Small peptide for promoting osteoblast proliferation
A technology for osteoblasts and osteoblast activity, applied in the field of small peptides that promote osteoblast proliferation, can solve problems such as side effects of calcitonin
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Embodiment 1
[0025] Embodiment 1 prepares the method for the active peptide that promotes osteoblast proliferation
[0026] A method for preparing an active peptide that promotes osteoblast proliferation, comprising the steps of:
[0027] (1) Enzymatic hydrolysis of papain
[0028] Soybean protein isolate (Shanghai Yuanye Biotechnology Co., Ltd., product number S30914) was weighed and dissolved in ultrapure water to prepare a protein solution with a concentration of 4.5% by mass, and NaOH solution was added to adjust the pH to 7.0. According to the ratio of adding 3000U papain per gram of protein, papain was added to the protein solution, and then placed on a rotary mixer, and the enzymolysis reaction was carried out at 55°C for 5 hours. After testing, the hydrolysis degree of soybean protein isolate was 11.08± 0.31%. After the reaction, raise the temperature to 85°C and keep it warm for 20 minutes to inactivate papain, cool to room temperature, centrifuge at 3000×g for 15 minutes, take ...
Embodiment 7
[0044] The effect of embodiment 7 small peptide A on osteoblast ALP (alkaline phosphatase) activity
[0045] Alkaline phosphatase kit was used to detect the effect of small peptide A on ALP activity of osteoblasts. The specific method is as follows: after digesting osteoblast MC3T3-E1, use α-MEM culture solution to prepare the concentration of 1×10 5 cells / mL of cell suspension, inoculated in a 6-well plate, 2 mL per well. After the MC3T3-E1 cells grew to confluence, the culture medium was removed, and they were cultured in mineralization-inducing medium (ODM purchased from Sigma, USA, with a final concentration of 50 μg / mL ascorbic acid and 4 mM β-glycerophosphate) 7 and 10 days to induce differentiation, then replace the serum-free medium, and add small peptide A with a final concentration of 20 μM for 48 hours, and set 3 parallels. At the same time, a control group was set up, without small peptide A intervention, and the others remained unchanged. The ALP activity was d...
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