Proliferation ability of bone marrow mesenchymal stem cells in corticosteroid-induced osteonecrosis of femoral head

doi:10.3969/j.issn.2095-4344.2014.01.002

Abstract

Abstract:

BACKGROUND: Corticosteroid-induced osteonecrosis of femoral head is one of the major causes of the loss of hip joint function. More and more studies have shown that corticosteroid-induced osteonecrosis of femoral head may be associated with proliferation ability of bone marrow mesenchymal stem cells.
OBJECTIVE: To detect the proliferation and differentiation ability of bone marrow mesenchymal stem cells isolated from patients with steroid-induced osteonecrosis of femoral head, providing rational evidences for treatment of corticosteroid-induced osteonecrosis of femoral head with the transplantation of autologous bone marrow containing bone marrow mesenchymal stem cells into the necrotic area of femoral head.
METHODS: Bone marrow mesenchymal stem cells from the femoral heads were collected from patients with corticosteroid-induced osteonecrosis of femoral head, and new femoral neck fractures without osteonecrosis who were scheduled for total hip arthroplasty. In another group, bone marrow mesenchymal stem cells were collected from ilium bone marrow of the same steroid-induced osteonecrosis of femoral head patients. The femoral neck fracture was defined as fracture without preceding trauma or in response to minimal trauma. Cases with corticoid treatment were excluded from the femur neck fracture patients. All bone marrow mesenchymal stem cells were divided three groups: femoral neck fracture group; femoral head group of corticosteroid-induced osteonecrosis of femoral head; ilium group of corticosteroid-induced osteonecrosis of femoral head. The bone marrow mesenchymal stem cells were isolated by enzyme digestion or density gradient centrifugation from bone marrow blood of the three detecting area, and then selected by the adhesive method. Passage 3 bone marrow mesenchymal stem cells were selected for experiments.
RESULTS AND CONCLUSION: The results of methyl-thiazolyl-tetrazolium assay indicated that the bone marrow mesenchymal stem cells obtained from the femoral head group showed reduced proliferation ability compared with those obtained from the other two groups. The percentage of bone marrow mesenchymal stem cells was increased at G0/G1, but decreased significantly at S+G2/M in the femoral head group (P < 0.05). The bone marrow mesenchymal stem cells obtained from the ilium group were proliferated best. The decreased proliferation ability of bone marrow mesenchymal stem cells may play a role in the low repair capacity of corticosteroid-induced osteonecrosis of femoral head, and bone marrow mesenchymal stem cells from the ilium of patients with corticosteroid-induced osteonecrosis of femoral head have a better proliferative ability.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: femur head necrosis, femur head, transplantation, mesenchymal stem cell, bone marrow, femur

CLC Number:

R394.2

Wang Bai-liang, Li Tie-jun, Yue De-bo, Sun Wei. Proliferation ability of bone marrow mesenchymal stem cells in corticosteroid-induced osteonecrosis of femoral head[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(1): 7-13.

Figures/Tables 2

Identification of BMSCs from corticosteroid-induced ONFH patients There was marked difference in growth state of BMSCs between corticosteroid-induced ONFH femoral head group and the other two groups under an inverted microscope 24 hours after primary culture, and BMSCs from all groups adhered and extended in spindle or polygonal shape. Forty-eight hours later, the clone formed and adherent cells increased. However, the number of clones in the corticosteroid-induced ONFH femoral head group was much less than that in the other two groups. BMSCs resembled typical features of fibroblasts 3 days later in the other two groups and 5 days later in the corticosteroid-induced ONFH femoral head group. Clones enlarged and fused into monolayer 7 days later in the femoral neck fracture group and the corticosteroid-induced ONFH ilium group and 10 days later in corticosteroid-induced ONFH femoral head group. In the femoral neck fracture group and the corticosteroid-induced ONFH ilium group, BMSCs proliferated and distributed evenly after passage and adhered partially 2 hours later and completely in funicular shape 24 hours later. In contrast, BMSCs from corticosteroid-induced ONFH femoral head group proliferated very slowly and overgrew the bottle till 10-12 days later. The immunofluorescent staining showed that the cultured cells expressed typical BMSCs markers such as CD29 and CD44, but not typical haematopoietic cell markers including CD34, CD14 and CD133 (Figures 1 A-C). Proliferative activity of BMSCs from corticosteroid- induced ONFH patients The proliferative activities of BMSCs using MTT methods were obtained from the three groups and drawn into the growth curves. As shown in the figures, the proliferative activity of BMSCs in the femoral neck fracture group and the corticosteroid-induced ONFH ilium group were remarkably stronger than that in the corticosteroid-induced ONFH femoral head group. For the corticosteroid-induced ONFH femoral head group, BMSCs growth fell into the stagnant stage within 1-7 days after seeding, turned into the logarithmic stage at 8 days, and came to the plateau stage thereafter. However, the growth curve in the femoral neck fracture group and the corticosteroid-induced ONFH ilium group moved left obviously and the peak increased (Figure 2)."

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