KR-20260065679-A - Anisotropic magnetic nanorod-substrate complex for ligand interconnectivity control and manufacturing method of the same, regulation method of cell behavior using the same

KR20260065679AKR 20260065679 AKR20260065679 AKR 20260065679AKR-20260065679-A

Abstract

The present invention relates to an anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, a method for manufacturing the same, and a method for controlling cell behavior using the same. The composite comprises a substrate, gold nanoparticles bonded to the surface of the substrate and spaced apart, ligand nodes formed by binding a cell-adhesive ligand to each of the gold nanoparticles, and magnetic nanorods attached to a substrate where the ligand nodes are not formed, wherein the anisotropy of the physical shape can be controlled and a polymer linker is bonded for adhesion to the substrate. The invention relates to an anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, a method for manufacturing the same, and a method for controlling cell behavior using the same. According to the present invention, the interconnectivity of ligand clusters formed on the surface of a substrate can be controlled through the anisotropic control and magnetic control of magnetic nanorods attached to the surface of a substrate, thereby facilitating the control of cell behaviors such as the attachment and differentiation of stem cells and the attachment and polarization of macrophages.

Inventors

홍현식
강희민
강나연
김초원
김성열
김다희
나현지

Assignees

고려대학교 산학협력단

Dates

Publication Date: 20260511
Application Date: 20241101

Claims (20)

Substrate; Gold nanoparticles bonded to the surface of the above substrate and spaced apart; Ligand nodes formed by binding a cell-adhesive ligand to each of the above gold nanoparticles; and An anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, comprising: a magnetic nanorod attached to a substrate where the above-mentioned ligand node is not formed, capable of controlling the anisotropy of the physical shape, and having a polymer linker coupled for adhesion to the substrate.
In claim 1, The above magnetic nanorods Anisotropic magnetic nanorod-substrate composite having different length and width sizes and capable of controlling ligand interconnectivity.
In claim 1, The above magnetic nanorods Anisotropic magnetic nanorod-substrate composite capable of controlling ligand interconnectivity, with length increasing and width decreasing as anisotropy increases.
In claim 1, The above magnetic nanorods An anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, wherein when anisotropy increases, the interconnection between spaced ligand nodes arranged on the substrate is proportionally blocked according to the increase in anisotropy.
In claim 1, The above magnetic nanorods The length is 380 nm to 1000 nm, and Anisotropic magnetic nanorod-substrate composite having a width of 70 nm to 250 nm and capable of controlling ligand interconnectivity.
In claim 1, The above magnetic nanorods An anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, wherein an amine group ( NH₂ ) is bonded for bonding with the above-mentioned polymer linker.
In claim 1, The above magnetic nanorods Magnetite core; and It includes a silica outer shell surrounding the magnetite core; and The above magnetite core is an anisotropic magnetic nanorod-substrate composite capable of controlling ligand interconnectivity, which is phase-transformed from acaraganite to magnetite by annealing treatment.
In claim 6, The above polymer linker is An anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, wherein the magnetic nanorod to which the above amine group ( NH₂ ) is bonded is bonded to the magnetic nanorod through amide bonding.
In claim 1, The above polymer linker is An anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of a ligand, which is a compound comprising any one of n-hydroxysuccinimide, maleimide, thiol, methoxy, hydroxyl, amine, azide, carboxylic acid, acrylate, cyanine, aldehyde, acrylamide, epoxide, hydrazide, halide, methacrylate, and silane.
In claim 1, The above polymer linker is Anisotropic magnetic nanorod-substrate composite having a molecular weight of 1 kDa to 50 kDa and capable of controlling the interconnectivity of ligands.
In claim 1, The above gold nanoparticles are Anisotropic magnetic nanorod-substrate composite having a diameter of 10 nm to 30 nm and capable of controlling ligand interconnectivity.
In claim 1, The above ligand Anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, which are peptide ligands (CDD RGD).
In claim 1, The above substrate Anisotropic magnetic nanorod-substrate composite having a thiolized surface with thiol groups (SH) and capable of controlling the interconnectivity of ligands.
In claim 13, The above ligand Thiol groups (SH) are bonded to the above gold nanoparticles and substrate for bonding, and An anisotropic magnetic nanorod-substrate composite capable of controlling ligand interconnectivity, treated with a tris(2-carboxyethyl)phosphine (TCEP) solution to prevent abnormal binding of ligands by inducing disulfide bonds through self-oxidation of the thiol group on the surface of the substrate having the above thiol group (SH).
In claim 1, An anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, wherein the surface of the substrate where the ligand nodes and magnetic nanorods are not located is deactivated by washing with a compound having a methoxy(O- CH3 ) group.
In claim 1, When the intersecting intervals connecting the ligand nodes where the above gold nanoparticles and ligands are bound are called 'ligand cluster edges', An anisotropic magnetic nanorod-substrate composite capable of controlling ligand interconnectivity, wherein the number of 'ligand cluster edges' is controlled according to the anisotropic control of the magnetic nanorods.
In claim 1, An anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, wherein the interconnection between ligand nodes is controlled when the orientation of the magnetic nanorods on the substrate is aligned by applying a magnetic field to the magnetic nanorods.
In claim 1, An anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, wherein when a magnetic field is applied to the magnetic nanorod to control the movement of the magnetic nanorod in the upward direction of the substrate, the movement of cells downward of the magnetic nanorod is controlled.
Step of cleaning and thiolizing the surface of the substrate; A step of bonding and spaced arranging gold nanoparticles on the surface of the substrate; A step of coating the gold nanoparticles with a cell-adhesive ligand to form ligand nodes to which the ligand is bound to each gold nanoparticle; and A method for manufacturing an anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, comprising the step of attaching a magnetic nanorod with controlled anisotropy of physical shape onto a substrate on which the above-mentioned ligand nodes are not formed, wherein the magnetic nanorod is grafted onto the substrate via a polymer linker of the magnetic nanorod to which the polymer linker is attached.
In claim 19, A method for manufacturing an anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, further comprising the step of washing the surface of the substrate where the ligand node, to which a ligand is bound to the gold nanoparticle, and the magnetic nanorod are not located with distilled water in which a compound having a methoxy(O- CH3 ) group is dissolved, to remove residual thiol groups on the substrate and deactivate the substrate.

Description

Anisotropic magnetic nanorod-substrate complex capable of controlling ligand interconnectivity and manufacturing method of the same, regulation method of cell behavior using the same The present invention relates to an anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands, a method for manufacturing the same, and a method for controlling cell behavior using the same. Specifically, it relates to a technology capable of controlling the interconnectivity between ligand nodes formed on the substrate surface through the anisotropic control and magnetic control of magnetic nanorods attached to the surface of the substrate, thereby enabling the control of stem cell adhesion and differentiation and macrophage adhesion and polarization. The extracellular matrix (ECM) is a substance located outside the cell that surrounds it, playing a role that aids the cell structurally and biochemically. Its main components are glycoproteins, and representative examples include collagen, hydroxyapatite (a major component of bone), cellulose (a component of plant cell walls), and hyaluronic acid. In general, for multicellular organisms, the extracellular matrix acts as a support by filling the space outside the cell and also serves to create boundaries with other tissues. Additionally, it acts as a physical scaffolding that allows cells to adhere to the matrix. The extracellular matrix possesses diverse characteristics corresponding to its various types, one of which is stiffness and elasticity. The difference is immediately apparent when considering skin and bone. Accordingly, each cell prefers the optimal environment suited to it. Furthermore, in the case of stem cells, the direction of differentiation can vary depending on the characteristics of the extracellular matrix, while for macrophages, the direction of polarization can differ. Meanwhile, physical screens occurring in the extracellular matrix (ECM) separate various compartments of tissues to aid in the regulation of homeostasis and tissue regeneration by controlling biomolecule transport and cell infiltration. Certain tissues can act as physical screens to regulate tissue repair mechanisms involving the interactions of various cells. However, artificial materials that mimic the ECM and can dispersively and dynamically regulate bioactive surfaces are uncommon. For example, integrins dynamically form connections with the ECM on which bioactive ligands are expressed, and RGDs capable of attracting cells (Amino acid sequences of arginine, glycine, and aspartic acid) Ligands mediate focal adhesion and mechanotransduction within cells. Light or magnetic fields can regulate cell adhesion by remotely controlling the blocking of ligands. In this case, since magnetic fields have the characteristic of easily penetrating tissues in vivo, non-invasive control of physical screens is possible, such as remotely regulating cell adhesion by controlling the density of ligands through the control of particles with magnetic properties. In other words, cell behavior can be controlled through the regulation of ligand arrangements, etc. Meanwhile, when examining prior art literature, Korean Patent Publication No. 10-2024-0109638 discloses a "magnetic rod-substrate composite for controlling the attachment and differentiation of stem cells and a method for manufacturing the same." Although this utilizes nanorods and controls cell behavior through magnetic control, it differs in that it is a technology in which ligand coatings are formed on both "nanorods" and "gold particles" and they are physically bonded through magnetic control to control cell behavior. In particular, it differs in that it completely lacks the concept of anisotropic control that the present invention aims to implement. Furthermore, Korean Patent Publication No. 10-2022-0048697 discloses a "nanocoil-substrate composite for controlling the behavior of stem cells, a method for manufacturing the same, and a method for controlling the attachment and differentiation of stem cells using the same." This differs in that it utilizes a "nanocoil" with a ligand coating and controls cell behavior by physically stretching the "nanocoil" through magnetic control; in particular, it differs in that it does not use anisotropically controlled materials at all, as in the present invention. FIG. 1 is a diagram illustrating an anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands according to one embodiment of the present invention and a manufacturing process thereof. FIG. 2 is a diagram illustrating an anisotropic magnetic nanorod-substrate composite capable of controlling the interconnectivity of ligands according to an embodiment of the present invention, wherein a is a diagram illustrating ligand cluster modeling through ligand nodes, b is a diagram illustrating the correlation of ligand cluster edges according to the increase in anisotropy of a