Stem cell-loaded injectable bone repair adhesive for highly bionic active bone tissues and preparation method thereof

A bone repair and bone tissue technology, applied in the field of bone repair materials, can solve the problems of limited stability and compatibility of the combination of inorganic phase and organic phase, unfavorable cell adhesion and proliferation, agglomeration, uneven dispersion, etc., to achieve increased The effect of cell adhesion site, improvement of hygrothermal stability, and improvement of cross-linking degree

Active Publication Date: 2020-11-24
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, HAp is agglomerated and unevenly dispersed in composite materials, and the combination stability and compatibility of inorganic phase and organic phase are relatively limited, which is not conducive to cell adhesion and proliferation and limits its application.

Method used

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  • Stem cell-loaded injectable bone repair adhesive for highly bionic active bone tissues and preparation method thereof
  • Stem cell-loaded injectable bone repair adhesive for highly bionic active bone tissues and preparation method thereof
  • Stem cell-loaded injectable bone repair adhesive for highly bionic active bone tissues and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] In this example, the preparation of dopamine-modified hyaluronic acid (HAD), the steps are as follows:

[0071] (1) Add a concentration of 84mg / mL N-hydroxysuccinimide (NHS) solution dropwise to a sodium hyaluronate (Mw=2000kDa) aqueous solution with a concentration of 21mg / mL, and then add dropwise a concentration of 200mg / mL 1-Ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) solution, stirred for 4 hours, added dropwise an aqueous solution of dopamine hydrochloride with a concentration of 3 mmol / L, stirred for 24 hours, twice During the stirring reaction process, the pH value was controlled at 6.0, and the operation of this step was carried out under nitrogen protection. The molar ratio of EDCI, NHS, dopamine hydrochloride and carboxyl groups on sodium hyaluronate was 7:5:4:1;

[0072] (2) Dialyze the reaction solution obtained in step (1) with a dialysis membrane (MW=3.5-8kDa) in ultrapure water with a pH value of 4 for 72h, and vacuum freeze-dry to ob...

Embodiment 2

[0091] In this example, the unloaded bone repair adhesive (HCLH, that is, the hydrogel with multiple hybrid cross-linked network structures prepared in Comparative Example 1) was first prepared according to the operation of Comparative Example 1, and then infrared and Thermal analysis confirmed the formation of multiple hybrid cross-linked network structures.

[0092] Using infrared spectrometer (Nicolet 6700, Germany) at 500cm -1 ~2000cm -1 In the wavelength range, the chemical structures of Col I, sodium hyaluronate (HA), HAD and HCLM were characterized. Such as Figure 8 As shown, 1627cm -1 (Amide I band) place peak belongs to the characteristic amide peak of Col I, at 1550cm -1 (amide II band) and 1235cm -1 The peak (band of amide III) indicates amidation reaction between the amino group of dopamine and the carboxyl group of HA to form HAD. 1747cm -1 The peak at the position weakens, which is due to the fact that on the one hand, the phenolic hydroxyl group of the H...

Embodiment 3

[0096] In this example, the preparation of the injectable bone repair adhesive (HCLH-MSCs) loaded with stem cells highly imitating living bone tissue is as follows:

[0097] The HAD with a dopamine grafting rate of 10% prepared in Example 1 was dissolved in deionized water, vortexed until transparent and clear to obtain a HAD solution with a concentration of 25 mg / mL, and collagen type I (Col I) was dissolved in 0.5mol / L acid solution to obtain a concentration of 25mg / mL Col I solution.

[0098] Add the micron-sized spherical HAp slurry with a solid content of 35wt.% to the HAD solution under ice bath conditions, and use the probe of the cell disruptor to ultrasonically disperse for 15 minutes, then add the Col I solution dropwise, and use the probe of the cell disruptor to fully ultrasonically disperse, micron The add-on of grade spherical HAp slurry, HAD solution and Col I solution should make the mass ratio of micron grade spherical HAp, HAD and Col I be 2:5:5, and the pH v...

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Abstract

The invention provides a stem cell-loaded injectable bone repair adhesive for highly bionic active bone tissues. The stem cell-loaded injectable bone repair adhesive is composed of hydrogel with a multiple hybrid cross-linked network structure and stem cells distributed in the hydrogel with the multiple hybrid cross-linked network structure. The bone repair adhesive has an interpenetrating networkstructure after freeze drying. A high polymer material with carboxyl and catechol functional groups is subjected to oxidation self-crosslinking, the high polymer material with carboxyl and catechol functional groups and a high polymer material with amino and carboxyl are subjected to Michael addition reaction, phenolic hydroxyl groups on the high polymer material with carboxyl and catechol functional groups and calcium ions dissociated from micron-sized spherical hydroxyapatite are chelated to form the multiple hybrid cross-linked network structure, and the micron-sized spherical hydroxyapatite is uniformly distributed in the multiple hybrid cross-linked network structure. The bone repair adhesive can promote adhesion and proliferation of stem cells while improving the tissue adhesion, and enhance angiogenesis and osteogenesis of stem cells.

Description

technical field [0001] The invention belongs to the field of bone repair materials, and relates to an injectable bone repair adhesive highly imitating living bone tissue loaded with stem cells and a preparation method thereof. Background technique [0002] At present, the degradable bone-like repair materials for the treatment of extreme bone defects mainly include autologous bone, allogeneic bone and artificial bone. Although autologous bone and allogeneic bone have good osteogenesis and biocompatibility, they generally have disadvantages such as limited sources, not easy to degrade, potential risk of disease transmission and immune rejection, and long healing time. Degradable materials for artificial bone repair have become the most widely used bone repair materials in clinical practice because of their structure and performance, which can be artificially designed and adjusted in a variety of ways. [0003] Hydrogels derived from natural products are attractive three-dime...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/12A61L27/20A61L27/24A61L27/38A61L27/50A61L27/56A61L27/58
CPCA61L27/12A61L27/20A61L27/24A61L27/3834A61L27/50A61L27/56A61L27/58A61L2400/06A61L2430/02C08L5/08
Inventor 孙勇樊渝江卢恭恭徐杨
Owner SICHUAN UNIV
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