Peptide for repairing cartilage and/or treating osteoarthritis
A technology for osteoarthritis and cartilage, applied in the field of peptides and drugs containing the peptides, can solve the problems of difficulty in inducing activity, limited clinical application of BMP-2, difficulty in maintaining therapeutic concentration, etc., and achieve improved efficacy in vivo Duration, promotion of chondrocyte proliferation, and stability improvement effects
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Embodiment 1
[0037] The solid-phase synthesis of embodiment 1 BM23 peptide
[0038] BM23 peptide (SEQ ID NO: 1, in which amino acids 1, 7 and 23 use D-glutamine, homoarginine and amidated leucine respectively) commissioned by Shanghai Aobo Biotechnology Co., Ltd. The company adopts conventional solid-phase process synthesis, and the purity of the synthesized peptide is >98%, see figure 1 .
Embodiment 2
[0039] Example 2 Isolation and cultivation of mesenchymal stem cells (BMSCs)
[0040] Three 8-week-old SD rats were killed by dislocation and sterilized with 70% ethanol for 5 minutes. The layers of the abdominal wall are then cut with scissors, the incision is drawn to the femur, and the muscles on the femur are carefully separated.
[0041] Cut off the middle of the femur, draw low-sugar DMEM culture solution (containing 1 mL / L heparin) with a syringe and repeatedly flush the bone marrow cavity to flush out the bone marrow.
[0042] The bone marrow fluid was flushed repeatedly with a syringe, passed through No. 7 and No. 4 needles in turn to make a single-cell suspension.
[0043] The single cell suspension was centrifuged (1500r / min, 10 minutes), and the supernatant was discarded. Add DMEM-F12 complete medium containing 10% FBS to 10 7 The density of each / mL was inoculated in a culture flask, placed at 37°C, 5% CO 2cultured in an incubator.
[0044] Cells were purified...
Embodiment 3
[0048] Example 3 The effect of BM23 peptide on the differentiation of mesenchymal stem cells in vitro
[0049] chondrogenic induction
[0050] The BM23 peptide was dissolved in phosphate buffered saline (PBS).
[0051] The cells were resuspended in complete chondrogenic medium (purchased from Guangzhou Saiye Biotechnology Co., Ltd., product number: RASMX-90011) containing different concentrations of BM23 peptide (0.1, 0.3 and 1 μM), so that the concentration of BMSCs was 5.0× per ml. 10 5 cells. In addition, PBS was used as a negative control, and 1 μM human insulin growth factor (IGF-1, purchased from Beijing Sino Biological Technology Co., Ltd.) was used as a positive control.
[0052] The treated BMSCs were incubated at 37°C and 5% CO2 saturated humidity. Change the medium every 2-3 days and add 0.5 mL of fresh complete chondrogenic medium to each tube.
[0053] Generally, the samples can be collected after 28 days of continuous induction, and the cartilage balls can b...
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