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The purpose of preparation of acrylic bone cement contained alendronate and biocompatibility evaluation. This experiment according to the GB/T16886.1-2001 the provisions of the national standard of the People's Republic of China, according to the the biocompatibility evaluation project of surgical implants. 1, the preparation of the test module: according to design requirements are divided into five different mixture proportioning, 10mg per 50g PMMA powder, 50 mg, 100 mg, 500 mg, 1000 mg alendronate, made into the cured The standard set out Allen sodium phosphate bone cement module, packaging sterilization spare. Cytotoxicity test (in vitro) and cell proliferation: each test group module extracts of the culture liquid extraction 24h as spare. Take the proliferation of osteoblast-like cells MG-63, 1 × 10 ~ 4 per well were inoculated into 96-well plates, plus extracts co-cultured for 24 h, the MTT colorimetric assay absorbance values ??to calculate the relative cell proliferation rate and cytotoxicity grading (cell toxicity level 0 or 1 is passed). 3, the adhesion test: The osteoblast-like cells MG-63 to 10 to 5 / ml were seeded in 6-well plates containing adhesion specimens (each hole 1), cultured for 3 days to remove the specimen, washed 2 times with PBS, 2% cold glutaraldehyde and 1% osmium tetroxide fixed gradient alcohol dehydration, critical point drying, to spray gold underwent scanning electron microscope, and cell morphology was observed and the number. Local reactions after implantation test: selection of 15 adult white rabbits were randomly divided into 5 groups, each group of three modules, the relative position of implanted the blank acrylic bone cement were implanted into rabbit back muscles each test group bone cement mold. 1,4,12 weeks after the animals were sacrificed remove the specimen the observational material surrounding tissue inflammation and the formation of the fibrous capsule. 5, percutaneous stimulation test: Experiment Module with saline extraction 24h sterilized. 5 selection of adult white rabbits, spine area of ??skin preparation. Along the spine on both sides of a symmetrical position were injected with saline and each group extracts 0.2ml, 4h, 24h, 48h, 72h observed skin irritation and graded. 6, acute systemic toxicity test: Experiment Module with saline extraction 24h sterilized. The selection of 30 KM mice were randomly divided into six groups, each group of five. Experimental mice by tail vein injection of extracts of constant speed, dose 50ml/Kg, control group injected with normal saline, and 4h, 24h, 48h, 72h was observed in mice without toxic reactions and grading. Results of the cytotoxicity test results show, set five test groups cytotoxicity the classification are 0-1, to meet the requirements; visible adhesion test specimens on the cell-attached, the experimental group of osteoblast-like cells without toxicity meet biocompatibility requirements; intramuscular implant test one week after the capsule formation incomplete the inflammation grade three, four weeks and 12 weeks showed that encapsulated form, no obvious thickening, inflammation of level are 1-0, in line with the requirements of national standards; percutaneous stimulation test immediately after injection, 24, 48 and 72h, the control group and the experimental group at the injection site showed no erythema, edema, showed that the test group skin stimulation; acute toxicity test 72h observation period, the control group and the sample group of animals is generally in good condition and showed no toxicity performance, without the death of the animal, to meet the requirements. Conclusion alendronate amount of 50g of bone cement powder to exceed 1000mg prepared composite bone cement no osteoblasts toxicity, good biocompatibility, no acute systemic toxicity, biocompatibility.
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