IGF-1C polypeptide combined injectable hydrogel and application thereof to carrier material to enhance repair effect of stem cells in tissue damage

A stem cell and gel technology, applied in the field of preparation of active injectable chitosan hydrogels

Active Publication Date: 2017-05-17
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But in a previous study, we found that when stem cells were injected directl

Method used

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  • IGF-1C polypeptide combined injectable hydrogel and application thereof to carrier material to enhance repair effect of stem cells in tissue damage
  • IGF-1C polypeptide combined injectable hydrogel and application thereof to carrier material to enhance repair effect of stem cells in tissue damage
  • IGF-1C polypeptide combined injectable hydrogel and application thereof to carrier material to enhance repair effect of stem cells in tissue damage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Embodiment 1: IGF-1C modifies chitosan (CS) to obtain the preparation of CS-IGF-1C

[0100] Solid Phase Synthesis of IGF-1C Polypeptide

[0101] The IGF-1C polypeptide is synthesized by a well-established solid-phase synthesis (SPPS) procedure using 2-chlororityl chloride resin and amino acids whose side chains are protected by tert-buty groups and amino groups are protected by Fmoc. The steps are:

[0102] 1. The C-terminus of the first amino acid is grafted on the resin, and the loading rate is 0.6 mmol / g. 20% piperidine was dissolved in N, N'-dimethylformamide (DMF) for removing the Fmoc protecting group of the amino group;

[0103] 2. Under the joint action of the condensing agent BTU and the catalyst DIPEA, the carboxyl group of the next amino acid is combined with the free amino group; repeating the above steps, the polypeptide chain will continue to grow;

[0104] 3. Synthesis of azide caproic acid: react ethyl 6-bromohexanoate with sodium azide in DMF, then t...

Embodiment 2

[0124] Example 2: Preparation and performance evaluation of CS-IGF-1C hydrogel

[0125] Preparation of CS-IGF-1C hydrogel

[0126] (1) Dissolve CS-IGF-1C (1.5%, w / v) with acetic acid (0.1 mol / L), room temperature, overnight;

[0127] (2) Use distilled water to dialyze the CS solution in a dialysis membrane (MWCO: 8kDa–10kDa) for 1 week to remove residual acetic acid;

[0128] (3) obtaining CS-IGF-1C hydrochloride by lyophilization;

[0129] (4) Add the ice-bathed β-GP solution (2.29 mol / liter) dropwise to the CS solution, and stir for 0.5 hours under ice-bath conditions;

[0130] (5) Transfer the mixed solution to a 37°C incubator, and gelation can be seen in 5 minutes;

[0131] (6) The final concentration of β-GP in the mixed solution was 0.023 mol / L, and the pH value of the CS-IGF-1C / β-GP solution after dialysis was 7.2.

[0132] Scanning Electron Microscopy Observation of Hydrogel Morphology

[0133] After freeze-drying the hydrogels, put them into a desiccator to dry ...

Embodiment 3

[0140] Example 3: Biocompatibility of CS-IGF-1C

[0141] CCK-8 staining

[0142] We coated the material or hydrogel on a 96-well plate (5 μl / well), and placed the well plate in an incubator (37°C) for 30 minutes (and then used directly or stored at 4°C). On the one hand, in order to evaluate the effect of different concentrations of CS-IGF-1C material on cell viability, the material was applied to the well plate to culture ADSCs according to the gradient of 1%, 1.5%, 3%, 5% and 10%, and the ADSCs were cultured after 24 hours. CCK-8 staining. On the other hand, in order to compare the effect of CS-IGF-1C and CS hydrogel on cell proliferation, the two hydrogels (concentration: 3%) were coated on well plates to culture ADSCs, and CCK-8 staining was carried out after 24 hours. . Cells were plated at a concentration of 3×104 / well (4 wells / group). The specific dyeing method is as follows:

[0143] 1. Prepare CCK-8 staining solution (dilute according to CCK-8 stock solution: com...

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Abstract

The invention relates to insulin-like growth factor C (IGF-1C) structural domain polypeptide combined chitosan hydrogel, which has high cytocompatibility and biological activity. The active hydrogel has injectability and can serve as a carrier material to promote the treatment effect of the stem cells in tissue damage. A proper tissue regeneration micro-environment is provided for stem cell transplantation, so that the survival rate of the transplanted stem cells is increased. Survival and retention of the stem cells in the damage area are enhanced, survival and proliferation of the transplanted cells are promoted, apoptosis is reduced, and angiogenesis and functional recovery of the damage part are promoted, so that the treatment effect of stem cell transplantation on tissue damage is improved. In addition, the hydrogel after transplantation does not promote poor differentiation of the stem cells.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and in particular relates to a preparation method of an active injectable chitosan hydrogel combined with IGF-1C domain polypeptide, and also relates to the application of this type of material as a stem cell carrier material in tissue damage repair and enhancement of stem cell treatment effect. Background technique [0002] Stem cell-based cell therapy may offer promise in the treatment of tissue damage. Stem cells have self-renewal and multi-directional differentiation capabilities, and can participate in kidney repair through transdifferentiation and paracrine. Compared with drugs, a major advantage of stem cells in the treatment of tissue damage is that their diverse therapeutic effects can target multiple mechanisms in the pathophysiology of renal injury; endogenously mobilized or exogenously infused stem cells home to the damaged kidney , on the one hand, transdifferentiate into kidney...

Claims

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Application Information

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IPC IPC(8): A61K9/06A61K47/36A61K38/30A61K35/28A61P13/12A61P9/10A61P17/02
Inventor 赵强李宗金冯国伟张计敏孔德领
Owner NANKAI UNIV
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