Tissue engineered cartilage scaffold containing biologically active factors and its preparation method and application

A biologically active factor and tissue engineering technology, applied to medical preparations containing active ingredients, pharmaceutical formulas, prostheses, etc., can solve problems such as calcification, achieve the effect of improving the quality of cartilage repair and inhibiting cartilage calcification

Active Publication Date: 2019-11-12
ZHEJIANG XINGYUE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention provides a method for preparing a tissue engineering cartilage scaffold containing bioactive factors, that is, a silk fibroin collagen double-layer scaffold compounded with an active factor parathyroid hormone-related peptide, which can solve the problem of cartilage repair in the prior art. There will be shortcomings of cartilage calcification, which can effectively improve the repair effect of osteochondral damage

Method used

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  • Tissue engineered cartilage scaffold containing biologically active factors and its preparation method and application
  • Tissue engineered cartilage scaffold containing biologically active factors and its preparation method and application
  • Tissue engineered cartilage scaffold containing biologically active factors and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] (1) Preparation of silk fibroin solution: a) Degumming: Put 100g of silkworm silk (grade 5A) into 5L of 2M sodium carbonate aqueous solution, bathe in 98°C for 30 minutes, wash with pure water, repeat this process 3 times, degumming Remove sericin, leave silk fibroin, dry silk fibroin at 50°C, obtain 70g of dried silk fibroin, and set aside; b) dissolving: dissolve the above-mentioned dried silk fibroin with a mass concentration of 16%. In 9.3 M lithium bromide aqueous solution, 60 ℃ water bath for 90 minutes until the silk fibroin is fully dissolved to obtain a mixed solution containing silk fibroin and a small amount of insoluble particles; c) Dialysis: use the mixed solution with a regenerated cellulose dialysis bag (molecular weight cut-off 4000 Dalton) dialysis, dialyze 12 times in 3 days with sterile pure water of 10 times the volume of the mixed solution, remove lithium bromide ions in the solution, and obtain the retentate a; d) concentration: put the dialysis ba...

Embodiment 2

[0049] (1) Osteochondral injury modeling: Adult male New Zealand white rabbits weighing 2.5-3 kg were anesthetized with 10% chloral hydrate (dose 4 mL / kg) preoperatively by ear vein injection and intraperitoneal injection. The hair was cut within 5cm around the knee joint, the rabbit was placed supine on the operating table, the limbs were fixed, disinfected with povidone iodine, and a sterile drape was spread. Take the medial incision of the knee joint, cut the skin and subcutaneous fascia, cut the rabbit knee joint through the medial edge of the patella, retract the patellar tendon and quadriceps tendon to the outside, and expose the knee joint. A cylindrical osteochondral defect with a diameter of 4 mm and a depth of 3.5 mm was drilled, as shown in Figure 2.

[0050] (2) Application of tissue engineered cartilage scaffold: A tissue engineered cartilage scaffold containing biologically active factors in Example 1 was implanted into the osteochondral lesion formed in (1), and...

Embodiment 3

[0053] A preparation method of a tissue engineered cartilage scaffold containing biologically active factors, comprising the following steps:

[0054] (1) Spread type Ⅰ collagen solution with a concentration of 5mg / mL on the silk fibroin support at a volume ratio of 1:2, and vacuum freeze-dry. The freeze-drying condition is vacuum freeze-drying at -40~℃ for 10 days , to obtain a silk fibroin collagen double-layer scaffold;

[0055] (2) Soak the obtained silk fibroin collagen bilayer scaffold in 20 μg / mL parathyroid hormone-related peptide solution and incubate at 30° C. for 2 hours, and incubate at 30° C. for 2 hours.

[0056] The preparation method of the silk fibroin collagen double-layer scaffold is as follows:

[0057] (1.1) Using mulberry silk as raw material, the silk fibroin solution is obtained through degumming, dissolving and dialysis;

[0058] (1.2) Mix anhydrous hydroxyapatite and anhydrous sodium chloride particles 1:1, add the 5% silk fibroin solution obtained ...

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Abstract

The invention provides a tissue engineering cartilage stent containing a bioactive factor as well as a preparation method and application of the tissue engineering cartilage stent. The preparation method comprises the following steps: (1) flatly laying an I-type collagen solution on a silk fibroin stent in the volume ratio of 1 to 2, and freeze-drying in a vacuum environment, so that a silk fibroin and collagen dual-layer stent is obtained; and (2) soaking the obtained silk fibroin and collagen dual-layer stent in a parathyroid hormone related peptide solution with concentration being 20-200 [mu]g/ml, and breeding at 30-40 DEG C for 0.5-2h. According to the tissue engineering cartilage stent containing the bioactive factor disclosed by the invention, by virtue of the silk fibroin and collagen dual-layer stent, parathyroid hormone related peptide can be effectively relieved and a cartilage calcification phenomenon in a process of repairing cartilage injury can be inhibited, so that cartilage repair quality is improved.

Description

technical field [0001] The invention relates to the field of biomaterial preparation, in particular to a preparation method and application of a tissue engineering cartilage scaffold containing bioactive factors. Background technique [0002] Articular cartilage can reduce the friction between two adjacent bones and buffer the vibration generated during exercise, so cartilage damage caused by sports is extremely common. Statistics show that there are more than 400,000 cartilage-related surgeries in the United States alone every year. Because cartilage has no blood vessels and nerves, the traditional view is that cartilage damage cannot be repaired spontaneously. If no external intervention is taken, it will lead to further cartilage wear and even osteoarthritis, causing irreversible damage to its structure and function. Patient's quality of life work. The current conservative treatment methods for clinical treatment of superficial cartilage injuries include joint cavity la...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/29A61L27/48A61L27/54
Inventor 张薇陈隆坤徐国伟宣铁锋
Owner ZHEJIANG XINGYUE BIOTECH
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