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Tissue-engineered bone and preparation method thereof

A tissue-engineered bone and bone marrow mesenchymal technology, applied in medical science, prosthesis, etc., can solve the problems of affecting cell activity, high equipment requirements, inapplicability, etc., to achieve the effect of convenient method, reduce pollution, and avoid aging

Active Publication Date: 2014-12-24
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the conditions of temperature control are relatively complicated, and the requirements for equipment are high. Occasional changes in temperature also affect the activity of cells and accelerate the aging of some cells. It is not suitable for human osteoblast cells that are more sensitive to temperature (Fukumori K, Akiyama Y , Yamato M, Kobayashi J, Sakai K, Okano T. Temperature responsive glass coverslips with an ultrathin poly(N-isopropylacrylamide) layer. Acta Biomater 2009;5:470-476)

Method used

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  • Tissue-engineered bone and preparation method thereof
  • Tissue-engineered bone and preparation method thereof
  • Tissue-engineered bone and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0031] 1) Preparation of photosensitive cell culture dish

[0032] Titanium oxide nanoparticles were uniformly dispersed in water to form a dispersion with a mass concentration of 20%, and the dispersion was prepared into a precursor solution with methanol and tetrahydrofuran at a ratio of 0.06:9:3; then the above precursor solution was mixed at 48 μL / cm 2 Evenly drop it on the polystyrene culture dish, put it into a 65°C oven to dry, and after drying, a cell culture dish containing a photoresponsive nano-titanium oxide film with a grain size of 20-30nm and a thickness of 60nm-100nm can be obtained;

[0033] 2) Cultivate bone marrow mesenchymal stem cell sheets

[0034] Take human bone marrow mesenchymal stem cells from passage 3 to 5, and use 2×10 5 piece / cm 2 The density is implanted into a petri dish containing nano-titanium oxide with a diameter of 5cm. After culturing for 10 days, use a wavelength of 450nm and an intensity of 4mW / cm 2 Ultraviolet irradiation for 5 mi...

Embodiment 2

[0043] 1) Preparation of photosensitive cell culture dishes

[0044] Uniformly disperse the zinc oxide nanoparticles in water to form a dispersion liquid with a mass concentration of 10%, prepare the precursor solution with the dispersion liquid, ethanol and chloroform at a ratio of 0.02:6:1, and then mix the above precursor solution with 25 μL / cm 2 Evenly drop it onto a polystyrene culture dish, put it into a 65°C oven for drying, and then obtain a cell culture dish containing a light-responsive nano-zinc oxide film with a grain size of 10-20nm and a thickness of 40nm-80nm.

[0045] 2) Culture bone marrow mesenchymal stem cells

[0046] Take the homologous human bone marrow mesenchymal stem cells after several generations of culture, and use 5×10 4 piece / cm 2 The density is implanted in a petri dish containing nano zinc oxide with a diameter of 5cm. After culturing for 4 days, the 2 Ultraviolet irradiation for 30 minutes resulted in exfoliated sheets of human bone marr...

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Abstract

The invention discloses a tissue-engineered bone. The tissue-engineered bone is multilayer cell sheet lamination compound with a three-dimensional capillary network, the multilayer cell sheet lamination compound consists of n laminated bone marrow mesenchymal stem cell sheets and vascular endothelial cells among all bone marrow mesenchymal stem cell sheet layers, and n is in a range of 3-8. A preparation method of the tissue-engineered bone comprises steps as follows: firstly, a photosensitive semiconductor structural layer is prepared on the surface of a cell culture dish, so that a photosensitive cell culture dish is obtained; then bone marrow mesenchymal stem cells are cultured with the photosensitive cell culture dish, and the bone marrow mesenchymal stem cell sheets are obtained; finally, the multilayer cell sheet lamination compound consisting of multiple layers of the bone marrow mesenchymal stem cell sheets and the vascular endothelial cells is constructed and cultured to obtain the tissue-engineered bone. The tissue-engineered bone purely consists of homologous cells, so that pollution caused by immunological rejection and stent degradation is reduced, and the tissue-engineered bone is significant in bone defect repair and early vascularization; the method is simple, easy to implement and convenient to popularize.

Description

technical field [0001] The invention relates to a tissue engineering bone and a preparation method thereof, in particular to a tissue engineering bone composed of multilayer cell sheets and a preparation method thereof, belonging to the field of tissue engineering. Background technique [0002] Since bone tissue defects and loss caused by congenital malformations, trauma, tumors, etc. are common clinical diseases, the functional reconstruction and repair of bone tissue defects has always been an important topic in regenerative medicine. Traditional treatment methods often use autologous or allogeneic bone transplantation, but its clinical application is often limited due to limited supply sources or immune rejection. As one of the most widely used artificial bone substitute materials, tissue engineering technology has been gradually replaced by its biodegradability. For example, "An Artificial Bone Material and Its Preparation Method" disclosed in CN103285427A, the arti...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/50A61L27/58
Inventor 王慧明俞梦飞洪逸董灵庆
Owner ZHEJIANG UNIV
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