A Study On The Preparation And Efficacy Of Myocardial-Targeted Curcumin Liposomes

Myocardial infarction is one of the public health problems with high morbidity and mortality worldwide.Common treatment of myocardial infarction relies on surgery and antithrombotic drugs,but many patients cannot receive these traditional treatments for various reasons.Curcumin become a hit for natural anti-inflammatory effects,has been widely applied in certain countries as a traditional medicine in the treatment and prevention for types of diseases.Recent studies have also demonstrated the cardioprotective effects of curcumin against cardiovascular disease.However,the poor water solubility and low bioavailability of it have limited its clinical application.To break this limitation,biocompatible nanoformulations have been used as carrier agents to promote curcumin release in vivo.In this study,anti-cardiac troponin I antibody was used to modify liposomes to deliver curcumin targeting to the position of myocardial infarction,thereby achieving targeted therapy for myocardial infarction.In this study,blank liposomes(LIP)and curcumin liposomes(Cur-LIP)were prepared using a thin film dispersion method,and then they were treated by hydrated ultrasound.Anticardiac troponin I antibody-coupled phospholipid derivatives were embedded into the blank liposomes and curcumin liposomes to obtain liposomes modified with anti-cTnI antibody(cTLIP)and curcumin liposomes modified with anti-cTnI antibody(cT-Cur-LIP).A methodology for curcumin analysis was constructed and liposome encapsulation rates were calculated.The liposome preparation prescription was optimized by screening with single-factor experiments and response surface methodology.The morphology of liposomes was observed by Transmission Electron Microscope.Particle size and Zeta potential of liposomes were analyzed by Zetasizer Nano S90.The stability,dispersion and drug release of liposomes were investigated.Cytotoxicity of liposomes and drugs were evaluated by Cell Counting Kit-8.Flow cytometry to detect liposome uptake by cardiomyocytes.The drug uptake and distribution of cardiomyocyte was assessed by Live cell Imaging System.Fluorescence distribution of in vivo imaging in rat hearts of Ctl,DiR,Cur-LIP and cT-Cur-LIP by Living animal imaging system.TTC staining and HE staining showed the improvement of cT-Cur-LIP on myocardial infarction.The standard curves of curcumin samples were determined by UV spectrophotometer and proved that curcumin has good linearity within 0.22-56.25μg/mL.By optimizing the prescription through single-factor experiment and response surface method,the prepared liposomes with rounded surface,oval shape and good morphology,139.92±2.10 nm particle size,-28.39±3.11 mV potential,88.94±1.67%encapsulation rate and 62.68±5.73 24 h release rate,had excellent stability and dispersibility.The results of CCK-8 experiment showed that liposomes were non-toxic to cells and that curcumin at appropriate concentrations significantly increased the survival rate of cardiomyocytes under hypoxia.Flow cytometry and Live cell Imaging System showed that cardiomyocytes showed the strongest uptake of modified liposomes against cTnI antibodies.Living animal imaging system confirmed that cT-LIP can accumulate significantly in infarcted areas of the heart in MI rats.The successful construction of a rat model of myocardial infarction was verified by electrocardiography.By TTC staining and HE staining,it was confirmed that cT-Cur-LIP could treat myocardial infarction more effectively compared to Cur-LIP.