Multivalent glycopolymers containing three different glucose pendant spacer lengths in the polymer backbone having various weight percentages of covalently bonded glucose moieties were obtained by the deacetylation of acetylated polymers synthesized via reversible addition-fragmentation chain transfer (RAFT) process. This allowed us to control the packing density of the glucose moieties in one unit of the glycopolymer. The biological responses of these macromolecules in terms of cytotoxicity of glucose moieties at various spacer lengths and concentrations were investigated without perturbing the intrinsic chemical nature of functional moiety by employing in vitro osteoblast cells. Osteoblast cell adhesion, viability and proliferation response with synthesized glycopolymers revealed that the higher glycopolymer concentration tolerance limit was associated with an increase in pendant spacer length of glucose moiety in the glycopolymer.