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Preparation and application of pcl-pla tissue engineering composite scaffold against hepatocyte senescence

A tissue engineering and composite scaffold technology, which is applied in the field of preparation of PCL-PLA tissue engineering composite scaffolds, can solve the problems of cell culture vascularization, immune rejection, xenovirus infection, etc., and achieves improved cell growth state, good anti-aging effect, The effect of significant anti-aging effect on cells

Active Publication Date: 2017-05-03
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nowadays, liver tissue engineering research is still in its infancy, and there are still many problems, such as selection of heterogeneous liver cells will face immune rejection and heterogeneous virus infection; vascularization of cell culture in bioreactors; tissue and organ culture and construction in vitro, etc. problem

Method used

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  • Preparation and application of pcl-pla tissue engineering composite scaffold against hepatocyte senescence
  • Preparation and application of pcl-pla tissue engineering composite scaffold against hepatocyte senescence
  • Preparation and application of pcl-pla tissue engineering composite scaffold against hepatocyte senescence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Preparation of PCL-PLA Tissue Engineering Composite Scaffold Against Hepatocyte Aging

[0068] 1. Preparation of free IGF and TNF-a

[0069] Insulin-like growth factor (IGF) and tumor necrosis factor-alpha (TNF-α) were purchased from Zhongshan Third Hospital, and sterile PBS, pipette tips, droppers, reagent bottles, EP tubes, etc. were prepared.

[0070] Dissolve IGF (45 μg, 0.02 μmol) and TNF-α (40 μg, 0.02 μmol) in PBS solution respectively, and after adjusting to a concentration of 1 ng / μl, use a disposable filter membrane (a filter membrane with a pore size of 10,000 Mw) Filter and sterilize them separately to obtain free IGF solution and free TNF-a solution, which are divided into EP tubes and labeled for later use.

[0071] 2. Preparation of co-immobilized IGF and TNF-a

[0072] (1) Add DMF:PBS 4:1 buffer solution on the basis of free IGF and free TNF-a to help it dissolve, add a magnetic stirrer, put it in a magnetic stirrer and stir in an ice bath fo...

Embodiment 2

[0083] Example 2 Characterization of PCL-PLA Tissue Engineering Composite Scaffold Against Hepatocyte Aging

[0084] In this example, for the anti-hepatocyte aging PCL-PLA tissue engineering composite scaffold prepared in Example 1, Raman spectrum, infrared spectrum, scanning electron microscope, X-ray Photoelectron spectroscopy and other methods can be used to judge whether the synthesis is successful or not.

[0085] 1. Raman spectral analysis

[0086] (1) Analysis method

[0087] Spectra of Raman scattering. In 1928, C.V. Raman experiment found that when light passes through a transparent medium and is scattered by molecules, the frequency changes. This phenomenon is called Raman scattering , later in the same year at Soviet Union and France was also observed. Scattering in transparent media spectrum Among them, the component with the same frequency as the frequency of the incident light υ0 is called Rayleigh scattering; the spectral line or band υ0±υ1 that is sy...

Embodiment 3

[0106] Example 3 Application of PCL-PLA Tissue Engineering Composite Scaffold Against Hepatocyte Aging

[0107] 1. Culture of hepatocellular carcinoma cells (HepG2)

[0108] The source of the cells is the Medical College of Jinan University. The shape of the liver cancer cells (HepG2) is round under the microscope. The culture method of the liver cancer cells is basically the same as that of the human stellate hepatocytes. Use sterile PBS buffer to wash away the cells that are not firmly attached to the wall, digest with PBS(-) solution containing trypsin 0.25% (W / V) and EDTA 0.02% (W / V) for about 2 minutes, add an equal amount of Low-sugar serum-containing DMEM culture medium (containing 10% newborn calf serum, 0.06 mg / ml penicillin, 0.10 mg / ml streptomycin, pH 7.2-7.4), gently pipet with a sterilized dropper, and collect the cell suspension into a sterile centrifuge tube. Centrifuge at 1000 r / min for 5 min. Discard the supernatant and inoculate into culture flasks. Put a...

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Abstract

The invention discloses a PCL-PLA (polycaprolactone-polylactic acid) tissue engineering composite scaffold with a liver cell anti-aging function. The PCL-PLA tissue engineering composite scaffold is made of scaffold materials on the basis of PCL-PLA, and polyacrylamide, IGF (insulin-like growth factors), TNF-alpha (tumor necrosis factors-alpha) are subjected to photo-grafting to carry out synergetic joint modification to form the PCL-PLA tissue engineering composite scaffold. The PCL-PLA tissue engineering composite scaffold has the advantages that materials with positive charge and dual growth factors are jointly modified on the basis of the three-dimensional scaffold materials, accordingly, obvious cell anti-aging effects can be realized by the prepared PCL-PLA tissue engineering composite scaffold [PCL-PLA-PAAm (polyacrylamide)-IGF / TNF-alpha], cells can be attached to the scaffold to grow advantageously, and tissues and even organs can be possibly formed by the cells; the cells are cultivated on the three-dimensional scaffold, the PCL-PLA tissue engineering composite scaffold effectively conforms to internal human cell life environments, growth states of the cells can be obviously improved, the PCL-PLA tissue engineering composite scaffold has a practical significance on clinical application, positive promotion effects can be realized for liver tissue engineering research, and directions can be provided for cell anti-aging research in tissue engineering.

Description

technical field [0001] The invention belongs to the technical field of biomaterials. More specifically, it relates to the preparation and application of an anti-aging PCL-PLA tissue engineering composite scaffold. Background technique [0002] The liver is the largest solid organ in the human body and one of the most important vital organs of the body. It plays important physiological functions in the body, including digestion, detoxification, synthesis, secretion, etc., and is also responsible for a series of complex physiological functions such as blood coagulation, immunity, metabolism, heat production, and water and electrolyte balance. my country is an area with a high incidence of chronic liver disease, and its morbidity and mortality are high. Acute and chronic liver diseases, especially liver failure and liver cancer, seriously threaten human life and health. [0003] At present, liver transplantation is still the only feasible cure for many liver diseases, but in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/18A61L27/22A61L27/54
Inventor 关燕清杨润财
Owner SOUTH CHINA NORMAL UNIVERSITY
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