Cultured, full-thickness integument substitutes based on three-dimensional matrix membranes

A surgical kit comprising a sealed container providing a sterile environment, and contained therein a sterilized surgical implant device comprising a cultured, full-thickness or split-thickness integument substitute to be implanted in a patient requiring the same, wherein said three-dimensional mark membrane comprises essentially two surfaces, in that the length and width thereof are substantially greater than the thickness thereof which is from about 0.1 mm to about 5 mm, said matrix membrane being non-reactive with respect to calls and components of said integument;

• in which either pores in the area of a first surface having a mean pore diameter in the range of from about 50 .mu.m to about 900 .mu.m, have immobilized therein cells and components of a dermal layer of said integument, or in which pores in the area of a second, opposite surface having a mean pore diameter in the range of from about 0.5 .mu.m to about 100 .mu.m, have immobilized therein cells and components of an epidermal layer of said integument, or in which pores in the area of both said first and second surfaces have immobilized therein cells and components of said dermal layer and said epidermal layer, respectively;
• said membrane also having a lateral pore structure in the internal space thereof interconnecting said pores which pass from one surface to the other of said membrane, permitting co-planar flow of a liquid or suspension therethrough.

A method of preparing a cultured, full-thickness integument substitute implanted in or to be implanted in a patient requiring the same, comprising:

• A) establishing a three-dimensional matrix membrane with essentially two surfaces, in that the length and width thereof are substantially greater than the thickness thereof which is from about 0.1 mm to about 5 mm, said matrix membrane being non-reactive with respect to cells and components of said integument;
• said membrane having pores in the area of a first surface thereof which are directly connected to, but substantially larger on average than those in the area of a second, opposite surface of said membrane, said first surface pores having a mean pore diameter in the range of from about 50 .mu.m to about 900 .mu.m;
• said membrane having pores in said area of said second surface thereof which are directly connected to but substantially smaller on average than those in said area of said first, opposite surface of said membrane, said second surface pores having a mean pore diameter in the range of from about 0.5 .mu.m to about 100 .mu.m;
• said membrane also having a lateral pore structure in the internal space thereof interconnecting said pores which pass from one surface to the other of said membrane, permitting co-planar flow of a liquid or suspension therethrough;
• B) bringing said first surface of said matrix membrane with said larger pores in contact with cells and components of said integument whereby they pass through said matrix membrane, with larger cells and components of a dermal layer of said integument becoming immobilized within said pores in said area of said first surface of said matrix membrane, and with smaller cells and components of an epidermal layer of said integument becoming immobilized within said pores in said area of said second surface of said matrix membrane;
• C) culturing said matrix membrane thus seeded with said cells and components of said integument immobilized in different pore areas thereof under conditions favorable to growth of said integument cells, whether in vivo or in vitro, whereby there results a cultured, full-thickness integument substitute already implanted in, or ready for implantation in said patient.

A method of preparing a cultured, split-thickness or full-thickness integument substitute implanted in or to be implanted in a patient requiring the same, comprising:

• A) establishing a three-dimensional matrix membrane with essentially two surfaces, in that the length and width thereof are substantially greater than the thickness thereof which is from about 0.1 mm to about 5 mm, said matrix membrane being non-reactive with respect to cells and components of said integument;
• said membrane having pores in the area of a first surface thereof which are directly connected to, but substantially larger on average than those in the area of a second, opposite surface of said membrane, said first surface pores having a mean pore diameter in the range of from about 50 .mu.m to about 900 .mu.m;
• said membrane having pores in said area of said second surface thereof which are directly connected to but substantially smaller on average than those in said area of said first, opposite surface of said membrane, said second surface pores having a mean pore diameter in the range of from about 0.5 .mu.m to about 100 .mu.m;
• said membrane also having a lateral pore structure in the internal space thereof interconnecting said pores which pass from one surface to the other of said membrane, permitting co-planar flow of a liquid or suspension therethrough;
• and then carrying out either or both of the following steps B) and C):
• B) bringing said first surface of said matrix membrane with said larger pores in contact with cells and components of a dermal layer of said integument whereby they become immobilized within pores in the area of said first surface of said matrix membrane;
• and/or
• C) bringing said second surface of said matrix membrane with said smaller pores in contact with cells and components of an epidermal layer of said integument whereby they become immobilized within pores in the area of said second surface of said matrix membrane;
• and
• D) culturing said matrix membrane thus seeded with said cells and components of said dermal and/or epidermal layers of said integument immobilized in different pore areas thereof under conditions favorable to growth of said integument cells, whether in vivo or in vitro, whereby there results a cultured, split thickness and/or full-thickness integument substitute already implanted in, or ready for implantation in said patient.

A method of treating a patient requiring integument replacement as the result of burns, trauma or other condition that leaves significant areas of said integument destroyed, so badly damaged that normal recovery is not possible, or vulnerable to invasion by pathogens, comprising applying to an area requiring integument replacement a substitute integument which will enhance the rate of healing and will grow and become integrated into surrounding natural integument layers of said patient which still remain, wherein said integument substitute comprises a three-dimensional matrix membrane with essentially two surfaces, in that the length and width thereof are substantially greater than the thickness thereof which is from about 0.1 mm to about 5 mm, said matrix membrane being non-reactive with respect to cells and components of said integument;

• said membrane having pores in the area of a first surface thereof which are directly connected to but substantially larger on average than those in the area of a second, opposite surface of said membrane, said first surface pores having a mean pore diameter in the range of from about 50 .mu.m to about 900 .mu.m, said pores having immobilized therein cells and components of a dermal layer of said integument;
• said membrane having pores in the area of a second surface thereof which are directly connected to but substantially smaller on average than those in said area of said first, opposite surface of said membrane, said second surface pores having a mean pore diameter in the range of from about 0.5 .mu.m to about 100 .mu.m, said pores having immobilized therein cells and components of an epidermal layer of said integument;
• said membrane also having a lateral pore structure in the internal space thereof interconnecting said pores which pass from one surface to the other of said membrane, permitting co-planar flow of a liquid or suspension therethrough.