| [Objective] Investigate the preparation of molecular targeted gold nanorods, observation of gold nanorods into the process of cell, conjugated of gold nanorods before and after the change of cell toxicity and detection of the conjugated gold nanorods to improve the effect of infrared hyperthermia, for the treatment of basal cell carcinoma provides a new way.[Methods] Cultured A-431 human epidermal cancer cells and normal human skin fibroblasts in vitro, at the same time synthesis of gold nanorods and also to conjugated anti-epidermal growth factor receptor monoclonal antibody with molecular targeted. Conducted the following experiment:(1)Observe the characteristics of gold nanorods was prepared. (2) Improved MTT Colorimetric assay with different concentrations of gold nanorods and gold nanorods conjugated the toxicity of the two cells. (3)Gold bars and A-431 cells were incubated for different time transmission electron microscopy to observe the process of entering cells. (4) Divided into four groups of two cells:①egative control group,②aser irradiation alone,③25% concentrations of gold nanorods,④25% concentrations of anti-EGFR/ Au nanorods,②③④By different power of infrared laser photo-thermal therapy, then Modified MTT assay cell viability, cell survival rate according to formula, Trypan blue staining observed changes in cell morphology. Experimental data obtained with (x±s). Express, Application of statistical package with SPSS 16.0 statistical analysis, Data were statistically analyzed using a one-way analysis of variance, P<0.05 determined statistical significance. [Results] (1) Preparation of gold nanoparticles are bat, Average diameter is 35-45nm, Optical density around 0.8, there were two plasma resonance absorption peak in the vicinity of 520nm and 820nm. The 820nm absorption peak in the vicinity of near-infrared region (800-1200nm) is the organization's security body the transmission window. It can be used for near infrared laser thermotherapy. (2) Improved MTT Colorimetric assay cell-cytotoxic:Concentration of less than 25% of the gold nanorods and concentration of less than 50% of the anti-EGFR/Au nanorods were the safe concentration for the two cell. (3) Gold nanorods and A-431 cells were incubated after 1h, gold nanorods was also found primarily in extracellular.4h after the cell membrane out of swallowing gold nanorods wrapped pseudopodia.8-24h within the cytoplasm shows a large number of gold nanorods wrapped in bubble phagocytosis, gold nanorods are not seen in the cell nucleus.48h after the large number of gold nanorods have appeared in the extracellular. (4) Two kinds of cells respectively and 25% of the concentration of gold nanorods and 25% of the concentration of the anti-EGFR/Au nanorods incubated, then different power of infrared laser photo-thermal therapy. Improved MTT Colorimetric assay the cell growth activity, Trypan blue staining observed changes in cell morphology. A-431 cell:②③④roup irradiated by the same laser power,④roup cells relative survival rate was significantly lower than②③group(P< 0.05). Also③group cells relative survival rate was significantly lower than②roup(P<0.05). Trypan blue staining②③④group found:④roup a large number of cells were killed need a laser power is much smaller than②③group(P<0.05).③roup to kill a large number of cells used in laser power is also far less than②roup. Comparison of normal fibroblast②③④roup can get the same results. But the comparison group data③④(P>0.05). Comparison of A-431 cells and normal fibroblasts of the two cell②③④group found:After the same laser power irradiation, group②③④the A-431 cells of the relative survival rat are much smaller than normal fibroblasts(P<0.05).[Conclusions]1. Photochemically synthesized gold nanorods. Average diameter is 35-45nm, Optical density around 0.9. The 822nm absorption peak in the vicinity of near-infrared region (800-1200nm) is the organization's security body the transmission window. It can be used for near infrared laser thermotherapy. 2. Prepared into the anti-epidermal growth factor receptor monoclonal antibody/ conjugated gold nanorods in the cell body of the safe concentration is much higher than pure gold nanorods. Enhance its biocompatibility. At the same time combined with the antibody also increased the stability of gold nanorods and dispersion, Making it in the biomedical field has more potential applications.3. Gold nanorods can form nanoparticles and phagocytosis way can a large number of gold nanorods entering A-431 cells. Into the cells shape of gold nanorods have not been destroyed.4. Anti-EGFR/Au nanorods prepared by a combination with a high molecular target-specific to the A-431 cells, Compared with pure gold nanorods, Anti-EGFR/Au nanorods just 14w/cm~2 near-infrared laser irradiation on the A-431 of human skin cancer cell growth inhibition rate of 87% or more can. While the laser power irradiation on the growth of normal fibroblasts very little inhibition.5. After infrared laser photo-thermal therapy, A-431 cells than normal human skin fibroblasts more sensitive to heat.
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