Materials and method for the immobilization of bioactive species onto polymeric subtrates

• US 5,874,165 A
• Filed: 05/27/1997
• Issued: 02/23/1999
• Est. Priority Date: 06/03/1996
• Status: Expired due to Term

• Current Assignee: W. L. Gore & Associates (W.L. Gore & Associates)
• Original Assignee: Gore Enterprise Holdings Incorporated
• Inventors: Drumheller, Paul D.
• Primary Examiner(s): Richter, Johann
• Assistant Examiner(s): MURRAY, JOSEPH H
• Application Number:
  US08/863,263
• Time in Patent Office: 637 Days
• Field of Search: 428/308.4
• US Class Current: 428/308.4
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: C07K 17/08 the carrier being a synthetic polymer
• CPC Class Codes: C12N 11/084 Polymers containing vinyl alcohol units
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2533/50 Proteins
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: Y10T 428/249958 Void-containing component is synthetic resin or natural rubbers

Abstract

The present invention is directed to support members having surfaces that are rendered hydrophilic for use as a substrate for the immobilization of bioactive species thereon. The hydrophilic surfaces are chemically stable on the support member. The surfaces are also chemically variable to provide a variety of chemically functional groups for immobilization of bioactive species thereto. The surfaces comprise polymeric surfactants attached onto the surfaces of a support member and covalently cross-linked thereon to form a first layer. Hydrophilic polymers are then attached to the first layer on the support member to form a second layer thereon. The second layer is used to enhance the hydrophilicity of a support member, as well as, to provide a substrate for immobilizing bioactive species thereto. Methods for forming the first and second layers are also provided. In addition, methods for attaching bioactive substances to the hydrophilic polymers are provided.

Claims

1. A vascular graft having bioactive species immobilized thereto, the vascular graft comprising:

a porous polytetrafluoroethylene material;

a first layer comprised of at least one species of a polymeric surfactant non-covalently attached to the porous polytetrafluoroethylene material and cross-linked to itself in situ;

a second layer comprised of at least one species of a hydrophilic polymer covalently attached to the first layer; and

at least one type of bioactive species attached to the second layer, the second layer and immobilized bioactive species serving as a blood contact surface.

2. The vascular graft of claim 1 wherein the bioactive species comprises at least one isomer of lysine.

3. The vascular graft of claim 1 wherein the bioactive species is an amino acid sequence comprising Gly-Arg-Gly-Asp-Ser.

4. The vascular graft of claim 1 wherein cells are immobilized to the bloactive species.

5. The vascular graft of claim 3 wherein cells are immobilized to the Gly-Arg-Gly-Asp-Ser.

6. The vascular graft of claim 1 wherein the polymeric surfactant comprises a multifunctional copolymer comprised of at least one domain having a physicochemical affinity for the porous polytetrafluoroethylene material and at least one domain that is chemically reactive with a cross-linking agent.

7. The vascular graft of claim 1 wherein the hydrophilic polymer comprises chemically functional groups to which the first layer is covalently attached and additional chemically functional groups to which are attached the bioactive species.

8. The vascular graft of claim 1 wherein a spacer compound is interposed between the first layer and the bioactive species.

9. The material of claim 8 wherein the spacer compound is selected from a member of a group consisting of succinic acid, diaminohexane, glyoxylic acid, short chain polyethylene glycol, and glycine.

10. The material of claim 8 wherein the spacer compound is cleavable.

11. The material of claim 10 wherein the cleavable spacer compound is selected from the group consisting of polyhydroxyacids, polyanhydrides, polyamino acids, tartarates, and cysteine-linkers.

12. A vascular graft having bioactive species immobilized thereto, the vascular graft comprising:

a porous polytetrafluoroethylene material;

a first layer of polyvinyl alcohol non-covalently attached to the porous polytetrafluoroethylene;

a second layer of polyvinyl alcohol covalently attached to the first layer; and

a bioactive species comprising Gly-Arg-Gly-Asp-Ser attached to the second layer.

13. The vascular graft of claim 12 further comprising cells immobilized to the bioactive species.

14. The vascular graft of claim 13 wherein the cells are endothelial cells.