Porous polymer scaffold useful for tissue engineering in stem cell transplantation

• US 9,925,298 B2
• Filed: 10/27/2016
• Issued: 03/27/2018
• Est. Priority Date: 10/27/2015
• Status: Active Grant

• Current Assignee: Council of Scientific & Industrial Research
• Original Assignee: Council of Scientific & Industrial Research
• Inventors: Das, Amitava, Basak, Pratyay, Geesala, Ramasatyaveni, Bar, Nimai, Dhoke, Neha Raghuvir, Kaushik, Komal
• Primary Examiner(s): Arnold, Ernst V
• Application Number:
  US15/336,187
• Publication Number:
  US 20170112960A1
• Time in Patent Office: 516 Days
• CPC Class Codes: A61L 27/18 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: A61L 27/58 Materials at least partially resorbable by the body
• CPC Class Codes: C08G 18/14 Manufacture of cellular products
• CPC Class Codes: C08G 18/36 Hydroxylated esters of higher fatty acids
• CPC Class Codes: C08G 18/4833 Polyethers containing oxyethylene units
• CPC Class Codes: C08G 18/7671 containing only one alkylene bisphenyl group
• CPC Class Codes: C08G 18/8051 with compounds of C08G18/36
• CPC Class Codes: C08G 65/33351 acyclic
• CPC Class Codes: C08J 2201/0462 using organic solvents
• CPC Class Codes: C08J 2205/044 Micropores, i.e. average diameter being between 0,1 micrometer and 0,1 millimeter
• CPC Class Codes: C08J 2205/05 Open cells, i.e. more than 50% of the pores are open
• CPC Class Codes: C08J 2207/10 Medical applications, e.g. biocompatible scaffolds
• CPC Class Codes: C08J 2300/16 Biodegradable polymers
• CPC Class Codes: C08J 2375/08 from polyethers
• CPC Class Codes: C08J 9/26 by elimination of a solid phase from a macromolecular composition or article, e.g. leaching out
• CPC Class Codes: C08L 2203/02 for biomedical use
• CPC Class Codes: C08L 2205/04 containing interpenetrating networks
• CPC Class Codes: C08L 71/02 Polyalkylene oxides
• CPC Class Codes: C08L 75/08 from polyethers
• CPC Class Codes: C12N 2533/30 : Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 2537/10 : Cross-linking

Abstract

The present invention relates to the synthesis of porous polymer scaffold from polyethyleneglycol-polyurethane having castor oil linkages under controlled conditions and their use as stem cell delivery vehicles thereby accelerating the tissue regeneration process. The present invention further studies the biodegradability, stability and biocompatibility of porous polymer scaffolds in various cell lines and primary bone marrow stem cells. Particularly the present invention further relates to the physio-chemical characterization of the porous polymer scaffolds.

Claims

1. A porous polymer scaffold for tissue engineering in stem cell transplantation consisting of a crosslinker, where the crosslinker comprises castor oil, polyether backbone, an isocyanate containing compound, and a secondary component, wherein the scaffold has a pore size that ranges from 50 nm-5 μ

m.

2. The porous polymer scaffold of claim 1, wherein the crosslinker is a triglyceride of castor oil.

3. The porous polymer scaffold of claim 1, wherein the polyether backbone is selected from the group consisting of di-hydroxyl, di-amine, and di-carboxyl terminated compounds.

4. The porous polymer scaffold of claim 1, wherein the polyether backbone is selected from the group consisting of polyethylene glycol (PEG), polypropylene glycol (PPG), polytetramethylene glycol (PTMG), block copolymers thereof, branched/graft copolymers thereof, and combinations thereof.

5. The porous polymer scaffold of claim 4, wherein the polyether backbone is polyethylene glycol (PEG) with molecular weight of 400-10000 Daltons.

6. The porous polymer scaffold of claim 1, wherein the isocyanate containing compound is selected from the group consisting of methylene diphenylene diisocyanate (MDI), polymeric methylene diphenylene diisocyanate (p-MDI), toluene diisocyanate (TDI), hexamethylene diisocyanate (HMDI), dicyclohexane methylene diisocyanate (H12MDI), isophoronediisocyanate (IPDI), xylene diisocyanate, hydrogenated xylene diisocyanate, and Desmodur-N.

7. The porous polymer scaffold of claim 1, wherein the secondary component is polyethylene glycol dimethylether of average molecular weight 250, 500, 750, 2000 or 5000 Daltons.

8. The porous polymer scaffold of claim 7, wherein the secondary component is polyethylene glycol dimethylether of average molecular weight 500 Daltons.

9. A process to prepare the porous polymer scaffold of claim 1, wherein the process comprises:

(a) reacting Castor oil (10 wt % to 60 wt % of total reactant weight) with diphenylmethane-4,4′

-diisocyanate (with total —

NCO/—

OH ratio in the range of 0.8-2.5) in tetrahydrofuran (THF) as solvent for 1 hour to form a pre-polymer (stage-I);

(b) charging the pre-polymer (stage-I) as obtained in step (a) with polyether macromonomer, N, N-dimethylaniline, and additional THF to obtain charged pre-polymer;

(c) adding a catalyst to the charged pre-polymer obtained in step (b) at room temperature to initiate the formation of a polyethylene glycol-polyurethane (PEG-PU), component-I (stage-II) and to obtain a growing polymer network;

(d) adding polyethylene glycol dimethylether (PEGDME) to the growing polymer network of step (c) to obtain a reaction mixture;

(e) degassing and vigorously mixing the reaction mixture as obtained in step (d) under inert atmosphere to obtain a uniformly homogeneous viscous mix;

(f) casting the uniformly homogeneous viscous mix as obtained in step (e) onto a teflon petri-dish to obtain a polymeric product;

(g) drying the polymeric product as obtained in step (f) at room temperature for 24 h followed by curing at higher temperature and inert atmosphere at 60-90°

C. for 48 h-96 h forming a semi-IPN matrix;

(h) wrapping free standing films of the semi-IPN matrix as obtained in step (g) in Whatman filter paper bag and exposing to a repeated soxhlet extraction process to obtain processed films;

(i) subjecting the processed films as obtained in step (h) to repeated swelling and drain cycles for 4-7 days against THF to extract out the PEGDME from the semi-IPN matrix completely, leaving behind a porous polymer network scaffold with impurities; and

(j) continuing extraction on the porous polymer network scaffold with impurities for 2 days using deionized millipore water (18MΩ

) to obtain an impurity free and sterile porous polymer scaffold.

10. The process of claim 9, wherein the castor oil in step (a) is 40% of the total reactant weight.

11. The process of claim 9, wherein the —

NCO/—

OH ratio of diphenylmethane-4,4′

-diisocyanate is in the range of 1.2 to 1.4.

12. The process of claim 9, wherein the polyether macromonomer in step (b) is polyethylene glycol (PEG).

13. The process of claim 12, wherein the polyethylene glycol (PEG) in step (b) is in the range of 70 wt % to 30 wt % of total weight.

14. The process of claim 9, wherein the THF in steps (a) and (b) is in the range of 20 wt % to 30 wt % of solids during reaction.

15. The process of claim 9, wherein the N, N-dimethylaniline in step (b) is in the range of 0.1 wt % to 2 wt % of solid content.

16. The process of claim 9, wherein the catalyst in step (c) is a tertiary amine.

17. The process of claim 16, wherein the tertiary amine is dimethylaniline (DMA).

18. The process of claim 9, wherein the polyethylene glycol dimethylether (PEGDME) in step (d) has a non-reactive end group and is used in the range of 20 wt % to 70 wt % of total weight of component-I.

19. The process of claim 18, wherein the polyethylene glycol dimethylether (PEGDME) is used in the weight ratio (50:

50).

20. A method of treating tissue damage and expediting wound tissue regeneration and repair, wherein the method comprises administering to a subject a composition comprising the porous polymer scaffold of claim 1.