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This study investigates the long-term effects of PEG-PLA nano-artificial cells containing hemoglobin (nanoRBCs) on renal function and renal histology after a one-third blood volume toploading in rats. The experimental rats received one of the following infusions: NanoRBCs in Ringer’s lactate solution, stroma-free hemoglobin (SFHb), polyhemoglobin (polyHb) or autologous rat whole blood (rat RBCs). Blood samples were taken before infusions and on Days 1, 7 and 21 after infusions for biochemistry analysis. Rats were sacrificed on Day 21 after the infusions and their kidneys were excised for histology examination. Infusion of SFHb induced significant decrease in renal function damage as evidenced by elevated serum urea, creatinine and uric acid throughout the 21 days. Kidney histology in the SFHb infusion group revealed focal tubular necrosis and intraluminal cellular debris in the proximal tubules, whereas the glomeruli were not observed to be damaged. There was no abnormalities in renal biochemistry or histology in all the other groups (nanoRBCs, polyHb, Ringer’s lactate solution and rat RBCs). In conclusion, the injections of nanoRBCs did not have adverse effects on renal function or remal histology.