| Objective: This study analyzed the effectt of Carboxymethyl chitosanCMCS combined with Methotrexate MTX killing lung cancer cells by integratedmicrofluidic chip. In according to the basic theory, we added CMCS into bone cementsystem to modify antitumous bone cement properties and explored its antitumous effectand physico-chemical properties, In order to used in the treatment of malignant tumorbone metastasesMethod: Firstly, We cultured human lung cancer H460cells by microfluidic chip to observe theeffect of different concentration of carboxymethyl chitosan combined with methotrexate killing lungcancer H460cell, and select the optimal concentration ratio of the two drugs. Secondly, we drawnthree H460growth curves respectively, which are under the influence of two drugs combined witheach other and separately, and analyzed the effect of different concentration of carboxymethylchitosan combined with methotrexate killing lung cancer H460cell. Thirdly, we measured thebending modulus, bending strength and compressive strength of bone cement which containeddifferent doses of drugs (CMCS: MTX).High resolution scanning electron microscope to observedifferent composition of bone cement microstructure. Fourthly, by liquid chromatography-massspectrometry/mass spectrometry we analyzed the different release rate and release amount of MTXdrugs which combined with CMCS or not in bone cement. Fifthly, antitumous bone cement withCMCS and pure antitumor bone cement were implanted in the guinea pig femur respectively. Then,we gave four times X-ray to these guinea pig femurs in0month,1.5months,3months and6monthsin chronological order. At the same time we observed the morphology of two kinds of bone cementand their relationship with the surrounding bone tissue. Finally, the implanted bone cement with theirsurrounding bone tissue were cut out. High resolution scanning electron microscope observation ofbone cement microstructure of bone cement and bone fusion lineResult: Firstly, MTX combined with CMCS showed strongger killing effect to tumorcell, and the CMCS: MTX optimal concentration ratio is57μg/mL:21μg/mL. Secondly,by analyzing these growth curves, we found that the two drugs’ combination had a stronger killing effect on tumor cell than one drug in bone cement. CMCS can makeMTX tumor kill rate increased by33.4%. And both of them have synergy which issignificant at the beginning of the cell growth especially. Thirdly, mechanics testrevealed that adding1.6%CMCS to MTX bone cement could improve the mechanicalstrength significantly, flexural modulus increased by5%, the flexural strength increasedby2.8%, and the compressive strength increases by5.2%. If the percent of CMCS wasover3.2%, the mechanical strength of bone cement began to decline. Furthermore,scanning electron microscopy (sem) presented the result was that1%to2%ofmedicated bone cement was better others: its surface structure was more uniform andsmooth and the crystal structure was more conform. When the percent of CMCSreached5%, the gap on the surface of the bone cement become larger and intertwined,which was bad for stable structure of bone cement. Fourthly, liquidchromatography-mass spectrometry/mass spectrometry measured that bone cementsystem with CMCS prolonged release of MTX28.5%than pure bone cement, and drugrelease rate of the former one increased by64.8%. Fifthly, over time, the X rayexamination results indicated that the bone cement with CMCS had more blurringboundaries between itself and its surrounding bone tissue than the normal bone cement.And the former one had higher radiation rate. Above all, the bone cement with CMCShad better integration. Finally, by scanning electron microscopy (sem), the resultshowed that surface of the bone cement with CMCS formed pore and bone cement andbone fusion boundary is better six months later.Conclusion: Firstly, CMCS and can improve the lung cancer of MTX H460cell killingrate, and joint application show the synergy between the two choices. Secondly, CMCScan improve the mechanical strength of bone cement and the rate of drug release, andprolong drug release time significantly. Finally, surface of the implanted bone cementforms pore in late period that is good for new bone ingrowth, and can reinforce the bonecement. This modification scheme for bone cement has the clinical feasibility andextensive developing space. |
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