RESUMO
BACKGROUND: A superior composite scaffold hopes be constructed to resolve adhesion between seed cell and scaffoldmaterial.OBJECTIVE: To construct composite scaffolds with fibrin glue and xenogeneic inorganic bone and to explore the three-dimensional culture of rabbit marrow stromal cells (MSCs).DESIGN, TIME AND SETTING: Observational experiment was performed at the Department of Orthopedics of China-Japan Union Hospital and the Department of Mechanical Engineering, College of Mechanical Engineering of Jilin University from November 2007 to March 2008.MATERIALS: Fibrin glue was made by a certain ratio of fibrinogen and thrombin; bovine cancellous bone following defatting and deproteinization was mixed with fibrin glue to establish composite scaffold.METHODS: Rabbit MSCs were cultured in v#ro and transferred, and the MSCs were collected for three-dimensional culture withcombined scaffolds made of xenogeneic inorganic bone and fibrin glue.MAIN OUTCOME MEASURES: The growth and proliferation of MSCs were examined by phase-contrast microscope andtransmission electron microscopy.RESULTS: Phase contrast microscope showed that the MSCs could spread evenly in the combined scaffolds. After cultured 4 weeks, the MSCs formed into densely three-dimensional net. It could be observed under the transmission electron microscopethat there were micro-protrusions in local stromal cells at 4 weeks after culture, and the mitochondrion as well as ribosomes wasobserved in the cytoplasm with rough endoplasmic reticulum.CONCLUSION: The MSCs cultured in the combination of fibrin glue and xenogeneic inorganic bone scaffolds show a betteractivity, and they can proliferate rapidly.
RESUMO
BACKGROUND: In tissue engineering, three-dimensional biodegradable scaffolds are generally used as a basic structure for cell anchorage, proliferation. Currently, no perfect scaffold is available. OBJECTIVE: To observe the growth of rabbit bone marrow stromal cells (BMSCs) cultured in different-intensity three-dimensional fibrin glue in vitro, and to discuss the feasibility of fibrin glue used as a scaffold material of bone tissue engineering. DESIGN, TIME AND SETTING: The single sample observational study was performed at the China-Japan Union Hospital of Jilin University and School of Mechanical Engineering of Tianjin University of Technology from September 2007 to January 2008. MATERIALS: Fibrinogen and thrombin were mixed at various proportions, and prepared into different intensity fibrin glue. A month-old male New Zealand white rabbits, weighing 0.25 kg was used in this study. METHODS: Rabbit BMSCs were cultured and serial subcultivation in a CO2 incubator. And then the amplified BMSCs were collected and continue to be cultured in different intensity fibrin glue for 4 weeks. MAIN OUTCOME MEASURES：Observation of growing BMSCs is performed using the phase contrast microscope. The activity of BMSCs in fibrin glue at different stages was observed using hematoxylin-eosin staining. The ultrastructural changes of BMSCs were observed which had been cultivated in fibrin glue for 4 weeks. RESULTS: After growing in fibrin glue for 4 weeks, BMSCs showed strongly active status in low intensity fibrin glue and growing slowly or dying in high intensity fibrin glue. Under the electron microscope, BMSCs following 4 weeks culture in fibrin glue (proportation of fibrinogen and thrombin was 4:1) were found, with visible cellular organs, and BMSCs had good activities. CONCLUSION: BMSCs can spread and proliferate quickly in low intensity fibrin glue. The optimal proportion of fibrinogen and thrombin is 4: 1.
RESUMO
Objective To observe the change endoplasmic reticulum stress in renal injury of fluorosis rats,and explore the effect of endoplasmic reticulum stree in the mechanism of renal injury of fluorosis.Methods 48 Wistar rats were divided into 4 groups:control,fluoride-treated,low calcium,and low calcium plus fluoride-treated.The rats in fluoride-treated and low calcium plus fluoride-treated groups were treated with sodium fluoride (NaF,221 mg?L-1) through drinking water for 3 months.The pathological slice of rat kidney was made and the morphological changes were observed under optical microscope.The transcription levels of Grp78,Xbp1,CHOP and PDI in the kidney tissues were determined by reverse transcription-polymerase chain reaction (RT-PCR).Results Edema and vacuolar degeneration of the proximal tubule and distal tubule cells in the kidney tissues in fluoride-treated group were observed;and cell edema,vacuolar degeneration,scattered necrosis,minor regeneration repair and interstitial hyperemia in the kidney tissues in low calcium plus fluoride-treated group were found.The mRNA expressions of Xbp1 in the kidney tissues in fluoride-treated and low calcium plus fluoride-treated groups were markedly higher than that in the responding control groups(P