Mallick Sarada Prasanna, Rastogi Amit, Tripathi Satyavrat, Srivastava Pradeep
School of Biochemical Engineering, Indian Institute of Technology (Banaras Hindu University), Varanasi, 221005, India.
Department of Orthopaedics, Institute of Medical Sciences, Banaras Hindu University, Varanasi, 221005, India.
Bioprocess Biosyst Eng. 2017 Apr;40(4):601-610. doi: 10.1007/s00449-016-1724-4. Epub 2016 Dec 19.
The current work is an attempt to study the strategies for cartilage tissue regeneration using porous scaffold in wavy walled airlift bioreactor (ALBR). Novel chitosan, poly (L-lactide) and hyaluronic acid based composite scaffold were prepared. The scaffolds were cross-linked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, N-hydroxysuccinimide and chondroitin sulfate to obtain interconnected 3D microstructure showing excellent biocompatibility, higher cellular differentiation and increased stability. The surface morphology and porosity of the scaffolds were analyzed using scanning electron microscopy (SEM) and mercury intrusion porosimeter and optimized for chondrocyte regeneration. The study shows that the scaffolds were highly porous with pore size ranging from 48 to 180 µm and the porosities in the range 80-92%. Swelling and in vitro degradation studies were performed for the composite scaffolds; by increasing the chitosan: HA ratio in the composite scaffolds, the swelling property increases and stabilizes after 24 h. There was controlled degradation of composite scaffolds for 4 weeks. The uniform chondrocyte distribution in the scaffold using various growth modes in the shake flask and ALBR was studied by glycosaminoglycans (GAG) quantification, MTT assay and mixing time evaluation. The cell culture studies demonstrated that efficient designing of ALBR increases the cartilage regeneration as compared to using a shake flask. The free chondrocyte microscopy and cell attachment were performed by inverted microscope and SEM, and from the study it was confirmed that the cells uniformly attached to the scaffold. This study focuses on optimizing strategies for the culture of chondrocyte using suitable scaffold for improved cartilage tissue regeneration.
当前的工作旨在研究在波浪壁气升式生物反应器（ALBR）中使用多孔支架进行软骨组织再生的策略。制备了新型的基于壳聚糖、聚（L-丙交酯）和透明质酸的复合支架。这些支架用1-乙基-3-（3-二甲基氨基丙基）碳二亚胺、N-羟基琥珀酰亚胺和硫酸软骨素进行交联，以获得具有优异生物相容性、更高细胞分化和更高稳定性的相互连接的三维微观结构。使用扫描电子显微镜（SEM）和压汞仪分析了支架的表面形态和孔隙率，并针对软骨细胞再生进行了优化。研究表明，支架具有高度多孔性，孔径范围为48至180 µm，孔隙率在80 - 92%之间。对复合支架进行了溶胀和体外降解研究；通过增加复合支架中壳聚糖：HA的比例，溶胀性能增加，并在24小时后稳定下来。复合支架在4周内有可控的降解。通过糖胺聚糖（GAG）定量、MTT分析和混合时间评估，研究了在摇瓶和ALBR中使用各种生长模式时支架中软骨细胞的均匀分布。细胞培养研究表明，与使用摇瓶相比，ALBR的有效设计增加了软骨再生。通过倒置显微镜和SEM进行了游离软骨细胞显微镜检查和细胞附着研究，从研究中证实细胞均匀地附着在支架上。本研究着重于优化使用合适支架培养软骨细胞的策略，以促进软骨组织再生。
