Chondrogenic potential of canine articular cartilage derived cells (cACCs)

Abstract

influence the cartilage phenotype of cACCs during long term culture.Keywords: canine articular cartilage derived cells, chondrogenic differentiation, morphology, ultrastructure, gene expression, chondrogenic markers IntroductionOsteochondral defects are a serious clinical problem both in human as well as in veterinary medicine. The avascular nature of articular cartilage and its limited capacity for spontaneous regeneration are the reasons why cartilage injuries have poor prognosis for repair [1].Currently, cell-based therapies provide real promise for the treatment of musculoskeletal diseases involving osteochondrotic articular cartilage defects [2]. Due to the fact that articular cartilage lesions in dogs are very common, the treatment of these is a great challenge for orthopaedics. The naturally-occurring pathology of articular cartilage in dogs means that these animals represent a clinically important group, and the canine model is often used in translational research. Current studies in canine cartilage regenerative medicine are focusing on the application of multipotent stromal cells for treatment of defects, however the clinical application of chondrocytes in autologous transplantation (ACT) is also noted [1][2][3][4].Nevertheless, a small percentage of the transplanted chondrocytes have sufficient ability to produce components of the extracellular matrix. Therefore, in considering autologous transplantation of chondrocytes, studies regarding optimal methods for the in vitro expansion of chondrocytes are essential , due to the fact that chondrocyte culture is an integral part of the ACT procedure [3].The extracellular matrix (ECM) of functional articular cartilage is rich in proteoglycans and type II collagen (COL2A), constituting respectively 40% and 60% of the dry weight of cartilage. The main proteoglycan in Abstract: In the present paper, the potential of canine articular cartilage-derived cells (cACCs) for chondrogenic differentiation was evaluated. The effectiveness of cACCs' lineage commitment was analyzed after 14 days of culture in chondorgenic and non-chondrogenic conditions. Formation of proteoglycan-rich extracellular matrix was assessed using histochemical staining -Alcian Blue and Safranin-O, while elemental composition was determined by means of SEM-EDX. Additionally, ultrastructure of cACCs was evaluated using TEM. The expression of genes involved in chondrogenesis was monitored with quantitative Real Time PCR. Results obtained indicate that the potential of cACCs for cartilagous extracellular matrix formation may be maintained only in chondrogenic cultures. The formation of specific chondro-nodules was not observed in a non-chondrogenic culture environment. The analysis of cACCs' ultrastructure, both in non-chondrogenic and chondrogenic cultures, revealed well-developed rough endoplasmatic reticulum and presence of mitochondria. The cACCs in chondrogenic medium shed an increased number of microvesicles. Furthermore, it was shown that the extracellular matrix of cACCs in chondrogenic cultu...

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