Preparation Of Compound Ornidazole-Berberine Buccal Film And Its Effect On Periodontitis In SD Rats

Objective:To prepare oral ornidazole berberine membrane on the basis of antibacterial activity test in vitro,to study the biological properties of the film,release in vivo and in vitro,and to evaluate its initial effect on periodontitis in SD rats with experimental periodontitis.Methods:?1?The microbiological dilution method was used to study the in vitro antibacterial activity of ornidazole and berberine on common pathogens of periodontitis and determine the minimum inhibitory concentration.Combined susceptibility testing was performed using a trace checker dilution method.?2?Using the L₉?3⁴?orthogonal test method,taking the appearance of the drug film and its in vitro release as the inspection index,the proportion of each main drug and auxiliary material of the compound ornidazole berberine oral film was selected and the prescription was optimized.Membrane agents were used for quality control studies.Inhibitory effect of compound ornidazole berberine oral membranes on common pathogens of periodontitis was observed by inhibition zone method.The desorption of oral membranes to Sm was observed by the attached plate method.Cup-disc method was used to study the sensitivity of the drug film dilution to Aa.?3?The animal model of periodontitis in SD rats was established successfully and drug treatment was performed to observe the change of clinical indexes of rat periodontium before and after administration;the rat buccal,temporal mucosa and maxilla were taken.Routine HE staining was used to observe the pathological changes.The expression of TNF-?in the alveolar bone and its surrounding tissues was detected by immunohistochemistry.The levels of TNF-?and IL-1?in serum of each group were detected by enzyme-linked immunosorbent assay?ELISA?.The degree of oral film release of compound ornidazole berberine was measured by UPLC.Results:?1?The MICs of Ornidazole to Pg,Fn,Aa,Av,and Sm were 0.64,0.000078,0.64,0.64,and 5.12 mg·mL^-1,respectively;the MICs of berberine were0.16,0.01,0.16,1.28,and 0.04 mg·mL^-1,respectively.The combination of ornidazole and berberine showed synergistic effect on Pg,additive on Fn,and irrelevant on Av.?2?Optimum prescriptions:1 mg ornidazole,0.5 mg berberine,1.5 g carboxymethyl cellulose sodium,3 g polyvinyl alcohol,1 g gelatin,2.5 g glycerol,0.5 mL Polysorbate 80,water 120 mL.The prepared compound ornidazole berberine oral film basically meets the quality requirement of the preparation.Compound ornidazole berberine oral membrane has obvious inhibition on common pathogens of periodontitis,and has anti-adhesive effect on Sm.The antibacterial effect of the oral membrane diluent was stronger than that of the control ornidazole and berberine monofilament serial dilutions.After serially diluting it for 4 h,the dilution solution still has a significant inhibitory effect on Aa.?3?The GI value in each treatment group decreased after treatment?P<0.05?,and PD and MD had a downward trend?P>0.05?.Compared with the model group,the value of GI,PD and MD in each treatment group had shown a downward trend in different degrees,and PD in ORN-BER group was significantly lower than that in model group?P<0.05?.The periodontal tissue and alveolar bone HE staining results showed that the ORN-BER group had less keratinocyte hyperplasia than the ORN and BER groups.The distribution of layers was normal,the basement membrane was intact,and the periodontal ligament tissue was increased,but the alignment direction was regular.Infiltration of inflammatory cells decreased.The expression of TNF-?in the periodontal tissue of periodontitis rats treated with ORN-BER film agent was significantly lower than that in model,ORN and BER group?P<0.01?.The content of TNF-?in serum of ORN-BER group was lower than that in BER group?P<0.05?;The content of IL-1?in serum of ORN-BER group was significantly lower than that in BER and ORN group?P<0.01?.The in vivo release test showed that the ORN concentration in saliva was 765.9±23.5?g·mL^-1 and the concentration of BER was 410.4±31.9?g·mL^-1 in the periodontal pocket of SD rats.All the concentrations are higher than common periodontal bacteria MIC.Conclusion:Berberine oral film based ornidazole,berberine periodontitis pathogens in a synergistic and complementary antibacterial effect,the preparation of compound ornidazole by prescription screening.The film agent is a quick-acting,sustained-release formulation with good process reproducibility and the quality standards meet the requirements of the Chinese Pharmacopoeia.In vitro tests showed that the film agent can more stably inhibit common pathogens of periodontitis.In the rat periodontitis experiment have shown that the film can rapidly reach the therapeutic concentration in the oral cavity of rats and can maintain effective antibacterial concentration at least 5 hours.After treatment,the levels of inflammatory cytokines in the serum and the periodontal tissue of periodontitis rats were significantly decreased,and the clinical and pathological parameters were improved.All of them were more obvious than ORN and BER single drug groups.In summary,considering the compound ornidazole berberine oral membrane has a good prospect for the treatment of periodontitis...