Advances in Mesenchymal Stem Cells and Tissue Engineering

Nguyen, Hue Thi-Ngoc, Vu, Ngoc Bich

The damaged articular cartilage has limited self-regeneration capacity because of the absence of blood vessels, lymphatics, and nerves. Cartilage transplantation is, hence, a popular method used to treat this disease. However, sources of autograft and allogenic cartilage for transplantation are limited. Therefore, this study aims to suggest a simple method to produce engineered cartilage from human adipose-derived mesenchymal stem cells (ADSCs) and poly (ε-caprolactone) (PCL) scaffolds. ADSCs were isolated and expanded from fat tissues according to published protocols. PCL-porous scaffolds were produced from PCL with 5 × 5 × 0.6 mm3 with 200-400 μ m pore sizes. ADSCs were seeded on the PCL scaffolds at three different densities (104, 105, 106 cells per scaffold). The adherence of ADSCs on the surface of PCL scaffolds was evaluated based on an immunostaining assay to determine the presence of ADSCs. The cell proliferation on PCL scaffolds was determined by MTT assay. The complexity in ADSCs and PCL scaffolds was induced to cartilage using a chondrogenesis medium. The engineered cartilage was characterized by the accumulation of proteoglycan and aggrecan by Safranin O staining assay. Their structures were evaluated using an H-E staining assay. Finally, these engineered cartilage tissues were transplanted into mice to assess cartilage maturation when compared to natural cartilage. The results showed that the engineered cartilage tissues could be successfully produced by cultures of ADSCs on poly ε-caprolactone scaffolds in combination with chondrogenesis medium. The suitable density of ADSCs was 106 cells/per scaffold of 5 × 6 × 0.6 mm3 with pore size from 200 to 400 μ m. The results showed that an in vitro cartilage tissue was created from ADSCs and PCL scaffold. The cartilage tissue exists in the mice for 6 months.