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Preparation method of microcarrier used for cell three dimensional multiplication culture

A technology of amplifying culture and microcarriers, which is applied in the field of tissue engineering materials to achieve the effect of good biological performance

Inactive Publication Date: 2018-02-09
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been reported that polylactic acid is composited with hydroxyapatite to construct microcarriers.

Method used

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  • Preparation method of microcarrier used for cell three dimensional multiplication culture
  • Preparation method of microcarrier used for cell three dimensional multiplication culture
  • Preparation method of microcarrier used for cell three dimensional multiplication culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation of L-polylactic acid microspheres and biocompatibility detection (comparative example, the concentration of L-polylactic acid in the dichloromethane solution of L-lactic acid is 10g / mL, and the amount of gelatin is: gelatin / L-lactic acid mass ratio 1:40, mixing speed 150rpm / min / min)

[0031] First, weigh 10.0g of poly-L-lactic acid (PLLA) and add it to 100mL of dichloromethane solution, stir in a fume hood for 4h until PLLA dissolves to obtain PLLA in dichloromethane solution; then weigh 0.25g of gelatin and add it to 100mL of distilled water, Seal and stir for 3 hours to obtain a gelatin dispersant; add the PLLA dichloromethane solution to the gelatin dispersant solution, and stir for 7 hours in a fume hood at a speed of 150 rpm / min. During the stirring process, the liquid microspheres gradually solidify as the organic solvent volatilizes , into balls, and precipitate at the bottom of the beaker; stand for 30 minutes to separate the phases, separa...

Embodiment 2

[0033] Example 2 Preparation of L-polylactic acid / nanometer hydroxyapatite (mass ratio 5:1) composite microspheres (polylactic acid dichloromethane solution concentration 5g / mL, gelatin dosage: gelatin / L-polylactic acid mass ratio 1:20, Mixing speed 200rpm / min)

[0034] Weigh 5.0g of L-polylactic acid (PLLA), add it into 100mL of dichloromethane, and stir magnetically for 3h under water bath conditions until PLLA is completely dissolved in dichloromethane to obtain PLLA dichloromethane solution; weigh 1.0g of nano Hydroxyapatite (nHAp), add it to the above PLLA dichloromethane solution, seal it, stir rapidly on a magnetic stirrer for premixing, and further ultrasonically disperse and mix for 30min in an ultrasonic oscillator until nHAp is evenly dispersed in the solution , to obtain PLLA / nHAp dichloromethane solution; weigh 0.25g gelatin, add it to 100mL distilled water, seal and stir for 2h until the gelatin dissolves, and obtain a gelatin dispersant; add the prepared PLLA / nH...

Embodiment 3

[0035] Example 3 Preparation of L-polylactic acid / nanometer hydroxyapatite (mass ratio 5:1) composite microspheres (polylactic acid dichloromethane solution concentration 10g / mL, gelatin dosage: gelatin / L-polylactic acid mass ratio 1:40, Mixing speed 200rpm / min)

[0036] Weigh 10.0g of L-polylactic acid (PLLA), add it to 100mL of dichloromethane, and stir magnetically for 4h under water bath conditions until PLLA is completely dissolved in dichloromethane to obtain PLLA dichloromethane solution; weigh 2.0g of nano Add hydroxyapatite (nHAp) to the above-mentioned PLLA dichloromethane solution, seal it, stir rapidly on a magnetic stirrer for premixing, and further ultrasonically disperse and mix in an ultrasonic oscillator for 50 minutes until nHAp is evenly dispersed in the solution. Obtain PLLA / nHAp dichloromethane solution; Weigh 0.25g of gelatin, add it to 100mL distilled water, seal and stir for 2h until the gelatin is dissolved, and obtain a gelatin dispersant; add the pre...

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Abstract

The invention discloses a preparation method of a microcarrier used for cell three dimensional multiplication culture, and belongs to the field of tissue engineering material. According to the preparation method, Poly-L-lactic acid (PLLA) is taken as a matrix, gelatin is taken as a dispersant, and composite microspheres are prepared; the particle size of the composite microspheres is adjusted viacontrolling the concentration of a PLLA dichloromethane solution, the mass ratio of nano-hydroxyapatite (nHAp) to PLLA, and the using amount of gelatin; the PLLA is taken as the matrix to ensure the mechanical strength and biological degradability of a support; composite nHAp is capable of improving the osteo inductivity and the conductivity of the support, and at the same time, assisting PLLA toimprove cell osteogenic capability, and avoiding damage of cells under an overly acidic environment; the existing of nHAp is capable of forming a large amount of small pores on the surfaces of the PLLA microspheres, the existing of the small pores is capable of improving the mass transfer capability of the support, and providing cells with more abundant attachment sites. The microcarrier possessesexcellent biological performance on osteoblasts, and is suitable to be used in the field of bone tissue engineering.

Description

technical field [0001] The invention belongs to the field of tissue engineering materials, in particular to a method for compounding L-polylactic acid / nanometer hydroxyapatite and constructing tissue engineering cell microcarriers. Background technique [0002] Tissue engineering is the preparation and cultivation of artificial tissue substitutes in vitro through cell biology, material molecular science and engineering principles. Substitute products are biologically active and perform many clinical functions, mainly in repairing and improving tissues and organs in the human body, and maintaining them. One of its methods is to prepare biological scaffolds on which cells are combined with growth factors to repair damaged tissues and organs in a new way. This alternative method has been widely used clinically. The three elements of tissue engineering are scaffold materials, growth factors and seed cells, and they coordinate and work closely with each other. In recent years, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/46A61L27/58A61L27/50
CPCA61L27/46A61L27/50A61L27/58A61L2400/12A61L2430/02C08L67/04C08L89/00
Inventor 宋克东李丽颖孔倩李文芳刘天庆
Owner DALIAN UNIV OF TECH
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