This paper deals with the preparation of novel magnetic materials made from tetraaza macrocyclic Schiff base bacterial cellulose ligands with magnetite nanoparticles (Fe3O4NPs) through a multi-step procedure for antimicrobial and cytotoxic activities and chemotherapy in cancer treatment. First, the 2,3-dialdehyde bacterial cellulose (DABC) was chemically modified by ethylenediamine (EDA) and benzil (Bzl) in the presence of ferrous ions. Then, the magnetite nanoparticles (Fe3O4NPs) was produced inside the complex [Fe(DABC-EDA-Bzl)Cl2] through a co-precipitation method. In nanobiotechnology, the magnetic [Fe3O4NP-INS-(DABC-EDA-Bzl)] material was showed moderate antimicrobial and cytotoxic activities against different species and cells, respectively. In particular, the magnetic [Fe3O4NP-INS-(DABC-EDA-Bzl)] material have not any cytotoxic activity towards peripheral blood mononucleocyte (PBMC) cells. Anti-tumor studies demonstrated that the magnetic [Fe3O4NP-INS-(DABC-EDA-Bzl)] material effectively inhibits the growth of the CT26 tumor model in BALB/c mice compared with other resulting materials throughout the experimental period and can be effective drug delivery in nanomedicine.