Method, device and system for spatially controlling the formation of a hydrogel electrochemically

Patent metadata

Field	Value
Publication number	US-2016230197-A1
Application number	US-201415025559-A
Country	US
Kind code	A1
Filing date	Sep 29, 2014
Priority date	Sep 29, 2013
Publication date	Aug 11, 2016
Grant date	—

Abstract

The invention relates to a method, a device and a system for producing particularly a hydrogel ( 200 ) and for controlling an enzymatically catalysed formation of a covalent bond in a solution, wherein said covalent bond is formed between a first compound ( 20 ) comprising a first moiety ( 21 ) and a second compound ( 22 ) comprising a second moiety ( 23 ), wherein the first and the second moiety ( 21, 23 ) are a substrate of an enzyme wherein said enzyme catalyzes the formation of a covalent bond between the first and the second moiety ( 21, 23 ), and wherein a voltage is applied to the solution for spatially controlling said formation, wherein said voltage is adjusted such that it induces electrolysis of said solution.

First claim

1 . A method for controlling a linking reaction in a solution in proximity of a first electrode ( 11 ),
characterized in that for controlling said linking reaction the pH of the solution is altered locally in the proximity of said electrode ( 11 ) by applying an electrolysis-inducing electrical current to the solution via said first electrode ( 11 ).

2 . The method according to claim 1 , characterized in that a spatially structured hydrogel ( 200 ) is formed by said linking reaction, wherein said hydrogel ( 200 ) is particularly formed by local inhibition of the linking reaction, wherein said inhibition is particularly controlled by altering said pH locally, particularly such that said altered pH affects the enzymatic activity of an enzyme in the solution, and wherein particularly said enzyme is converting a first and a second moiety ( 21 , 23 ) comprised by a first and a second compound ( 20 , 22 ) in the solution to said hydrogel ( 200 ) by means of forming a covalent bond between said first and second moiety ( 21 , 23 ).

3 . The method according to claim 1 , characterized in that the linking reaction is spatially and/or temporally confined by altering the electrical current flowing through the solution.

4 . The method, particularly according to claim 1 , for controlling an enzymatically catalyzed formation of a covalent bond in a solution, wherein said covalent bond is formed between a first compound ( 20 ) comprising a first moiety ( 21 ) and a second compound ( 22 ) comprising a second moiety ( 23 ), wherein the first and the second moiety ( 21 , 23 ) are a substrate of an enzyme wherein said enzyme catalyzes the formation of a covalent bond between the first and the second moiety ( 21 , 23 ),
characterized in that a voltage is applied to the solution for spatially controlling said formation, wherein said voltage is adjusted such that it induces electrolysis of said solution.

5 . The method according to claim 4 , characterized in that said enzyme is an aminoacyltransferase, particularly a transglutaminase, more particularly factor XIIIa or precursor thereof, the first moiety ( 21 ) is an acyl, particularly an amide, and the second moiety ( 23 ) is an amine.

6 . The method according to claim 4 , characterized in that the first and/or second compound consists of or comprises a polymer, wherein said polymer is a natural polymer, particularly one of the following polymers: fibrin, alginate, chitosan, hyaluronic acid, chondroitin sulfate, heparin; or a synthetic polymer, particularly one of the following polymers: polyethylene glycol (PEG), polyactic acid, SU-8; or any polymer consisting of—or including—a combination of monomers, e.g. of dopamine, amine-containing groups such as lysine, cathecols, phosphate containing groups, thiol containing groups, alcohol containing groups, active esters and any dendrimer containing any of said groups (e.g. Hybrane, Boltorn).

7 . The method according to claim 4 , characterized in that the first compound ( 20 ) and/or the second compound ( 22 ) comprises polyethylenglycol (PEG), particularly PEG with a molar weight in the range from 4000 Da to 100000 Da, particularly 40000 Da, and wherein said PEG is an unbranched or branched PEG and wherein the branched PEG comprises particularly eight arms.

8 . The method according to claim 4 , characterized in that the formation of the covalent bond is a condensation reaction, a linking reaction, a ligation reaction, a cross-linking reaction or a polymerization reaction.

9 . The method according to claim 4 , characterized in that the voltage is induced via electrodes ( 11 , 12 ) in said solution, and wherein the voltage is adjusted such that the pH of the solution ranges between 1 and 14.

10 . The method according to claim 4 , characterized in that the enzymatic activity of the enzyme, particularly the aminoacyltransferase, more particular the transglutaminase, is locally inhibited, reduced or promoted depending on the voltage applied to the solution.

11 . The method according to claim 4 , characterized in that the solution comprises a third compound ( 24 , 25 ) comprising a third moiety, particularly an amide and/or an amine, wherein said third moiety is convertible by said enzyme with the respective first or second moiety, and wherein said third compound ( 24 , 25 ) is particularly a bioactive molecule, particularly a growth factor or interleukin 4.

12 . A method for providing a spatially structured hydrogel ( 200 ), particularly using the method according to claim 1 , wherein a first compound ( 20 ) comprising a first moiety ( 21 ) and a second compound ( 22 ) comprising a second moiety ( 23 ) are linked into a spatially structured hydrogel ( 200 ) by formation of a covalent bond between the first and the second moiety ( 21 , 23 ) of the first and the second compound ( 20 , 22 ),
characterized in that a voltage is applied to a solution comprising the first and the second compound ( 20 , 22 ) for spatially controlling said formation, wherein said voltage is adjusted such that it induces electrolysis of said solution.

13 . The method according to claim 12 , characterized in that the linking reaction is an enzymatically catalyzed particularly cross-linking reaction, wherein the first moiety ( 21 ) and the second moiety ( 23 ) are a substrate of an enzyme, wherein the enzyme is particularly n aminoacyltransferase, particularly a transglutaminase, more particularly factor XIIIa or a precursor thereof, and wherein the first moiety ( 21 ) is particularly an amine and wherein the second moiety ( 23 ) is particularly an amide.

14 . The method according to claim 1 , characterized in that one or several parameters of the linking reaction are changed during the linking reaction.

15 . The method according to claim 14 , characterized in that at least one of the following parameters is changed during the linking reaction:
the composition of the solution, particularly at least one of the following: buffer, enzymes, substrate of the enzyme, the used polymers and/or their functional groups, the position of an electrode, particularly of the first and/or second electrode, the applied current or voltage, and/or the duration of the linking reaction,

16 . The method according to claim 1 , characterized in that the linking reaction is interrupted, particularly for changing a parameter of the linking reaction.

17 . A device for generating a hydrogel, particularly using the method according to claim 1 , wherein said device comprises:
a reaction chamber ( 100 ) that is designed to hold a solution, and a first and a second electrode ( 11 , 12 ) arranged at the reaction chamber ( 100 ), and particularly a voltage source being designed to apply a pre-defined voltage to said electrodes ( 11 , 12 ), so as to induces electrolysis in said solution, particularly so as to control a linking reaction in said solution by locally altering the pH of the solution.

18 . The device according to claim 17 , characterized in that the first and/or second electrode ( 11 , 12 ) is designed to be moved with respect to the reaction chamber ( 100 ) and/or released from the reaction chamber ( 100 ), particularly so as to remove the first and/or second electrode ( 11 , 12 ) from the hydrogel.

19 . The device according to claim 17 , characterized in that the first and/or second electrode ( 11 , 12 ) comprises a metal, particularly tungsten, a semiconductor, a conductive polymeric material, or a combination of these materials.

Assignees

• Univ Zuerich
• ETH Zürich
• Univ Zuerich
• Eth Zuerich

Inventors

• Milleret Vincent
• Simona Benjamin R
• Voros Janos
• Ehrbar Martin

Classifications

• C08G2210/00

  Compositions for preparing hydrogels · CPC title

• C08G65/2696

  characterised by the process or apparatus used · CPC title

• C08G65/32

  Polymers modified by chemical after-treatment · CPC title

• C25B3/00

  Electrolytic production of organic compounds · CPC title

• C12N11/04

  entrapped within the carrier, e.g. gel or hollow fibres · CPC title