The objective of this study was to evaluate the use of calcium oxide (CaO) on the nutritive profile, digestibility, and carbohydrate fractionation of three sugarcane (Saccharum officinarum hybrids) genotypes: IAC-862480, SP-791011, and CTC-3. Four CaO levels (0, 1.5, 3.0, and 4.5% on a fresh matter basis) were used in a 3 × 4 factorial scheme, whose factors were the three sugarcane genotypes and the four levels of calcium oxide. The chemical composition, carbohydrate fractionation, total digestible nutrients, and in vitro dry matter digestibility (IVDMD) were evaluated. The chemical composition of the treated sugarcane was affected by the genotype and the use of CaO. The CTC-3 genotype showed lower values of crude protein (CP), ether extract (EE), neutral detergent fiber (NDFap), acid detergent fiber (ADF), and phosphorus (p < 0.01) contents when compared to the IAC-862480 genotype. A decreasing linear effect (p < 0.01) of CP, EE, ADF, lignin, cellulose, non-fibrous carbohydrates, and total digestible nutrients was observed with increasing levels of CaO. A quadratic effect was observed for the contents of DM, organic matter, NDFap, and hemicellulose when the sugarcane was treated with CaO (p < 0.05). The treatment of sugarcane with CaO resulted in the reduction (p < 0.05) in the indigestible fraction of sugarcane. The IAC-862480 and SP-791011 genotypes showed a better nutritional profile compared to the CTC-3 genotype. On the other hand, CaO treatment improved IVDMD and decreased the non-digestible fraction of sugarcane.