Viability inhibition effect of gambogic acid combined with cisplatin on osteosarcoma cells via mitochondria-independent apoptotic pathway.

We previously demonstrated that gambogic acid (GA) is a promising chemotherapeutic compound for human osteosarcoma treatment. The aim of this study was to detect whether the combination of lower-dose GA (0.3 mg/L) and cisplatin (CDDP) (1 mg/L) could perform a synergistic effect on inhibiting tumor in four osteosarcoma cell lines. Our results showed that the combination between GA at lower dose and CDDP significantly exerts a synergistic effect on inhibiting the cellular viability in MG63, HOS, and U2OS cells. In contrast, an antagonistic character was detected in SAOS2 cells exposed to the combined use of lower-dose GA (0.3 mg/L) and CDDP (1 mg/L). Then, analysis of cell cycle showed the combination of both drugs significantly induced the G2/M phase arrest, without any difference relative to GA treatment alone, in MG63 cells. Flow-cytometric analysis of cell apoptosis displayed that the apoptotic rate in the combination group is higher than that in GA treatment alone in MG63, HOS, and U2OS cells. The combined use of both drugs had no effect on mitochondrial membrane potential, but promoted the apoptosis-inducing function through triggering of CDDP in the three cell lines. By measurement of mitochondrial membrane potential, the activity of caspase-3 and the expressions of caspase-8 and caspase-9, it was showed that the apoptosis-promoting effect of the combined use of both drugs could be dependent on the death receptor apoptosis pathway, not dependent on the mitochondria apoptosis mechanism. This research, for the first time, demonstrates that GA could increase the chemotherapeutic effect of CDDP in human osteosarcoma treatment through inducing the cell cycle arrest and promoting cell apoptosis.

[In vitro study of combination rhOPG-Fc and alendronate on inhibition osteoclast].

OBJECTIVE: To study the effect of combination rhOPG-Fc and alendronate on mature osteoclasts. METHODS: Recombinant human osteoprotegerin secretory expression in P. pastoris was performed. Osteoblasts were got from new born mouse skeletal bone and proved by ALP staining and incubated together with osteoclasts precursor cell line Raw 264.7 in 96 well plate. After 9 d, 10 micromol/L ALN, 10(-5) g/L rhOPG-Fc, 10 micromol/L ALN + 10(-5) g/L rhOPG-Fc, 5 micromol/L ALN + 5 x 10(-6) g/L rhOPG-Fc were added to these coculture systems. Osteoblasts cultured without the drugs mentioned above served as controls. TRAP stain positive cells counting and cortical bone pit formation counting were preformed in the following the 3rd and 7th d. RESULTS: SDS-PAGE and Western blot showed that molecular weight of the expressed protein was about 55 KD, and it could reach specifically with anti-IgG antibody. Many multi-nuclear TRAP stain positive cells were found in the coculture control group after 9 d incubation, and proved to be mature osteoclasts by TRAP stain. In the 3rd and 7th d after the addition of rhOPG-Fc, ALN or both, TRAP stain positive cells counting and cortical bone pit formation counting decreased significantly in the rhOPG-Fc, ALN or both groups than in the control group, and the combine group (10(-5) g/L rhOPG-Fc + 10 micromol/L ALN) decreased most significantly when compared with rhopG-FC or ALN single. CONCLUSIONS: rhOPG-Fc can decrease the number of osteoclasts and inhibit their function. The combination of both rhOPG-Fc and ALN shows the significant inhibition effect on mature osteoclasts.

Extensor tendon injury due to repetitive wrist dorsiflexion: morphological study of extensor retinaculum and extensor tendon.

Most etiological studies of extensor tendon injury were based on the normal anatomy of extensor tendon and extensor retinaculum of the wrist. Further understanding of the morphological changes of the extensor tendon and extensor retinaculum during wrist dorsiflexion might contribute to improved and more accurate understanding of the etiology. The morphology of the extensor tendon of the mid-finger and the fourth compartment of the wrist extensor retinaculum was studied by sonography, and the anatomy was studied in 15 extremities from 11 young male cadavers. Compared with anatomical images, ultrasonography provides similar morphological observations of the extensor retinaculum of the wrist and extensor tendon. Ultrasonography findings revealed that as the dorsiflexion angle changed, the extensor retinaculum of the wrist formed different shaped trochleas. The trochlea guides the rotation of the extensor tendon at the wrist, but it does not form a sharp corner with the extensor tendon; thus, the extensor tendon is not compressed. As the dorsiflexion angle increased from 0° to 60°, the length of the trochlea gradually decreases. The shortening of the trochlea length will lead to a smaller frictional contact area between the extensor tendon and the extensor retinaculum. Consequently, the friction is centralized. During wrist dorsiflexion, the extensor retinaculum provides a trochlea for the extensor tendon. Extensor tendon injury of repetitive wrist dorsiflexion might be caused by centralized friction at the small contact area.