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Left-handed chiral nanogel cell scaffold material and preparation method thereof

A nanogel and chirality technology, applied in the field of left-handed chiral nanogel cell scaffold material and its preparation, can solve the problems of difficult directional differentiation, poor cell adhesion, etc., and achieves easy regulation of chirality of fiber structure and stable gel Good performance, easy to mass industrial production effect

Active Publication Date: 2021-09-07
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] For the defects in the prior art, the object of the present invention is to provide a kind of left-handed chiral nanogel cell scaffold material and preparation method thereof, that is, a kind of left-handed chiral super nanogel cell culture scaffold material and preparation method thereof, with Solve the problem of poor adhesion and directional differentiation of cells on existing matrix materials

Method used

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  • Left-handed chiral nanogel cell scaffold material and preparation method thereof
  • Left-handed chiral nanogel cell scaffold material and preparation method thereof
  • Left-handed chiral nanogel cell scaffold material and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0057] Embodiment 1, the preparation of left-handed nanogel fiber scaffold material

[0058] This embodiment relates to a chiral nanogel scaffold material for cell culture, the chiral nanogel scaffold material is self-assembled by a small molecule gel factor with chiral amino acid units, the gel factor is Cyclohexane is the central core, the amide bond is the connecting key, and the 1 and 4 positions of cyclohexane are connected with L-methionine-ethylene glycol group.

[0059] 1.1 Preparation of L-methionine ester derivative gelling factor

[0060] Add 5.6g of L-methionine methyl ester hydrochloride into 20mL of dichloromethane and stir, then add 8.0mL of triethylamine dropwise. Under ice bath, dissolve 2.4g of 1,4-cyclohexanedioyl chloride in 10mL of dichloromethane and add dropwise to the above solution, remove the ice bath, stir and react at room temperature for 24 hours, and remove the dichloromethane solvent by rotary evaporation to obtain 1 , 4-cyclohexanedi-L-methi...

Embodiment 2

[0065] Example 2, the left-handed nanogel scaffold material promotes cell adhesion

[0066] 2.1 Left-handed chiral nanogel fiber scaffolds promote cell adhesion of mouse fibroblasts (NIH 3T3)

[0067] In this example, a left-handed nanogel scaffold was used to prepare a methionine derivative with a left-handed chiral unit for the aforementioned Example 1. The gel factor was self-assembled in water through non-covalent interactions such as hydrogen bonds. The right-handed nanogel scaffold is self-assembled by methionine derivative gel factor with right-handed chiral units through non-covalent interactions such as hydrogen bonds, and serves as an experimental control material.

[0068] Digest NIH 3T3 cells with trypsin for 2-3min, centrifuge, remove the digestive juice, wash with 1×PBS, add complete culture medium and blow the cells to form a uniform cell suspension, adjust the density of NIH 3T3 cells to 5.0×10 4 / mL, add 200 μL of NIH 3T3 cell suspension to each well of a 2...

Embodiment 3

[0071] Example 3, the left-handed nanogel scaffold material promotes directed differentiation of cells

[0072] Digest dental pulp stem cells (DPSC) with trypsin for 2-3min, centrifuge, remove the digestive juice, rinse with 1×PBS, add complete culture medium and blow the cells to form a uniform cell suspension, and adjust the cell density of DPSC to 1.0×10 5 / mL, add 200uL cell suspension to each well of a 24-well plate, place at 37°C, 5% CO 2 Cell differentiation was observed after 7 days of incubation in a cell culture incubator. DPSC cells clearly differentiated toward osteoblasts on the left-handed fibers, and the osteoblast signature gene (BMP2) was relatively expressed (see Figure 4 ) is 1.5 to 2.0 times that of cells on right-handed fibers.

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Abstract

The invention provides a left-handed chiral nanogel cell scaffold material and a preparation method thereof. The preparation method comprises the following steps: (1) dispersing the left-handed amino acid derivative gel factor in deionized water to obtain a mass volume concentration of 0.5 mg / mL~1.0mg / mL L-amino acid derivative gel factor dispersion solution; (2) heating the above dispersion to a clear solution; (3) adding the above clear solution to the orifice of the cell culture plate; (4) adding The prepared cell culture plate is left standing at room temperature for 20-30 minutes to form a left-handed hydrogel; (5) the cell culture plate is dried at 40-50°C, that is, the left-handed chiral nanogel is covered on the orifice plate Cell scaffold material. The invention provides a preparation method of a left-handed chiral nanogel cell scaffold material for promoting cell adhesion and osteogenic differentiation, which solves the problems of poor adhesion of cells on matrix materials and difficulties in directional differentiation in the existing cell culture process .

Description

technical field [0001] The present invention relates to the field of hydrogel cell culture scaffold materials, in particular to a left-handed chiral nanogel cell scaffold material and a preparation method thereof, in particular to a left-handed chiral nanogel that promotes cell adhesion and directional bone differentiation. Gel cell culture scaffold material and preparation method thereof. Background technique [0002] Human tissue damage and defect will lead to dysfunction. The traditional repair method is autologous tissue transplantation. Although it can achieve satisfactory curative effect, it is at the expense of autologous healthy tissue, which will lead to many complications and the source of extra body organs is extremely difficult. Limited, long-term use of immunosuppressants is required due to immune rejection, and the resulting complications are sometimes fatal. According to incomplete statistics, every year, many people suffer from skin burns, damaged corneas, o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/00C07C319/20C07C323/58
CPCC07B2200/07C07C319/20C07C323/58C12N5/0075C12N2531/00C12N2533/00C07C2601/14
Inventor 冯传良窦晓秋
Owner SHANGHAI JIAOTONG UNIV
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