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Collagen-based bilayer membrane material with directional release of loaded growth factors and its manufacturing method

A double-layer technology of growth factors and collagen, which is applied in pharmaceutical formulations, prostheses, and drug delivery, can solve the problems of disorganized bioactive delivery or poor controllability of cell targeting and release behavior, achieving good plasticity and preparation Simple process, low immunogenicity and good effect

Active Publication Date: 2020-06-09
INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention can overcome the lack of biological activity of existing collagen materials for tissue repair, and the current problems of unorganized delivery of growth factors or poor controllability of cell targeting and release behavior, etc., and provides a collagen-based bilayer for directional release of growth factors Membrane material, which can be used for the repair and treatment of diseased and defective tissues

Method used

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  • Collagen-based bilayer membrane material with directional release of loaded growth factors and its manufacturing method
  • Collagen-based bilayer membrane material with directional release of loaded growth factors and its manufacturing method
  • Collagen-based bilayer membrane material with directional release of loaded growth factors and its manufacturing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: the preparation of dense layer and loose layer collagen film

[0033] 1. Preparation of Bovine Tendon Type Ⅰ Collagen Swelling Solution

[0034] Fully wash the fresh bovine tendon purchased in the market, remove the connective tissue such as the tissue around the tendon, tendon sheath and fascia, and freeze it in a freezer after washing. Take out the frozen bovine tendon and cut it into slices about 1 mm thick, add ficin solution (concentration 0.05-0.25%) to treat it, stir it and place it in an incubator for 24 hours for enzymatic hydrolysis, then add excess H 2 o 2 Terminate the reaction, wash with distilled water 3-4 times, add 0.3% malonic acid solution to swell for 24 h after decanting. The swollen tendons were fully stirred for 6 hours, and the viscosity was adjusted with 0.3% malonic acid solution, filtered with 80-120 mesh stainless steel mesh under positive pressure to remove impurities and unswollen substances, and packaged.

[0035] 2. Prepar...

Embodiment 2

[0047] Example 2: Preparation of dense-loose collagen bilayer membrane

[0048] Add 80 mg / mL fibrinogen solution dropwise to one side of the cross-linked dense layer collagen membrane in the above example, and cover the entire surface; then drip 400 IU / mL thrombin solution to one side of the cross-linked loose layer collagen membrane, Also cover the entire surface; then overlap the two collagen films and glue them together, use the chemical reaction of thrombin and fibrinogen to quickly react to form fibrin glue to bond the collagen double-layer film, and air dry; then use the pressure film Machine flattening, that is, a dense-loose collagen double-layer membrane material. The average thickness of the bilayer film is 1 mm.

Embodiment 3

[0049] Example 3: Cy5.5-NHS ester labeled FGF2

[0050] 1. Weigh 0.46mg of Cy5.5-NHS ester and dissolve it in 100uL dimethyl sulfoxide solution.

[0051] 2. Prepare FGF2 into a 100ug / mL solution with a pH value above 8.0.

[0052] 3. Take 20uL FGF2 solution, add 1uL Cy-5.5-NHS ester solution, mix well, avoid light, and react at room temperature for at least 4 hours.

[0053] 4. Ultrafilter the reaction solution of FGF2 and Cy5.5-NHS ester in an ultrafiltration tube, 10000 rpm, 20 minutes, ultrafiltration 8 times, the reaction solution in the ultrafiltration tube turns from blue to colorless, collect the ultrafiltration tube medium reaction solution.

[0054] 5. For the detection of Cy-5.5-labeled FGF2 in the in vivo imager, see figure 2 .

[0055] 6. The FGF2 solution labeled Cy5.5 was added dropwise to the side of the loose layer in the dense-loose collagen bilayer membrane of Example 2, and it was observed in the live imager that the FGF2 solution would not penetr...

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Abstract

The invention relates to a collagen-based bilayer membrane material for directional release of growth factors and a manufacturing method of the collagen-based bilayer membrane material. The collagen-based bilayer membrane material is prepared from collagen, the growth factors, fibrinogen and thrombin; a dense layer collagen membrane and a loose layer collagen membrane are prepared, and the dense layer collagen membrane and the loose layer collagen membrane are bonded by virtue of fibrous protein, so that a bilayer membrane structure is constituted. The growth factors can be loaded on the surface of the membrane by a dosage of 1-10000ng of the growth factors per square centimeter; preferably, the growth factors are loaded on the bilayer membrane by a dosage of 30ng per square centimeter; and preferably, average thickness of the bilayer membrane is 1-5mm. The material has a bidirectional controlled-release effect on the loaded growth factors, so that directional release of the growth factors is achieved and the material can act on target tissues; and the material is applicable to restorative treatment of diseased and defective tissues. The growth factors, which are loaded on the bilayer collagen membrane prepared by the invention and have a targeted release effect, can be released to the target tissues in a directional mode and activity of the growth factors can be kept; and the bilayer membrane material can be used for guiding and inducing regeneration and repair of tissues.

Description

technical field [0001] The invention relates to a collagen-based double-layer membrane material loaded with growth factors for directional release and a manufacturing method thereof. The material has a directional and controlled release effect on the growth factors loaded on it, and the growth factors released directionally act on target tissues. The material can be used for Repair of diseased and defective tissue. Background technique [0002] Collagen is the most important water-insoluble fibrin outside the cell, and it is the skeleton of the extracellular matrix. Collagen provides tension and elasticity to cells in the extracellular matrix and plays a role in cell migration and development. It widely exists in the skin, bones, muscles, cartilage, joints, and hair tissues of the human body, and plays a triple anti-aging role of support, repair, and protection. Collagen is rich in content, almost accounting for more than 30% of the total protein weight. Glue is widely us...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/40A61L27/24A61L27/22A61L27/50A61L27/54A61L27/58C08J7/16C08L89/00
CPCA61L27/225A61L27/227A61L27/24A61L27/50A61L27/54A61L27/58A61L2300/252A61L2300/412A61L2300/414A61L2300/602A61L2400/12C08J7/16C08J2389/00
Inventor 李学敏张其清杨文裕曹一汀
Owner INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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