Compound growth factor as well as preparation method and application thereof

A compound growth factor, growth factor technology, applied in the field of gel, to achieve high safety, easy to use, and promote the effect of granulation tissue proliferation

Inactive Publication Date: 2013-03-27
GUANGZHOU GENERAL HOSPITAL OF GUANGZHOU MILITARY COMMAND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Tissue-engineered skin provides a new approach for the clinical treatment of various types of skin...

Method used

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  • Compound growth factor as well as preparation method and application thereof
  • Compound growth factor as well as preparation method and application thereof
  • Compound growth factor as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Determination of the concentration of growth factors suitable for clinical treatment

[0028] Platelet gel (PG) supernatant prepared from platelet-rich plasma (PRP) with different concentrations of platelets on ECV304 cell proliferation experiment: the concentration of platelets in the control group was 0×10 9 / L, the concentrations of the experimental groups were 500×10 9 / L, 1000×10 9 / L, 1500×10 9 / L, 2000×10 9 / L, 2500×10 9 / L, 3000×10 9 / L six concentrations; the activator selected 10% calcium gluconate solution containing 1000U / ml human blood thrombin, the ratio of PRP and activator was 10:1, and the cell proliferation was detected at 24h.

[0029] The results showed that in the platelet concentration range of 1500~2500×10 9 / L, with the increase of platelet concentration, the prepared platelet gel (PG) supernatant has a significant effect on promoting cell proliferation; when the number of platelets reaches 1000-2500×10 9 When t...

Embodiment 2

[0030] The preparation method of embodiment 2 composite growth factors

[0031](1) Virus inactivation of platelet concentrate: the platelet concentrate is packed in a container containing an organic solvent / detergent mixture (S / D solution) Choose a mixture of 1% TnBP and 1% TritonX-45, shake the mixture of organic solvent / detergent mixture (S / D solution) and platelet concentrate for one minute to mix thoroughly, and then place it in a water tank at 31°C , oscillating continuously for at least one hour;

[0032] (2) Oil extraction: add an appropriate amount (10% (v / v)) of soybean oil to the mixture of organic solvent / detergent mixture (S / D solution) and platelet concentrate after shaking in step (1), Shake well, then shake on a rotating shaker for 20 minutes, the platelet part (lower layer) is poured out from the oil (upper layer), and put into another container under the action of gravity for a second oil extraction; after oil extraction Reduce both TnBP and Tr...

Embodiment 3

[0036] Example 3 Determination of Growth Factors

[0037] Extract 1ml samples from the original platelet concentrate, the oil-extracted platelet concentrate, and the platelet concentrate after column chromatography, and carry out the following treatment: centrifuge the sample at 10,000×g for 15 minutes to obtain Platelet and cell lysates were then snap frozen in a -80°C freezer. Use an ELISA kit to measure, follow the instructions of the ELISA kit, melt at 37°C, and analyze within 1 hour. Standards and samples were determined in duplicate and average values ​​were calculated. The result is multiplied by the dilution factor of the sample.

[0038] The composite growth factor prepared in Example 2 was measured by the above method, and it was found that in the composite growth factor prepared in Example 2, the platelet growth factor (PDGF) contained in 1L of platelet concentrate was 7-33 μg, and the vascular endothelial growth Factor (VEGF) is 0.2~0.6μg, transforming growth fa...

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Abstract

The invention discloses a compound growth factor of blood platelet sources as well as a preparation method and application of the compound growth factor. According to the compound growth factor as well as the preparation method and the application of the compound growth factor, platelet-rich plasma as the raw material is firstly subjected to virus inactivation through an S/D (Solvent/Detergent) method to remove such reagents as organic solvents and eradicators in the platelet-rich plasma and then activated by adding calcium or/and thrombin, supernatant fluid after activation is extracted to obtain the compound growth factor fitting for the natural proportion of the human body; and the problems that single recombinant growth factors in clinic are high in price, and difficult to purify, is involved into gene recombination biological safety and on the like are overcome. The invention further discloses a compound growth factor gel obtained based on the compound growth factor by adding chitosan microspheres carriers, which can slowly release growth factors during use. The compound growth factor has remarkable treatment effects for acute wounds, burns, cosmetic treatment, and chronic hard-healing wounds, is convenient to use, and has high safety.

Description

technical field [0001] The invention relates to a composite growth factor and its preparation method, and also relates to a gel obtained by processing the composite growth factor. The invention also relates to the therapeutic effect of the composite growth factor and the composite growth factor gel in acute wounds, burns, plastic surgery, chronic difficult-to-heal wounds and the like. Background technique [0002] Chronic intractable ulcers are defined as wounds that fail to heal through the normal wound healing process under the influence of various internal or external factors, and enter a state of pathological inflammatory response, resulting in prolonged wound healing. As my country gradually enters an aging society, the elderly population is increasing, especially in the elderly population with diabetes, vasculitis and lower extremity venous reflux disorder. Such patients have slow wound healing, need long-term hospitalization, long-term dressing changes, and often und...

Claims

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Application Information

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IPC IPC(8): A61K38/48A61K38/30A61K38/18A61K9/00A61K35/14A61P17/02A61K31/191A61K35/19
Inventor 单桂秋马静李艳辉丘金浪谭美芳耿文艳
Owner GUANGZHOU GENERAL HOSPITAL OF GUANGZHOU MILITARY COMMAND
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