Injectable hydrogels incorporating igf-1c polypeptides

A technology of IGF-1C and CS-IGF-1C, applied in the field of preparation of active injectable chitosan hydrogel

Active Publication Date: 2020-11-10
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But in a previous study, we found that when stem cells were injected directly into the ischemic injury site, only ~1% of the cells survived after 1 week

Method used

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  • Injectable hydrogels incorporating igf-1c polypeptides
  • Injectable hydrogels incorporating igf-1c polypeptides
  • Injectable hydrogels incorporating igf-1c polypeptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Example 1: IGF-1C modified chitosan (CS) to obtain preparation of CS-IGF-1C

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

[0101] The IGF-1C polypeptide was synthesized by a well-established solid-phase synthesis (SPPS) procedure using 2-chlorotrityl chloride resin and an amino acid whose side chain was protected by a tert-buty group and the amino group was 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 in N,N'-dimethylformamide (DMF) was used to remove the Fmoc protecting group of the amino group;

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

[0104] 3. The synthesis of hexanoic acid azide: ethyl 6-bromohexanoate and sodium azide are reacted in DMF, and 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), overnight at room temperature;

[0127] (2) Use distilled water to dialyze the CS solution against a dialysis membrane (molecular weight cut-off of 8 kDa–10 kDa) for 1 week to remove residual acetic acid;

[0128] (3) obtain CS-IGF-1C hydrochloride by freeze-drying;

[0129] (4) β-GP solution (2.29 mol / L) in ice bath was added dropwise to CS solution, and stirred for 0.5 hour under ice bath condition;

[0130] (5) The mixed solution is transferred 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 / liter, and the pH value of the CS-IGF-1C / β-GP solution after dialysis was 7.2.

[0132] Scanning electron microscopy to observe the morphology of hydrogels

[0133] After the hydrogels were freeze-...

Embodiment 3

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

[0141] CCK-8 staining

[0142] We spread the material or hydrogel in a 96-well plate (5 μl / well) and placed the plate in an incubator (37°C) for 30 minutes (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 coated on the well plate according to the gradient of 1%, 1.5%, 3%, 5% and 10% to culture ADSCs, and the culture was carried out after 24 hours. CCK-8 staining. On the other hand, in order to compare the effects of CS-IGF-1C and CS hydrogels on cell proliferation, the two hydrogels (at a concentration of 3%) were coated on well plates to culture ADSCs, and CCK-8 staining was performed 24 hours later. . 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 (diluted 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 structural domain polypeptides, and also relates to the application of such materials as stem cell carrier materials for tissue damage repair, enhancing stem cells treatment effect. Background technique [0002] Stem cell-based cell therapy could offer promise for the treatment of tissue damage. Stem cells have the ability of self-renewal and multi-directional differentiation, and can participate in kidney repair through transdifferentiation and paracrine. A major advantage of stem cells for the treatment of tissue damage over drugs is that their diverse therapeutic effects can target multiple mechanisms of the pathophysiology of kidney injury; endogenously mobilized or exogenously infused stem cells home to the damaged kidney On the one hand, transdifferentiation into renal cells...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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