Construct for promoting absorption of molecules by a cell and methods of using the contstruct

Abstract

The present disclosure is directed to a construct for promoting absorption of molecules by a cell and the application thereof in drug and gene delivery. The present disclosure further describes topographical modulation of endocytosis for drug and gene delivery.

1 . A method of promoting absorption of molecules by cells, wherein the method comprises:
 providing a construct comprising a plurality of micro and/or nanoscale protrusions located at the surface of the construct; wherein the protrusions have a size and are spaced apart from each other at a distance that promotes absorption of molecules by said cell; and   seeding and culturing at least one cell at the surface of the construct under conditions suitable for absorption of molecules by the cells.   
     
     
         2 . The method of  claim 1 , wherein the absorption of molecules is via endocytosis, or pinocytosis, or phagocytosis, or
 wherein absorption of the molecules uses a non-viral carrier or a viral carrier.   
     
     
         3 . (canceled) 
     
     
         4 . The method of  claim 1 , wherein the molecules to be absorbed are selected from the group consisting of nucleic acid, nucleic acid vectors, siRNA, microRNA, magnetic nanoparticles, gold nanoparticles, fluorescent nanoparticles, quantum dots, aptamers (oligonucleic acid or peptide), peptides, growth factors, therapeutically active substances, biomarkers, colouring agents, and any one of the aforementioned molecules attached to a microparticles. 
     
     
         5 . A method of cell transfection, or drug-delivery or high-throughput screening arrays using a construct for promoting absorption of molecules by a cell located at the surface of the construct; wherein the construct comprises:
 a plurality of micro and/or nanoscale protrusions located at the surface of the construct;   wherein the protrusions have a size and are spaced apart from each other at a distance that promotes absorption of molecules by said cell.   
     
     
         6 . The method of  claim 1 , wherein the construct comprises a plurality of protrusions located at the surface of the construct;
 wherein the protrusions are in the form of pillars having a diameter of between about 200 nm to about 2 μm;   wherein the pitch between the protrusion from edge to edge is between 150 to 300 nm such that the pitch between the protrusions is less than the size of the cells to be located at the surface of the construct;   wherein the protrusions have a size and are spaced apart from each other at a distance that promotes absorption of molecules by said cell and wherein the absorption of molecules is facilitated by endocytosis, or receptor mediated endocytosis, or pinocytosis, or phagocytosis;   wherein the pillars are collapsed pillars lying at the surface of the construct; and   wherein the collapsed pillars have a length of between about 50 nm to about 5 μm.   
     
     
         7 . The method of  claim 1 , wherein the protrusions are arranged in an isotropic pattern. 
     
     
         8 . The method of  claim 1 , wherein the protrusions are arranged in an anisotropic pattern. 
     
     
         9 . The method of  claim 1 , wherein the protrusions are located at the surface of the construct in a detachable form. 
     
     
         10 . The method of  claim 1 , wherein the protrusions are located at the surface of the construct in a non-detachable form. 
     
     
         11 . The method of  claim 1 , wherein the protrusions are round. 
     
     
         12 . The method of  claim 1 , wherein the protrusions are polygonal. 
     
     
         13 . The construct of  claim 1 , wherein the pillars are extend about 50 nm to 4 μm, or about 100 nm to about 2 μm above the surface of the construct. 
     
     
         14 . The construct of  claim 1 , wherein the pitch between the pillars from edge to edge is between 200 nm to 250 nm. 
     
     
         15 . The construct of  claim 1 , wherein a residual layer is arranged between the pillars and the surface of the construct. 
     
     
         16 . The construct of  claim 1 , wherein the construct is obtained via nano-imprinting lithography. 
     
     
         17 . The construct of  claim 16 , wherein the nano-imprinting lithography is thermal nano-imprinting lithography or UV nano-imprinting lithography. 
     
     
         18 . The construct of  claim 16 , wherein the construct and/or the protrusions are made of a polymer. 
     
     
         19 . The construct of  claim 18 , wherein the polymer is a synthetic polymer, or a rigid synthetic polymer, or a bioresorbable polymer, or a biodegradable polymer. 
     
     
         20 . The construct of  claim 19 , wherein the synthetic polymer is selected from the group consisting of poly(methyl methacrylate) (PMMA), polydimethylsiloxane (PDMS), polystyrene (PS) and mixtures thereof, or
 wherein the biodegradable polymer is selected from the group consisting of chitosan, poly(ε-caprolactone), polyglycolic acid, poly(lactic acid), polyphosphoester (PPE) and mixtures thereof, or   wherein the rigid synthetic polymer is polystyrene, optionally the polystyrene is tissue-culture grade polystyrene (TCPS).   
     
     
         21 . The construct of  claim 18 , wherein during manufacture the polymer is mixed with molecules which are to be absorbed by the cell to be located on the construct, optionally wherein the molecules are attached to the surface of the protrusions.