Abstract
The present study aims to assess coculture of allogenic decalcified bone matrix (DBM) and bone marrow mesenchymal stem cells (BMSCs) in the knee joint cavity of rabbits for cartilage tissue engineering. Rabbits were assigned to an in vitro group, an in vivo group, and a blank control group. At the 4th, 8th, and 12th week, samples from all groups were collected for hematoxylin–eosin (HE) staining and streptavidin–peroxidase (SP) method. The morphological analysis software was used to calculate the average absorbance value (A value). SP and flow cytometry demonstrated that BMSCs were induced into chondrocytes. DBM scaffold showed honeycomb-shaped porous and three-dimensional structure, while the surface pores are interlinked with the deep pores. At the 4th week, in the blank control group, DBM scaffold structure was clear, and cells analogous to chondrocytes were scattered in the interior of DBM scaffolds. At the 8th week, in the in vivo group, there were a large amount of cells, mainly mature chondrocytes, and the DBM scaffolds were partially absorbed. At the 12th week, in the in vitro group, the interior of scaffolds was filled up with chondrocytes with partial fibrosis, but arranged in disorder. In the in vivo group, the chondrocytes completely infiltrated into the interior of scaffolds and were arranged in certain stress direction. The in vivo group showed higher A value than the in vitro and blank control groups at each time point. Allogenic DBM combined BMSCs in the knee joint cavity of rabbits could provide better tissue-engineered cartilage than that cultivated in vitro.
Highlights
Normal human articular cartilage is a relatively simple tissue without blood vessels or lymph, and it acquires the nutrients via the infiltration of synovial fluid, which in turn limits the proliferation of chondrocytes, it has limited repair capabilities during a cartilage injury [1, 2]
The original generation of rabbit bone marrow mesenchymal stem cell (BMSC) were adherent to the glass wall after 48-h culture, it was grown in colonies for 4–5 days with noticeably increasing cells, and formed plenty of cell clones in different sizes
The 3rd generation of BMSCs was observed under the microscope at high magnification: cells were long fusiform- or spindle-shaped with obvious nucleus, clear nucleolus, and high nucleus/cytoplasm ratio; vacuoles and lipid droplets were found in cytoplasm; and cells were closely adherent
Summary
Normal human articular cartilage is a relatively simple tissue without blood vessels or lymph, and it acquires the nutrients via the infiltration of synovial fluid, which in turn limits the proliferation of chondrocytes, it has limited repair capabilities during a cartilage injury [1, 2]. Cartilage tissue engineering offers a promising approach to repair cartilage injury via combining cells and biocompatible scaffold, so it is therapeutically possible to replace injured articular cartilage by tissue-engineered cartilage successfully [5]. Due to similar three-dimensional (3D) structure to autogenous bone, DBM possesses great biocompatibility mainly based on type I collagen (good scaffold for cell adhesion and growth) and high seed c 2017 The Author(s).
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