Abstract
The regeneration of cartilage lesions still represents a major challenge. Cartilage has a tissue-specific architecture, complicating recreation by synthetic biomaterials. A novel approach for reconstruction is the use of devitalised cartilage. Treatment with high hydrostatic pressure (HHP) achieves devitalisation while biomechanical properties are remained. Therefore, in the present study, cartilage was devitalised using HHP treatment and the potential for revitalisation with chondrocytes and mesenchymal stem cells (MSCs) was investigated. The devitalisation of cartilage was performed by application of 480 MPa over 10 minutes. Effective cellular inactivation was demonstrated by the trypan blue exclusion test and DNA quantification. Histology and electron microscopy examinations showed undamaged cartilage structure after HHP treatment. For revitalisation chondrocytes and MSCs were cultured on devitalised cartilage without supplementation of chondrogenic growth factors. Both chondrocytes and MSCs significantly increased expression of cartilage-specific genes. ECM stainings showed neocartilage-like structure with positive AZAN staining as well as collagen type II and aggrecan deposition after three weeks of cultivation. Our results showed that HHP treatment caused devitalisation of cartilage tissue. ECM proteins were not influenced, thus, providing a scaffold for chondrogenic differentiation of MSCs and chondrocytes. Therefore, using HHP-treated tissue might be a promising approach for cartilage repair.
Highlights
Result in unwanted donor side morbidity and additional long-term complications
Results indicated that MSCs and chondrocytes seeded on the high hydrostatic pressure (HHP)-treated cartilage differentiate into chondrogenic lineage
The proliferation on the surface was detected demonstrating that HHP-treated tissue is biocompatible
Summary
Result in unwanted donor side morbidity and additional long-term complications. the availability of healthy tissue is restricted and, in the case of ACI, a second operative procedure is required[4,7]. The use of allogenic cartilage could provide an alternative treatment option[8] This is a restorative procedure in which either chondrocytes or articular cartilage and its underlying subchondral bone obtained from a donor of the same species are transplanted into the defect. Used decellularisation and sterilisation procedures include chemical detergents, autoclaving and irradiation These techniques influence the hydration status and three-dimensional orientation of proteins, resulting in altered biomechanical properties of the cartilage tissue[10]. To overcome these limitations, a novel approach has been developed using high hydrostatic pressure (HHP) to achieve the devitalisation of cartilage tissue while maintaining the biomechanical characteristics[11,12]. The effect of HHP on the viability of chondrocytes within a cartilage matrix was tested and the potential of devitalised tissue for revitalisation using chondrocytes and mesenchymal stem cells was analysed
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