Cartilage tissue damage and diseases are the most common clinical situation that occurs because of aging and injury, thereby causing pain and loss of mobility. The inability of cartilage tissue to self-repair is instrumental in developing tissue engineered substitutes. To this effect, the present study aims to engineer cartilage construct by culturing umbilical cord blood-derived human mesenchymal stem cells (hMSCs) on novel 3D porous scaffolds developed from natural biopolymers, silk fibroin (SF) and chitosan (CS), with addition of cartilage matrix components, glucosamine (Gl) and chondroitin sulfate (Ch). The presence of Gl and Ch is expected to enhance cartilage regeneration. The developed SF/CS-Gl-Ch scaffolds possess desired pore size in the range 56.55–168.15 μm, 88–92% porosity, 44.7–46.8̊ contact angle, controlled swelling and biodegradability. Upon culturing under dynamic condition in a spinner flask bioreactor, the scaffold supported hMSCs attachment, proliferation, and further promoted chondrogenic differentiation. Cartilage-specific matrix and gene (Collagen II, Sox9 and aggrecan) expression analyses by histology, immunophenotype, immunofluorescence and quantitative PCR studies showed superiority of cell-scaffold construct generated in dynamic culture towards cartilage tissue generation as compared to cell aggregates formed by pellet culture. This study demonstrates the potentiality of SF/CS-Gl-Ch porous scaffold for the development of tissue construct for cartilage regeneration under dynamic culture condition.

软骨组织损伤和疾病是由于衰老和受伤而发生的最常见的临床情况，从而引起疼痛和活动能力丧失。软骨组织无法自我修复有助于开发组织工程替代品。为此，本研究旨在通过在由天然生物聚合物，丝素蛋白（SF）和壳聚糖（CS）开发的新型3D多孔支架上培养脐血来源的人间充质干细胞（hMSC），从而工程化软骨构造。软骨基质成分，葡萄糖胺（G1）和硫酸软骨素（Ch）。预期G1和Ch的存在可增强软骨再生。已开发的SF / CS-Gl-Ch支架具有理想的孔径，范围为56.55–168.15μm，孔隙率为88–92％，接触角为44.7–46.8̊，控制溶胀和生物降解性。在动态条件下在旋转瓶生物反应器中培养后，支架支持hMSCs的附着，增殖并进一步促进软骨分化。通过组织学，免疫表型，免疫荧光和定量PCR研究的软骨特异性基质和基因（胶原II，Sox9和聚集蛋白聚糖）表达分析表明，与通过沉淀培养形成的细胞聚集体相比，动态培养中生成的细胞支架构建体对软骨组织生成具有优越性。这项研究证明了SF / CS-Gl-Ch多孔支架在动态培养条件下发展用于软骨再生的组织结构的潜力。并进一步促进软骨分化。通过组织学，免疫表型，免疫荧光和定量PCR研究的软骨特异性基质和基因（胶原II，Sox9和聚集蛋白聚糖）表达分析表明，与通过沉淀培养形成的细胞聚集体相比，动态培养中生成的细胞支架构建体对软骨组织生成具有优越性。这项研究证明了SF / CS-Gl-Ch多孔支架在动态培养条件下发展用于软骨再生的组织结构的潜力。并进一步促进软骨分化。通过组织学，免疫表型，免疫荧光和定量PCR研究的软骨特异性基质和基因（胶原II，Sox9和聚集蛋白聚糖）表达分析表明，与通过沉淀培养形成的细胞聚集体相比，动态培养中生成的细胞支架构建体对软骨组织生成具有优越性。这项研究证明了SF / CS-Gl-Ch多孔支架在动态培养条件下发展用于软骨再生的组织结构的潜力。免疫荧光和定量PCR研究表明，与通过沉淀培养形成的细胞聚集体相比，动态培养中生成的细胞支架构建体对软骨组织生成的优越性。这项研究证明了SF / CS-Gl-Ch多孔支架在动态培养条件下发展用于软骨再生的组织结构的潜力。免疫荧光和定量PCR研究表明，与通过沉淀培养形成的细胞聚集体相比，动态培养中生成的细胞支架构建体对软骨组织生成的优越性。这项研究证明了SF / CS-Gl-Ch多孔支架在动态培养条件下发展用于软骨再生的组织结构的潜力。