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Preparation method of artificial alveoli

An alveolar and artificial technology, used in pharmaceutical formulations, prostheses, drug delivery, etc., to solve problems such as limited treatment methods and insufficient lung transplant donors

Active Publication Date: 2020-12-04
XUZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the problems of irreversible repair of lung tissue caused by lung diseases, lack of donors for lung transplantation, and extremely limited treatment methods, the present invention provides a simple and easy preparation method of artificial alveoli with controllable size. Co-culture composition of inverse opal scaffold, growth factor VEGF, lung cells, endothelial cells and epithelial cells

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  • Preparation method of artificial alveoli
  • Preparation method of artificial alveoli
  • Preparation method of artificial alveoli

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Effect of Gelatin Concentration

[0035] 1. The acquisition of gelatin microspheres: gelatin particles are dissolved in ultrapure water in a water bath environment greater than 45°C, and gelatin solutions with concentrations of 3%, 5%, 10%, 15%, and 20% (wt / v) are prepared respectively . Collect gelatin microspheres with a microfluidic device, by controlling the flow rate of the aqueous phase solution (gelatin solution) to be 3ml / h, the flow rate of the organic phase (based on toluene, adding 3wt% Span 80 as a surfactant) is 18ml / h, The diameter of the capillary is 0.7 mm as the organic phase channel, and the inside of the capillary uses a syringe needle (with an inner diameter of 0.3 mm) as the channel for the aqueous phase solution to obtain gelatin microspheres, which are collected in methanol solution for later use (attached figure 1 .A).

[0036] 2. Preparation of gelatin template: put the collected gelatin microspheres into the mold, slightly vibrate the mold to...

Embodiment 2

[0043] Effect of gelatin solution flow rate

[0044]This example is basically the same as Example 1, except that the gelatin concentration is 10%, and the gelatin solution flow rate is controlled to be 0.5ml / h, 1ml / h, 3ml / h, 5ml / h and 7ml / h respectively.

[0045] When the flow rate of the gelatin solution is 0.5ml / h, the flow rate is too slow, and spherical gelatin cannot be obtained, but irregular. The mean diameters of the gelatin microspheres prepared by the flow rate of 1ml / h and 3ml / h were 120 μm and 300 μm respectively (attached image 3 D and C). At flow rates of 5ml / h and 7ml / h no microspheres were obtained but columnar long lines. Therefore, when the gelatin solution flow rate is 1-3ml / h, the size of the prepared gelatin microspheres is suitable for preparing artificial alveoli, and the optimum gelatin solution flow rate is 3ml / h.

Embodiment 3

[0047] Effect of organic phase flow rate

[0048] This example is basically the same as Example 1, except that the concentration of gelatin is 10%, and the flow rate of the organic phase is controlled to be 10ml / h, 18ml / h, 30ml / h and 35ml / h respectively.

[0049] The average diameter of the gelatin microspheres prepared by the organic phase flow rate of 10ml / h, 18ml / h and 30ml / h was respectively 400 μm (attached image 3 E), 300μm (with image 3 C), 120μm (with image 3 D). When the flow rate was 35ml / h, gelatin microspheres with smaller diameter but inhomogeneous were obtained. Therefore, when the flow rate of the organic phase is 10-30 ml / h, the size of the prepared gelatin microspheres is suitable for preparing artificial alveoli, and the optimum flow rate of the organic phase is 18 ml / h. If the flow rate of the organic phase is too low, the size of the prepared gelatin microspheres is too large; if the flow rate is too high, the size of the prepared gelatin microsphere...

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Abstract

The invention discloses a preparation method of artificial alveoli. The method first prepares gelatin microspheres with uniform diameter through a gelatin aqueous solution through a microfluidic device, obtains regularly arranged gelatin scaffolds through a self-assembly method, heats them to make them closely arranged, and then perfuses the PU solution. After removing the solvent by freeze drying, the water bath method The gelatin template was removed to obtain a three-dimensional porous PU scaffold with an inverse opal structure. Then the PU scaffold was treated with ammonia plasma to graft amino groups, heparin was connected to the amino groups by EDC / NHS, and then VEGF was added to bind VEGF to the heparin. Finally, the MRC-5 cells were rotated and inoculated on the VEGF-modified PU scaffold, and the mixed cell suspension of HUVECs and NL20 cells was inoculated in proportion, and the tissue-engineered artificial alveoli were obtained after co-culture. The invention effectively regulates the diameter of the obtained artificial alveoli to be about 300 microns by regulating the gelatin solution and polyurethane concentration, the flow rate and channel diameter of the water phase and the organic phase, and the self-assembly temperature and time.

Description

technical field [0001] The invention relates to a method for preparing artificial alveoli, belonging to the technical field of tissue engineering. Background technique [0002] As an important organ of the human body, the lung is responsible for the function of gas exchange. The lung tissue is composed of 150-400 million alveoli, with a surface area of ​​70-80m 2 , which plays an important role in providing oxygen to the body and expelling carbon dioxide. [0003] The size of alveoli is usually around 250 microns, of which 90% of the surface area is occupied by type I alveolar cells, about 5% by type II alveolar cells, and the rest is jointly occupied by macrophages and vascular endothelial cells. However, many diseases such as lung cancer, chronic obstructive pulmonary disease, and pulmonary hypertension can cause irreversible damage to the lungs. Lung transplantation is currently the most widely used treatment technique, but the extremely limited number of donors, expens...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/18A61L27/38A61L27/54A61L27/56
CPCA61L27/18A61L27/3804A61L27/3808A61L27/3813A61L27/3839A61L27/3886A61L27/54A61L27/56A61L2300/252A61L2300/414A61L2300/602A61L2400/08C08L75/04
Inventor 吕兰欣杨膺许铁胡书群燕宪亮
Owner XUZHOU MEDICAL UNIV