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A method for preparing hydroxyapatite using red blood cells as a bioreactor

A hydroxyapatite, bioreactor technology, applied in chemical instruments and methods, phosphorus compounds, inorganic chemistry, etc., to achieve the effects of temperature controllable reaction conditions, environmental friendly reaction conditions, and simple operation

Inactive Publication Date: 2019-09-13
HENAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of the relevant reports use microorganisms as reactors, such as: yeast, Escherichia coli, Shewanella, etc., but there are few reports on the synthesis of inorganic nanomaterials using red blood cells as reactors

Method used

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  • A method for preparing hydroxyapatite using red blood cells as a bioreactor
  • A method for preparing hydroxyapatite using red blood cells as a bioreactor

Examples

Experimental program
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Effect test

Embodiment 1

[0021] The upper layer of plasma was removed by centrifugation from pig blood whole blood and washed with normal saline for 3 times to obtain red blood cell pellet, and 0.5mL red blood cell pellet was suspended in 5mL containing 40mM CaCl 2 Add 0.5g of glucose to normal saline, react in an ice-water bath for 3 hours, then centrifuge and wash to fully wash away the CaCl outside the cells 2 , the centrifuged cell pellet was collected and resuspended in 5 mL of 24 mM PBS buffer solution with pH=9, placed in a shaker (oscillating speed of 90 rpm) at 10 ° C for 6 h, centrifuged and washed, and finally obtained the hydroxyapatite-containing Functionalized red blood cells are pulverized, separated and purified to obtain hydroxyapatite nanoparticles with an average particle size of 5.6 nm.

[0022] figure 1 It is the XRD spectrum of the hydroxyapatite nanoparticles prepared in this example, which is consistent with the standard spectrum of hydroxyapatite (JCPDS NO.09-0432), and each ...

Embodiment 2

[0024] The upper layer of plasma was removed by centrifugation from sheep blood whole blood and washed 3 times with normal saline to obtain red blood cell pellet, and 0.5mL red blood cell pellet was suspended in 5mL containing 30mM CaCl 2 Add 0.5g of glucose to normal saline, react in an ice-water bath for 3 hours, then centrifuge and wash to fully wash away the CaCl outside the cells 2 , the centrifuged cell pellet was collected and resuspended in 5 mL of 18 mM PBS buffer solution with pH=9, placed in a shaker (oscillating speed of 50 rpm) at 4°C for 6 h, and then washed by centrifugation to finally obtain hydroxyapatite-containing Functionalized erythrocytes are pulverized, separated and purified to obtain hydroxyapatite nanoparticles with an average particle size of about 5.6 nm.

Embodiment 3

[0026] The upper layer of plasma was removed by centrifugation from whole rabbit blood and washed three times with normal saline to obtain the red blood cell pellet, and 0.5mL red blood cell pellet was suspended in 5mL containing 20mM CaCl 2 Add 0.5g of glucose to normal saline, react in an ice-water bath for 2 hours, then centrifuge and wash to fully wash away the CaCl outside the cells 2 , the centrifuged cell pellet was collected and resuspended in 5mL of 12mM PBS buffer solution with pH=9, placed in a shaker (oscillation speed: 120rpm) at 25°C for 4h, then centrifuged and washed to obtain hydroxyapatite-containing Functionalized red blood cells are pulverized, separated and purified to obtain hydroxyapatite nanoparticles with an average particle size of 5.6 nm.

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Abstract

The invention discloses a preparation method of hydroxyapatite by using red blood cells as bioreactor, which belongs to the technical field of synthesis of inorganic nano materials. The technical proposal of the invention comprises the following main points that the upper plasma is removed from animal blood or human blood by centrifugation and the remainder is washed with physiological saline several times to obtain erythrocyte sedimentation, the erythrocyte sedimentation is added to the physiological saline containing soluble barium salt and the reaction is carried out under the condition of ice-water bath, then the mixture obtained by the reaction is centrifuged and washed after the reaction, the erythrocyte sedimentation obtained by the centrifugation is collected and resuspended in the PBS buffer solution(PH=9) to carry out the reaction, and functional red blood cells containing hydroxyapatite are obtained by centrifugation after the reaction, then the hydroxyapatite nanoparticles with an average particle size of 5.6 nm are prepared by the crush, separation and purification of the functional red blood cells. The invention has a series of advantages of green environmental protection, simple operation, low cost and good biocompatibility.

Description

technical field [0001] The invention belongs to the technical field of synthesis of inorganic nanometer materials, and in particular relates to a method for preparing hydroxyapatite by using red blood cells as a bioreactor. Background technique [0002] At present, the existing chemical synthesis methods of inorganic nanomaterials have the disadvantages of poor stability and easy agglomeration; or environmental pollution, high synthesis cost, complex and difficult to control synthesis process, poor repeatability, and harsh reaction conditions. . With the development of science and technology, the focus of chemical synthesis has changed from traditional chemical methods to a new green synthesis method, and biosynthesis methods have emerged as the times require. Nano-biosynthesis technology mainly utilizes the biologically active molecules of organisms (such as proteins, polysaccharides, etc.) to synthesize new nanomaterials. It has the characteristics of low energy consumpt...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C01B25/32B82Y40/00
Inventor 杨林谢晓晓陈会锋茹祥利
Owner HENAN NORMAL UNIV
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