[An experimental study on treatment of steroid-associated femoral head necrosis with simvastatin and BMSCs transplantation].
OBJECTIVE
To approach the possibility of combination of simvastatin and BMSCs transplantation for steroid-associated osteonecrosis of femoral head.


METHODS
The BMSCs harvested from 24 rabbits were prepared for cell suspension at a concentration of 1 x 107/mL, and combined with gelatin sponge. Seventy New Zealand white rabbits received one intravenous injection of lipopolysaccharide (10 microg/kg). After 24 hours, three injections of 20 mg/kg of methylprednisolone were given intramuscularly at a time interval of 24 hours. Forty-eight rabbits diagnosed as having femoral head necrosis by MRI were divided into 4 groups randomly, group A: no treatment; group B: only decompression; group C: decompression and BMSCs transplantation; and group D: simvastatin drench (10 mg/kg.d) decompression and BMSCs transplantation. The general information of animals were recorded; after 4 and 8 weeks of operation, 6 rabbits of each group were chosen randomly to do MRI scan, and femoral heads were harvested to do histopathology and scanning electron microscope examination.


RESULTS
After 8 weeks, rabbits became more active than before treatment, and walking way became normal gradually in groups C and D. Four weeks after operation, the MRI low signal region of all groups had no obvious changes, but 8 weeks later, the necrosis signal region of group A magnified while it reduced obviously in group D. Histopathological observation: 4 weeks after operation, diffuse presence of empty lacunae and pyknotic nuclei of osteocytes were found in the trabeculae, and few newborn micrangium could been seen in group A; lots of empty lacunae and a small quantity of newborn micrangium could been found in group B; and large amounts of osteoblats and newborn micrangium were found around the necrosis regions in groups C and D. The positive ratio of empty lacunae and microvessel density in group D were 19.30 +/- 1.52 and 7.08 +/- 1.09, showing significant difference compared with other groups (P < 0.05). After 8 weeks of treatment, the bone trabecula collapsed in many regions in group A; there was fibra callus formation along the decompression channel in group B; few empty lacunae was in the bone trabecular, but the shape of marrow cavity was not normal in group C; and it showed almost normal appearance in group D. The positive ratio of empty lacunae and microvessel density in group D were 11.31 +/- 1.28 and 12.37 +/- 1.32, showing significant differences compared with other groups (P < 0.05), meanwhile, showing significant difference compared with that of 4 weeks after operation(P < 0.05). Scanning electron microscope: 8 weeks after operation, the bone trabecula collapsed in many regions, and few osteoblasts could be found on the surface, a great quantity of fat cells cumulated in the bone marrow in group A; cracked bone trabecula could be found occasionally in group B; the density of bone trabecula was lower than the normal in group C; and the shape of the marrow cavity and the density of bone trabecula were similar to the normal in group D.


CONCLUSIONS
Simvastatin can promote the differentiation of osteocyte and vascular endothelial cell from MSCs, the combination of simvastatin and marrow stem cells transplantation for the treatment of steroid-associated osteonecrosis of femoral head have good application prospects.
Yang
Jianping
J
Department of Orthopeadics, Nanjing First Hospital Affiliated to Nanjing Medical University, Nanjing Jiangsu, 210006, P. R. China.
Wang
Liming
L
Xu
Yan
Y
Wang
Jinsong
J
Wang
Yan
Y
D000818:Animals; D016026:Bone Marrow Transplantation / Q000379:methods*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005271:Femur Head Necrosis / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology / Q000601:surgery*; D008297:Male; D008775:Methylprednisolone / Q000633:toxicity; D011817:Rabbits; D011897:Random Allocation; D019821:Simvastatin / Q000008:administration & dosage / Q000627:therapeutic use*; D033581:Stem Cell Transplantation / Q000379:methods*; D016896:Treatment Outcome
D019821:Simvastatin; D008775:Methylprednisolone