Method for making a porous scaffold suitable for use in repair of osseous, chondral, or osteochondral defects in a mammal

A method for making a porous scaffold suitable for use in repair of osseous, chondral, or osteochondral defects in a mammal, the method comprising the steps of:

• A) providing a gel comprising solubilized extracellular matrix (ECM) tissue;
• B) chemically crosslinking the extracellular matrix (ECM) tissue utilizing glyoxal as the chemically crosslinking agent;
• C) freeze-drying the gel to provide the porous scaffold;
• D) dehydrothermally (DHT) crosslinking the scaffold to produce a first layer;
• and
• E) repeating steps A-D to make a second layer, wherein the first layer is attached to the second layer;
• wherein the extracellular matrix is treated to reduce the GAG content to less than 90% of the GAG content of untreated ECM, and wherein the ECM tissue in at least one layer is cartilage ECM.