Polypeptide, polypeptide derivative and nanofiber capable of inducing differentiation and mineralization of dentin of human dental pulp cells and application of polypeptide, polypeptide derivative and nanofiber
A technology of dental pulp cells and peptide derivatives, applied in the field of biomedical engineering, can solve the problems of short half-life, low bioavailability, lack of space structure similar to natural protein, etc. simple effect
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Embodiment 1
[0051] Peptide derivative Nap-G D f D f D Synthesis and preparation of YGKWYQNMIR and its nanofibers
[0052] (1) Nap-G D f D f D Solid Phase Synthesis of YGKWYQNMIR
[0053] Specific steps are as follows:
[0054] 1) Weigh 0.5 mmol of 2-Cl-Trt resin into a solid-phase synthesizer, add 10 mL of anhydrous dichloromethane (hereinafter referred to as DCM), and place it on a shaker to swell for 10 min;
[0055] 2) Remove the DCM from the solid-phase synthesizer with an ear wash ball;
[0056] 3) Dissolve 0.75mmol Fmoc-Arg in 20mL of anhydrous DCM, add 1.5mmol of DIPEA, transfer to the above solid-phase synthesizer after fully dissolved, and react at room temperature for 1h;
[0057] 4) Sealing: Remove the reaction solution in the solid-phase synthesizer with ear wash balls, then wash with 10 mL of anhydrous DCM, 1 min each time, for a total of 4 washes, and add the prepared volume ratio of anhydrous DCM: DIPEA: methanol = 17:1:2 solution 20mL, react at room temperature fo...
Embodiment 2
[0065] (1) With reference to Preparation Example 1, the polypeptide derivative Nap-GFFYGKWYQNMIR was synthesized by Fmoc-short peptide solid-phase synthesis method, Figure 4 It is the high-resolution mass spectrum of Nap-GFFYGKWYQNMIR: the difference is only that (1) the D configuration Fmoc-Tyr, D configuration Fmoc-Phe, and D configuration Fmoc-Phe added in step 7 of solid phase synthesis are changed to L configuration .
[0066] (2) The preparation of the nanofibers of the polypeptide derivative Nap-GFFYGKWYQNMIR refers to the preparation example 1, the only difference is that the color of the formed hydrogel is not completely the same as that of the example 1. Figure 5 with Image 6 Respectively, the optical photograph of Nap-GFFYGKWYQNMIR and the lower picture of electron transmission microscope.
Embodiment 3
[0068] (1) Referring to Preparation Example 1, the polypeptide KWYQNMIR was synthesized by Fmoc-short peptide solid-phase synthesis method: the only difference is that (1) Fmoc-Met, Fmoc-Asn, Fmoc-Gln, Fmoc-Asn, Fmoc-Gln, The synthesis of Fmoc-Tyr, Fmoc-Trp, and Fmoc-Lys is completed, and the crude polypeptide product is obtained by cleavage. Figure 7 is the mass spectrum of the peptide.
[0069] (2) Take 1.1 mg of the purified polypeptide KWYQNMIR and place it in a 2 mL glass bottle, add 500 μL PBS solution (pH=7.4), blow and suck several times, the compound is completely dissolved, and adjust its pH value to 7.4 with sodium carbonate solution to obtain clarification transparent peptide solution, Figure 8 with Figure 9 The optical photographs and transmission electron microscope images of the peptides are respectively.
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