Preparation method of artificial alveoli

An alveolar and artificial technology, applied in the field of preparation of artificial alveoli, can solve the problems of limited treatment methods, insufficient lung transplant donors, etc.

Active Publication Date: 2018-09-21
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 provide

Method used

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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 comprises the following steps: firstly, preparing gelatin micro-spheres with uniform diameters from a gelatin aqueous solution through a microfluidic device, obtaining gelatin brackets which are regularly arranged through a self-assembly method, heating the gelatin brackets to be closely arranged, then filling a PU solution, performing freeze drying to remove the solvent, and removing a gelatin template by a water bath method to obtain a three-dimensional porous PU bracket with an inverse opal structure; then, performing ammonia plasma treatment on the PU bracket to graft the amino group, connecting heparin to the amino group through EDC/NHS, then adding VEGF, and combining the VEGF to the heparin; and finally,rotationally inoculating MRC-5 cells to the VEGF-modified PU bracket, proportionally inoculating mixed cell suspensions of HUVECs and NL20 cells, and performing co-culture to obtain tissue engineeringartificial alveoli. The diameters of the prepared artificial alveoli can be effectively regulated and controlled to be about 300 microns by regulating and controlling the concentration of the gelatinsolution and polyurethane, the flow rate and the passage diameter of a water phase and an organic phase as well as 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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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
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