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Objectives: In order to assess the biosafety of HAuNS using zebrafish models and the cancer cell lines HepG2, HEK293, and A549, this study prepared HAuNS in a variety of sizes and alterations. Methods: By oxidizing cobalt nanoparticles encased in gold shells, HAuNS were created. In the meantime, PEG- and PEI-coated HAuNS were created. The diameters of the HAuNS that were produced were 30~40 nm, 50~60 nm, and 70~80 nm. MTT assay was used to assess the toxicity of HAuNS on HepG2, HEK293, and A549 cells. For the investigation of their toxicities, HAuNS (50~60 nm) of various concentrations were incubated with zebrafish embryos. Then, cell death was determined using acridine orange staining. Results: In a cell line model, it was demonstrated that purified HAuNS exhibit lower toxicity than unpurified HAuNS. Meanwhile, it was discovered that surface-modified HAuNS was less hazardous than unmodified HAuNS. Unpurified HAuNS (50-60 nm) exposure to embryos caused deformity and increased mortality. Moreover, embryos exposed to HAuNS displayed an increase in cell death, showing that HAuNS can put zebrafish under physiological stress. Conclusion: The possible toxicity of HAuNS is now more understood thanks to this investigation. The details could improve our comprehension of the nanotoxicity of medication delivery systems. Comparing HAuNS (50~60 nm) to the other two particle sizes, its toxicity was quite low. Compared to unpurified HAuNS, purified HAuNS displayed less toxicity. Comparing PEI-HAuNS and HAuNS to PEG-HAuNS, cytotoxicity was found to be lower. Our data support the use of pure HAuNS, HAuNS-PEG, and HAuNS (50~60 nm) as possible photothermal conductors when seen as a whole.