Cartilage Tissue Engineering in Plasma and Whole Blood Scaffolds
Cartilage Tissue Engineering in Plasma and Whole Blood Scaffolds
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DOI：
10.1002/adma.200701344
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发表时间：
2008-06
影响因子：
29.4
通讯作者：
M. Haberhauer;Göran Zernia;A. Deiwick;C. Pösel;A. Bader;D. Huster;R. Schulz
中科院分区：
文献类型：
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作者：
M. Haberhauer;Göran Zernia;A. Deiwick;C. Pösel;A. Bader;D. Huster;R. Schulz
生物组织是一种纳米/微米结构的材料，具有优异的力学性能。例如，关节软骨由高度组织化的细胞外分子结构构建，所述细胞外分子结构包括相对刚性的胶原支架和柔性的蛋白聚糖部分，其以水含量为80重量%的凝胶状结构组织。[1]这种结构组件为软骨提供其承载和减震能力。[2]虽然细胞外基质（ECM）组分的分解和再合成的连续过程恢复了组织中所需的材料性质，但各种退行性疾病如关节炎或创伤等病症可严重分解和损伤软骨层。由于软骨是无血管组织，较大缺损的自愈能力非常有限。[3]因此，在组织工程领域中正在进行人工生长用于置换手术的软骨构造的若干尝试。[4]三维（3D）软骨组织工程不是一项简单的任务。在体外，培养系统和生长条件刺激分离的软骨细胞（软骨形成细胞）改变它们的表达模式、表型或生物化学性质。[5]由于在单层培养中缺乏3D微环境，软骨细胞在体外合成不同类型和少量的ECM组分。[6]经过几次传代培养和更长的培养期后，细胞去分化为成纤维细胞样表型，其特征在于II型胶原蛋白（COL 2）和蛋白聚糖产生减少，I型胶原蛋白（COL 1）表达增加，这对关节软骨是非特异性的。[7]相反，当接种到3D环境中时，例如基于胶原的水凝胶，软骨细胞开始重新分化并保持其原始表型。[6a目前的研究集中在合成或生物支架的开发上，其中新鲜分离的软骨细胞增殖并保持其表型。[9]该主要支撑基质的组成和纳米/微米结构是所得组织质量的关键决定因素。适用于软骨修复的基质偶联自体软骨细胞移植物的要求包括生物相容性、生物降解性、弹性和稳定性。[10]几种材料，已经被应用于生长人工软骨。[4a例如，全血或血浆的级分已经用于几种应用中，例如纤维蛋白胶、纤维蛋白-软骨细胞构建体、当接种到胶原支架上时装载细胞的纤维蛋白凝胶，以及用于在体外自体补充培养基的富血小板血浆（PRP）或至少以含有软骨细胞或间充质干细胞的PRP支架的形状。[11]大多数报告的是有益的结果。发表了一项有前景的体内研究，其中将软骨细胞负载的自体纤维蛋白移植物植入绵羊股骨外侧和内侧髁的全层缺损中，根据ICRS分类，3个月后软骨修复得到改善。[11b]全血成分如纤维蛋白由于免疫相容性、非细胞毒性、生物降解性和机械稳定性而在再生医学中提供了巨大的应用潜力。在这里，我们描述了两种生物基质作为纳米/微支架材料，首次用于软骨组织工程。在这项研究中，我们利用收集的全血和血浆作为软骨细胞支架材料后，直接必要的冻融程序，但省略了进一步的制备步骤，如分离和分馏。骨关节炎新组织的形成...
Biological tissues are nano/micro-structured materials of remarkable mechanical properties. For instance, articular cartilage is built from a highly organized extracellular molecular architecture comprising a relatively rigid collagen scaffold and ffexible proteoglycan moieties, organized in a gel like structure with a water content of $80 wt%.[1] This structural assembly provides cartilage its load bearing and shock adsorbing capacity.[2] While a continuous process of decomposition and resynthesis of extracellular matrix (ECM) components restores the desired material properties in the tissue, various degenerative diseases such as arthritis or conditions like trauma can severely decompose and damage the cartilage layer. Since cartilage is an avascular tissue, the self healing capacity of larger defects is very limited.[3] Therefore, several attempts to artificially grow cartilage constructs for replacement surgery are being undertaken in the field of tissue engineering.[4] Three-dimensional (3D) cartilage tissue engineering is not a trivial task. In vitro, both the culture system and the growth conditions stimulate isolated chondrocytes, the cartilage forming cells, to change their expression pattern, phenotype or biochemical properties.[5] Due to the lacking 3D microenvironment in monolayer cultivation chondrocytes synthesize different types and smaller amounts of ECM components in vitro.[6] After a few subcultures and longer cultivation periods the cells de-differentiate to a fibroblast-like phenotype, characterized by a decrease in type II collagen (COL2) and proteoglycan production and increased expression of type I collagen (COL1), which is unspecific for articular cartilage.[7] In contrast, when seeded into 3D environments such as collagen based hydrogels chondrocytes start to re-differentiate and remain their original phenotype.[6a, 8] Current research is focused on the development of synthetic or biological scaffolds, in which freshly isolated chondrocytes proliferate and maintain their phenotype.[9] The composition and nano/micro-structure of this primary support matrix is a key determinant for the quality of the resulting tissue. The requirements for a matrix coupled autologous chondrocyte transplant applicable for cartilage repair include biocompatibility, biodegradability, elasticity and stability.[10] Several materials, have already been applied to grow artificial cartilage.[4a, 6a]For instance, fractions of whole blood or blood plasma have already been used in several applications, such as fibrin glue, fibrin-chondrocyte constructs, cell loaded fibrin gel when seeded onto a collagen scaffold, as well as platelet-rich plasma (PRP) for autologous supplementation of media in vitro or at least in the shape of a PRP scaffold containing chondrocytes or mesenchymal stem cells,[11] mostly reporting beneficial results. A promising in vivo study was published, where chondrocyte loaded autologous fibrin grafts were implanted into fullthickness defects of lateral and medial femoral condyles of sheep and in conclusion resulted in improved cartilage repair after 3 months according ICRS classification.[11b] Whole blood components such as fibrin offer great potential for the application in regenerative medicine due to immuno-compatibility, non-cytotoxity, biodegradability and mechanical stability. Here we describe two biological matrices as nano/microscaffold materials that are used for the first time for cartilage tissue engineering. In this study we utilized the collected whole blood and plasma as chondrocyte scaffolding material directly after an essential freeze-thawing procedure but omitting further preparative steps such as separation and fractionation. Cartilage neo-tissue formation in …