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Lacunaris bladder acellular matrix preserving biological activity factor and preparation

A bioactive factor, acellular matrix technology, applied in medical science, prosthesis and other directions, can solve the problems of poor bladder smooth muscle cell regeneration, destruction of bioactive factors, disordered and irregular arrangement, etc. Compatibility and easy access to materials

Inactive Publication Date: 2008-12-24
THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In previous studies, after bladder replacement with these prepared BAMs, bladder smooth muscle cells regenerated poorly. In the central area of ​​BAM replacements, smooth muscle cells were rare and irregularly arranged, which may be due to the destruction of endogenous bioactive factors

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Preparation of acellular matrix of porcine bladder, the preparation steps are as follows:

[0056] a) Acquisition of pig bladder: The bladder and part of the urethra of a pig weighing 20 kg were all taken out, placed in 10 mM phosphate buffer, pH 7.2-7.4 pre-cooled at 4°C, quickly brought to the laboratory and carefully removed Remove the fat and serosal tissue on the surface, insert an infusion leather catheter through the urethra and fix it on the urethra with silk threads, and then rinse the bladder with 10 mM phosphate buffer, pH 7.2-7.4, pre-cooled at 4°C through the infusion leather catheter. 3 times, rinse the urine in the bladder cavity;

[0057] b) Infuse 100ml of digestive juice containing 0.25% / 0.038% trypsin / tetrasodium EDTA into the bladder cavity through the infusion thong catheter, the pH is 7.2-7.4, clamp the infusion thong catheter, and then put the entire bladder Completely immerse in the digestion solution, shake and digest at room temperat...

Embodiment 2

[0071] The preparation of embodiment 2 rabbit bladder acellular matrix, preparation steps are as follows:

[0072] a) Acquisition of rabbit bladder: The bladder and part of the urethra of a New Zealand white rabbit weighing 4 kg were taken out, placed in 10 mM phosphate buffer, pH 7.2-7.4 pre-cooled at 4°C, and quickly brought to the laboratory. Carefully remove the fat and serosal tissue on the outer surface, insert an infusion leather catheter through the urethra and fix it on the urethra with silk threads, and then rinse the infusion leather catheter with 10mM phosphate buffer, pH 7.2-7.4, pre-cooled at 4°C. Bladder 3 times to flush out the urine in the bladder cavity;

[0073] b) Infuse 40ml of digestive juice containing 0.25% / 0.038% trypsin / tetrasodium EDTA into the bladder cavity through the infusion thong catheter, the pH is 7.2-7.4, clamp the infusion thong catheter, and then put the entire bladder Completely immerse in the digestion solution, shake and digest at room...

Embodiment 3

[0087] Example 3. Isolation, culture and identification of seed cells

[0088] 3.1 Isolation, culture and identification of HBSMC

[0089] With the consent of bladder cancer patients who underwent radical total cystectomy, a small piece of bladder tissue without obvious tumor growth was taken, and the urothelial layer was completely removed by mechanical dissection, using trypsin / ethylenediamine with a concentration of 0.25% / 0.038% Tetrasodium tetraacetate and 0.1% collagenase type I were sequentially digested, and the resulting cell suspension was inoculated in DMEM / F-12 complete medium containing 10% FBS for primary culture and subculture. Inverted microscope was used for morphological observation, and the expression of smooth muscle actin and desmin were observed by immunostaining of cell slides.

[0090] Observed under an inverted microscope, the human bladder smooth muscle cells cultured in vitro showed a long spindle shape and a typical "peak-valley" shape after fusion;...

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PUM

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Abstract

The invention discloses a porous bladder acellular matrix retaining bioactive factors and a preparation method thereof. A bladder of a pig, a rabbit, a dog or cattle is handled by digestive juice, hypotonic buffer solution, hypertonic buffer solution with surfactants, buffer solution with nuclease, and deionized water. An abacterial bladder acellular matrix is obtained by cooling, freezing out and sterilizing. The bladder acellular matrix is not provided with cells, an urinary tract epithelial layer, a submucous layer or a blood circulatory system, thus having good biocompatibility and appropriate ultrastructural structure. Bioactive factors are retained, including cell adhesive factors, growth factors and chemokines. Therefore, the bladder acellular matrix is an ideal scaffold material for a bladder in tissue engineering.

Description

technical field [0001] The invention relates to a biological material and a preparation method thereof, in particular to a porous bladder acellular matrix retaining biologically active factors and a preparation method thereof. Background technique [0002] Congenital malformations, infections, traumas, tumors and other diseases of the bladder can lead to structural damage and dysfunction of the bladder and require bladder replacement surgery. The gastrointestinal tract is currently the main alternative used for bladder replacement. However, because the gastrointestinal tract is in the environment of the urinary system, it may cause a series of complications such as metabolic disturbance, infection, stone formation and even malignant transformation. Therefore, finding a reasonable bladder substitute is a challenge before us, and tissue engineering technology undoubtedly provides us with a powerful tool. [0003] The basic principle of tissue-engineered bladder is to inocula...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/56
Inventor 杨斌孙则禹周六化戴玉田
Owner THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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