In this paper, the action of suet oil in the preparation of self-assembled micelles of the active flavonoids in Epimedium in the simulated human environment was researched. Twelve suet oil samples were collected from different growing areas and different positions of sheep or goat to simulate the formation of micelles. Then the effects of the fatty acids in suet oil on the preparation of self-assembled micelles were studied furthermore. The results showed that the micelles had a dispersed state and spherical smooth surface. To compare the diameter, potential, encapsulation efficiency and drug loading of the 12 batches micelles, the micelles prepared by the suet oil from Qinghai were more stable and had a higher encapsulation efficiency. The fatty acids in suet oil could promote the formation of self-assembled micelles, but the whole suet oil had a better effect. Above all the study, we confirmed that the suet oil promoted the formation of self-assembled micelles of the flavonoids in Epimedium, it laid foundation for further research about increasing the efficacy of Epimedium and improved the absorption of the active flavonoids in Epimedium.
Animals ; Chemistry, Pharmaceutical ; methods ; China ; Drug Carriers ; chemistry ; Electric Conductivity ; Epimedium ; chemistry ; growth & development ; Fatty Acids ; chemistry ; Flavonoids ; chemistry ; Geography ; Goats ; Humans ; Micelles ; Oils ; chemistry ; Sheep

The present paper is aimed to observe the effects of basic fibroblast growth factor (bFGF) on bone marrow mesenchymal stem cell (BMSCs) differentiation. The bFGF was used to stimulate BMSCs and histology, immunohistochemistry and enzyme linked immunosorbent assay (ELISA) were used to examine the extracellular matrix produced by induced BMSCs, evaluated the feasibility of BMSCs being the seeding cells of temporomandibular joint (TMJ) disc tissue engineering. The results showed that having been induced with bFGF, the BMSCs could differentiate into fibroblast-like cells, which could synthesize GAG and collagen type I matrix. So it is feasible for BMSCs as seeding cells for engineered TMJ disc.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Fibroblast Growth Factor 2 ; pharmacology ; Glycosaminoglycans ; biosynthesis ; Goats ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Temporomandibular Joint Disc ; cytology ; Tissue Engineering

OBJECTIVETo present that Nickel-Titanium (NT) memory alloy staples in fusionless controlling the growth of the vertebrates in the sagittal plane.

METHODSEighteen infant female goats were selected and equally divided into 3 random groups: long staple group, short staple group and blank control group. Five long staple (the legs' length = 7 mm) and five short staple (the legs' length = 4 mm) were implanted into each goat in long and short staple groups respectively by anterior approach, right on the front of the thoracic vertebrae from T(6) to T(11). The control group was not given any treatment. X-ray examination was performed pre-operatively and post-operatively. Cobb angle of lateral radiograph was measured and the data of Cobb angle were statistically analyzed. At the end of the experiment, whether the staples implanted spinal columns were fused or not were evaluated by gross observation.

RESULTSFinally, all of the goats were included in the final results. Before the operations, T(6-11) sagittal Cobb angle was 7.0° ± 2.3° in short staple group, and 6.2° ± 4.0° in long staple group. And after the operation, the T(6-11) Cobb angle was increased to 12.7° ± 4.7° in short staple group with the increased rate of 81.4%, and 14.0° ± 4.9° in long staple group with the increased rate of 125.8%, respectively. Before and after the surgery, there were no significant differences between long staple group and short staple group in terms of Cobb angle (pre-operation P = 0.655, post-operation P = 0.596). Before the surgery, there were no differences in terms of Cobb angle, between long staple groups and control group (P = 0.929), and short staple groups and control group (P = 0.720). At the end of the experiment, there were significant differences between long staple group and control group in terms of Cobb angle (P = 0.007), and between short staple group and control group (P = 0.021). The staples implanted spinal columns were not fused which was proved by gross observation.

CONCLUSIONSThe memory alloy staple implantation by anterior approach, right on the front of the thoracic vertebrae of goats, can control the growth of thoracic vertebrates leading to kyphosis.

Animals ; Bone Nails ; Female ; Goats ; Nickel ; Thoracic Vertebrae ; growth & development ; surgery ; Titanium

OBJECTIVETo examine the effects of high and low concentrations of transforming growth factor (TGF) β(1) and insulin-like growth factor-I (IGF-I) on the extracelluar matrix synthesis of the self-assembled constructs of temporomandibular joint (TMJ) disc.

METHODSThe experimental groups of self-assembled constructs were exposed to IGF-I (10, 100 µg/L) and TGF-β(1) (5, 50 µg/L), the control groups were not added with any growth factors. All groups were examined at 3 and 6 weeks for gross morphological, histological, and biochemical changes. Safranin-O/fast green staining was used to examine glycosaminoglycan (GAG) distribution, picrosirius red and immunohistochemical staining to observe type I collagen distribution. Type I collagen contents were tested by ELISA assay kit, GAG contents were measured by Blyscan GAG assay kit, and the cell numbers were quantified with a Picogreen reagent kit.

RESULTSThe growth factor groups all upregulated the matrix synthesis of the self-assembled constructs compared with control groups. TGF-β(1) (5 µg/L) and IGF-I (10 µg/L) were the two most potent concentration in increasing type I collagen and GAG synthesis and cells proliferation. IGF-I group (10 µg/L) produced nearly 2 times (109.16 ± 5.12 µg) as much type I collagen as the control group (69.13 ± 5.94 µg) at 3 weeks. The matrix contents and the number of the proliferated cells in control group and all GF groups at 6 weeks were more than those at 3 weeks.

CONCLUSIONSIGF-I (10 µg/L) is the most beneficial growth factor and can be applied in tissue-engineering stratigies of the temporomandibular joint disc. At the same time, the exposure time of growth factors is another key factor that affects matrix synthesis of TMJ disc constructs.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Extracellular Matrix ; metabolism ; Glycosaminoglycans ; biosynthesis ; Goats ; Insulin-Like Growth Factor I ; pharmacology ; Temporomandibular Joint Disc ; cytology ; metabolism ; Tissue Engineering ; methods ; Transforming Growth Factor beta1 ; pharmacology

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 effects that shape memory alloy (SMA) staples implanted to the lateral aspect of the thoracic vertebrae on spinal growth in goats.

METHODSSixteen goats (age 2 - 3 months) were divided into 3 groups: six in single staple group; six in double staples group and four in control group. Single staples group underwent right-side thoracotomy for exposing the thoracic spine through the eighth rib. Five SMA staples were placed laterally into vertebral bodies of T(6 - 11) spanning discs. Double staples group underwent the same operation. Laterally directed 10 SMA staples were placed into vertebrae of T(6 - 11) spanning discs and two staple spanning each disc. The last four goats in control groups just only underwent right-side thoracotomy. In the next 4 months after operation, radiographs were taken to observe the spinal growth every month.

RESULTSThe radiographic analysis demonstrated scoliosis of 12.83 degrees +/- 12.17 degrees in single staple group and 12.00 degrees +/- 3.22 degrees in double staple group after 2 months of the operation. Cobb angle of 6.00 degrees +/- 4.94 degrees and 25.17 degrees +/- 3.71 degrees were observed in the two groups respectively after 4 months of operation, as compared with 0 degrees in the control groups. Only 2 goats developed kyphosis.

CONCLUSIONSCompression between vertebral bodies by SMA staples can depress spinal growth in the same side and greater compression result in larger curves.

Alloys ; Animals ; Bone Nails ; Female ; Goats ; Spine ; growth & development ; Thoracic Vertebrae ; surgery ; Thoracotomy ; instrumentation ; methods

Animal Feed ; Animals ; Cysteamine ; Female ; Goats ; Mammary Glands, Animal ; growth & development ; physiology

The morphological changes in the anterior horn of the L4 and L5 spinal segments were observed following anterior root avulsion in the adult male Sprague-Dawley rat (300~350 gm) at 5 days, 1 week, 2 weeks and 3 weeks postlesion. The animals were perfused with 4% paraformaldehyde, 0.15% picric acid in 0.1 M phosphate buffer solution and cryostat sections were prepared. Immunohistochemistry was used to identify changes of the phenotype in the anterior horn cells. Primary antibodies, goat anti-choline acetyltransferase (ChaT, 1 : 500, Chemicon), mouse antirat ED-1 (1 : 200, Serotec), rabbit anti-glial fibrillary acidic protein (GFAP, 1 : 200, DAKO) and rabbit anti-vascular endothelial growth factor (VEGF, 1 : 500, Santa Cruz Biotechnology) were used. Avidin-Biotin complex method was performed for immunohistochemical reaction and color reaction was developed with DAB-H2O2. Following results were observed in the anterior horn of lumbar spinal cord; 1. The number of ChaT-immunoreactive (ir) cells were reduced 20% level of control animals at 3 weeks after avulsion. 2. ED-1-ir microglia were significantly increased at 1 week and processes of ED-1-ir microglia surrounded around the axotomized neuronal cell bodies. 3. Gliosis defined by extensive GFAP immunoreactivity was observed both ipsilateral and contralateral side of lesion but the VEGF-ir cells were significantly increased in the ipsilateral side of lesion. Therefore, this study suggested that the majority of axotomized motor neurons were degenerated and the cellular proliferation and phenotype changes including glial cell activation were observed in the lumbar spinal cord after anterior root avulsion of adult rats.
Adult* ; Animals ; Anterior Horn Cells* ; Antibodies ; Cell Proliferation ; Choline O-Acetyltransferase ; Endothelial Growth Factors ; Gliosis ; Goats ; Horns ; Humans ; Immunohistochemistry ; Male ; Mice ; Microglia ; Motor Neurons ; Neuroglia ; Neurons ; Phenotype ; Rats* ; Rats, Sprague-Dawley ; Spinal Cord* ; Vascular Endothelial Growth Factor A

OBJECTIVETo evaluate the osteogenic potential of bone morphogenetic protein (BMP)-2 gene transfected goat bone marrow-derived mesenchymal stem cells (MSCs).

METHODSGoat bone marrow-derived MSCs were transfected by Adv-human bone morphogenetic protein (hBMP)-2 gene (Group 1), Adv-beta gal transfected MSCs (Group 2) and uninfected MSCs (Group 3). Western blot analysis, alkaline phosphatase staining, Von Kossa staining and transmission electron microscopy were adopted to determine the phenotype of MSCs. Then the cells were injected into thigh muscles of the nude mice. Radiographical and histological evaluations were performed at different intervals.

RESULTSOnly Adv-hBMP-2 transfected MSCs produced hBMP-2. These cells were positive for alkaline phosphatase staining at the 12th day and were positive for Von Kossa staining at the 16th day after gene transfer. Electron microscopic observation showed that there were more rough endoplasmic reticulum, mitochondria and lysosomes in Adv-hBMP-2 transfected MSCs compared to MSCs of other two groups. At the 3rd and 6th weeks after cell injection, ectopic bones were observed in muscles of nude mice of Group 1. Only fibrous tissue or a little bone was found in other two groups.

CONCLUSIONSBMP-2 gene transfected MSCs can differentiate into osteoblasts in vitro and induce bone formation in vivo.

Animals ; Blotting, Western ; Bone Marrow Cells ; cytology ; metabolism ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; genetics ; Cell Differentiation ; Genetic Therapy ; Goats ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Nude ; Osteogenesis ; physiology ; Staining and Labeling ; Tissue Engineering ; Transfection ; Transforming Growth Factor beta ; genetics

OBJECTIVETo investigate the effectiveness of human insulin-like growth factor I (hIGF-I) gene transferred into the cultured goat bone marrow mesenchymal stem cells with liposome, and find a new method of cell-mediated gene therapy.

METHODSBone marrow was extracted from adult goats and cultured in vitro by monolayer. Then the recombinant eukaryotic expression plasmid pIRES2-EGFP-hIGF was transfected into cells by FuGene 6. After transfection, the marker gene coding enhanced green fluorescent protein (EGFP) was observed at different time points. The hIGF concentration in the supernatant fluids was measured by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry stain of hIGF was performed to detect the target protein. Besides, reverse transcription polymerase chain reaction and flow cytometry were also adopted in order to find out the changes of cells after transfection.

RESULTSBone marrow stem cells were all in the long shuttle-like shape and adhered to the disk. The expression of EGFP was first found at 4 h after transfection. The amount and intensity of EGFP increased gradually during the period of detection and got to the peak degree at 72 h, after that decreased slowly. EGFP was also seen in the second generation cells, but the intensity was relatively faint. The IGF-I concentration secreted into the supernatant was in accordance with the EGFP observed with the peak concentration at 34.75 ng/ml. The outcome of RT-PCR and immunohistochemistry was positive. The morphology of many stem cells was changed into triangular or irregular forms under the circumstance of the secreted hIGF-I. Percentage of stem cells in the S stage increased after transfection.

CONCLUSIONThe hIGF-I gene can be transfected efficiently and safely into BMSCs by FuGene 6, and the hIGF-I protein can be secreted into the supernatant in a relatively high level during a long period, therefore accelerate the proliferation and differentiation of the transfected cells.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Genetic Vectors ; Goats ; Humans ; In Vitro Techniques ; Insulin-Like Growth Factor I ; biosynthesis ; genetics ; Male ; Mesenchymal Stromal Cells ; cytology ; Plasmids ; genetics ; Transfection