| Backgrounds and ObjectivePeriodontitis is one of prevalent, deleterious oral infection diseases. This disease has serious effect on the function of mastication and digestion and results in the defluvium of the teeth. It is a dangerous factor which will induce the diseases of the whole body. so periodontitis is the focus of scholars both domestic and international in the long run. Periodontitis is a chronic, proof-chemical disease and it has close relation with other apparatus, so up to now, the therapeutics is not so ideal. As a result, we pay more attention to the research of new method and medicine. In the prophases, according to the periodontal pathogeny and its pathologic characteristics, we reasonably mix Compound Ornidazole and pefloxacinmesylat. According to the outer characteristics of the periodontal physiology, we work out periodontal suppository which is fit to periodontal pocket with different teeth, different depth and fit to different periodontal configuration. By adopting new sustained-release material, we made great contribution to the bacteriostasis in vitro, overcoming many disadvantages such as the easily loss of the liquor medicine, the easily contamination of the gel medicine and trimming of solid medicine and so on.In order to make further development and application, we did a series of animal toxicological research in the spirit of the related new medical regulation. When Periodontal Suppository bacteriostatic and anti-inflammatory effect exists in the periodontal pocket, in the meanwhile, it will impact the periodontal tissue. The HPDLF is the main functional cell of periodontal tissue, which plays great role in the renovation of the wound and the rebirth of the tissue. We did related analysis on the biological active influence that Periodontal Suppository to HPDLF, which can supply foundation to further animal and clinical trial.Material and method1. Take use of cold compression to manufacture Periodontal Suppository, cooled naturally, using ultraviolet radiation to irradiate 30min, sterilizing befor use, 10mg each piece, contained raw medicine 1 mg.2. Using the highest concentration dosage (210g/L) in 1L distilled water sustained-release periodontal suppository to produce blending suspending solution. Pouring the highest dimension (0.4ml/10g) solution into stomach of mouse at one time. Obserived in the following 14 days. Using the same capability NS to administrate for control mouse. Observing the acute intoxication reflection of the mouse after taking up overdose reagent, Choose the appropriate dose the periodontal suppository repeated administration toxic experiment, analyzing the primary target organs of the toxic effect and the potential toxic reaction when human body are in overdose.3. 20 male and 20 female SD rats weigh 135±10 g are divided into 4 groups according to the experiment design.①low dosage group(5g/kg per day).②middle-dose group(10g/kg per day).③high-dose group(20g/kg).④control group (same capability NS). Observing the toxic reaction and primary target organs and reversibility damage of rats after repeated administration of the reagent. Offering nontoxic reaction dose and primary inspected index and referenced dose for human being.4. Using 40 guinea pigs dividing into 4 groups evenly to carry out skin scratch test:①control group.②sustained-release Periodontal Suppository group (10g/kg).③complement substrate group.④positive control group. Shearing 3cm×3cm hairless area on both sides back of guinea pig 24 hours before test. Then observing the effect of medicine on skin.5. The ectolabium of 40 SD rats being sewed into blind bags. Then the rats divided into 2 groups randomly. Pouring separately sustained-release Periodontal Suppository (210g/L) and the same volume substrate into bags and daubing the lower Gingival mucous membrane. Finding out the toxicity and stimulating effect of medicine to periodontal mucous membrane.6. Choosing six healthy and no cornea injury proved by 2% fluorescence New Zealand rabbits. Dripping 0.2ml sustained-release Periodontal Suppository (210g/L) into conjunctival sac of right eye, dripping the same quantity NS into the left eye for comparison. Observing the stimulate effect of medicine for eyes.7. Taking use of immunohistochemistry ABC method to human periodontal ligament fibroblast to colour the undee silk protein and cell keratin monoclone antibody coloration and identify the origin of cells. 8. Choosing the fourth generation well growth ligament fibroblast and then confect hang cell solution at 4×10~7 /L concentration. Putting DMEM which contained and sustained-released periodontal suppository 0.5 percent FBS into each hole. The cultivation of sustained-release Periodontal Suppository is divided into seven concentration groups as follows 0, 0.1, 0.5, 2.5, 10, 50, 200 g/L,in which the former acts as control group. Each group will be cultivated for different time, putting 5 g/L MTT 20ul into each hole in one culture board. Taking use of enzyme mark apparatus to test the OD value at 470 nm wavelength ,which is to evaluate effects of the drug on multiplication of cells .9. Chosing a culture board with 96 holes cultivated for 24 hours under standard condition, and putting 0,0.1,0.5,2.5,10,50,200 g/L concentration DMEM sustained-release periodontal suppository into holes,and putting 1μCi3H-TdR into each holes. Testing glittering count in one minute by glitter counting apparatus to evaluate effects of drug on HPDLF DNA synthesis.10. Taking use of 96 holes full of different concentration sustained-release periodontal suppository. Putting 100ul 0.2% TritonX-100 into each hole. Putting over 20ul solution from each hole into 200ul dying fluid, keeping on surging 10 minutes, testing the OD value at 570 wavelength.Results1. None of the Jimpy mice died from poisoning,even at maximum concentration (210g/L) and the maximum intragastric administration volume (0.4ml/10g)of sustained-release periodontal suppository reagent,the MTD of retarder is 8.4g/kg for Jimpy mice.This is more than 1,300 times of the adult's(50kg) dose per day( provided each of the thirty two teeth is placed a suppositorium).2. The rat of Medication's group act normally, behavior lively and the pelage is gloss, and none of them died during experimental stage of the sixty days, which is coincident with those of control group; There is no significant difference as for hemo-rout, test of Hepatic and renal function, partly remarkably heighten than control group, though these changes are within normal limits, without statistical difference,To check the pathological section by microscope,the transverse striation of cardiac muscle fiber is clear, cellular differentiation is well; the structure of hepatic lobules is clear; cellular differentiation of liver is well; there is no denaturation , and atosis for the nephric tubule, renal interstitium is non-haemorrhage or inflammatory; spleen didn't hydrops, there are no congestion, hydroncus and inflammatory reaction for pulmo.3. The mucous membrane of mouth is integrity, there is no difference between each group.4. The animal of sustained-release periodontal suppository group didn't appear erythema and dropsy, which has no difference with the control group; substrate group has no sensibilization; the skin of animal ,received test ,blonging to positive control group, obviously emerge erythema slightly, with no dropsy, sensitize rate for 100%.5. It's reported that there are conjunctival congestion, light hydroncus and little secreta by stimulation test of ocular region, with scoring no more than three points. These symptoms are reversible change and vanish totally after medication for 48 hours. Majority of people think that hare's eye is more sensitive than human's, so we can presume that periodontal suppository has no stimulation to human's eyes.6. The primary cell is long fusiform shape, and it's cytoplam is uniform, with nucleus round or ellipse, chromatospherite clear. Immunohistochemistry coloretur indicated that the undee silk protein is masculine, keratin negative.7. By analysis of variance of single factor, It's showed that the OD value (0.1,0.5,2.5 10,50,200 g/L) of sustained-release Periodontal Suppository has no significant difference compared with control group(P>0.05).8. When sustained-release Periodontal Suppository is at the concentration of 0,0.1,0.5,2.5 10,50,200 g/L, contrast to the control group, both OD value of protein and DNA's synthesis has no significant difference. (P>0.05)Conclusions1. The preparation artwork is simple for Compound Ornidazole and pefloxacinmesylat sustained-release Periodontal Suppository, nodule equation is little, the extraorgan sustained-release stable, which is consistent with the regulation for suppositories in the book《dispensatory of China》in 2005.2. The result of acute and chronic toxicity test manifests that sustained-release Periodontal Suppository has no overt toxicity, the general condition of rat, behavior activity, drink and food, urine and stool, growing of body weight, hematology and blood biochemics, coefficient of major organ as well, have no obviously difference with the control group, the check of pathology for each organ has no manifest medicinal impairment patho-alter, no part stimulation and allergic response, diversity of animals can tolerance this sustained release Periodontal Suppository greatly, which means using this medicine is safe.3. Immunohistochemistry proved that cultured cytes come from collagenoblast of mesoderm.4. When the concentration of sustained-release is within 200 g/L, the sustained-release Periodontal Suppository has better biocompatibility, without cyto-inhibition of HPDLF.
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