Preparation method of animal decellularized lung biological scaffold material

A decellularization and bio-scaffold technology, applied in tissue regeneration, medical science, prosthesis, etc., can solve problems such as unstable effect, low decellularization efficiency, and large damage to extracellular matrix components, and achieve enhanced tolerance , weaken the effect of contradiction

Active Publication Date: 2020-08-07
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the current methods for obtaining decellularized lung bioscaffold materials at home and abroad are mainly suitable for small animals such as rats, and when applied to large animals, the decellularization efficiency is low, the effect is unstable, or the extracellular matrix components are damaged (refer to Literature Petersen TH, Calle EA, Zhao L, Lee EJ, Gui L, Raredon MB, Gavrilov K, Yi T, Zhuang ZW, Breuer C, Herzog E, Niklason LE. Tissue-engineered lungs for invivoimplantation. Science. 2010; 329: 538–541; Ott HC, Clippinger B, Conrad C, Schuetz C, Pomerantseva I, Ikonomou L, Kotton D, Vacanti JP. Regeneration and orthotopic transplantation of abioartificial lung. Nat Med. 2010; 329:538-541), decellularized The conflict between efficiency and ECM protection is more prominent

Method used

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  • Preparation method of animal decellularized lung biological scaffold material
  • Preparation method of animal decellularized lung biological scaffold material
  • Preparation method of animal decellularized lung biological scaffold material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of decellularized porcine lung bioscaffold material

[0043]Animals were anesthetized with Zytex, and after systemic heparinization, the lungs were removed. Insert and secure catheters from the pulmonary artery and trachea. Perfuse 2L of PBS solution containing heparin (enough dose is sufficient for anticoagulation) from the cannula of the pulmonary artery (it can also be other isotonic solutions containing anticoagulant substances, such as: aqueous solution containing calcium ion chelating agent, low molecular weight heparin , heparin analogs, etc.), the perfusion time is 0.5 hours, and then placed in a -20°C refrigerator for at least 24 hours. After the pig lungs were fully thawed, 0.9% m / v normal saline and 0.2% m / v dextrose solution (osmotic pressure 10mOsm / L) were perfused successively from the trachea at a flow rate of 100 mL / min, and the perfusion time was 1 hour and 3 hours, respectively. Hour. Afterwards, 1% v / v SLES (or other ionic det...

experiment example 1

[0052] Experimental Example 1 Comparison of pig lung bio-scaffold materials with and without dextrose solution perfusion

[0053] In this experimental example, on the basis of Example 1, the step of treating the ECM protective agent dextrose was selectively omitted, and as a control, the difference between it and the biological scaffold material obtained in Example 1 was compared.

[0054] The result is as Figure 4-Figure 9 Shown:

[0055] Figure 4 Decellularized porcine lung bioscaffold material treated with dextrose solution perfusion ( Figure 4 A) and pig lung bioscaffold material without dextrose solution perfusion treatment ( Figure 4 B), the results show that the texture of the decellularized porcine lung bioscaffold treated with dextrose solution perfusion is firmer and better maintains the shape of the original organ.

[0056] Figure 5 with Image 6 It is the results of the collagen and glycosaminoglycan GAG content of the porcine lung bioscaffold treated wi...

experiment example 2

[0059] Comparison of Experimental Example 2 Degradability and Immunogenicity

[0060] In order to test the biodegradability and immunogenicity of bioscaffolds, pig lung bioscaffolds treated with and without dextrose solution perfusion were implanted into the omentum of rats. At the 1st and 4th week of implantation, the material was removed and fixed with 4% formalin, and stained after sectioning.

[0061] The result is as Figure 9 Shown: the extracellular matrix components of the decellularized porcine lung bioscaffold treated with dextrose solution perfusion degrade more slowly, the number of neutrophils and lymphocytes infiltrated around the scaffold material is less, and the immunogen caused by Sex is lower.

[0062] Conclusion: The decellularized porcine lung bioscaffold treated with dextrose solution perfusion degrades slowly in animals, and can provide a more stable and long-lasting scaffold for stem cell attachment, propagation, and differentiation in tissue engineer...

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Abstract

The invention discloses a preparation method of an animal decellularized lung biological scaffold material, and belongs to the field of tissue engineering and regenerative medicine. According to a traditional decellularized lung biological scaffold material, cells are removed by pouring a detergent, extracellular matrix (ECM) can be damaged while the detergent removes the cells, and then the scaffold structure is affected. According to the method disclosed by the invention, the ECM protective agents such as a dextrose solution are creatively used for filling the trachea, so that the damage ofthe detergent to the ECM is reduced, the contradiction between ECM retention and cell component removal is more easily balanced, and a stent material with more thorough cell removal and more intact ECM is obtained.

Description

technical field [0001] The invention belongs to the field of tissue engineering and regenerative medicine. Background technique [0002] Pulmonary failure caused by chronic obstructive pneumonia, pulmonary fibrosis and other diseases has become a major problem in the medical field. So far, lung transplantation is the only effective radical cure in clinical practice, but its development is restricted by factors such as shortage of donor lungs, difficulty in storing donor lungs during transport, and complicated surgical procedures. As a potential alternative treatment for orthotopic organ transplantation, tissue engineered organ technology has the advantages of diverse sources and simple treatment methods, and is gradually being valued. [0003] A number of studies have shown that biological scaffolds play a key role in supporting the distribution, differentiation, regeneration, and maintenance of cell functions, and are one of the cores of research in the fields of tissue en...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3633A61L27/3687A61L27/3691A61L27/50A61L2430/40
Inventor 李奕李为民
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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