Cell culture is one of the usual methods for studying living cell and tissue, the method presented in this paper is based on image processing and analyzing technology for activity estimation of mesenchymal stem cells (MSCs). The existing activity estimation methods are costly, complex and invasive. In this method, thresholding is used to preprocess image and to separate out the growth hallow. Then the area is calculated by counting the pixels of the growth hallow. The changes of the activity estimated by this method are similar to those by corresponding cellular experiments. Compared with the existing methods in biology, medicine or medical cellular science, this method is easier, faster, cost-effective and non-invasive. The proposed method has been proved to be efficient by primary experiments of MSCs.
Animals ; Cells, Cultured ; Femur ; cytology ; Image Processing, Computer-Assisted ; methods ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Sprague-Dawley

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 alpha-minimum essential medium (alpha-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(TM) 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(TM) 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.
Animals ; Bone Marrow Cells/cytology/drug effects ; Bone Resorption/*physiopathology ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Female ; Femur/cytology ; Osteoclasts/cytology ; Rats ; Rats, Sprague-Dawley ; Stem Cells/cytology/metabolism ; Stroke/*metabolism/pathology ; Tartrates/pharmacology ; Tibia/cytology

OBJECTIVETo develop an epiphyseal slide-traction plate in child, which can supply the fracture a sufficient internal fixation, and will not restrain the growth of epiphyses. Animal experiments were carried out with the plates to compare the slide-traction with traditional plate.

METHODSDevelop a slide-traction plate for the configuration of the femur condylus of children. Thirty adolescent goats in the experiment were divided into control group (12 goats) and plate group (18 goats). In plate group, right femurs of goats were fixed with common plates and the left femurs with slide-traction plates. All the goats were given X-ray examination at different time after surgery. And the goats were sacrificed at 3 and 6 month, histological method and electron microscopy were performed to evaluate the development of epiphyseal plate.

RESULTSThe both femurs of the goats in control group have no difference in evidence in length at all time we examined. And the both femurs of the goats fixed with plates have no difference in evidence in length at 1 day after surgery. However, the both femurs of the goats fixed with plates have difference in evidence in length at 1 month, 2 month, 3 month, 6 month after surgery. The increased length of the femurs at I month, 2 month, 3 month, 6 month after surgery was also compared with the length at 1 day after surgery, there was difference in evidence between the right femurs of the control group and the femurs were fixed with common plates, but no difference in evidence between the left femurs of the normal control group and the femurs were fixed with slide-traction plate (P > 0.05). More thicker epiphyseal plate were found in the left femurs than the right femurs of the group fixed with plates at 3 and 6 month after surgery (P < 0.01). In the plate group, safranine O staining showed epiphyseal plates at the left femurs had more fuscous staining than the right femurs at 3 and 6 month after surgery and electron microscopy also found that the cells of the epiphyseal plates of left femurs were more eugenic than the right femurs at 3 and 6 month after surgery.

CONCLUSIONThe epiphyseal slide-traction plate can slide with the growth of epiphyses, which is suitable for fixation of the fracture in this part.

Animals ; Bone Plates ; Female ; Femur ; cytology ; diagnostic imaging ; growth & development ; surgery ; Goats ; Growth Plate ; cytology ; diagnostic imaging ; growth & development ; surgery ; Humans ; Internal Fixators ; Male ; Orthopedic Procedures ; Radiography ; Traction

OBJECTIVETo observe the osteoinductive activity of demineralized bone matrix (DBM) and deproteinized bone (DPB) made from human avascular necrotic femoral head.

METHODSThe femoral head was cut into pieces with the size of 3 mm x 3 mm x 5 mm, which were made into DBM and DPB. These two kinds of biomaterials were cocultured with human bone mesenchymal stem cells (hBMSCs). Monolayer cells without biomaterials were cultured as control. Proliferative activity of hBMSCs was evaluated on days 1, 3, 5, 7 and 14. The concentration of alkaline phosphatase (ALP), osteocalcin (OC), and Ca(2+) were detected on days 1, 7, 14 and 21.

RESULTSCells cultured in DBM showed higher proliferative activity than did in DPB and monolayer cells (F =39.773, P <0.01). DBM and DPB also had osteoinductive activity. The concentrations of ALP (F=93.162, P <0.01), OC (F =236.852, P < 0.01), Ca(2+)(F =80.711, P <0.01) of DBM group were significantly higher than that of DPB and control groups.

CONCLUSIONSIn vitro, DBM and DPB made from avascular necrotic femoral head have osteoinductive activity when cocultured with hBMSCs, and the former is stronger than the latter.

Alkaline Phosphatase ; analysis ; Biocompatible Materials ; Bone Demineralization Technique ; Bone Matrix ; cytology ; Bone and Bones ; cytology ; Calcium ; analysis ; Cell Proliferation ; Cells, Cultured ; Femur Head Necrosis ; Humans ; Mesenchymal Stromal Cells ; cytology ; Osteocalcin ; analysis ; Osteogenesis

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Femur Head Necrosis ; therapy ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Osteoblasts ; cytology ; Phytotherapy

OBJECTIVETo fabricate bone grafts by bone marrow stromal cell combined with modified PLGA/Type-I collagen compound scaffold using tissue engineering method.

METHODSThe modified PLGA/Type-I collagen compound scaffold was fabricated. The rabbit primary cultured osteoblasts were identified and seeded onto the modified compound scaffold for one week in vitro. The adhesion and growth of cells were observed with scanning electron microscope. The complex of cells and scaffold was implanted into the subcutaneous region of rabbits and new bone formation was evaluated.

RESULTSThe rabbit bone marrow stromal cells were induced and differentiated into osteoblasts. The adhesion and growth of osteoblasts in cluster were observed on the surface of scaffolds. New bone formation was observed at one month postoperatively and active osteoblasts were found on the surface of the newly formed bone in vivo.

CONCLUSIONThe complex of PLGA and type-I collagen is an appropriate biodegradable scaffold and can be applied in bone tissue engineering.

Absorbable Implants ; Animals ; Biocompatible Materials ; Cells, Cultured ; Collagen Type I ; therapeutic use ; Female ; Femur ; cytology ; Lactic Acid ; therapeutic use ; Male ; Osteoblasts ; cytology ; Polyglycolic Acid ; therapeutic use ; Polymers ; therapeutic use ; Prostheses and Implants ; Rabbits ; Stents ; Stromal Cells ; cytology ; transplantation ; Tissue Engineering

OBJECTIVE: To study the effect of electro- acupuncture at Zusanli acupoint in regulating perioperative cell immune functions in rats. METHODS: Forty-two SD rats were divided into blank control group (=6), model group (=18), and electroacupuncture group (=18). The rats in the latter two groups underwent thigh incision and femoral dissection under anesthesia; the rats in electro-acupuncture group received electro-acupuncture at bilateral Zusanli acupoint for 15 min before anesthesia and 1 h after the surgery. The rats in the model group and electro-acupuncture group were sacrificed at 6 h, 24 h, and 72 h after the operation and blood samples were taken from the ventricle for analyzing CD3, CD4, and CD8 T cell subpopulations and calculation of CD4/CD8 using flow cytometry. ELISA was used to detect the levels of interleukin-1 (IL-1) and IL-6. RESULTS: The CD3 T cell subpopulation was significantly lower in the model group and electro-acupuncture group than in the blank group at 6 h and 24 h after the operation. At 72 h after the operation, CD3 subpopulation levels still remained low in the model group, but recovered the control level in electro-acupuncture group. At each time point of measurement, CD3 level was significantly lower in the model group than in the electro-acupuncture group. CD4 level in the model group was significantly lowered at 6 h and 24 h after the operation, and recovered the control level at 72 h. In the electro-acupuncture group, CD4 level was significantly lowered at 6 h after the operation, but recovered the control level at 24 h. At 24 h and 72 h, the levels of CD4 were significantly lower in the model group than in the electro-acupuncture group. CD8 level underwent no significant changes after the operation in either the model group or electro-acupuncture group. CD4/CD8 was significantly lowered at 24 h and 72 h after the operation in the model group but showed no significant variation in the electro-acupuncture group. Compared with that in the control group, IL-1 level was significantly lowered in both the model group and electroacupuncture group at 6 h, 24 h, and 72 h after the operation, and was significantly lower in the model group than in the electroacupuncture group at these time points. IL-6 level increased significantly in the model group and the electro- acupuncture group at 6 h and 24 h. at 72 h, IL-6 level was obviously lowered in the electro-acupuncture group but remained elevated in the model group. CONCLUSIONS Electro-acupuncture alleviates postoperative immune suppression and promotes recovery of the immune function in rats, suggesting a protective effect of electro-acupuncture at Zusanli acupoint on cellular immune function after surgery.
Acupuncture Points ; Animals ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Electroacupuncture ; methods ; Femur ; surgery ; Flow Cytometry ; Humans ; Immunity, Cellular ; Perioperative Period ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; cytology ; immunology

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

This study was aimed to evaluate the biocompatibility and mechanical property of carbon/carbon composites. At first, carbon/carbon composites were prepared by chemical vapor deposition, and the mechanical property of carbon/carbon composites was tested. The biocompatibility of carbon/carbon composites was evaluated by cytotoxicity test, sensitization test, micronucleus test and implantation test. Mechanical property test showed such carbon/carbon composites are of good compression property and tension property. Cytotoxicity test showed that the leaching liquor of samples has no effect on the growth and proliferation of L-929 cells. The medullary micronucleus frequency of mouse was 2.3 per thousand +/- 0.7 per thousand in experiment group. The sensitization test showed that the skin of the subjects of experiment group had slight erythema and edema, which was 0.188 +/- 0.40 according to Magnusson and Kligman classification. Implantation test revealed that there was slight inflammation around the tissue after the implantation of sample. At 12 weeks, scanning electron microscopy and histopathological exam indicated that the samples of experiment group were of good histocompatibility; and in comparison with control group, there was no significant differences (P > 0.05). So these kinds of samples have good biocompatibility, mechanical property and prospects of clinical application.
Animals ; Biocompatible Materials ; chemistry ; Bone Substitutes ; Carbon ; chemistry ; Cell Line ; Female ; Femur ; surgery ; Fibroblasts ; cytology ; Implants, Experimental ; Male ; Mice ; Micronucleus Tests ; Rabbits ; Random Allocation

OBJECTIVETo review the recent developments in the mechanisms of glucocorticoids induced osteonecrosis of femoral head (ONFH) and introduce a new theory of ONFH.

DATA SOURCESBoth Chinese- and English-language literatures were searched using MEDLINE (1997 - 2011), Pubmed (1997 - 2011) and the Index of Chinese-language Literature (1997 - 2011).

STUDY SELECTIONData from published articles about mechanisms of glucocorticoids induced ONFH in recent domestic and foreign literature were selected. Data extraction Data were mainly extracted from 61 articles which are listed in the reference section of this review.

RESULTSGlucocorticoids are steroid hormones secreted by the adrenal cortex that play a pivotal role in the regulation of a variety of developmental, metabolic and immune functions. However, high dose of exogenous glucocorticoids usage is the most common non-traumatic cause of ONFH. Glucocorticoids can affect the metabolisms of osteoblasts, osteoclasts, bone marrow stromal cells and adipocytes which decrease osteoblasts formation but increase adipocytes formation and cause ONFH finally.

CONCLUSIONSGlucocorticoids affect the differentiation of mesenchymal stem cells, through activating or inhibiting the related transcript regulators of osteogenesis and adipogenesis. At last, the size and volume of mesenchymal stem cells derived adipocytes will increase amazingly, but the osteoblasts will be decreased obviously. In the meantime, the activity of the osteoclasts will be activated. So, these mechanisms work together and lead to ONFH.

Animals ; Bone Marrow Cells ; drug effects ; metabolism ; Femur Head ; cytology ; Glucocorticoids ; metabolism ; pharmacology ; Humans ; Osteonecrosis ; etiology ; metabolism ; Stromal Cells ; drug effects ; metabolism