Composite used for curing urethra, preparation method and application thereof

A composite and urethral technology, applied in the fields of medicine and biomedical engineering, can solve the problems of large surgical trauma and cumbersome steps

Inactive Publication Date: 2010-05-05
SHANGHAI SIXTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method can ensure the homology of tissue-engineered urothelial cells, its disadvantages are also very obvious, that is, the operation trauma is large and the steps are cumbersome

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Microseparation, decellularization, and preparation of collagenous fascia from bladder submucosal tissue of allogenic rabbits

[0040] The bladder of the rabbit was cut, and the submucosa was separated under a microscope, which was placed in the decellularization solution composed of double distilled water, 0.2% Triton X-100 and ammonium hydroxide, processed and rinsed to remove the cellular components in the tissue, In order to avoid the occurrence of immune reaction, part of the submucosal tissue was excised for identification to judge the effect of decellularization.

[0041] After the bladder submucosal tissue was treated with decellularized solution, it was in the shape of a loose film. After random sampling, the fascia was stained with HE, and the light microscope observed that the fascia was loose collagen. The part is collagen fibers with loose structure, and no cells exist, indicating that the decellularization effect is good, and avoiding the possible rejectio...

Embodiment 2

[0043] Isolation, Culture and Proliferation of Rabbit Foreskin Epidermal Cells

[0044] Under ketamine anesthesia, a small piece of rabbit foreskin tissue was dissected and treated with DSBS2 and various proteases (trypsin) to separate epidermal cells, which were planted on 3T3 trophoblast, and cells composed of KSFM+EGF+bovine pituitary extract were added Culture medium, in vitro culture, proliferation, subculture, to make it reach a certain cell density (2.7×10 6 piece / cm 2 ). The expanded epidermal cell markers (anti-AE1 / AE3) were identified by immunohistochemistry.

[0045] The isolated and digested epithelial cells grew well, and were determined to be normal epithelial cells by immunohistochemistry, and were passaged for 3 generations. The number of cells reached the requirement of implanting and growing the biological scaffold.

Embodiment 3

[0047] Compound preparation and use

[0048] Materials and methods

[0049]One, the amplified epidermal cells obtained in Example 2 are compounded with BAMG, using immersion and gas-liquid interface culture technology (air-liquid interface culture technology method: get male young rabbits, body weight 0.3kg, adopt 3% ketamine anesthesia, after the anesthesia takes effect , local disinfection, cut 1 × 0.5cm of foreskin tissue, put the removed skin into a centrifuge tube with 20 ml of KSFM solution, and put the centrifuge tube into an ice bucket for storage. Take out the skin and put it into 0.25% chloramphenicol solution at 40C + penicillin and streptomycin, rinse for 5 minutes / time, three times in a row. Prepare a small ophthalmic scissors and two small forceps, trim the subcutaneous tissue to a size of 1 cm * 2 mm. Rinse three times with PBS solution. Insert Dispase II Digested in 40C for 14 hours. Abandon the DSBS, peel off the dermis and epidermis with tweezers and scissor...

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PUM

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Abstract

The invention discloses a composite used for curing urethra, a preparation method and an application thereof. The composite used for curing the urethra contains a cell-free BAMG and epidermal cells. The invention also provides the preparation method and the application of the composite.

Description

technical field [0001] The invention relates to the fields of medicine and biomedical engineering, and more particularly relates to a compound using decellularized bladder submucosal collagen fascia (BAMG) as a scaffold, a preparation method and application thereof. Background technique [0002] Hypospadias and urethral stricture caused by different reasons are common diseases in urology, and the incidence of hypospadias is 1 / 250-1 / 350. However, due to the limited tissue of the urethral mucosa, it has brought great difficulties to the repair of the urethra. [0003] At present, natural or artificial bio-scaffold materials are mostly used for urethral repair, but due to the severe inflammatory reaction caused by the degradation process, it affects the differentiation and proliferation of cells and the repair of the urethra, which can easily cause urethral stricture and decellularization. The Bladder Submucosal Collagen Fascia (BAMG) is taken from bladder tissue. Because it c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/36
Inventor 傅强邓辰亮徐月敏宋小飞廖国龙
Owner SHANGHAI SIXTH PEOPLES HOSPITAL
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