Stimuli responsive nanofibers

Abstract

A stimuli responsive nanofiber that includes a stimuli responsive polymer, such as a thermally responsive polymer, and a cross-linking agent having at least two latent reactive activatable groups. The nanofiber may also include a biologically active material or a functional polymer. The stimuli responsive nanofiber can be used to modify the surface of a substrate. When the nanofiber includes a thermally responsive polymer, the physical properties of the surface can be controlled by controlling the temperature of the system, thus controlling the ability of the surface to bind to a biologically active material of interest.

1 .- 17 . (canceled) 
     
     
         18 . A cell culture article comprising a surface and a thermally responsive polymer, wherein the thermally responsive polymer is associated with the surface via a crosslinking agent having at least two latent reactive groups. 
     
     
         19 . The article according to  claim 18  wherein the thermally responsive polymer comprises a polymer which has a first physical property at a first predetermined temperature range and a second physical property at a second predetermined temperature range. 
     
     
         20 . The article according to  claim 18  wherein at least one of the latent reactive groups comprises a photochemical reactive group. 
     
     
         21 . The article according to  claim 20  wherein the photochemical reactive group is an aryl ketone. 
     
     
         22 . The article according to  claim 21  wherein the aryl ketone is selected from the group acetophenone, benzophenone, anthraquinone, anthrone, acridone, xanthone, and thioxanthone. 
     
     
         23 . The article according to  claim 20  wherein all of the latent reactive groups of the crosslinking agent are photochemical reactive groups. 
     
     
         24 . The article according to  claim 23  wherein all of the photochemical reactive groups are the same. 
     
     
         25 . The article according to  claim 18  wherein at least one of the latent reactive groups is an aryl azide. 
     
     
         26 . The article according to  claim 18  wherein at least one of the latent reactive groups comprises a thermally-reactive group. 
     
     
         27 . The article according to  claim 23  wherein the thermally-reactive group includes a heat labile bond selected from oxygen-oxygen bonds, nitrogen-oxygen bonds, and nitrogen-nitrogen bonds. 
     
     
         28 . The article according to  claim 26  wherein all of the latent reactive groups of the crosslinking agent comprise thermally-reactive groups. 
     
     
         29 . The article according to  claim 28  wherein all of the thermally-reactive groups are the same. 
     
     
         30 . The article according to  claim 18  wherein the crosslinking agent is a monomeric material. 
     
     
         31 . The article according to  claim 18  wherein the crosslinking agent is a compound having a formulae selected from (a)-(g):
   L-(D-T-C(R 1 )(XP)CHR 2 GR 3 C(═O)R 4 ) m   (a)   wherein L is a linking group;   D is CR 6 R 7 ;   T is (—CH 2 —) x , (—CH 2 CH 2 —O—) x , (—CH 2 CH 2 CH 2 —O—) x , (—CH 2 CH 2 CH 2 CH 2 —O—) x  or forms a bond;   R 1  is a hydrogen atom, an alkyl, alkyloxyalkyl, aryl, aryloxyalkyl or aryloxyaryl group;   X is O, S, or NR 8 R 9 ;   P is a hydrogen atom or a protecting group, with the proviso that P is absent when X is NR 8 R 9 ;   R 2  is a hydrogen atom, an alkyl, alkyloxyalkyl, aryl, aryloxyalkyl or aryloxyaryl group;   G is O, S, SO, SO 2 , NR 10 , (CH 2 ) t —O— or C═O;   R 3  and R 4  are each independently an alkyl, aryl, arylalkyl, heteroaryl, or an heteroarylalkyl group or optionally, R 3  and R 4  can be tethered together via (—CH 2 —) q , (—CH 2 —) r C═O(—CH 2 —) s , (—CH 2 —) r S(—CH 2 —) x , (—CH 2 —) r S═O(—CH 2 —) s  or (—CH 2 —) r S(O) 2 (—CH 2 —) s , (—CH 2 —) r NR(—CH 2 —) s ;   R 10  is a hydrogen atom or an alkyl, aryl or arylalkyl group;   R 6  and R 7  are each independently a hydrogen atom, an alkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl group;   R 8  and R 9  are each independently a hydrogen atom, an alkyl, aryl, or arylalkyl group;   R is a hydrogen atom, an alkyl or an aryl group;   q is an integer from 1 to about 7;   r is an integer from 0 to about 3;   s is an integer from 0 to about 3;   m is an integer from 2 to about 10;   t is an integer from 1 to about 10; and   x is an integer from 1 to about 500;
   L-(T-C(R 1 )(XP)CHR 2 GR 3 C(═O)R 4 ) m   (b) 
   wherein L is a linking group;   T is (—CH 2 —) x , (—CH 2 CH 2 —O—) x , (—CH 2 CH 2 CH 2 —O—) x , (—CH 2 CH 2 CH 2 CH 2 —O—) x  or forms a bond;   R 1  is a hydrogen atom, an alkyl, alkyloxyalkyl, aryl, aryloxyalkyl or aryloxyaryl group;   X is O, S, or NR 8 R 9 ;   P is a hydrogen atom or a protecting group, with the proviso that P is absent when X is NR 8 R 9 ;   R 2  is a hydrogen atom, an alkyl, alkyloxyalkyl, aryl, aryloxyalkyl or aryloxyaryl group;   G is O, S, SO, SO 2 , NR 10 , (CH 2 ) t —O— or C═O;   R 3  and R 4  are each independently an alkyl, aryl, arylalkyl, heteroaryl, or an heteroarylalkyl group, or optionally, R 3  and R 4  can be tethered together via (—CH 2 —) q , (—CH 2 —) r C═O(—CH 2 —) s , (—CH 2 —) r S(—CH 2 —) s , (—CH 2 —) r S═O(—CH 2 —) s  or (—CH 2 —) r S(O) 2 (—CH 2 —) s , (—CH 2 —) r —NR(—CH 2 —) s ;   R 10  is a hydrogen atom or an alkyl, aryl or arylalkyl group;   R 8  and R 9  are each independently a hydrogen atom, an alkyl, aryl, or arylalkyl group;   R is a hydrogen atom, an alkyl or aryl group;   q is an integer from 1 to about 7;   r is an integer from 0 to about 3;   s is an integer from 0 to about 3;   m is an integer from 2 to about 10;   t is an integer from 1 to about 10; and   x is an integer from 1 to about 500;
   L-(GTZR 3 C(═O)R 4 ) m   (c) 
   wherein L is a linking group;   T is (—CH 2 —) x , (—CH 2 CH 2 —O—) x , (—CH 2 CH 2 CH 2 —O—) x , (—CH 2 CH 2 CH 2 CH 2 —O—) x  or forms a bond;   G is O, S, SO, SO 2 , NR 10 , (CH 2 ) t —O— or C═O;   Z is C═O, COO, or CONH when T is (—CH 2 —) x ;   R 3  and R 4  are each independently an alkyl, aryl, arylalkyl, heteroaryl, or an heteroarylalkyl group, or optionally, R 3  and R 4  can be tethered together via (—CH 2 —) q , (—CH 2 —) r C═O(—CH 2 —) s , (—CH 2 —) r S(—CH 2 —) s , (—CH 2 —) r S═O(—CH 2 —) s  or (—CH 2 —) r S(O) 2 (—CH 2 —) s , (—CH 2 —) r NR(—CH 2 —) s ;   R is a hydrogen atom or an alkyl or aryl group;   q is an integer from 1 to about 7;   r is an integer from 0 to about 3;   s is an integer from 0 to about 3;   m is an integer from 2 to about 10;   t is an integer from 1 to about 10; and   x is an integer from 1 to about 500;
   L-(TGQR 3 C(═O)R 4 ) m   (d) 
   wherein L is a linking group;   T is (—CH 2 —) x , (—CH 2 CH 2 —O—) x , (—CH 2 CH 2 CH 2 —O—) x , (—CH 2 CH 2 CH 2 CH 2 —O—) x  or forms a bond;   G is O, S, SO, SO 2 , NR 10 , (CH 2 ) t —O— or C═O;   Q is (—CH 2 —) p , (—CH 2 CH 2 —O—) p , (—CH 2 CH 2 CH 2 —O—) p  or (—CH 2 CH 2 CH 2 CH 2 —O—) p ;   R 3  and R 4  are each independently an alkyl, aryl, arylalkyl, heteroaryl, or an heteroarylalkyl group, or optionally, R 3  and R 4  can be tethered together via (—CH 2 —) q , (—CH 2 —) r C═O(—CH 2 —) s , (—CH 2 —) r S(—CH 2 —) s , (—CH 2 —) r S═O(—CH 2 —) s  or (—CH 2 —) r S(O) 2 (—CH 2 —) s , (—CH 2 —) r NR(—CH 2 —) s ;   R 10  is a hydrogen atom or an alkyl, aryl, or an arylalkyl group;   R is a hydrogen atom or an alkyl or aryl group;   q is an integer from 1 to about 7;   r is an integer from 0 to about 3;   s is an integer from 0 to about 3;   m is an integer from 2 to about 10;   p is an integer from 1 to about 10;   t is an integer from 1 to about 10; and   x is an integer from 1 to about 500;
   L-(—CH 2 —) xx C(R 1 )(GR 3 C(═O)R 4 ) m   (e) 
   wherein L is a linking group;   R 1  is a hydrogen atom, an alkyl, alkyloxyalkyl, aryl, aryloxyalkyl, or aryloxyaryl group;   each G is G is O, S, SO, SO 2 , NR 10 , (CH 2 ) t —O— or C═O;   each R 3  and R 4  is independently an alkyl, aryl, arylalkyl, heteroaryl, or an heteroarylalkyl group, or optionally, R 3  and R 4  can be tethered together via (—CH 2 —) q , (—CH 2 —) r C═O(—CH 2 —) s , (—CH 2 —) r S(—CH 2 —) s , (—CH 2 —) r S═O(—CH 2 —) s  or (—CH 2 —) r S(O) 2 (—CH 2 —) s , (—CH 2 —) r NR(—CH 2 —) s ;   each R 10  is a hydrogen atom or an alkyl, aryl, or an arylalkyl group;   each R is a hydrogen atom or an alkyl or aryl group;   each q is an integer from 1 to about 7;   each r is an integer from 0 to about 3;   each s is an integer from 0 to about 3;   m is an integer from 2 to about 10;   each t is an integer from 1 to about 10; and   xx is an integer from 1 to about 10;
   L-(-C(R 1 )(XP)CHR 2 GR 3 C(═O)R 4 ) m   (f) 
   wherein L is a linking group;   R 1  is a hydrogen atom, an alkyl, alkyloxyalkyl, aryl, aryloxyalkyl or aryloxyaryl group;   X is O, S, or NR 8 R 9 ;   P is a hydrogen atom or a protecting group, with the 27. that P is absent when X is NR 8 R 9 ;   R 2  is a hydrogen atom, an alkyl, alkyloxyalkyl, aryl, aryloxyalkyl or aryloxyaryl group;   G is G is O, S, SO, SO 2 , NR 10 , (CH 2 ) t —O— or C═O;   R 3  and R 4  are each independently an alkyl, aryl, arylalkyl, heteroaryl, or an heteroarylalkyl group, or optionally, R 3  and R 4  can be tethered together via (—CH 2 —) q , (—CH 2 —) r C═O(—CH 2 —) s , (—CH 2 —) r S(—CH 2 —) s , (—CH 2 —) r S═O(—CH 2 —) s  or (—CH 2 —) r S(O) 2 (—CH 2 —) s , (—CH 2 —) r NR(—CH 2 —) s ;   R 8  and R 9  are each independently a hydrogen atom, an alkyl, aryl, or arylalkyl group;   R 10  is a hydrogen atom or an alkyl, aryl, or an arylalkyl group;   R is a hydrogen atom, an alkyl or an aryl group;   q is an integer from 1 to about 7;   r is an integer from 0 to about 3;   s is an integer from 0 to about 3;   m is an integer from 2 to about 10; and   t is an integer from 1 to about 10; and
   L-(GR 3 C(═O)R 4 ) m ;  (g) 
   wherein L is a linking group;   G is O, S, SO, SO 2 , NR 10 , (CH 2 ) t —O— or C═O;   R 3  and R 4  are each independently an alkyl, aryl, arylalkyl, heteroaryl, or an heteroarylalkyl group, or optionally, R 3  and R 4  can be tethered together via (—CH 2 —) q , (—CH 2 —) r C═O(—CH 2 —) s , (—CH 2 —) r S(—CH 2 —) s , (—CH 2 —) r S═O(—CH 2 —) s  or (—CH 2 —) r S(O) 2 (—CH 2 —) s , (—CH 2 —) r NR(—CH 2 —) s ;   R is a hydrogen atom, an alkyl or an aryl group;   q is an integer from 1 to about 7;   r is an integer from 0 to about 3;   s is an integer from 0 to about 3;   m is an integer from 2 to about 10; and   t is an integer from 1 to about 10.   
     
     
         32 . The article according to  claim 31  wherein the crosslinking agent is a compound of formula: 
       
         
           
           
               
               
           
         
       
     
     
         33 . The article according to  claim 18  wherein the cell culture article comprises a nanofiber or nanofiber mesh. 
     
     
         34 . The article according to  claim 18  wherein the cell culture article is selected from slides, multi-well plates, Petri dishes, tissue culture plates, tissue culture flasks, and coverslips. 
     
     
         35 . The article according to  claim 33  wherein the nanofiber or nanofiber mesh is provided on a surface of a slide, multi-well plate, Petri dish, tissue culture plate, tissue culture flask or coverslip. 
     
     
         36 . The article according to  claim 18  wherein the surface comprises a polyolefin, polystyrene, poly(methyl)methacrylate, polyacrylonitrile, poly(vinylacetate), poly(vinyl alcohol), chlorine-containing polymer, polyoxymethylene, polycarbonate, polyamide, polyimide, polyurethane, phenolic, amino-epoxy resin, polyester, silicone, cellulose-based plastic, fluoropolymer or rubber-like plastics. 
     
     
         37 . The article according to  claim 18  wherein the surface comprises pyrolytic carbon, parylene coated surface, or a silylated surface of glass, ceramic or metal. 
     
     
         38 . The article according to  claim 18  wherein the thermally responsive polymer comprises poly(isopropylacrylamide), copolymers of polyethylene glycol and poly(isopropylacrylamide), or mixtures thereof. 
     
     
         39 . A cell culture article comprising a surface and a stimuli responsive polymer which undergoes a physical or chemical change when exposed to external stimuli, wherein the stimuli responsive polymer is associated with the surface via a crosslinking agent having at least two latent reactive groups. 
     
     
         40 . A kit for preparing a cell culture article, the kit comprising: a stimuli responsive polymer suitable for cell culture, which polymer undergoes a physical or chemical change when exposed to external stimuli; and a crosslinking agent having at least two latent reactive groups, the crosslinking agent being adapted for associating the stimuli responsive polymer to a surface of a cell culture article. 
     
     
         41 . A method for preparing a cell culture article, the method comprising steps of:
 (a) contacting a surface of the cell culture article with a coating composition, the coating composition comprising a stimuli responsive polymer which undergoes a physical or chemical change when exposed to external stimuli, and a crosslinking agent having at least two latent reactive groups; and   (b) treating the coating composition to activate latent reactive groups of the crosslinking agent, thereby coupling the stimuli responsive polymer to the surface of the cell culture article in a manner in which at least some of the latent reactive groups remain in an inactive state.   
     
     
         42 . The method according to  claim 41  further comprising a step of coupling a biologically active material to the surface of the cell culture device by activating remaining latent reactive groups of step (b). 
     
     
         43 . A method for preparing a cell culture article, the method comprising steps of:
 (a) contacting a surface of the cell culture article with a coating composition comprising a stimuli responsive polymer which undergoes a physical or chemical change when exposed to external stimuli, a crosslinking agent having at least two latent reactive groups, and a biologically active material; and   (b) treating the coating composition to activate latent reactive groups of the crosslinking agent to form a coating on the cell culture device surface.   
     
     
         44 . A method for preparing a cell culture article, the method comprising steps of:
 (a) contacting a surface of the cell culture article with a crosslinking agent having at least two latent reactive groups;   (b) treating the crosslinking agent to activate less than all of the latent reactive groups of the crosslinking agent, thereby coupling the crosslinking agent to the surface of the cell culture and allowing at least some of the latent reactive groups to remain in an inactive state;   (c) contacting the surface of the cell culture article with a stimuli responsive polymer which undergoes a physical or chemical change when exposed to external stimuli;   (d) treating the surface of the cell culture article to activate latent reactive groups which are present in an inactive state subsequent to step (b), thereby coupling the stimuli responsive polymer to the cell culture article surface.   
     
     
         45 . A method for preparing a cell culture article, the method comprising steps of:
 (a) preparing a composition comprising a stimuli responsive polymer which undergoes a physical or chemical change when exposed to external stimuli, and a crosslinking agent having at least two latent reactive groups;   (b) forming a cell culture article from the composition of step (a); and   (c) treating the cell culture article formed in step (b) to activate latent reactive groups of the crosslinking agent.   
     
     
         46 . The method according to  claim 45 , wherein the step of forming a cell culture article comprises forming a nanofiber. 
     
     
         47 . A method for preparing a cell culture article, the method comprising steps of:
 (a) preparing a composition comprising a polymer and a crosslinking agent having at least two latent reactive groups;   (b) forming a cell culture article from the composition of step (a);   (c) contacting a surface of the cell culture article with a stimuli responsive polymer which undergoes a physical or chemical change when exposed to external stimuli; and   (d) treating the surface of the cell culture article to activate latent reactive groups of the crosslinking agent, thereby coupling the stimuli responsive polymer to the cell culture article surface.   
     
     
         48 . The method according to  claim 47  wherein the polymer of step (a) comprises a polyolefin, polystyrene, poly(methyl)methacrylate, polyacrylonitrile, poly(vinylacetate), poly(vinyl alcohol), chlorine-containing polymer, polyoxymethylene, polycarbonate, polyamide, polyimide, polyurethane, phenolic, amino-epoxy resin, polyester, silicone, cellulose-based plastic, fluoropolymer or rubber-like plastic. 
     
     
         49 . The article according to  claim 47  wherein the polymer of step (a) comprises pyrolytic carbon, or a silylated surface of glass, ceramic or metal.