Wound dressing nanomesh impregnated with human umbilical cord Wharton's jelly stem cells

Patent metadata

Field	Value
Publication number	US-10413574-B2
Application number	US-201314421676-A
Country	US
Kind code	B2
Filing date	Aug 15, 2013
Priority date	Aug 15, 2012
Publication date	Sep 17, 2019
Grant date	Sep 17, 2019

Abstract

The invention is directed to a method of treating a wound (e.g., to suppress scar formation) in an individual in need thereof comprising contacting the wound with an effective amount of a composition comprising (i) Wharton's jelly stem cells (WJSCs), (ii) a cell culture medium that has been conditioned with WJSCs, (iii) a lysate of WJSCs, (iv) a cell culture medium that has been conditioned with WJSCs exposed to apoptotic skin cells and keloid cells, or (v) a combination thereof. The invention is directed to a medical dressing (e.g., pharmaceutical compositions) comprising Wharton's jelly stem cells (WJSCs), a cell culture medium that has been conditioned with WJSCs, a lysate of WJSCs, a cell culture medium that has been conditioned with WJSCs exposed to apoptotic skin cells and keloid cells, or a combination thereof.

First claim

What is claimed is:

1. A medical dressing comprising:
a) (i)Wharton's jelly stem cells (WJSCs), (ii) a cell culture medium that has been conditioned with WJSCs, (iii) filtered supernatant from a lysate of living WJSCs, (iv) a cell culture medium that has been conditioned with a co-culture of WJSCs and one or more of living skin cells, dying skin cells or keloid cells, for a period of time from about 1 hour to about 14 days to release biologically active components into the medium, wherein the cells are then removed from the medium, or (v) a combination thereof, and
b) a scaffold formed from a mixture comprising polycaprolactone (PCL) and aloe vera powder, wherein the scaffold is impregnated with the cells, culture medium and/or lysate of a).

2. The medical dressing of claim 1 wherein the scaffold formed from a mixture comprising PCL and aloe vera powder is a nanofibrous scaffold.

3. The medical dressing of claim 1 wherein the scaffold has a porosity of about 85% to 95%, a pore size of about 0.5 μm to about 10 μm, a fiber diameter of about 250 nm to about 650 nm, a fiber thickness of about 250 nm to about 650 nm, or combinations thereof.

4. The medical dressing of claim 1 wherein the lysate of WJSCs is obtained by contacting the WJSCs with a lysis buffer that lyses the WJSCs.

5. The medical dressing of claim 1 wherein for (iv) the co-culturing is under a normoxic or hypoxic environment.

6. The medical dressing of claim 1 , wherein the period of time in (a)(iv) is from about 24 hours to about 72 hours.

7. A pharmaceutical composition comprising the medical dressing of claim 1 .

8. A medical dressing comprising:
a) (i) a cell culture medium that has been conditioned with Wharton's jelly stem cells (WJSCs) for a period of time to release biologically active components into the medium, wherein the WJSCs are then removed from the medium, (ii) filtered supernatant from a lysate from WJSCs, (iii) a cell culture medium that has been conditioned with a co-culture of WJSCs and one or more of living skin cells, dying skin cells or keloid cells, for a period of time from about 1 hour to about 14 days to release biologically active components into the medium, wherein the cells are then removed from the medium, or (iv) a combination thereof, and
b) (i) a scaffold formed from a mixture comprising polycaprolactone (PCL) and aloe vera powder; or (ii) a scaffold consisting of PCL, wherein the scaffold is impregnated with the culture medium, lysate, and/or combination of a),
wherein the WJSCs are a homogeneous population of WJSCs from Wharton's jelly.

9. The medical dressing of claim 8 , wherein the period of time in (a)(iii) is from about 24 hours to about 72 hours.

10. A sterile pharmaceutical composition comprising the medical dressing of claim 8 and a physiologically acceptable carrier or excipient.

11. A method of treating a wound or suppressing scar formation at a wound, or both, in an individual in need thereof comprising contacting the wound with an effective amount of:
A) a composition comprising:
a) (i)Wharton's jelly stem cells (WJSCs), (ii) a cell culture medium that has been conditioned with WJSCs, (iii) filtered supernatant from a lysate of WJSCs, (iv) a cell culture medium that has been conditioned with a co-culture of WJSCs and one or more of living skin cells, dying skin cells or keloid cells, for a period of time from about 1 hour to about 14 days to release biologically active components into the medium, wherein the cells are then removed from the medium, or (v) a combination thereof, and
b) a scaffold formed from a mixture comprising polycaprolactone (PCL) and aloe vera powder, wherein the scaffold is impregnated with the cells, culture medium and/or lysate of a); or

B) a composition comprising:
a) (i) a cell culture medium that has been conditioned with Wharton's jelly stem cells (WJSCs) for a period of time to release biologically active components into the medium, wherein the WJSCs are then removed from the medium, (ii) filtered supernatant from a lysate from WJSCs, (iii) a cell culture medium that has been conditioned with a co-culture of WJSCs and one or more of living skin cells, dying skin cells or keloid cells, for a period of time from about 1 hour to about 14 days to release biologically active components into the medium, wherein the cells are then removed from the medium, or (iv) a combination thereof, and
b) (i) a scaffold formed from a mixture comprising polycaprolactone (PCL) and aloe vera powder; or (ii) a scaffold consisting of PCL, wherein the scaffold is impregnated with the culture medium, lysate, and/or combination of a),
wherein the WJSCs are a homogeneous population of WJSCs from Wharton's jelly.

12. The method of claim 11 wherein the (i) scaffold formed from a mixture comprising PCL and aloe vera powder; or (ii) scaffold consisting of PCL is a nanofibrous scaffold.

13. The method of claim 1 wherein the scaffold has a porosity of about 85% to 95%, a pore size of about 0.5 μm to about 10 μm, a fiber diameter of about 250 nm to about 650 nm, a fiber thickness of about 250 nm to about 650 nm, or combinations thereof.

14. The method of claim 1 wherein the cell culture medium that has been conditioned with WJSCs is obtained by culturing WJSCs in one or more cell culture media for about 24 hours to 72 hours.

15. The method of claim 11 wherein the lysate of WJSCs is obtained by contacting the WJSCs with a lysis buffer that lyses the WJSCs.

16. The method of claim 11 wherein in A(a)(iv) or B(a)(iii) the co-culturing is under a normoxic or hypoxic environment.

17. The method of claim 11 , wherein the period of time in A(a)(iv) and B(a)(iii) is from about 24 hours to about 72 hours.

18. A method of treating a scar in an individual in need thereof comprising contacting the scar with an effective amount of:
A) a composition comprising:
a) (i) Wharton's jelly stem cells (WJSCs), (ii) a cell culture medium that has been conditioned with WJSCs, (iii) filtered supernatant from a lysate of WJSCs, (iv) a cell culture medium that has been conditioned with a co-culture of WJSCs and one or more of living skin cells, dying skin cells or keloid cells, for a period of time from about 1 hour to about 14 days to release biologically active components into the medium, wherein the cells are then removed from the medium, or (v) a combination thereof, and
b) a scaffold formed from a mixture comprising polycaprolactone (PCL) and aloe vera powder, wherein the scaffold is impregnated with the cells, culture medium and/or lysate of a); or

B) a composition comprising:
a) (i) a cell culture medium that has been conditioned with Wharton's jelly stem cells (WJSCs) for a period of time to release biologically active components into the medium, wherein the WJSCs are then removed from the medium, (ii) filtered supernatant from a lysate from WJSCs, (iii) a cell culture medium that has been conditioned with a co-culture of WJSCs and one or more of living skin cells, dying skin cells or keloid cells, for a period of time from about 1 hour to about 14 days to release biologically active components into the medium, wherein the cells are then removed from the medium, or (iv) a combination thereof, and
b) (i) a scaffold formed from a mixture comprising polycaprolactone (PCL) and aloe vera powder; or (ii) a scaffold consisting of PCL, wherein the scaffold is impregnated with the culture medium, lysate, and/or combination of a),
wherein the WJSCs are a homogeneous population of WJSCs from Wharton's jelly.

19. The method of claim 18 wherei

Assignees

• Nat Univ Singapore

Inventors

• Fong Chui Yee
• Choolani Mahesh
• Biswas Arijit
• Bongso Tuan Ariffeen
• Ramakrishna Seeram

Classifications

• A61P17/02

  for treating wounds, ulcers, burns, scars, keloids, or the like · CPC title

• C12N5/0605

  Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly · CPC title

• A61L2400/12

  Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces · CPC title

• A61L2300/64

  Animal cells · CPC title

• A61L2300/418

  Agents promoting blood coagulation, blood-clotting agents, embolising agents · CPC title