The monodispersed spherical MSN with different mesoporous structure (MCM 48) was developed and employed as a template for the synthesis of mesoporous carbon nanoparticles (MCN) via nanocasting. MCN was demonstrated for the delivery of membrane impermeable chemical agents inside the cells. The cellular uptake efficiency and biocompabtibility of MCN with human cervical cancer cells were also investigated. In addition to the biocompabtibility of MCN, MCN was demonstrated to support Rh-Mn nanoparticles for catalytic reaction. Owing to the unique mesoporosity, Rh-Mn nanoparticles can be well distributed inside the mesoporous structure and exhibited interesting catalytic performance on CO hydrogenation. In addition, the synthesis route of the aforementioned MSN (MCM 48) was discussed and investigated in details and other metal oxide nanoparticles were also developed via nanocasting by using MSN (MCM 48) as a template.