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Devitalisation of human cartilage by high hydrostatic pressure treatment: Subsequent cultivation of chondrocytes and mesenchymal stem cells on the devitalised tissue

View Article: PubMed Central - PubMed

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.

No MeSH data available.


FESEM images of untreated and HHP-treated cartilage.High pressure treatment did not affect the ECM structure of hyaline cartilage (upper panel: bar = 20 μm; lower panel: bar = 5 μm).
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f4: FESEM images of untreated and HHP-treated cartilage.High pressure treatment did not affect the ECM structure of hyaline cartilage (upper panel: bar = 20 μm; lower panel: bar = 5 μm).

Mentions: Moreover, FESEM analyses demonstrated no damaged areas in HHP-treated tissue, indicating the preserved integrity of the tissue (Fig. 4). The collagen fibres of pressure-loaded cartilage were in the native conformation and showed no signs of disruption or damage.


Devitalisation of human cartilage by high hydrostatic pressure treatment: Subsequent cultivation of chondrocytes and mesenchymal stem cells on the devitalised tissue
FESEM images of untreated and HHP-treated cartilage.High pressure treatment did not affect the ECM structure of hyaline cartilage (upper panel: bar = 20 μm; lower panel: bar = 5 μm).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037397&req=5

f4: FESEM images of untreated and HHP-treated cartilage.High pressure treatment did not affect the ECM structure of hyaline cartilage (upper panel: bar = 20 μm; lower panel: bar = 5 μm).
Mentions: Moreover, FESEM analyses demonstrated no damaged areas in HHP-treated tissue, indicating the preserved integrity of the tissue (Fig. 4). The collagen fibres of pressure-loaded cartilage were in the native conformation and showed no signs of disruption or damage.

View Article: PubMed Central - PubMed

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.

No MeSH data available.