Biomatrix scaffolds for industrial scale dispersal

• US 9,944,896 B2
• Filed: 10/13/2015
• Issued: 04/17/2018
• Est. Priority Date: 07/02/2010
• Status: Active Grant

• Current Assignee: University of North Carolina At Chapel Hill (University of North Carolina System)
• Original Assignee: University of North Carolina At Chapel Hill (University of North Carolina System)
• Inventors: Roach, Marsha Lynn, Malavarca, Richard Harold, Wang, Yunfang, Reid, Lola Cynthia McAdams
• Primary Examiner(s): Paguio Frising, Michelle F.
• Application Number:
  US14/813,993
• Publication Number:
  US 20160024463A1
• Time in Patent Office: 917 Days
• Field of Search: None
• CPC Class Codes: A61L 2430/28 for liver reconstruction
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3683 subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
• CPC Class Codes: A61L 27/3691 characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2501/20 Cytokines; Chemokines
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/14 from hepatocytes
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/067 Hepatocytes
• CPC Class Codes: C12N 5/0671 Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: C12N 5/0693 Tumour cells; Cancer cells
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

The present invention provides biomatrix scaffolds for industrial scale dispersal.

Claims

1. A plurality of biomatrix particles produced according to a method comprising:

comprising;

(a) perfusing or homogenizing the biological tissue with a buffer comprising a salt concentration from about 3.5 M NaCl to about 4.5 M NaCl;

(b) perfusing the biological tissue or extracting the homogenate of step (a) with a delipidating buffer comprising lipases and detergents in a serum-free first medium, having a neutral pH and an osmolality from about 250 mOsm/kg to about 350 mOsm/kg, wherein the detergents do not comprise sodium dodecyl sulfate or Triton X-100;

(c) perfusing the biological tissue or extracting the homogenate of step (b) with a buffer at a neutral pH and comprising a salt concentration ranging from about 2.0 M NaCl to about 5.0 M NaCl, the concentration chosen to not lose insoluble collagens identified in the biological tissue;

(d) perfusing the biological tissue or extracting the homogenate of step (c) with RNase and DNase in a buffer;

(e) rinsing the biological tissue or homogenate of step (d) with a second medium having a neutral pH, is serum-free and having an osomolality ranging from about 250 mOsm/kg to about 350 mOsm/kg, thereby producing an intact or homogenized biomatrix scaffold from the biological tissue;

(f) pulverizing the intact or homogenized biomatrix scaffold into a plurality of biomatrix particles ranging in size from about 1 nm to about 100 μ

m.

2. The plurality of biomatrix scaffold particles of claim 1, wherein the biological tissue is selected from the group consisting of liver, lung, thyroid, skin, pancreas, blood vessels, bladder, kidneys, brain, biliary tree, duodenum, abdominal aorta, iliac vein, heart and intestines.

3. A composition comprising a plurality of biomatrix particles of claim 1, wherein the plurality of biomatrix particles are obtained from a biomatrix scaffold comprising at least 95% of the collagens and most collagen-associated matrix components, matrix bound growth factors, hormones, and cytokines of a biological tissue.

4. The composition of claim 3, wherein the biological tissue is selected from the group consisting of liver, lung, thyroid, skin, pancreas, blood vessels, bladder, kidneys, brain, biliary tree, duodenum, abdominal aorta, iliac vein, heart and intestines.

5. The plurality of biomatrix scaffold particles of claim 1, wherein the method further comprises diluting the biomatrix scaffold in a basal medium and/or freezing the biomatrix scaffold at about −

80°

C. prior to step (f).