Mechanical stimulation plays an important role in maintaining the growth and normal function of the skeletal system. Mechanical unloading occurs, for example, in astronauts spending long periods of time in space or in patients on prolonged bed rest, and causes a rapid loss of bone mass. Casein kinase 2‑interacting protein‑1 (CKIP‑1) is a novel negative bone regulation factor that has been demonstrated to reduce bone loss and enhance bone formation. The aim of this study was to investigate the effect of constrained dynamic loading (Loading) in combination with CKIP‑1 gene knockout (KO) on unloading‑induced bone loss in tail‑suspension mice. The blood serum metabolism index [alkaline phosphatase (ALP) activity and osteocalcin (OCN) levels], tibia mechanical behavior (including bone trabecular microstructure parameters and tibia biomechanical properties), osteoblast‑related gene expression [ALP, OCN, collagen I and bone morphogenetic protein‑2 and osteoprotegerin (OPG)] and osteoclast‑related gene expression [receptor activators of NF‑kB ligand (RANKL)] were measured. The results demonstrated that mice experienced a loss of bone mass after four weeks of tail suspension compared with a wild type group. The mechanical properties, microarchitecture and mRNA expression were significantly increased in mice after Loading + KO treatment (P<0.05). Furthermore, compared with loading or KO alone, the ratio of OPG/RANKL was increased in the combined treatment group. The combined effect of Loading + KO was greater than that observed with loading or KO alone (P<0.05). The present study demonstrates that Loading + KO can counter unloading‑induced bone loss, and combining the two treatments has an additive effect. These results indicate that combined therapy could be a novel strategy for the clinical treatment of disuse osteoporosis associated with space travel or bed rest.