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EP-4741008-A2 - STERILE HUMAN PLACENTAL ALLOGRAFTS AND METHODS OF MAKING THEREOF

EP4741008A2EP 4741008 A2EP4741008 A2EP 4741008A2EP-4741008-A2

Abstract

A method of preparing sterile human placental allografts by providing a human placental tissue from a donor within 24 hours to 72 hours post-childbirth; removing any visible blood, blood clots, and/or blood components from the human placental tissue without scraping or scrubbing the human placental tissue to preserve structural integrity of the human placental tissue; washing the human placental tissue in an isotonic solution while maintaining the structural integrity of the human placental tissue; dehydrating the human placental tissue thereby forming the dehydrated human placental tissue; resizing the dehydrated human placental tissue into dehydrated human placental tissue portions having predetermined sizes; and sterilizing the dehydrated human placental tissue portions of step (e) thereby forming the sterile human placental allograft. Also disclosed is a sterile human placental allograft produced by the method.

Inventors

  • MATUSZEWSKI, Jason, V.
  • WESTON, Wendy, W.

Assignees

  • Biostem Technologies, Inc.

Dates

Publication Date
20260513
Application Date
20230628

Claims (13)

  1. A method of preparing a sterile human placental allograft comprising: (a) providing a human placental tissue from a donor within 24 hours to 72 hours post-childbirth; and either before step (a) or during step (a) disinfecting the human placental tissue with at least one of a bactericidal composition, a tuberculocidal composition, a fungicidal composition, a virucidal composition, or any combination thereof, wherein the at least one of a bactericidal composition, tuberculocidal composition, fungicidal composition, virucidal composition, or any combination thereof includes an alcohol solution; (b) removing any visible blood, blood clots, and/or blood components from the human placental tissue without scraping or scrubbing the human placental tissue to preserve structural integrity of the human placental tissue; (c) washing the human placental tissue in an isotonic solution while maintaining the structural integrity of the human placental tissue; wherein each wash step (c) is at a temperature ranging from 4 to 20°C for a time period from 5 minutes to 15 minutes; (d) dehydrating the human placental tissue thereby forming the dehydrated human placental tissue, wherein the dehydrating or drying of step (d) is (d-1) at a temperature ranging from 20 to 40 °C for a time period of from 60 minutes to 4.5 hours; and/or (d-2) to a residual water content of 10 to 15%; (e) resizing the dehydrated human placental tissue into dehydrated human placental tissue portions having predetermined sizes; and (f) sterilizing the dehydrated human placental tissue portions of step (e) thereby forming the sterile human placental allograft wherein the sterilizing step (f) comprises sterilizing with e-beam irradiation.
  2. The method of claim 1, wherein the alcohol solution is an isopropyl alcohol at a concentration of 70% to 100%.
  3. The method of claim 1 or 2, wherein step (b) comprises manual hematopoietic reduction by manually removing any visible blood, blood clots, and/or blood components from the human placental tissue.
  4. The method of any one of the preceding claims, wherein the isotonic solution comprises at least one of 1x phosphate buffered saline, isotonic saline, lactated ringers, Plasma-Lyte ® (NaCl 5.26 g/L, KCI 0.37 g/L, Magnesium Chloride hexahydrate 0.30 g/L, Sodium Acetate trihydrate 3.68 g/L, Sodium Gluconate 5.02 g/L at pH 7.4), Normosol ® (NaCl 5.26 g/L, KCI 0.37 g/L, Magnesium Chloride 0.30 g/L, Sodium Acetate anhydrous 2.22 g/L, Sodium Gluconate 5.02 g/L at pH 7.4), or any combination thereof.
  5. The method of any one of the preceding claims, wherein the predetermined sizes of the placental tissue portions comprise from 1 cm x 1 cm to 8 cm x 8 cm for rectangular and/or square shaped placental tissue portions and from 8mm diameter to 50 mm diameter for circular shaped placental tissue portions and/or wherein the placental tissue portions comprise a predetermined shape and/or wherein the human placental tissue comprises intact placental tissue.
  6. The method of any one of the preceding claims, wherein step (f) comprises e-beam irradiating the dehydrated human placental tissue portions to a sterility assurance level (SAL) of 10-6, as determined by dose mapping, to form the sterile human placental allograft.
  7. The method of any one of the preceding claims, wherein the dehydrated human placental tissue of step (d) and the dehydrated human placental tissue portions having predetermined sizes of step (e) are intact in cross-section in which a human amnion layer and a human chorion layer have an intact human intermediate spongy layer positioned there between connecting the human amnion layer to the human chorion layer and/or wherein sterile human placental allografts of step (f) are intact in cross-section in which the human amnion layer and the human chorion layer have an intact human intermediate spongy layer positioned there between connecting the human amnion layer to the human chorion layer and/or wherein the sterile human placental allograft is configured for dental, cosmetic, and/or wound healing applications.
  8. A sterile human placental allograft produced by the method of any one of the preceding claims.
  9. The sterile human placental allograft of claim 6, wherein the sterile human placental allograft comprises at least two of the following: (a) interleukin-1 receptor antagonist (IL-1ra) ranging from 200 to 7800 pg/cm2; (b) hepatocyte growth factor (HGF) ranging from 500 to 8500 pg/cm2; (c) vascular endothelial growth factor receptor 1 (VEGFR1) ranging from 800 to 2750 pg/cm2; (d) hyaluronic acid (HA) ranging from 5.0 x 106 to 1.5 x 108 pg/cm2; (e) platelet derived growth factor subunit B homodimer (PDGF-BB) ranging from 150 to 400 pg/cm2; (f) glycosaminoglycans (GAGs); (g) collagen; and (h) exosomes.
  10. The sterile human placental allograft of claim 8 or 9, wherein the sterile human placental allograft is intact in cross-section in which the human amnion layer and the human chorion layer have an intact human intermediate spongy layer positioned there between connecting the human amnion layer to the human chorion layer and/or wherein the sterile human placental allograft is configured for dental, cosmetic, and/or wound healing applications and/or wherein the sterile human placental allograft comprises a predetermined size of from 1 cm x 1 cm to 8 cm x 8 cm for rectangular and/or square shaped sterile human placental allografts and from 8mm diameter to 50 mm diameter for circular shaped sterile human placental allografts and/or wherein the sterile human placental allograft comprises a predetermined size of from 1 cm x 1 cm to 8 cm x 8 cm for rectangular and/or square shaped sterile human placental allografts and from 8mm diameter to 50 mm diameter for circular shaped sterile human placental allografts and/or wherein the sterile human placental allograft comprises a circular shape, a square shape, a rectangular shape, an ovoid shape, a triangular shape, or any combination thereof and/or wherein the sterile human placental allograft comprises a circular shape, a square shape, a rectangular shape, an ovoid shape, a triangular shape, or any combination thereof and/or wherein an upper and/or lower surfaces of the sterile human placental allograft is planar and/or wherein the upper and lower surfaces of the sterile human placental allograft are planar and/or wherein the sterile human placental allograft consists essentially of the amnion membrane.
  11. The sterile human placental allograft of any one of claims 8-10, wherein the sterile human placental allograft comprises at least two of the following: (a) interleukin-1 receptor antagonist (IL-1ra) ranging from 150 to 1800 pg/cm2; (b) hepatocyte growth factor (HGF) ranging from 75 to 400 pg/cm2; (c) vascular endothelial growth factor receptor 1 (VEGFR1) ranging from 50 to 250 pg/cm2; (d) hyaluronic acid (HA) ranging from 1.1 x 106 to 2.0 x 107 pg/cm2; (e) platelet derived growth factor subunit B homodimer (PDGF-BB) ranging from 200 to 500 pg/cm2; (f) glycosaminoglycans (GAGs); (g) collagen, and (h) exosomes.
  12. A kit comprising the sterile human placental allograft of any one of claims 8-11 or produced by the method of any one of claims 1-7 packaged in a sterile container.
  13. A sterile human placental allograft of any one of claims 8-11 for use in a method of treating a wound in a human subject in need thereof comprising contacting the wound with a sterile human placental allograft for a predetermined time period to facilitate wound healing in the human subject in need thereof, preferably wherein the sterile human placental allograft is implanted in the subject in need thereof, and/or preferably wherein the wound is an internal wound within the subject in need thereof and/or a dental wound and/or preferably wherein the sterile human placental allograft is topically applied to the wound in the human subject in need thereof and/or preferably wherein the wound is a diabetic ulcer, preferably a diabetic foot ulcer.

Description

TECHNICAL FIELD The present invention relates generally to the field of human placental tissues, and more particularly, to growth factor rich sterile human placental allografts and methods of making thereof in which the allografts have various dental, cosmetic, and wound healing applications. BACKGROUND The use of human placenta for medical purposes dates back to 1593, when Li Shizhen wrote about the use of "zi he chi" (human placenta) in the Compendium of Materia Medica. However, it wasn't until 1910 that the advantages of placental membranes for skin transplantation were formally documented in the medical literature. Use of placental membranes for clinical purposes continues to date. To date, placental membrane processing has primarily focused on the preservation of cells within a tissue or removal of all non-solid components for the purpose of delivering stem cells and/or providing a substrate for regenerative growth. These placental membrane processing techniques often result in loss of endogenous growth factors found within the human placental tissue and/or result in endogenous degradative processes that further result in a loss of endogenous growth factors found in the human placental tissue thereby resulting in reduced end use efficacy (e.g., reduced wound healing) for any allografts obtain from these processes. SUMMARY It is an object of the invention to provide safe, biocompatible, growth factor-rich sterile human placental allografts grafts. The methods disclosed herein achieve these biocompatible, growth factor-rich sterile human placental allografts grafts while maintaining a growth factor profile that mimics the human placenta in vivo. These methods clean and prepare the human placental tissues in a gentle and effective manner, while minimizing the risk of comprising biologic tissue and/or growth factors for subsequent allograft implantation/transplantation. In particular, the disclosed methods utilize various mild cleaning, preservation, and mild sterilization techniques using cold physiological buffers, gentle handling, dehydration at ambient or physiological temperatures, and mild sterilization conditions of the human placental allografts disclosed herein to obtain sterile human placental allografts that maintain a growth factor profile that mimics the human placenta in vivo. These sterile human placental allografts have various different clinical and cosmetic purposes and may achieve, for example, improved wound healing when compared with conventional human placental allografts because of its growth factor profile. Furthermore, the allografts disclosed herein meet or exceed all requirements set by the Food and Drug Administration (FDA) and American Association of Tissue Banks (AATB). In certain aspects, disclosed herein is a method of preparing a sterile human placental allograft comprising (a) providing a human placental tissue from a donor within 24 hours to 72 hours post-childbirth; and either before step (a) or during step (a) disinfecting the human placental tissue with at least one of a bactericidal composition, a tuberculocidal composition, a fungicidal composition, a virucidal composition, or any combination thereof;(b) removing any visible blood, blood clots, and/or blood components from the human placental tissue without scraping or scrubbing the human placental tissue to preserve structural integrity of the human placental tissue;(c) washing the human placental tissue in an isotonic solution while maintaining the structural integrity of the human placental tissue; wherein each wash step (c) is at a temperature ranging from 4 to 20°C;(d)dehydrating the human placental tissue thereby forming the dehydrated human placental tissue; wherein the dehydrating step (d) is at a temperature ranging from 20 to 40 °C;(e) resizing the dehydrated human placental tissue into dehydrated human placental tissue portions having predetermined sizes; and(f) sterilizing the dehydrated human placental tissue portions of step (e) thereby forming the sterile human placental allograft. In certain aspects, the at least one of a bactericidal composition, tuberculocidal composition, fungicidal composition, virucidal composition, or any combination thereof includes an alcohol solution and more preferably an isopropyl alcohol at a concentration of 70% to 100%. In preferred embodiments the alcohol concentration ranges from 70% to 75%, and in most preferred embodiments, the alcohol is 70% isopropyl alcohol. Isopropyl alcohol is preferred over other commercially available lab and/or pharmaceutical grade alcohols, such as ethanol, because isopropyl alcohol advantageously disinfects and cleans the human placental tissue without damaging (e.g., unduly dehydrating, initiating apoptotic processes, and/or necrotic processes) the placental tissue. In certain aspects, predetermined volumes of the at least of a bactericidal composition, a tuberculocidal composition, a fungicidal composition, a virucidal composition, or any