Method for preparation of neo-cartilage constructs

A method for fabrication of a three-dimensional neo-cartilage construct for in situ implantation into a cartilage lesion, said method comprising steps:

• a) preparing a support matrix structure;
• b) harvesting a piece of cartilage and isolating chondrocytes, or obtaining chondrocytes or cells capable of differentiation into chondrocytes, from a donor;
• c) culturing and expanding said chondrocytes or said cells;
• d) suspending the expanded chondrocytes or cells in a suspension fluid;
• e) incorporating said suspended chondrocytes or cells into said support matrix structure;
• and
• f) propagating said chondrocytes into a three-dimensional neo-cartilage construct using an algorithm of the invention,
• wherein said algorithm comprises subjecting said chondrocytes or cells to a cyclic or constant hydrostatic pressure from about zero to about 10 MPa at from about 0.01 to about 1 Hz, to atmospheric pressure or to non-pressure conditions, for from about 1 to about 24 hours a day, for about 1 to about 90 days at a rate of a medium perfusion from about zero to about 500 μL/min, at an oxygen concentration from about zero to about 20% and a carbon dioxide concentration of about zero to about 5%.

A method for preparation of a neo-cartilage construct suitable for in situ implantation into a cartilage lesion, said construct comprising a cultured differentiated autologous or heterologous chondrocytes or cells which could be differentiated into chondrocytes, incorporated into a support matrix and subjected to an algorithm of the invention,

• said construct prepared by a method comprising steps
• a) preparing a support matrix structure;
• b) harvesting a piece of cartilage and isolating chondrocytes, or obtaining chondrocytes or cells capable of differentiation into chondrocytes, from a donor;
• c) culturing and expanding said chondrocytes or said cells;
• d) suspending the expanded chondrocytes or cells in a suspension fluid;
• e) incorporating said suspended chondrocytes or cells into said support matrix structure;
• and
• f) propagating said chondrocytes into three-dimensional neo-cartilage construct using an algorithm comprising subjecting said propagated chondrocytes or cells within the neo-cartilage construct to a cyclic or constant hydrostatic pressure from about zero to about 10 MPa at from about 0.01 to about 1 Hz, preceded or followed by a period of time where said cells are subjected to an atmospheric pressure or to non-pressure conditions, for from about 1 to about 24 hours a day for about zero to about 90 days at a rate of a medium perfusion from zero to about 500 μL/min at an oxygen concentration from about zero to about 20% and a carbon dioxide concentration of zero to about 5% wherein a cell density of suspended chondrocytes or cells in the suspension fluid is from below 3 millions to about 60 millions.