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Preparation method for microvesicle composite regenerative medical material

A technology of microbubbles and scaffold materials, applied in medical science, prostheses, etc., can solve the problems of seed cell growth and premature apoptosis, and achieve the effect of promoting angiogenesis, simple cryopreservation method, and less immune rejection

Inactive Publication Date: 2015-10-14
XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a preparation method of microbubble composite regenerative medical materials to solve the problem of premature apoptosis of seed cells due to the failure of blood vessels to grow in time during the tissue engineering construction process. provide new ideas

Method used

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  • Preparation method for microvesicle composite regenerative medical material
  • Preparation method for microvesicle composite regenerative medical material
  • Preparation method for microvesicle composite regenerative medical material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of microbubble / decalcified bone matrix composite regenerative medical material

[0026] 1. Extraction of rat bone marrow MSC-derived MV

[0027] SD rat bone marrow-derived MSCs were cultured in primary culture, and MSCs from the 3rd to 5th passages were used to extract MVs. Before extracting MVs, the MSC medium was replaced with a serum-free medium to continue culturing for 24 h. Collect the cell supernatant, centrifuge at 2,000×g for 20 minutes, discard the pellet and take the pellet, centrifuge at 100,000×g, 4°C for 1 hour, discard the supernatant, resuspend the pellet in PBS, and freeze at minus 80°C for later use.

[0028] 2. Preparation of MV / DBM composite regenerative medical materials

[0029] 1) The biological material used in this example is the decalcified bone matrix (DBM) commonly used in tissue engineering bone. The scaffold material was cut into a size of 3×3×3 mm, soaked in 75% alcohol for 2 hours, Wash 3 times with PBS and dry at...

Embodiment 2

[0033] Example 2 Preparation of the composite regenerative medical material of microbubble / polycaprolactone

[0034] 1. Extract mouse bone marrow MSC-derived MV

[0035]BALB / c mouse bone marrow-derived MSCs were cultured in primary culture, and MSCs from the 3rd to 5th passages were used to extract MVs. Before extracting MVs, the MSC medium was replaced with a serum-free medium to continue culturing for 24 h. Collect the cell supernatant, centrifuge at 2,000×g for 20 minutes, discard the pellet and take the pellet, centrifuge at 100,000×g, 4°C for 1 hour, discard the supernatant, resuspend the pellet in PBS, and freeze at minus 80°C for later use.

[0036] 2. Preparation of MV / PCL composite regenerative medical materials

[0037] 1) The biological material used in this example is polycaprolactone (polycaprolactone, PCL), which is commonly used in tissue engineering bone. The scaffold material was cut into a size of 3×3×3 mm, soaked in 75% alcohol for 2 hours, and washed with...

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Abstract

The invention belongs to the technical field of regenerative medicine and tissue engineering and discloses a preparation method for a microvesicle composite regenerative medical material. The preparation method comprises the following steps: (1), extracting mesenchymal stem cell derived microvesicles; and (2), inoculating microvesicles extracted in the step (1) to a tissue-engineered bone scaffold material, so as to obtain the microvesicle composite regenerative medical material through incubation. According to the invention, the mesenchymal stem cell derived microvesicles are inoculated to the tissue-engineered bone scaffold material to obtain a novel composite material, experiment results show that the composite material can increase the survival rate of seed cells in vivo, and the difficulty of early apoptosis of seed cells because blood vessels cannot grow in timely in a tissue engineering establishment process is solved. In addition, the mesenchymal stem cell derived microvesicles do not cause immunological rejection in vivo easily, have good biological safety, and can cross species to take a biological effect, therefore, the problem that a donor source is limited in the regenerative medicine engineering is solved.

Description

technical field [0001] The invention relates to the technical fields of regenerative medicine and tissue engineering, in particular to a preparation method of a microbubble composite regenerative medical material. Background technique [0002] The repair and functional reconstruction of tissue and organ defects are still difficult problems faced by modern medicine. Autologous or allogeneic tissue and organ transplantation are currently the most commonly used treatment methods, but there are shortcomings such as a serious shortage of donor sources. The concept of tissue engineering is expected to solve this problem. Tissue engineering is dedicated to the research on the technology of constructing human organs or tissues in vitro, using biodegradable and compatible materials to prepare 3D scaffolds that meet the shape of the required organs or tissues, so that the stem cells adsorbed on the materials gradually grow in a specific way. It grows into the required organs and tis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/54
Inventor 谢慧陈智超张文杰汪振星
Owner XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
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