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Conjugate of collagen material and growth factor as well as preparation method thereof

A growth factor, collagen material technology, applied in chemical instruments and methods, medical science, carrier binding/immobilization of peptides, etc., can solve the problems of side effects and reduced effect of PDGF-BB

Active Publication Date: 2013-04-10
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using traditional methods to directly inject PDGF-BB into the body or smear it on the wound not only greatly reduces the effect of PDGF-BB, but also PDGF-BB may be transported to other tissues through blood circulation, causing side effects

Method used

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  • Conjugate of collagen material and growth factor as well as preparation method thereof
  • Conjugate of collagen material and growth factor as well as preparation method thereof
  • Conjugate of collagen material and growth factor as well as preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Preparation and activity detection of the cross-linking product of demineralized bone matrix and platelet-derived growth factor

[0031] 1. Preparation of cross-linked compounds

[0032] Preparation of DBM: The DBM was cut into small pieces of 4 mm × 4 mm × 1 mm, and then placed in a 96-well plate for radiation sterilization. The sterile DBM was then soaked overnight in phosphate buffered saline (PBS, pH 8.0) containing 4 mM ethylenediaminetetraacetic acid (EDTA).

[0033] 1. Chemical cross-linking to prepare the cross-linking product of DBM and platelet-derived growth factor

[0034] Chemical cross-linking preparation methods such as figure 1As shown, 1 represents DBM, 2 represents 2-iminosulfane hydrochloride, 3 represents DBM modified with 2-iminosulfane hydrochloride; I represents PDGF-BB, and II represents sulfosuccinimidyl4-(N-maleimidomethyl)cyclohexanehexane -1-carboxylate (Sulfo-SMCC), III represents PDGF-SMCC; IV represents the cross-linker of de...

Embodiment 2

[0072] Example 2. Preparation and activity detection of the cross-linking product of demineralized bone matrix and platelet-derived growth factor

[0073] 1. Preparation

[0074] Preparation of DBM: as described in Example 1.

[0075] 1. Chemical cross-linking to prepare the cross-linking product of DBM and platelet-derived growth factor

[0076] (1) Preparation of DBM modified with 2-iminosulfane hydrochloride: the same as the method described in Example 1, the difference is that the soaking temperature is 20°C and the time is 1.5 hours. The feed ratio of alkane hydrochloride is 0.2 μmol 2-iminosulfane hydrochloride / 16mm 2 DBM.

[0077] (2) Preparation of PDGF-SMCC: the same as the method described in Example 1, except that the reaction temperature was 20° C., the reaction time was 0.7 hours, and the molar ratio of PDGF-BB to Sulfo-SMCC was 1:40.

[0078] (3) Crosslinking of 2-iminosulfane hydrochloride-modified DBM and PDGF-SMCC: the method was the same as that describe...

Embodiment 3

[0095] Example 3. Preparation and activity detection of the cross-linking product of demineralized bone matrix and platelet-derived growth factor

[0096] 1. Preparation

[0097] Preparation of DBM: as described in Example 1.

[0098] 1. Chemical cross-linking to prepare the cross-linking product of DBM and platelet-derived growth factor

[0099] (1) Preparation of DBM modified with 2-iminosulfane hydrochloride: the same as the method described in Example 1, the difference is that the soaking temperature is 30°C and the time is 2.5 hours. The feed ratio of alkane hydrochloride is 0.7 μmol 2-iminosulfane hydrochloride / 16mm 2 DBM.

[0100] (2) Preparation of PDGF-SMCC: the same as the method described in Example 1, except that the reaction temperature was 30° C., the reaction time was 1 hour, and the molar ratio of PDGF-BB to Sulfo-SMCC was 1:80.

[0101] (3) Crosslinking of 2-iminosulfane hydrochloride-modified DBM and PDGF-SMCC: the method was the same as that described i...

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Abstract

The invention discloses a conjugate of a collagen material and a growth factor as well as a preparation method thereof. The method comprises the steps of: crosslinking the collagen material and 2-iminosulfane hydrochloride to obtain a conjugate I; crosslinking the growth factor and sulfosuccinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate to obtain a conjugate II; and mixing the substance I and the conjugate II for reaction to obtain the conjugate of the collagen material and the growth factor. Testes prove that by crosslinking the growth factor on the collagen material with a chemical crosslinking method, the invention ensures that the surface activity of the collagen material is effectively improved, the cellularization and vascularization degree of the collagen material can be obviously improved, and the collagen material can be more effectively applied to the medical fields of tissue engineering and the like. Therefore, the conjugate and the preparation method therefore in the invention have broad application prospect in the fields of tissue engineering and the like.

Description

technical field [0001] The present invention relates to a cross-linking product of collagen material and growth factor and a preparation method thereof. Background technique [0002] Collagen, demineralized bone matrix (DBM), is a kind of bioscaffold material, which has high pore size, good mechanical plasticity and mechanical resistance, biodegradability and good biological properties. Compatibility, it has been widely used in tissue repair such as bone injury repair. However, in large-area tissue repair, DBM alone is not enough, so it is generally combined with growth factors to enhance the surface activity of DBM, promote the cellularization and vascularization of DBM itself, and achieve tissue repair and reconstruction. [0003] Platelet-derived growth factor-BB (PDGF-BB) is a multifunctional growth factor, it binds to receptor proteins on the cell membrane, promotes its phosphorylation for signal transmission, and exerts its biological function . PDGF-BB is a chemota...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K17/06A61L27/44A61L27/54A61L27/24A61L27/22
Inventor 陈磊肖志峰陈冰戴建武
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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