Preparation method and application of ossein polypeptide for promoting human osteoblast cell proliferation

A technology of osteoblasts and collagen, applied in the field of bone processing, can solve the problems of not mentioning the difference of proliferation effect, and achieve the effect of small residue, good decalcification effect and good degreasing effect.

Inactive Publication Date: 2017-12-29
INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology

However, there is no mention of the difference in the proliferation effect of polypeptides with molecular weights

Method used

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  • Preparation method and application of ossein polypeptide for promoting human osteoblast cell proliferation
  • Preparation method and application of ossein polypeptide for promoting human osteoblast cell proliferation
  • Preparation method and application of ossein polypeptide for promoting human osteoblast cell proliferation

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Embodiment 1

[0049] The present invention provides a preparation method of collagen polypeptide that promotes the proliferation of human osteoblasts, comprising:

[0050]Step 1, degreasing and decalcifying the bovine bone raw material, then dissolving with acetic acid, and collecting the supernatant;

[0051] Step 2, adding 0.8mol / L NaCl solution to the supernatant obtained in step 1 for salting out, centrifuging, and collecting the precipitate;

[0052] Step 3. Dissolve the precipitate obtained in step 3 with 0.5mol / L acetic acid, put it into a dialysis bag with a flux of 12kDa, first dialyze with 0.1mol / L acetic acid for 2 days, then dialyze with distilled water for 2 days, and change the acetic acid every 12 hours or distilled water to collect the collagen in the dialysis bag;

[0053] Step 4: Add 2 mg / mL alkaline protease to the bone collagen obtained in Step 3, perform enzymatic hydrolysis for 1 hour under the conditions of pH 8 and temperature 40° C., and collect the enzymatic hydro...

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Abstract

The invention discloses a preparation method of an ossein polypeptide for promoting human osteoblast cell proliferation. The preparation method comprises the following steps: step one, carrying out degreasing and decalcification on a bone raw material, then dissolving the bone raw material with acetic acid, and collecting a supernate; step two, adding 0.7 to 1.2 mol/L of an NaCl solution into the supernate obtained in the step one for salting-out, centrifuging the solution, and collecting a sediment; step three, dissolving the sediment obtained in the step three with acetic acid, and putting the solution into a dialysis bag with the flux of 10 to 14 kDa for dialysis, and collecting ossein protein inside the ossein bag; step four, adding the ossein protein obtained in the step three into 2 to 5 mg/Ml of alkaline protease, and collecting an enzymatic hydrolysate; and step five, loading the enzymatic hydrolysate obtained in the step four into a dialysis bag with the flux of 1 kDa, putting the dialysis bag in water for dialysis, collecting a dialysis fluid, and freeze-drying. The invention also provides application of the ossein polypeptide. The preparation method and the application have a remarkable promotion effect on the proliferation of human osteoblast cells.

Description

technical field [0001] The invention relates to the technical field of bone processing. More specifically, the present invention relates to a preparation method of collagen polypeptide that promotes the proliferation of human osteoblasts. Background technique [0002] my country is rich in edible bone resources. At present, my country's annual output of livestock and poultry bones is as high as 12 million tons, accounting for 1 / 3 of the world's output, equivalent to about 2 million tons of animal protein, which can meet the annual protein demand of 75 million people. Our country is about to enter an aging society, and osteoporosis has become the biggest chronic threat to the elderly. Under the conditions of population pressure and limited carrying capacity of breeding environment, strengthening the comprehensive development and utilization of bone resources has important economic, social and environmental benefits. [0003] Studies have shown that collagen peptides can sig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K14/78C07K1/34
CPCC07K14/78C12P21/06
Inventor 李侠张春晖贾伟
Owner INST OF AGRO FOOD SCI & TECH CHINESE ACADEMY OF AGRI SCI
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