Biosafety Evaluation Of EGFR Monoclonal Antibody Modified Gold Nanorods For Photothermal Treatment Of Head And Neck Squamous Cell Carcinoma In Mice

Objectives:Gold nanorods are easy to prepare,have good biological stability,and have unique physical and chemical properties and biological affinity effects,and are typical representatives in the field of nanobiotechnology research.At present,a lot of research has been carried out in the fields of cell imaging,drugs and tumor photothermotherapeutics.Our previous work showed that the photothermal effect of gold nanorods can induce the death of human head and neck squamous carcinoma cells.By controlling the parameters of NIR laser irradiation(time and power),the photothermal effect of EGFR monoclonal antibody and gold nanorods was optimized to target head and neck squamous cell carcinoma in nude mice.The conditions and procedures of the tumor have obtained the best therapeutic effect.The nano-gold photothermal therapy mode greatly improves the efficacy of killing tumor cells due to the high-efficiency light-heat conversion characteristics and strong stability of gold nanoparticles.Based on this,we can provide the theoretical basis for biosafety in the future to enable gold nanorods to conduct medical research.Based on the parameters and therapeutic effects obtained from the optimization part,we use the optimized experimental parameters to evaluate the safety of gold nanorods based on the parameters and treatment effects obtained from the optimization part,and provide new and effective treatment methods for the treatment of head and neck squamous cell carcinoma in the future,as well as for future clinical research theoretical basis.Method:1.Different concentrations of AuNRs were used to detect the proliferation of Hep-2,CNE-2,BEAS-2B and NP-69 cells.2.Constructing two transplanted tumor models of larynx squamous cell carcinoma and nasopharyngeal squamous cell carcinoma in nude mice3.Conduct behavioral evaluation from CNE-2 and Hep-2 tumor-bearing nude mice with spontaneous activity,balance ability,grip strength and endurance.4.Physiological tests were carried out on CNE-2 and Hep-2 tumor-bearing nude mice for body temperature,respiration,heart rate,metabolism,eating,drinking,defecation,and urination5.The serum of CNE-2 and Hep-2 tumor-bearing nude mice were tested for biochemical indicators such as liver,kidney function and electrolytes.6.The liver,kidney,spleen,lung and other tissues of CNE-2 and Hep-2 tumor-bearing nude mice were taken for pathological preparation for histopathological observation.7.High-throughput sequencing technology was used to sequence the tumor tissue of CNE-2 tumor-bearing nude mice.Results:1.The effective killing concentration(IC50)of EGFR mAb-AuNRs on two tumor cells is almost unaffected by normal human epithelial cells.The cell inhibition rate increases as the concentration of gold nanorods is increased.2.Throughout the experiment period,different concentrations of EGFRmAb-AuNRs were injected through the tail vein and the tumor body.Compared with the control group,the CNE-2 and Hep-2 tumor-bearing nude mice were in ormal posture,with more active,balanced and grip The holding performance is good,the reaction is sensitive,no abnormalities are found in daily activities of eating and drinking,and no death is found.3.Throughout the experimental period,compared with the control group,no abnormalities were found in the diet and drinking water of the CNE-2 group and Hep-2 tumor-bearing nude mice,and there was no difference in defecation and urination from the normal control group.At the same time,in terms of body temperature and breathing,no increase in body temperature and rapid breathing were found,and physiological functions were normal.4.In the CNE-2 group,EGFRmAb-AuNRs showed that hepatic cell edema,extensive mononuclear lymphocyte infiltration,punctate necrosis,focal necrosis and fusion necrosis were observed in the high-injection tumor group,and inflammation was obvious.In other groups,hepatocytes were found to have mild edema,no point necrosis,and no infiltration of inflammatory cells.Renal histopathology showed infiltration of neutrophils and monocytes in the middle and high groups,and the endothelial cells were slightly swollen.No obvious abnormalities were seen in the other groups.The spleen and lung tissue showed no obvious changes under pathological observation.The pathology of liver tissue in the Hep-2 group showed that the liver cells in the high-concentration tumor injection group had mild edema and a small amount of inflammatory cell infiltration,while the other groups found obvious pathological changes.Renal histopathology showed no obvious pathological changes.There were no obvious pathological changes in the spleen and lung tissue under pathological observation.5.In the serum biochemical test results,the serum GOT in the CNE-2 nude mice treatment group generally increased,while the serum GPT content was normal compared with the normal value.The contents were all increased,and the contents of serum albumin,glycated serum protein and chloride ion were not significantly different from the normal values.In nude mice transplanted with Hep-2 tumors,the serum GOT in the treatment group was within the normal range,and the content of serum GPT was normal compared with the normal value.The content of high urea nitrogen in the tumor injection group was slightly increased.Elevated,serum albumin,glycated serum protein and chloride ion content were not significantly different from normal values.6.In the gene sequencing results,the ABCA3 and TMEM238 genes in the CNE-2 group of tumor-bearing nude mice with high-concentration tail vein injection of gold nanorods were highly expressed genes,and the LOC102724843,GRIP2 and FOSB genes were low-expressed genes.The FOSB,GRIP2,BIRC7,SLC47A2,FOS and other genes in the medium-concentration tumor-injected gold nanorod group were highly expressed genes,while the ASS1,ABCA3,TMEM238,REM1 genes were low-expressed genes.The CXCL2,NPIPB15,and SELENOM genes are highly expressed genes,and the C14or132 and DMC1 genes are low expressed genes in the high-concentration tail vein injection gold nanorod group.Conclusions:1.EGFR-AuNRs photothermal treatment will not cause damage to normal cells,but has a killing effect on tumor cells.2.Within the recommended reasonable concentration range,different concentrations of EGFR/AuNR had no effect on the behavior and physiology of Hep-2 and CNE-2 tumor-bearing nude mice.3.EGFRmAb-AuNRs can cause liver damage,may cause slight damage to the kidneys,and no damage to the lungs and spleen.4.Gold nanorods can cause up-regulation of ABCA3,TMEM238,FOSB,GRIP2,BIRC7,SLC47A2,FOS,CXCL2,NPIPB15,and SELENOM genes.