五种不同比例的半月板纤维软骨细胞(MFCs)和关节软骨细胞(ACs)的高密度共培养物接种到新型的半月板特异性环形琼脂糖孔中。使用了以下MFCs与ACs的比例:0% MFC、25% MFC、50% MFC、75% MFC和100% MFC。在4周的时间里,所有比例的细胞都自组装成具有不同机械生物学和形态学特性的三维构建体。所有组均对Ⅱ型胶原(Col II)进行染色,除0% MFC组外的所有组均对Ⅰ型胶原(Col I)进行染色。结果发现,拉伸模量与所用MFCs的百分比成正比。100% MFC组产生了最大的机械刚度,拉伸模量为432.2±47 kPa,极限拉伸强度为23.7±2.4 kPa。大体检查时,50% MFC构建体与我们理想化的半月板形状最为相似,这是我们的主要标准。进行了第二项实验,以检查构建体的各向异性,并将无支架的SA方法与基于聚乙醇酸(PGA)支架的构建体进行直接比较。与PGA构建体相比,SA组在拉伸时的硬度和强度高2至4倍。此外,在8周时,SA组呈现出与天然组织相似的周向纤维束。当沿周向拉动时,SA组的拉伸模量(226±76 kPa)明显高于沿径向拉动时(67±32 kPa)。PGA构建体既没有定向的胶原纤维取向,在径向或周向的机械性能也没有差异。有人提出,环形非粘性模具施加的几何约束引导胶原纤维的方向性,从而改变机械性能。以这种方式共培养ACs和MFCs似乎是一种用于组织工程纤维软骨组织的有前途的新方法,该组织具有一系列机械和生物力学特性。

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