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Chondroitin Sulfate- and Decorin-Based Self-Assembling Scaffolds for Cartilage Tissue Engineering

机译:软骨组织工程用硫酸软骨素和Decorin基自组装支架。

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摘要

Cartilage injury and degenerative tissue progression remain poorly understood by the medical community. Therefore, various tissue engineering strategies aim to recover areas of damaged cartilage by using non-traditional approaches. To this end, the use of biomimetic scaffolds for recreating the complex in vivo cartilage microenvironment has become of increasing interest in the field. In the present study, we report the development of two novel biomaterials for cartilage tissue engineering (CTE) with bioactive motifs, aiming to emulate the native cartilage extracellular matrix (ECM). We employed a simple mixture of the self-assembling peptide RAD16-I with either Chondroitin Sulfate (CS) or Decorin molecules, taking advantage of the versatility of RAD16-I. After evaluating the structural stability of the bi-component scaffolds at a physiological pH, we characterized these materials using two different in vitro assessments: re-differentiation of human articular chondrocytes (AC) and induction of human adipose derived stem cells (ADSC) to a chondrogenic commitment. Interestingly, differences in cellular morphology and viability were observed between cell types and culture conditions (control and chondrogenic). In addition, both cell types underwent a chondrogenic commitment under inductive media conditions, and this did not occur under control conditions. Remarkably, the synthesis of important ECM constituents of mature cartilage, such as type II collagen and proteoglycans, was confirmed by gene and protein expression analyses and toluidine blue staining. Furthermore, the viscoelastic behavior of ADSC constructs after 4 weeks of culture was more similar to that of native articular cartilage than to that of AC constructs. Altogether, this comparative study between two cell types demonstrates the versatility of our novel biomaterials and suggests a potential 3D culture system suitable for promoting chondrogenic differentiation.
机译:软骨损伤和退化性组织进展仍未被医学界所理解。因此,各种组织工程策略旨在通过使用非传统方法来恢复受损软骨区域。为此,仿生支架在重建复杂的体内软骨微环境中的用途已成为该领域中越来越多的兴趣。在本研究中,我们报告了两种具有生物活性基序的新型用于软骨组织工程(CTE)的生物材料的开发,旨在模拟天然软骨细胞外基质(ECM)。我们利用RAD16-I的多功能性,将自组装肽RAD16-I与硫酸软骨素(CS)或Decorin分子的简单混合物。在评估双组分支架在生理pH下的结构稳定性后,我们使用两种不同的体外评估方法对这些材料进行了表征:人关节软骨细胞(AC)的再分化和人脂肪来源的干细胞(ADSC)诱导成a软骨承诺。有趣的是,在细胞类型和培养条件(对照和软骨形成)之间观察到了细胞形态和活力的差异。另外,两种细胞类型都在诱导性培养基条件下经历了软骨形成的承诺,而在对照条件下却没有发生。值得注意的是,通过基因和蛋白质表达分析以及甲苯胺蓝染色证实了成熟软骨的重要ECM成分的合成,例如II型胶原蛋白和蛋白聚糖。此外,培养4周后ADSC构建体的粘弹性行为与天然关节软骨相比,与AC构建体更相似。总而言之,这项在两种细胞类型之间的比较研究证明了我们新型生物材料的多功能性,并提出了一种潜在的适合促进软骨分化的3D培养系统。

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