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US-20260124243-A1 - COMPOSITIONS AND METHODS FOR WOUND HEALING

US20260124243A1US 20260124243 A1US20260124243 A1US 20260124243A1US-20260124243-A1

Abstract

A method of promoting healing of a wound generally includes administering to the wound an amount of a PEP exosome preparation effective to promote healing of the wound. In one or more embodiments, the PEP exosome preparation includes a tissue sealant or surgical glue. In one or more embodiments, the wound is an ischemic wound, a puncture wound, a laceration, an abrasion, a surgical wound, a skin graft, or a traumatic wound.

Inventors

  • Atta Behfar
  • Steven L. Moran
  • Brooke Paradise
  • Laura Becher
  • Christopher Paradise

Assignees

  • RION INC.
  • MAYO FOUNDATION FOR MEDICAL EDUCATION AND RESEARCH

Dates

Publication Date
20260507
Application Date
20221025

Claims (20)

  1. 1 . A method of promoting healing of a wound, the method comprising administering to the wound an amount of a PEP preparation effective to promote healing of the wound.
  2. 2 . The method of claim 1 , wherein the PEP preparation comprises a hydrogel comprising a basement membrane protein.
  3. 3 . The method of claim 1 , wherein the PEP preparation comprises a hydrogel comprising a thrombin sealant or a fibrin sealant.
  4. 4 . The method of claim 1 , wherein the wound comprises an ischemic wound, a puncture wound, a laceration, an abrasion, a surgical wound, a skin graft, or a traumatic wound.
  5. 5 . The method of claim 1 , wherein the amount of PEP preparation is effective to increase angiogenesis, increase migration of fibroblasts into the wound, or increase migration of keratinocytes into the wound compared to a comparable untreated wound.
  6. 6 . The method of claim 1 , wherein the amount of PEP preparation is effective in donating TGF-β to increase expression COL1A or COL3A compared to a comparable untreated wound.
  7. 7 . The method of claim 1 , wherein the amount of PEP preparation is effective to decrease Wagner Ulcer Classification grade of the wound compared to a comparable untreated wound.
  8. 8 . The method of claim 1 , wherein the amount of PEP preparation is effective to decrease reaction force variation (R c ) or increase resistance to tensile force compared to a comparable untreated wound.
  9. 9 . The method of claim 1 , wherein the amount of PEP preparation is effective to increase expression of SMAD2, RAS, MKK3, RHOA, P38, or periostin in keratinocytes compared to untreated keratinocytes.
  10. 10 . The method of claim 1 , wherein the amount of PEP preparation is effective to increase expression of SMAD2, RAS, MKK3, ERK1, or TAK1 in fibroblasts compared to untreated fibroblasts.
  11. 11 . The method of claim 2 , wherein the wound comprises an ischemic wound, a puncture wound, a laceration, an abrasion, a surgical wound, a skin graft, or a traumatic wound.
  12. 12 . The method of claim 2 , wherein the amount of PEP preparation is effective to increase angiogenesis, increase migration of fibroblasts into the wound, or increase migration of keratinocytes into the wound compared to a comparable untreated wound.
  13. 13 . The method of claim 2 , wherein the amount of PEP preparation is effective to decrease Wagner Ulcer Classification grade of the wound compared to a comparable untreated wound.
  14. 14 . The method of claim 3 , wherein the wound comprises an ischemic wound, a puncture wound, a laceration, an abrasion, a surgical wound, a skin graft, or a traumatic wound.
  15. 15 . The method of claim 3 , wherein the amount of PEP preparation is effective to increase angiogenesis, increase migration of fibroblasts into the wound, or increase migration of keratinocytes into the wound compared to a comparable untreated wound.
  16. 16 . The method of claim 3 , wherein the amount of PEP preparation is effective to decrease Wagner Ulcer Classification grade of the wound compared to a comparable untreated wound.
  17. 17 . The method of claim 4 , wherein the amount of PEP preparation is effective to increase angiogenesis, increase migration of fibroblasts into the wound, or increase migration of keratinocytes into the wound compared to a comparable untreated wound.
  18. 18 . The method of claim 4 , wherein the amount of PEP preparation is effective to decrease Wagner Ulcer Classification grade of the wound compared to a comparable untreated wound.
  19. 19 . The method of claim 11 , wherein the amount of PEP preparation is effective to increase angiogenesis, increase migration of fibroblasts into the wound, or increase migration of keratinocytes into the wound compared to a comparable untreated wound.
  20. 20 . The method of claim 11 , wherein the amount of PEP preparation is effective to decrease Wagner Ulcer Classification grade of the wound compared to a comparable untreated wound.

Description

CROSS-REFERENCE TO RELATED APPLICATION This application claims the benefit of U.S. Provisional Patent Application No. 63/271,486, filed Oct. 25, 2021, which is incorporated herein by reference in its entirety. SEQUENCE LISTING This application contains a Sequence Listing electronically submitted to the United States Patent and Trademark Office via Patent Center as an XML file entitled “0560-000014WO01” having a size of 31.8 kilobytes and created on Oct. 25, 2022. Due to the electronic filing of the Sequence Listing, the electronically submitted Sequence Listing serves as both the paper copy required by 37 CFR § 1.821(c) and the CRF required by § 1.821(e). The information contained in the Sequence Listing is incorporated by reference herein. SUMMARY This disclosure describes, in one aspect, a method of promoting healing of a wound. Generally, the method includes administering to the wound an amount of a PEP preparation effective to promote healing of the wound. In one or more embodiments, the PEP preparation includes a hydrogel that includes a basement membrane protein. In one or more embodiments, the PEP preparation includes a hydrogel that includes a thrombin sealant or a fibrin sealant. In one or more embodiments, the wound is an ischemic wound, a puncture wound, a laceration, an abrasion, a surgical wound, a skin graft, or a traumatic wound. In one or more embodiments, the amount of PEP preparation administered to the wound is effective to increase angiogenesis, increase migration of fibroblasts into the wound, or increase migration of keratinocytes into the wound compared to a comparable untreated wound. In one or more embodiments, the amount of PEP preparation administered to the wound is effective to donate TGF-β to increase expression COL1A or COL3A compared to a comparable untreated wound. In one or more embodiments, the amount of PEP preparation administered to the wound is effective to decrease Wagner Ulcer Classification grade of the wound compared to a comparable untreated wound. In one or more embodiments, the amount of PEP preparation administered to the wound is effective to decrease reaction force variation (Rc) or increase resistance to tensile force compared to a comparable untreated wound. In one or more embodiments, the amount of PEP preparation administered to the wound is effective to increase expression of SMAD2, RAS, MKK3, RHOA, P38, or periostin in keratinocytes compared to untreated keratinocytes. In one or more embodiments, the amount of PEP preparation administered to the wound is effective to increase expression of SMAD2, RAS, MKK3, ERK1, or TAK1 in fibroblasts compared to untreated fibroblasts. The above summary is not intended to describe each disclosed embodiment or every implementation of the present invention. The description that follows more particularly exemplifies illustrative embodiments. In several places throughout the application, guidance is provided through lists of examples, which examples can be used in various combinations. In each instance, the recited list serves only as a representative group and should not be interpreted as an exclusive list. BRIEF DESCRIPTION OF THE FIGURES FIG. 1. PEP extracellular vesicles display exosomal characteristics. (A) A representative transmission electron microscopic image of PEP exosomes. Scale bar, 200 nm. (B) Representative western blot of CD63, CD9, and Alix in PEP exosomes. GAPDH was used as a loading control. (C) Size distribution of PEP exosomes as measured by Nanoparticle Tracking Analysis (NTA), peaking at a diameter of 105.4 nm. (D) Size distribution of PEP exosomes as measured by nano flow cytometry (NanoFCM), with a mean of 123.49 nm. FIG. 2. PEP extracellular vesicles display exosomal surface markers. (A) Representative simple western blot (Jess, ProteinSimple) shows the presence of CD41 (platelet marker), CD9, CD63, and Flotillin-1 (EV markers) in PEP. (B) Representative plot from nano flow cytometry of PEP demonstrates the presence of CD41a (platelet marker) on PEP extracellular vesicles (MemGlow488+, lipid bilayer stain). (C) Pie chart showing affinity-based capture of CD41a+ vesicles with subsequent fluorescent antibody staining of CD9, CD63, or CD81 surface markers (NanoView). (D) Bar graph showing affinity-based capture of CD41a+ vesicles with subsequent fluorescent antibody staining of CD9, CD63, or CD81 surface markers (NanoView). FIG. 3. PEP microvesicles promote angiogenesis in vitro. (A) In vitro angiogenesis assay using co-culture of human dermal fibroblast (HDFB) with GFP-tagged human umbilical vascular endothelial cells (HUVEC) in presence of VEGF, PEP, or suramin (angiogenesis inhibitor). Scale bar, 200 μm. (B) Graph displaying quantification of tube formation in six hour increments. (C) 3D organoid differentiation assay of human keratinocytes treated with PEP or serum free media. Collection of organoids was performed at day 24 and sections were prepped for microscopy. Hematoxylin and eosin (H&E) sta