Preparation method for injection for treating diabetic wounds and product thereof
A technology of diabetes and injection, which is applied in the field of medicine, can solve the problems of weakened activity, weakened therapeutic effect, poor healing effect of diabetic refractory wounds, etc., and achieve the effect of enhanced effect
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Embodiment 1
[0027] 1) Cultivate umbilical cord mesenchymal stem cells in a culture dish up to passage 6, then add 1 ml of trypsin with a mass fraction of 0.25% to digest for 1 min, add an equal volume of serum after digestion to remove the activity of trypsin, and then carry out the first For the first centrifugation, the centrifugal force is 800g, and the time is 5min. After the centrifugation is completed, the supernatant is discarded, and then the cells are resuspended with 10ml PBS solution, and the second centrifugation is performed. The centrifugal force is 800g, and the time is 5min. supernatant, and then add 3ml PBS solution to resuspend the cells to obtain a concentration of 5×10 6 cells / ml solution containing mesenchymal stem cells (MSCs).
[0028] 2) Dissolve 200ug type I collagen in 1ml of 0.5% acetic acid aqueous solution by mass volume ratio to obtain a type I collagen solution; dissolve 100ug laminin in 1ml PBS solution to obtain a laminin solution; dissolve the type I coll...
Embodiment 2
[0030] Example 2 Application of Injection for Treating Diabetic Wounds in Treating Diabetic Wounds in Rats
[0031] Rats fed with high sugar and high fat were taken, and 35 mg / kg streptozotocin (STZ) was injected intraperitoneally to establish a diabetic rat model. The modeled diabetic rats were randomly divided into three groups. A full-thickness skin defect wound with a diameter of 1 cm was created on the upper part, and each group was injected with local subcutaneous injection around the wound. The control group was injected with 1ml of PBS solution, the MSCs treatment group was injected with 1ml of the MSCs solution prepared in Example 1, and the ECM-MSCs treatment group was injected with 1ml The ECM-MSCs injection prepared in Example 1.
[0032] The size of the wound was measured at 2, 4, 6, 8, 10, and 12 days after the injection to observe the healing situation; HE and immunohistochemical staining were performed on the healed diabetic wound to observe the wound repair an...
Embodiment 3
[0040] 1) Cultivate umbilical cord mesenchymal stem cells in a culture dish to passage 7, then add 1ml of trypsin with a mass fraction of 0.1% to digest for 2 minutes, add an equal volume of serum after digestion to remove the activity of trypsin, and then carry out the first For the first centrifugation, the centrifugal force is 800g, and the time is 5min. After the centrifugation is completed, the supernatant is discarded, and then the cells are resuspended with 10ml PBS solution, and the second centrifugation is performed. The centrifugal force is 800g, and the time is 5min. supernatant, and then add 5ml PBS solution to resuspend the cells to obtain a concentration of 8×10 6 cells / ml solution containing mesenchymal stem cells (MSCs).
[0041] 2) Dissolve 100ug type I collagen in 1ml of 1.0% acetic acid aqueous solution by mass volume ratio to obtain a type I collagen solution; dissolve 38ug laminin in 1.5ml PBS buffer solution to obtain a laminin solution; The collagen sol...
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