Compositions derived from placenta and methods of producing the same
Compositions comprising perforated, unseparated amnion/chorion; perforated, separated amnion; and perforated, separated chorion derived from the placenta and methods of preparing and using those compositions are provided. Perforation of placental tissue before or during processing may allow for one or more benefits such as more efficient removal of blood remnants, retention of wound healing and tissue regeneration components, better handling characteristics, or improved healing capacity. The present invention also includes methods of healing a wound of the skin, eye, nerve, tendon, or dura comprising applying the perforated compositions of the invention to the wound.
1. A method of cleansing an unseparated amnion/chorion sheet or composition thereof comprising:
(a) obtaining an unseparated amnion/chorion tissue dissected away from the placental disc and umbilical cord;
(b) perforating the unseparated amnion/chorion tissue; and
(c) washing the unseparated amnion/chorion tissue with at least one washing medium over two or more washing steps to remove blood remnants from the unseparated amnion/chorion tissue,
wherein perforating occurs before or between two washing steps.
2. The method of claim 1 , wherein the unseparated amnion/chorion tissue comprises an intermediate layer.
3. The method of claim 1 , comprising performing a first washing step comprising washing the unseparated amnion/chorion tissue with at least one washing medium before perforating the unseparated amnion/chorion tissue.
4. The method of claim 1 , further comprising pretreating the unseparated amnion/chorion tissue before perforating, before washing, and/or between any washing steps, wherein pretreating comprises picking material away from the tissue, or dabbing, rubbing, or massaging the tissue.
5. The method of claim 1 , wherein perforating comprises making one or more holes that partially traverse the thickness of the unseparated amnion/chorion tissue.
6. The method of claim 1 , wherein perforating comprises making one or more holes that traverse the thickness of the unseparated amnion/chorion tissue.
7. The method of claim 1 , wherein at least one of the washing steps is conducted under vacuum; under pressure; by agitating using a rocker, shaker, stir plate, or rotating mixer; or by soaking.
8. The method of claim 1 , wherein (c) comprises performing at least three washing steps, at least four washing steps, at least five washing steps, at least six washing steps, at least seven washing steps, at least eight washing steps, at least nine washing steps, or at least ten washing steps.
9. The method of claim 1 , wherein the method comprises exchanging the at least one washing medium at least once, at least twice, at least thrice, at least four times, at least five times, at least six times, at least seven times, at least eight times, at least nine times, or at least ten times between any washing steps.
10. The method of claim 1 , wherein at least one of the washing steps comprises heating or cooling the unseparated amnion/chorion tissue or the at least one washing medium.
11. The method of claim 1 , wherein at least one of the washing steps comprises washing the unseparated amnion/chorion tissue in the washing medium in an environment having a temperature of below, at, or above room temperature for about 30 minutes to about 64 hours.
12. The method of claim 11 , wherein the temperature of the environment does not change over time.
13. The method of claim 1 , wherein the at least one washing medium for the two or more washing steps is selected from water, a sodium chloride solution, a sodium hypochlorite solution, a hydrogen peroxide solution, Lactated Ringer's solution, Ringer's solution, phosphate-buffered saline (PBS), tris-buffered saline (TBS), Hank's balanced salt solution (HBSS), Dulbecco's phosphate-buffered saline (DPBS), Earle's balanced salt solution (EBSS), standard saline citrate (SSC), HEPES-buffered saline (HBS), Gey's balanced salt solution (GBSS), and a cell culture medium.
14. The method of claim 1 , wherein the at least one washing medium for at least one of the two or more washing steps is a sodium chloride solution, wherein the sodium chloride solution contains a w/v concentration of NaCl of about 0.45% to about 30%.
15. The method of claim 14 , wherein the sodium chloride solution comprises one or more ingredients selected from phosphate, potassium, magnesium, calcium, dextrose, glucose, citrate, lactate, tris, and HEPES.
16. The method of claim 14 , wherein the sodium chloride solution contains a w/v concentration of NaCl of about 18% to about 30%.
17. The method of claim 1 , wherein the method comprises exchanging the washing medium between a sodium chloride solution and water at least once, at least twice, at least thrice, or at least four times between any washing steps.
18. The method of claim 1 , wherein the method comprises a washing step comprising washing the unseparated amnion/chorion tissue in a sodium chloride solution containing a w/v concentration of NaCl of about 26% 18% to about 30%.
19. A method of preparing a dehydrated unseparated amnion/chorion sheet or composition thereof comprising the method of claim 1 , further comprising: (d) dehydrating the unseparated amnion/chorion tissue.
20. The method of claim 19 , wherein the dehydrating comprises lyophilizing, heat-drying, oven-drying, air-drying, and/or chemically dehydrating the unseparated amnion/chorion tissue.
21. The method of claim 1 , further comprising layering on a surface of the unseparated amnion/chorion tissue at least one separated amnion sheet, at least one perforated, separated amnion sheet, at least one unperforated, separated amnion sheet, at least one separated chorion sheet, at least one perforated, separated chorion sheet, at least one unperforated, separated chorion sheet, or at least one perforated, unseparated amnion/chorion sheet.
22. The method of claim 1 , wherein the hemoglobin content of dry mass of the unseparated amnion/chorion sheet is reduced in an amount greater than about 50%, 60%, 70%, 80%, 90%, or 95% compared to the unseparated amnion/chorion tissue prior to perforating and washing.
23. The method of claim 22 , further comprising sterilizing the tissue or the sheet.
24. The method of claim 22 , wherein the method comprises layering on a surface of the unseparated amnion/chorion at least one sheet selected from a separated amnion sheet, a perforated, separated amnion sheet, an unperforated, separated amnion sheet, a separated chorion sheet, a perforated, separated chorion sheet, an unperforated, separated chorion sheet, and a perforated, unseparated amnion/chorion sheet, or combinations thereof.
25. A method of cleansing an unseparated amnion/chorion sheet or composition thereof comprising:
a. obtaining an unseparated amnion/chorion tissue dissected away from the placental disc and umbilical cord;
b. pretreating the unseparated amnion/chorion tissue, wherein pretreating comprises picking material away from the tissue, or dabbing, rubbing, or massaging the tissue;
c. a first washing step comprising washing the unseparated amnion/chorion tissue with a first washing medium;
d. perforating the unseparated amnion/chorion tissue;
e. washing the unseparated amnion/chorion tissue with at least one additional washing medium over two or more additional washing steps to remove blood remnants from the unseparated amnion/chorion tissue;
f. dehydrating the unseparated amnion/chorion tissue; and
g. sterilizing the tissue or the sheet.
26. The method of claim 25 , wherein the method comprises cutting the tissue or the sheet into one or more sheets before, during, or after any step or combination of steps.