A Preliminary Study On The Preparation Of Targeted Anticancer Drug System Combined With Near-infrared Hyperthermia

ObjectivesTo prepare a Graphene Oxide?GO?drug carrier targeted to anti-cancer,to study its properties of drug delivery when it carry CDDP,CBP and OXA and anti-tumor activity in vitro.Methods1.GO and GO-Fe₃O₄ are prepared by Hummer's method and by chemical precipitationmethod,respectively.GO-Fe₃O₄-PANIcarrierand GO-Fe₃O₄-PANI-FA carrier are prepared by modification of aniline monomer and amide reaction,separately.2.Structure characterizations are determined by Fourier Transform Infrared?FTIR?,Ultraviolet visible spectrophotometry?UV-vis?,X-ray diffraction?XRD?,Scanning Electron Microscopy?SEM?,Thermogravimetric Analyzer?TGA?and ZETA potential?ZETA?methods.3.The biocompatibility of GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA are evaluated by MTT test in vitro and BSA adsorption method.4.The properties of the drug delivery,GO-Fe₃O₄-PANI-FA loaded CDDP,CBP and OXA are tested in different conditions?pH,temperature and mass of anti-cancer drugs?.5.The inhibitiones of cell growth are determined separately by MTT assay,which is added the targeted drug delivery system GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA loaded CDDP,CBP or OXA.6.The inhibitiones of cell growth are determined in different experimental groups of GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA with CDDP,CBP or OXA combining therapy of near-infrared laser irradiation hyperthermia is detected by MTT assay in vitro.Results1.Structure characterizations showed that GO,GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA can be successfully prepared.2.MTT assay and BSA adsorption assay results:?1?The relative proliferation rates for HL-7702 cells,SMMC-7721 cells,Hep G-2 cells and RAW264.7 cells were>75%when the concentration of GO-Fe₃O₄-PANI was between 0?g/mL to 125?g/mL and the relative proliferation rates were>80%when the concentration of GO-Fe₃O₄-PANI-FA within 0?g/mL to 30?g/mL.?2?The time that GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA reached the maximum adsorption of BSA were 120 min and 270 min,separately.?3?Changing the initial concentration of BSA can change the effect of adsorption.?4?Changing the p H environment,the adsorption of BSA on the carrier also changes.3.Drug load study showed that:?1?The loading efficiency rate and entrapment efficiency of GO-Fe₃O₄-PANI carrier loaded CDDP,CBP and OXA were 83.28%and 4.16%,38.38%and 27.74%,84.90%and 18.37%,respectively.?2?The loading efficiency rate and entrapment efficiency of GO-Fe₃O₄-PANIFA loaded CDDP,CBP and OXA were 32.78%and 49.38%,25.93%and 70.00%,24.27%and 64.11%,respectively.4.The drug release performances study showed that:?1?When surrounding is pH8.0,37?,the release rate of GO-Fe₃O₄-PANI*CDDP reach 75%;When surrounding is pH6.0,37?,the release rate of GO-Fe₃O₄-PANI*CBP reach98%;When surrounding is pH6.0,47?,the release rate of GO-Fe₃O₄-PANI*OXA reach 75%.?2?When surrounding is pH6.0,37?,the release rate of GO-Fe₃O₄-PANI-FA*CDDP reach 70%;When surrounding is pH6.0,37?,the release rate of GO-Fe₃O₄-PANI-FA*CBP reach 80%;When surrounding is pH7.4,47?,the release rate of GO-Fe₃O₄-PANI-FA*OXA reach75%.5.The anti-tumor activity of the drug-loaded system in vitro showed that:?1?the inhibitory rate of CDDP on SMMC-7721 cells and Hep G-2 cells increased have a dose-dependence.When the CDDP concentration reached 8?g/mL,CDDP inhibited SMMC-7721 cells and HepG-2 cells more than 80%.The inhibitory rates of GO-Fe₃O₄-PANI*CDDP and GO-Fe₃O₄-PANI-FA*CDDP were 25%and 35%,20%and 40%for SMMC-7721 cells and HepG-2 cells when CDDP concentration was 8?g/mL,respectively.?2?The inhibitory effects on Hep G-2 within range of 0¹50?g/mL:GO-Fe₃O₄-PANI*CBP and GO-Fe₃O₄-PANI-FA*CBP>CBP;The inhibition rate went up as the concentration of CBP increased which have a dose-dependent manner.?3?When the OXA concentration reached 30?g/mL,OXA inhibited the Hep G-2 cells and SMMC-7721 cells were about 75%and65%,separately.Drug-containing systems GO-Fe₃O₄-PANI*OXA and GO-Fe₃O₄-PANI-FA*OXA were lower than OXA in the studied drug concentration range of 0⁶0?g/mL.?4?The inhibitory rate of GO-Fe₃O₄-PANI-FA*OXA to HepG-2 cells was higher than that of GO-Fe₃O₄-PANI*OXA drug-loading system in the range of 0⁶0?g/mL.6.In vitro near-infrared hyperthermia experiments showed that when the CDDPconcentrationwas16.00?g/mL,theinhibitionratesof GO-Fe₃O₄-PANI*CDDP and GO-Fe₃O₄-PANI*CDDP+hyperthermia on SMMC-7721 cells and HepG-2 cells were 23%and 25%,35%and 35%,respectively.The inhibitory rates of GO-Fe₃O₄-PANI-FA*CDDP and GO-Fe₃O₄-PANI-FA*CDDP+hyperthermia on SMMC-7721 cells and HepG-2cells were 32%and 36%,40%and 42%,separately.When the CBP concentration was 150?g/mL,the inhibition rates of GO-Fe₃O₄-PANI*CBP and GO-Fe₃O₄-PANI*CBP+hyperthermia on SMMC-7721 cells and HepG-2 cells were 45%and 40%,50%and 70%,respectively.The inhibitory rates of GO-Fe₃O₄-PANI-FA*CBP and GO-Fe₃O₄-PANI-FA*CBP+hyperthermia against SMMC-7721 cells and HepG-2 cells were 15%and 22%,44%and 60%,respectively.When the concentration of OXA was 60?g/mL,the inhibitory rates of GO-Fe₃O₄-PANI*OXA and GO-Fe₃O₄-PANI*OXA+hyperthermia on SMMC-7721 cells and HepG-2 cells were 30%and 80%,80%and 85%,respectively.Theinhibitoryrates ofGO-Fe₃O₄-PANI-FA*OXAand GO-Fe₃O₄-PANI-FA*OXA+hyperthermia against SMMC-7721 cells and HepG-2 cells were 37%and 86%,40%and 92%,respectively.Conclusions1.The potentials of GO,GO-Fe₃O₄,GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA are successfully prepared by FTIR,UV-vis,XRD,SEM,TGA and ZETA potential analysis.2.The results of MTT assay show that the GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA with a concentration range of 0¹25?g/mL and 0³0?g/mL were safe and non-toxic to model cells,respectively.3.The results of BSA adsorption experiments combine with the ZETA potential of the support indicated that pH>pI,the amount of BSA adsorbed by GO-Fe₃O₄-PANI was higher than GO-Fe₃O₄-PANI-FA,the biocompatibility of GO-Fe₃O₄-PANI was better than GO-Fe₃O₄-PANI-FA.4.The GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA carriers loaded with CDDP,CBP and OXA under different environmental conditions show that GO-Fe₃O₄-PANI was suitable as drug carrier for CDDP and CBP;GO-Fe₃O₄-PANI-FA is suitable as a drug carrier for OXA.5.The results of in vitro anti-tumor activity of the drug-loading system showed that the presence of GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA carriers had a sustained release effect on the release of CDDP;GO-Fe₃O₄-PANI-FA*CDDP and GO-Fe₃O₄-PANI-FA*OXA drug delivery system exhibits FA-targeting potential and is expected to be further studied as a targeted drug carrier.The presence of GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA carriers for the release of CBP has a pronounced slow-release effect,but it does not show a targeted nature and needs further study to explain why.6.GO-Fe₃O₄-PANI and GO-Fe₃O₄-PANI-FA loaded with CDDP,CBP and OXA as drug loading system combined with near-infrared hyperthermia have stronger anti-tumor effect than simple drug chemotherapy and have a potential targeted chemotherapy combination.Infrared hyperthermia is applied to new targeted tumor therapies.