Effect of Multiple Intraperitoneal Injections of Human Bone Marrow Mesenchymal Stem Cells on Cuprizone Model of Multiple Sclerosis

Abstract

Background: Bone marrow mesenchymal stem cells (BM-MSCs) elicit neuroprotective effects, and their repair ability has been investigated in different experimental models. We aimed to investigate the effect of multiple i.p. BM-MSCs injections in the cuprizone model of multiple sclerosis in mice.

Methods: Adult male C57BL/6 mice (n = 40) were fed a regular diet or a diet containing cuprizone (0.2% w/w) for 6 six weeks. Bone marrow samples were taken from patients with spinal cord injury. BM-MSCs (2 × 106 in 1 milliliter medium) were administered intraperitoneally for two consecutive weeks at the end of the forth weeks of cuprizone administration. Animals (n = 12) were perfused with 10% paraformaldehyde at the end of sixth week. The brains were sectioned coronally in 6-8-μm thickness (-2.3 to 1.8 mm from bregma). The sections were stained by luxol fast blue-cresyl violet, and images were captured via a microscope. Demyelination ratio was estimated in corpus callosum in a blind manner. A quantitative real-time PCR was used to measure the myelin basic protein gene expression at sixth week.

Results: Histologically, cuprizone induced demyelination in the corpus callosum. Demyelinated area was diminished in the corpus callosum of cell-administered group. Cuprizone could decrease myelin-binding protein mRNAs expression in corpus callosum, which was significantly recovered after BM-MSCs injections.

Conclusion: Our data indicated a remyelination potency of multiple i.p. BM-MSCs in the cuprizone model of multiple sclerosis in mice.

Keywords: Cuprizone; Mesenchymal stem cells; Mice; Multiple sclerosis.

Fig. 1

Stem cells from the bone marrow. Appearance and growth of fibroblastoid cells or bone marrow stromal stem cells at primary culture, passage 1 on days 3 (A), 7 (B), and 10 (C); adipose differentiation of BM-MSCs (D); osteogenic differentiation of BM-MSCs (E); Q-banding of human chromosomes (F).

Fig. 2

Flow cytometry histogram of the immunophenotype of BM-MSCs population. Expressions of five markers (CD90, CD49e, CD73, CD13, and CD105) and negative markers (CD34 and CD45) are shown.

Fig. 3

Histological examination of luxol fast blue myelin staining in corpus collasum. (A): Normal control; (B): cuprizone-induced demyelination (C): cuprizone-induced demyelination with PBS; (D): BM-MSCs induced remyelination. The scale bar is 300 μm

Fig. 4

The effect of BM-MSCs i.p. administration on remyelination in cuprizone-induced demyelination. Each value represents mean ± SD. ⁺p < 0.01 compared to control group. *p < 0.01 compared to model and sham group.

Fig. 5

Expression level of MBP in the corpus collasom. MBP mRNA levels was studied by real-time PCR. Results were expressed as relative gene expression referred to the control and sham groups from data obtained using the equation: 2^-∆∆C_T. *p < 0.01 compared to the model and sham groups.