OBJECTIVESTo analyze retrospectively the formation and histological changes of sclerosis rim in patients with osteonecrosis of the femoral head (ONFH), and to study the relationship between bone morphogenetic proteins (BMP4) and sclerosis rim, so as to acquire experimental and theoretical basis on individualized treatment for ONFH patients.

METHODSFrom November 2005 to November 2007, 184 hips of steroid-induced ONFH inpatients were collected. The mean age was (47 ± 7) years, the patients were divided into high (more than 54 years old), middle (40 - 54 years old) and low (less than 40 years old) age groups. Their clinical data were analyzed retrospectively according to gender and age. Parts of the femoral heads were selected for the study, including 18 hips in high age group, 11 hips in low age group and 20 hips in middle age group. Each 10 hips were selected with or without sclerosis rim. The femoral heads were cut along middle coronal plane, their weight-bearing and non-weight-bearing areas were used for the study. The specimens were processed by routine HE staining and picric acid-Sirius red staining and electron microscopy preparation and immunohistochemistry stain. The average optical density of BMP4 protein was calculated by image analysis software.

RESULTSThe trabecular of sclerosis rim was thickening and disorder. But its osteocytes were normal and with high secretion. The ratio of sclerosis rim was 71.4% (105/147) in middle age ONFH patients, which was significantly higher than the low age group patients (45.5%, 5/11) and high age group patients (38.5%, 10/26) (P < 0.01). The optical density of BMP4 in middle age ONFH patients was 0.32 ± 0.14, which was significantly higher than the low age group 0.20 ± 0.17 and high age patients 0.19 ± 0.27 (P < 0.05). The optical density was 0.16 ± 0.11 in ONFH patients without sclerosis rim, which was significantly lower than with sclerosis rim (0.28 ± 0.13) (P < 0.01). The time from hip pain to joint replacement in patients with sclerosis rim was (49 ± 11) months, and (15 ± 2) months without sclerosis rim. There was significant difference between the two groups (P < 0.01).

CONCLUSIONSThe formation of sclerosis rim is positively related to the expression of BMP4, and high expression of BMP maybe promote the formation of sclerosis rim.

Adult ; Bone Morphogenetic Protein 4 ; metabolism ; Female ; Femur Head ; metabolism ; pathology ; Femur Head Necrosis ; metabolism ; pathology ; Humans ; Male ; Middle Aged ; Retrospective Studies

Osteonecrosis of the femoral head is a common clinical orthopedic disease. The progression of the disease is rapid and the prognosis is poor. All the medical profession recognize it as one of the unresolved medical problems. Biomechanical factors play an important role in the course of progression and treatment of osteonecrosis of the femoral head. In this paper, we review the literature and introduce the advanced biomechanical studies on the symptoms, image, collapse, collapse prediction, preserving femoral head surgery of osteonecrosis of the femoral head.
Biomechanical Phenomena ; Femur Head Necrosis ; diagnosis ; metabolism ; pathology ; surgery ; Humans ; Prognosis

PURPOSE: Pathologic changes in the growth plate remain unknown in Legg-Calve-Perthes (LCP) disease. Spontaneously hypertensive rats have proven to be a good model for studying LCP disease. This study investigated the histopathologic changes and the expression of vascular endothelial growth factor in the growth plate of spontaneously hypertensive rats (SHR). MATERIALS AND METHODS: Sixty SHR rats were divided into two groups: those showing osteonecrosis (SHR+n group: 32), and those showing normal ossification (SHR-n group: 28). Thirty Wister Kyoto rats served as a control. For histomorphological measurement, the length of each zone of the growth plate was measured. Cell kinetics was measured by 5-bromo-2'-deoxyuridin (BrdU) immunohistochemistry and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assays. Vascular endothelial growth factor (VEGF) immunohistochemistry was used to identify of expression of VEGF. RESULTS: The lengths of growth plates of the SHR+n group were significantly shorter in the initial growth period than those of the other groups. The lowest proliferative rate and the highest apoptosis rate were observed in the SHR+n group at the initial growth period. The expression of VEGF in the growth plate of the SHR group was lower than the control group, and it was lower in the SHR+n group than in the SHR-n group. CONCLUSION: The growth plate of the SHR+n group was found to be affected by disease process of ischemic necrosis of the femoral head, and this might explain the relative overgrowth of the greater trochanter in the later stages of LCP disease.
Animals ; Apoptosis ; Femur Head/metabolism/*pathology ; Femur Head Necrosis/metabolism/pathology ; Growth Plate/*cytology/metabolism ; Osteogenesis/physiology ; Rats ; Rats, Inbred SHR ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A/metabolism

Adult ; Arthroplasty, Replacement, Hip ; Bone Cysts, Aneurysmal ; pathology ; Bone Neoplasms ; metabolism ; pathology ; surgery ; Chondroblastoma ; pathology ; Chondrosarcoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Femur Head ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Humans ; Osteosarcoma ; pathology ; S100 Proteins ; metabolism

OBJECTIVETo study the effects of Osteoking in promoting gene expression of core binding factor alpha 1 (cbfalpha 1) in necrotic femoral head of rabbits.

METHODSRabbit model of femoral head necrosis (FHN) was induced by intravenous injection of lipopolysaccharide (LPS) 10 microg/kg body weight twice with an interval of 24 h and intramuscular injection with methyl prednisone (MPS) 20 mg/kg body weight 3 times. The dynamic changes of cbfalpha 1 gene expression in the femoral head were observed with immunohistochemistry and real-time quantitative PCR.

RESULTSThe protein expression of cbfa 1 gene was negative in both model and treatment groups at the 4th week, it turned to weakly positive in the treatment group at the 8th and 12th week but still negative in the model group. The mRNA expression of cbfalpha 1 in the treatment group was 2.87 times that in the model group at the 12th week. There was no significant difference in cbfalpha 1 expression between the normal rabbits with or without Osteoking treatment.

CONCLUSIONOsteoking could promote the endogenous cbfalpha 1 expression in the FHN, the effect is better along with the prolonging of the time applied. But it showed no affection on cbfalpha 1 expression in the normal femoral head of rabbits.

Animals ; Core Binding Factor alpha Subunits ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Female ; Femur Head ; drug effects ; metabolism ; pathology ; Femur Head Necrosis ; chemically induced ; genetics ; Gene Expression ; genetics ; Immunohistochemistry ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits ; Random Allocation

OBJECTIVETo observe the regulation of Youguiyin (YGY, ) on the gene expression profile of the rat with steroid-induced femoral head necrosis (sFHN), for the sake of investigating its molecular mechanism of sFHN prevention and treatment.

METHODSAll the 30 rats were randomly divided into three groups, the normal control group (A), the model control group (B), and the YGY treated group (C), 10 in each group. After rats in Groups B and C were being made into FHN models with steroid injection, they received a daily intragastric administration of saline and YGY respectively in equal volume for a total of 6 weeks, while to the unmodeled normal rats in Group A, saline was administered instead. The rats were sacrificed at the terminal of administration; their mRNA from femoral head tissue was extracted and prepared to cDNA probe through inverse transcription for detecting gene expression profile by microarray, outcomes of which was passing fluorescence quantitative PCR verification, and the differential expressed genes were analyzed adopting gene ontology (GO) method.

RESULTSCompared with Group A, the numbers of differential genes found in Groups B and C were 190 and 92, respectively, but the changing trend in the two groups was opposite, mainly manifested as down-regulating in Group B/Group A (GB/GA) and up-regulating in Group C/Group B (GC/GB). The analysis showed that these differential genes were mainly assigned to cell apoptosis, signal transduction, metabolism, cell proliferation and differentiation, cell cycle, blood coagulation, antioxidant activity, etc.

CONCLUSIONSsFHN was regulated by various genes; the regulation of YGY on expressions of these genes and the intra/extra-cellular signaling processes was possibly the molecular mechanism of YGY for preventing/treating sFHN. This study gave an explanation to the effectiveness of Chinese medicine in preventing/treating FHN from aspects of gene expression and enriched the Chinese medicine theory of "Kidney (Shen) governing bones".

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Femur Head ; drug effects ; metabolism ; pathology ; Femur Head Necrosis ; chemically induced ; drug therapy ; genetics ; Gene Expression Profiling ; Male ; Rats ; Rats, Wistar ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; Steroids ; Treatment Outcome

OBJECTIVETo study the relationships among magnetic resonance imaging (MRI), histological findings, and insulin-like growth factor-I (IGF-I) in steroid-induced osteonecrosis of the femoral head in rabbits.

METHODSThirty rabbits were randomly divided into experimental Group A (n=15) and control Group B (n=15). The 7.5 mg/kg (2 ml) of dexamethasone (DEX) and physiological saline (2 ml) were injected into the right gluteus medius muscle twice at one-week intervals in animals of Groups A and B, respectively. At 4, 8 and 16 weeks after obtaining an MRI, the rabbits were sacrificed and the femoral head from one side was removed for histological study of lacunae empty of osteocytes, subchondral vessels, and size of fat cells under microscopy, and the femoral head from the other side was removed for enzyme-linked immunoadsorbent assay (ELISA) for IGF-I.

RESULTSAt 4, 8 and 16 weeks after treatment, no necrotic lesions were detected in Group B, while they were detected in Group A. Light microscopy revealed that the fat cells of the marrow cavity were enlarged, subchondral vessels were evidently decreased, and empty bone lacunae were clearly increased. The IGF-I levels in Group A were significantly higher than those in Group B. At 8 weeks after the DEX injection, the MRI of all 20 femora showed an inhomogeneous, low signal intensity area in the femoral head, and at 16 weeks, the findings of all 10 femora showed a specific "line-like sign". The MRI findings of all femora in Group B were normal.

CONCLUSIONMRI is a highly sensitive means of diagnosing early experimental osteonecrosis of the femoral head. However, the abnormal marrow tissues appeared later than 4 weeks when the expression of IGF-I increased. This reparative factor has an early and important role in response to steroid-induced osteonecrosis of the femoral head, and provides a theoretical foundation for understanding the pathology and designing new therapies.

Animals ; Dexamethasone ; Disease Models, Animal ; Femur Head Necrosis ; chemically induced ; metabolism ; pathology ; Insulin-Like Growth Factor I ; metabolism ; Magnetic Resonance Imaging ; Rabbits ; Steroids

OBJECTIVETo investigate the effect of two treating principles and formulas, which are named 'invigorating spleen to remove phlem and promoting blood circulation to remove meridian obstruction' (Jianpi) and 'invigorate the kidney and promoting blood circulation to remove meridian obstruction' (Bushen), on the expression of osteogenic factors in steroid-induced osteonecrosis of the femoral head (SONFH), such as bone morphogenetic protein 2 (BMP2) and transforming growth factor beta1 (TGFbeta1) and Smads, as well as to explore and compare their mechanisms of prevention and treatment of SONFH.

METHODAnimal model of SONFH was established by injection with methylprednisolone in chest muscle on chickens. 48 SONFH chickens were randomly assigned to model, Jianpi and Bushen group. Another 16 normal chickens served as control group. At the 8th and 16th week, the expression of BMP2, TGFbeta1, Smad4 and Smad7 of bilateral femoral heads were detected with immunohistochemistry.

RESULTThe expression of BMP2, TGFbeta1 and Smad4 decreased, and Smad7 increased significantly in model group compared with control group. The expression of BMP2, TGFbeta1, Smad4 increased and Smad7 decreased significantly in Jianpi group at the 8th week compared with model group, and the same changes in Bushen group at the 16th week.

CONCLUSIONBoth Jianpi and Bushen formulas exerted preventive and therapeutic activity on SONFH through regulating the expression of BMP2, TGFbeta1, Smad4 and Smad7 to promote bone repair. Notably Jianpi formula took effect earlier than Bushen formula

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Chickens ; Drugs, Chinese Herbal ; pharmacology ; Female ; Femur Head Necrosis ; chemically induced ; metabolism ; pathology ; Methylprednisolone ; Osteogenesis ; drug effects ; Random Allocation ; Smad4 Protein ; metabolism ; Smad7 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

PURPOSE: To explore the value of transplanting peripheral blood-derived mesenchymal stem cells from allogenic rabbits (rPBMSCs) to treat osteonecrosis of the femoral head (ONFH). MATERIALS AND METHODS: rPBMSCs were separated/cultured from peripheral blood after granulocyte colony-stimulating factor mobilization. Afterwards, mobilized rPBMSCs from a second passage labeled with PKH26 were transplanted into rabbit ONFH models, which were established by liquid nitrogen freezing, to observe the effect of rPBMSCs on ONFH repair. Then, the mRNA expressions of BMP-2 and PPAR-γ in the femoral head were assessed by RT-PCR. RESULTS: After mobilization, the cultured rPBMSCs expressed mesenchymal markers of CD90, CD44, CD29, and CD105, but failed to express CD45, CD14, and CD34. The colony forming efficiency of mobilized rPBMSCs ranged from 2.8 to 10.8 per million peripheral mononuclear cells. After local transplantation, survival of the engrafted cells reached at least 8 weeks. Therein, BMP-2 was up-regulated, while PPAR-γ mRNA was down-regulated. Additionally, bone density and bone trabeculae tended to increase gradually. CONCLUSION: We confirmed that local transplantation of rPBMSCs benefits ONFH treatment and that the beneficial effects are related to the up-regulation of BMP-2 expression and the down-regulation of PPAR-γ expression.
Animals ; Blood Cells/*cytology ; Bone Morphogenetic Protein 2/genetics ; *Cell- and Tissue-Based Therapy ; Femur Head Necrosis/metabolism/*pathology/*therapy ; Gene Expression Regulation ; *Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells/*cytology ; Osteonecrosis/*pathology/*therapy ; PPAR gamma/genetics ; Rabbits ; Transplantation, Homologous

OBJECTIVETo investigate the effect of Huogu II Formula (II) with medicinal guide Radix Achyranthis Bidentatae (Ach) on bone marrow stem cells (BMSCs) homing to necrosis area after osteonecrosis of the femoral head (ONFH) frozen by liquid nitrogen in rabbit as well as to explore the mechanism of prevention and treatment for ONFH.

METHODSThe animal model of ONFH was established by liquid nitrogen frozen on the rabbit left hind leg. Forty-eight Japanese White rabbits were randomly assigned to sham-operated group, model group, Huogu II group, and Huogu II plus Ach group, with 12 rabbits in each. During the course of ONFH animal model establishment, all rabbits were subcutaneously injected with recombinant human granulocyte colony-stimulating factor [rhG-CSF, 30 μg/(kg·day) for continuous 7 days]. Meanwhile, normal saline and decoction of the two formulae were administrated by gavage, respectively. White blood cells (WBC) were counted in peripheral blood before and after injection of rhG-CSF. Materials were drawn on the 2nd and 4th weeks after model built; bone glutamine protein (BGP) and bone morphogenetic protein 2 (BMP2) levels in serum were tested. Histopathologic changes were observed by hematoxylin and eosin (HE) staining. BMP2 mRNA levels were detected with in situ hybridization (ISH) staining. 5-Bromo-2'-deoxyuridine (BrdU) and stromal cell derived factor 1 (SDF-1) were measured by immunohistochemical assay in femoral head of the left hind leg.

RESULTSCompared with the shamoperated group, the ratio of empty lacuna, serum BGP, and SDF-1 level in the model group increased significantly, and BMP2 in both serum and femoral head decreased significantly. However, in comparison with the model group, the empty lacuna ratio of Huogu II group and Huogu II plus Ach group decreased obviously in addition to the levels of serum BGP and BMP2, and the expressions of BMP2 mRNA, BrdU, and SDF-1 increased significantly. Above changes were particularly obvious in Huogu II plus Ach group. BGP and SDF-1 on the 2nd week and empty lacuna rate and serum BMP2 level on the 4th week in Huogu II group significantly exceeded their counterparts. On the 2nd week, only in Huogu II plus Ach group that the BrdU counting rose significantly. On the 4th week, empty lacuna rate and serum BMP2 level in Huogu II plus Ach group exceeded those in Huogu II group distinctively.

CONCLUSIONSTo a certain extent, the medicinal guide Ach improves the preventive and therapeutic effects of Huogu II Formula on experimental ONFH model. The possible mechanism of this is related to its promoting effect on directional homing of BMSCs to the necrosis area.

Achyranthes ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; Bone Morphogenetic Protein 2 ; blood ; genetics ; Bromodeoxyuridine ; metabolism ; Cell Movement ; Chemokine CXCL12 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Femur Head ; drug effects ; pathology ; Femur Head Necrosis ; blood ; genetics ; pathology ; therapy ; Gene Expression Regulation ; drug effects ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Humans ; Leukocyte Count ; Male ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Radioimmunoassay ; Stem Cell Transplantation ; Stem Cells ; cytology ; drug effects