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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.


High hydrostatic pressure device HDR-100 for devitalisation of cartilage tissue.Samples were placed in the hermetically sealed pressure chamber and treated for 10 min at 480 MPa.
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f6: High hydrostatic pressure device HDR-100 for devitalisation of cartilage tissue.Samples were placed in the hermetically sealed pressure chamber and treated for 10 min at 480 MPa.

Mentions: The parameters chosen for HHP treatment based on previous studies involving investigations using articular and meniscal cartilage and ossicles11121830. The devitalisation of cartilage segments was performed 24 to 48 h after sample preparation using HHP. Samples were placed in 2 ml- cryo-tubes completely filled with cell culture medium to avoid air bubbles. Additionally, the vials were sealed with parafilm. The tubes were placed into the glycol-filled pressure chamber of the HHP device (HDR-100, RECORD GmbH, Koenigsee, Germany) and treated for 10 min at approximately 480 MPa and 20 °C (Fig. 6). Untreated cartilage samples were equally handled and served as untreated control.


Devitalisation of human cartilage by high hydrostatic pressure treatment: Subsequent cultivation of chondrocytes and mesenchymal stem cells on the devitalised tissue
High hydrostatic pressure device HDR-100 for devitalisation of cartilage tissue.Samples were placed in the hermetically sealed pressure chamber and treated for 10 min at 480 MPa.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6: High hydrostatic pressure device HDR-100 for devitalisation of cartilage tissue.Samples were placed in the hermetically sealed pressure chamber and treated for 10 min at 480 MPa.
Mentions: The parameters chosen for HHP treatment based on previous studies involving investigations using articular and meniscal cartilage and ossicles11121830. The devitalisation of cartilage segments was performed 24 to 48 h after sample preparation using HHP. Samples were placed in 2 ml- cryo-tubes completely filled with cell culture medium to avoid air bubbles. Additionally, the vials were sealed with parafilm. The tubes were placed into the glycol-filled pressure chamber of the HHP device (HDR-100, RECORD GmbH, Koenigsee, Germany) and treated for 10 min at approximately 480 MPa and 20 °C (Fig. 6). Untreated cartilage samples were equally handled and served as untreated control.

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.