Ischemic stroke in rats enhances bone resorption in vitro

Abstract

We hypothesized that the formation and differentialtion of osteoclasts are accelerated and the potential of bone resorption is increased in the hemiplegic bone marrow in the early stage of stroke. We randomly divided white female Sprague-Dawley (SD) rats (n = 30) into two groups, stroke (n = 15) and sham group (n = 15). On the 7th day after stroke, after cutting away the epiphyses of the femurs and tibias, diaphyseal channels were flushed using α-minimum essential medium (α-MEM) and bone marrow cells were collected. Bone marrow stem cells, which were extracted from the femur and tibia, were cultured on the 7th day after middle cerebral artery occlusion. We then estimated the ratio of non-adherent cells to total bone marrow cells that included osteoclast precursor cells. After culturing these cells separately, cells that tested positive on the tartrate resistant acid phosphatase (TRAP) were counted and bone resorption was evaluated by using the OAAS™ plate. In comparison to the control group, the stroke group showed a higher increase of non-adherent cells in the hemiplegic side bone marrow. In addition, after the primary culture, the stroke group showed an increased number of TRAP positive cells and a higher degree of bone resorption estimated by OAAS™ plate. As a result, osteoclastogenesis and osteoclast differentiation are accelerated and the potential of bone resorption is increased in the hemiplegic bone marrow and these changes are detected as early as within the first week after middle cerebral artery occlusion in SD rats.

Keywords: Bone Resorption; Osteoclasts; Osteoporosis; Stroke.

Fig. 1

Percentage of non-adherent cells from bone marrow. CTR, controls; S-HS, hemiplegic side of the stroke group; S-NHS, non-hemiplegic side of the stroke group. Data are the mean ± SEM. ^*P < 0.05 control.

Fig. 2

Tartrate resistant acid phosphatase (TRAP) positive cells from bone marrow. CTR, controls; S-HS, hemiplegic side of the stroke group; S-NHS, non-hemiplegic side of the stroke group. Data are the mean ± SEM. ^*P < 0.05 control.

Fig. 3

Photomicrographs of osteoclasts in primary culture obtained from bone marrow of rats, TRAP stain, magnification ×100. (A) Control. (B) Hemiplegic side of the stroke rats. (C) Non-hemiplegic side of the stroke rats.

Fig. 4

The percentage of mineral surface resorbed by osteoclasts on OAAS™ plate. CTR, controls; S-HS, hemiplegic side of the stroke group; S-NHS, non-hemiplegic side of the stroke group. Data are the mean ± SEM. ^*P < 0.05 control.

Fig. 5

Photomicrographs of pits formed by osteoclasts from rats, magnification ×25. (A) Control. (B) Hemiplegic side of the stroke group. (C) Non-hemiplegic side of the stroke group.