Application of native copper native copper microbial extracting liquid in preparing medicaments for treating bone fracture
A technology of microbial leaching and natural copper, which is applied in the direction of drug combination, bone disease, heavy metal active ingredients, etc., can solve the problems that natural copper and its related products are difficult to enter the international market, and the quality of natural copper is uneven.
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Embodiment 1
[0025] Example 1 Preparation of natural copper microbial leaching solution
[0026] Weigh 0.5 g of natural copper, grind it through a 200-mesh sieve, and obtain a powder. In a clean 250mL Erlenmeyer flask, add 100mL of sterilized 9K iron-free liquid medium, adjust the pH to 1.60 with 5M sulfuric acid, and inoculate the volume fraction of 10% Thiobacillus ferrooxidans after the pH is stable, and do not inoculate the blank control bacteria liquid. After weighing the mass, it was leached at 25°C and 130r / min for 15 days. During the experiment, 5M sulfuric acid was used to adjust the pH value, and distilled water was used to supplement the evaporated water. Shake the flask for 15 days to stop leaching, let it stand overnight, collect the supernatant, adjust the pH of the solution to about 7, pass through a 0.22 μm filter membrane to sterilize, and set aside.
Embodiment 2
[0027] Example 2 Effect of Natural Copper Leach Solution on Serum Calcium Concentration and Alkaline Phosphatase Activity in Fractured Rats
[0028] Experimental animals: healthy SD rats, female, 6-8 weeks old, weighing (200±20) g, SPF grade, purchased from the Animal Experiment Center of Lanzhou University.
[0029] Fracture model: SD rats were anesthetized with pentobarbital sodium, the right hind limb was depilated, iodophor was disinfected, the skin was cut straight with a scalpel, and the tibia was exposed, and part of the bone was resected at the upper 1 / 3 (about 1mm deep and 2mm wide) , Not completely sawed off, keep connected, suture the skin, not fixed after operation. Postoperative intramuscular injection of penicillin, 40,000 units per mouse, intramuscular injection for three days. On the fourth day after the operation, intragastric administration was started, and the rats with fractures were divided into three groups: the positive group (n=9, traumatology bone joi...
Embodiment 3
[0035] Example 3 Effect of natural copper leaching solution on the growth of callus at the fracture site of rats with fracture
[0036] The fracture model is the same as in Example 2. Animals in each group were taken X-rays at 7d, 14d, and 21d after operation. The radiographs were evaluated according to the five-level evaluation standard of fracture healing:
[0037] Grade I: The edge of the broken end is slightly blurred. The periosteal reaction was mild without callus.
[0038] Grade II: The edge of the broken end is obviously blurred, the density of internal and external callus is higher than that of soft tissue, and callus appears in more than 1 / 2 of the fracture gap.
[0039] Grade III: Most of the edge of the broken end disappears, the internal and external callus is similar to the medullary cavity, and more than 1 / 2 of the fracture gap is connected with the callus.
[0040] Grade IV: The edge of the broken end tends to disappear, the internal and external callus is si...
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