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Objective:Observing the effect of exosomes derived from hypoxic Bone marrow mesenchymal stem cells (BMSCs) on the function of chondrocytes, and exploring the role and mechanism of exosomal miR-196b-5p. Evaluating the application prospects of hypoxic BMSCs exosomes and miR-196b-5p for cartilage regeneration.Methods:Chondrocytes were cultured in the supernatant of BMSCs cultured under normoxia or hypoxia, respectively. The proliferation of chondrocytes was detected by CCK-8 assay and the expressions of Collagen type 2 (Col2), Col1, Aggrecan and SOX9 were detected by qPCR to evaluate the effect of hypoxic BMSCs paracrine on chondrocyte functions. Obtaining normoxic and hypoxic exosomes through ultracentrifugation, and testing their effects on the proliferation and anabolic-related genes of chondrocytes through CCK-8 assay and qPCR. Verifying the expression of miR-196b-5p in hypoxic exosomes based on exosomal miRNA array. Knocking out miR-196b-5p in hypoxic BMSCs, and detecting the effect of hypoxic exosomal miR-196b-5p on the functions of chondrocytes by loss-of-function assay. Predicting the downstream of miR-196b-5p through bioinformatics tools, and exploring the mechanism of hypoxic exosomal miR-196b-5p by gain-of-function assays. Hypoxic exosomes and miR-196b-5p-knockout hypoxic exosomes were loaded on silk fibroin hydrogel and subcutaneously into nude mice. After 4 weeks of culture, histological staining of saffron O, Masson and biochemical content of sGAG and collagen were performed to assess the application prospect of hypoxic exosomes and hypoxic exosomal miR-196b-5p on cartilage regeneration. Results:The results of CCK-8 assay and qPCR indicated that the supernatant of hypoxic BMSCs significantly promoted the proliferation of chondrocytes 1.20±0.07 and the expression of cartilage-related markers (Col2 2.95±0.17, Aggrecan 2.45±0.27, SOX9 2.92±0.29) compared to normoxic BMSCs (0.94±0.04, 1.89±0.09, 1.67±0.21, 1.76±0.16), the differences were statistically significant ( P<0.05). The result of CCK-8 assay showed that hypoxic exosomes (1.28±0.04) promoted the proliferation of chondrocytes compared to normoxic exosomes 1.05±0.06, the differences were statistically significant ( P<0.05). CCK-8 assay revealed that the down-regulation of miR-196b-5p in hypoxic exosomes 0.99±0.06 attenuated the proliferation of chondrocytes compared to control group 1.20±0.07, the differences were statistically significant ( P<0.05); the expression of Col2 0.56±0.04, Aggrecan 0.74±0.09, and SOX9 0.45±0.05 in chondrocytes was reduced in the miR-196b-5p knockdown group compared to the control group (1.00±0.09, 1.00±0.12, 1.00±0.07), the differences were statistically significant ( P<0.05). Co-transfection of pmirGLO-BACH1-WT reporter vector with miR-196b-5p mimics decreased the luciferase activity 0.73±0.06, the differences were statistically significant ( P<0.05). Co-transfection of pmirGLO-BACH1-MUT reporter vector with miR-196b-5p mimics showed no change in luciferase activity. BACH1 is the target of miR-196b-5p. Subcutaneous culture in nude mice showed that hypoxic exosomes significantly promoted the deposition of sGAG 383.2±21.54 and collagen 67.40±3.45, while reducing the expression of miR-196b-5p in hypoxic exosomes weakened the deposition of sGAG 258.4±19.50 and collagen 57.15±4.95, the differences were statistically significant ( P<0.05). Conclusion:Hypoxic exosomes promoted the functions of chondrocytes by inhibiting the expression of BACH1 through miR-196b-5p. Hypoxic exosomes can be applied in cartilage regeneration.
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【Objective】 To investigate the effects of miR-126-3p targeting chemokine receptor 1 (CCR1) in exosomes derived from bone marrow mesenchymal stem cells (BMSC) on the proliferation, migration, and invasion of lung cancer cells. 【Methods】 BMSC cells were cultured; exosomes were extracted and identified by the exosomal marker proteins CD63 and TSG101. After exosome culture of A549 cells for different durations (0, 24, 48, and 72 h), cell survival rate was detected by CCK-8, mRNA levels of miR-126-3p and CCR1 were detected by qRT-PCR, and cell migration and invasion abilities were detected by Transwell assay. The relative expressions of CCR1, epithelial cadherin (E-cad), neural cadherin (N-cadherin), and Vimentin were detected by Western blotting. 【Results】 Exosomes had round or oval cup-shaped structures with bright edges and dark middle, with a particle size distribution of about 152 nm, expressing CD63 and TSG101 proteins. The expression of miR-126-3p in exosomes was higher than that in A549 cells. The expression of miR-126-3p was low in A549 cells and that of CCR1 mRNA was high. However, after co-culture with exosomes, the expression of miR-126-3p in A549 cells was increased, while the expression of CCR1 was decreased. A549 cells were cocultured with exosomes for 0, 24, 48, and 72 h. The survival rate, migration and invasion abilities, CCR1 gene and protein expression levels, and N-cad and Vimentin protein expression levels of A549 cells decreased gradually with the extension of culture time. The level of miR-126-3p and the expression of E-cad protein increased gradually with the extension of culture time. 【Conclusion】 The co-culture of exosomes derived from bone marrow mesenchymal stem cells with A549 cells can increase the expression level of miR-126-3p, and miR-126-3p can reduce the proliferation, migration, and invasion of A549 cells by targeting the inhibition of CCR1 expression.
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Myeloma bone disease (MBD) is one of the most common complications of multiple myeloma (MM). MBD is considered to be caused by the activation of osteoclasts and suppression of osteoblasts resulting from the involvement of neoplastic plasma cells and the change of bone marrow microenvironment. It may be a feasible way to improve the treatment of MBD by promoting osteogenic differentiation of bone marrow mesenchymal stem cell (BMSC), from which the osteoblasts mainly originate. Resveratrol (RES), a naturally occurring polyphenolic flavonoid compound, was reported to function in the modulation of bone metabolism. But the effects of RES on osteogenic differentiation of MM derived BMSC (MM-BMSC) and its underlying mechanism remains unknown. Totally 10 cases of MM-BMSCs were isolated, cultured and identified successfully in the present study. RES was found to promote osteogenic differentiation of MM-BMSC by alkaline phosphatase activity assay, qRT-PCR and alizarin red staining. SIRT1 was predicted to be the target gene of RES in promoting osteogenic differentiation with bioinformatic analysis. RES upregulated the expression of silent information regulator 1 (SIRT1) in MM-BMSC (P<0. 001) and its osteogenic differentiation was inhibited in the SIRT1 small interfering RNA (si-SIRT1) transfected group. Furthermore, the mRNA (P<0. 001) and protein (P<0. 01) expression of runt related transcription factor 2 (RUNX2) was increased in the RES treated group and decreased (mRNA P < 0. 01, protein P < 0. 05) in si-SIRT1 transfected group, respectively. In conclusion, resveratrol promotes osteogenic differentiation of MM-BMSCs via upregulating SIRT1/RUNX2 and seems to be a potential therapeutic agent to counteract bone disease in MM patients.
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@#[Abstract] Objective: To investigate the effects of exosome originated from bone marrow mesenchymal stem cell (BMSCs) on proliferation, migration and invasion of prostate cancer PC-3 cell and its mechanism. Methods: qPCR was used to detect the expression level of miR-21-5p in prostate cancer cell lines. The morphology of exosomes isolated from BMSCs was observed with an electron microscope. Western blotting was used to detect the expressions of exosome surface markers and the epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin and Vimentin). Dual luciferase reporter gene experiment was used to detect the targeted regulation relationship between miR-21-5p and PH domain leucine-rich repeat protein phosphatase 2 (PHLPP2). PC-3 cells were co-cultured with 10 μl BMSCs exosomes suspension (Exo group), transfected with sh-PHLPP2 or antagomiR, then CCK-8 and Transwell experiments were used to detect changesinproliferation,migrationandinvasionofPC-3cell.Results: miR-21-5p was highly expressed in prostate cancer PC-3 cell line. The exosomes in the supernatant of BMSCs culture fluid were successfully isolated, and the typical vesicle-like structures of exosomes were observed under transmission electron microscope. Exosomes expressed specific proteins such as CD9, CD63 and CD81. In the Exo group, the proliferation, invasion, migration, as well as the expressions of N-cadherin, Vimentin and miR-21-5p in PC-3 cells were significantly higher than those in the control group (all P<0.05). PHLPP2 is a target gene of miR-21-5p. Compared with the control group, the expression of PHLPP2 in PC-3 cells of Exo group and sh-PHLPP2 group was significantly reduced (0.66±0.09, 0.42±0.05 vs 1.09±0.08, all P<0.01); cell viability, invasion and migration were significantly improved (all P<0.01); and E-cadherin expression level was significantly reduced while N-cadherin and Vimentin expressions were significantly increased (both P<0.05). Conclusion: miR-21-5p is highly expressed in prostate cancer PC-3 cell line. BMSC exosome miR-21-5p can increase the proliferation, migration and invasion ability of PC-3 cells through targeted down-regulation of PHLPP2.
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Objective To compare the actions of Longbie Capsules and bone marrow mesenchymal stem cell (BMSC)transplantation in repairing the damaged cartilage of knee osteoarthritis(KOA)rats. Methods Thirty-six rats aged 4-6 weeks old were induced into KOA model(bilateral knees)by collagenase injection method. All of the modeled rats were randomly divided into model group(intragastric administration of normal saline), BMSC transplantation group(giving tail vein injection of 1 ×106 of BMSCs per time, 2 times every week), and Chinese medicine group (intragastric administration of Longbie Capsules of 7.5 g·kg-1·d-1),12 rats in each group. Six weeks later,the cartilage of rat bilateral knees was taken out. The pathological changes of cartilage were observed by hematoxylin-eosin(HE) staining method, and the protein and mRNA expression levels of Col2a1, X-linkedinhibitor of apoptosis protein (XIAP), HuR in rat knee cartilage were detected by immunohistochemistry and real-time quantitative polymerase chain reaction (qPCR), respectively. Results The HE staining results showed that the cartilage tissue surface was rough with more cracks, and the cartilage cells gathered with the cytoplasm collapsed and arranging disorderly in the model group. The number of chondrocytes was increased and cell surface was flat,and the cracks of the cartilage were decreased with the chondrocytes arranging uniformly in Chinese medicine group and BMSC transplantation group compared with the model group. The results of immunohistochemistry and qPCR detection showed that in Chinese medicine dosage group and BMSC transplantation group, the protein and mRNA expression levels of Col2a1,XIAP and HuR were significantly higher than those in the model group (P<0.05 or P<0.0001), but there was no significant difference between the two medication groups(P>0.05). Conclusion Longbie Capsules and BMSC transplantation can promote the secretion of Col2a1 in the cartilage tissue of KOA rats,improve cartilage, and their mechanism may be related with up-regulating apoptosis-related proteins HuR and XIAP.
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Bone marrow-derived mesenchymal stem cells (BMSCs) are the most promising seed cells for cell transplant. The proliferation of BMSCs is one of the most important determinants of the efficiency of MSC-based transplant therapy. It has been reported that transforming growth factor-β1 (TGF-β1) activates Wnt/β-catenin signaling and regulates cell proliferation. In this study, we investigated the effect of low concentrations of TGF-β1 on proliferation of BMSCs and the related mechanisms. BMSCs were treated with 0, 1, 5 and 10 ng/L recombinant mouse TGF-β1 for 12 h. Cell proliferation was measured by cell counting and MTT assay, and the proliferation-related signals p53, Mdm2, Akt1, Wnt3, phospho-Akt and β-catenin were measured by quantitative polymerase chain reaction (qPCR) and/or Western blot. Our results showed that TGF-β1 at low concentrations induced BMSC proliferation and expression of Mdm2, Akt1, phospho-Akt, Wnt3 and β-catenin, and inhibited p53 expression in dose dependent manner. Importantly, β-catenin siRNA significantly inhibited TGF-β1-induced BMSC proliferation. These findings suggest that low concentrations of TGF-β1 can stimulate proliferation of BMSCs, which is at least partially dependent on the activation of Wnt/β-catenin pathway.
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ObjectiveTo study the effect of bone mesenchymal stem cells (BMSC) transplantation into traumatic brain injury(TBI) rats by the external carotid artery on neurological function and learning and memory.MethodsTen adult SD rats were randomly divided into TBI group ( n =5 ) and BMSC transplantation group ( n=5).Feeney free falling method was used to establish TBI models.The experimental rats were administrated with BMSC via external carotid artery (ECA),while TBI rats were injected with sterile liquid medium of equal volume via right ECA.Neurological function were evaluated according to the modified neurological severity score (NSS) at 1,3,7,15 days.Morris water maze test was used to observe the animal capabilities of place navigation and space exploration at 15 days,then animals were sacrificed.Survival and migration of implanted BMSC in brains under fluorescence microscope. ResultAfter traumatic brain,varying degrees convulsions,paralysis,loss of balance function in rats were found.Compared with TBI group,BMSC transplantation decreased significantly NSS (P ＜0.01 ).BMSC transplantation significantly decreased on escape latency ( ( 20.48 ± 2.29 ) s ) than the TBI group ( ( 85.93 ± 47.48 ) s) (P ＜ 0.01 ).Moreover,BMSC group in the target quadrant dwell time ( ( 28.62 ± 1.72) ％ )and distance ( (29.05 ± 3.08 )％ ) as well as the number of passing the platform (8.00 ± 2.45 ) were significantly higher than the TBI group ( ( 19.37 ± 2.81 ) ％,(21.78 ± 3.06) ％,(2.00 ± 1.87) respectively,P ＜ 0.01 ).Transplanted BMSC could survive and migrate around injury brain through Hochest mark immunofluorescence.ConclusionBMSC can survive and migrate around injury brain by transplantation of external carotid artery,which results in a significant neurological function improvement and learning and memory increase in rats with traumatic brain injury.
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Objective To investigate the effect of intermittent tensile strain on the proliferation and osteogenic differentiation of rBMSCs (rat bone mesenchymal stem cells). Methods Intermittent tensile strain was applied on rBMSCs in vitro by Flexcell 4 000 Tension System (10% elongation amplitude, 0.5 Hz, twice every day, 4 h every time), then effects of the strain after 1, 3, 5, 7 d on cell morphology, cell proliferation, and the relative expression of Cbfα1（core binding factor α1），ALP and collagen I mRNA as well as Cbfα1 protein were measured. Results Intermittent tensile strain slowed the proliferation of rBMSCs from the first day to the seventh day. The relative expression of ALP and collagen I mRNA increased by 3~6 times from the third day（P＜0.05）, meanwhile the expression of Cbfα1 mRNA and protein was up-regulated under the mechanical stimulation. Conclusions Mechanical stretch plays an important role in the proliferation and differentiation of rBMSC, and approprite intermittent tensile strain can slow the proliferation of rBMSC and promot its osteogenic differentiation.
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[Objective] To investigate the effects of preconditioning with low concentration of hydrogen peroxide (H2O2) on oxidative stress-induced BMSC apoptosis.[Methods] In vitro separation,purification,culture,and amplification of bone marrow mesenchymal stem cells were performed.BMSC were insulted with 0,50,100,200,300,400,500 μmol/L H2O2 and the effect of different consentration of H2O2 on BMSC was detected by Flow cytometry (FCM).And then cells were preconditioned with different consentraion of H2O2.(FCM) was used to determine the protective role of H2O2 preconditioning on BMSC apoptosis,BMSC chromatin distribution changes were observed by Hoechst33324;BMSC Caspase-3 and Bcl-2 gene changes were detected by RT-PCR.[Results] Analysis of BMSC apoptosis by flow cytometry showed that H2O2 induced BMSC apoptosis in a dose-dependent manner,and pretreatment of the cells with low concentration of H2O2 prevented subsequent stimulation with high H2O2.RT-PCR results showed that preconditioning with low concentration of H2O2 reduced the BMSC Caspase-3 gene expression but increased Bcl-2 gene expression.[Conclusion] Preconditioning with low concentration of H2O2 has an adaptive role in BMSC,and its mechanism may be related to inhibit abnormal gene expression of Caspase-3 and increase the gene expression of Bcl-2.
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Bone mesenchymal stem cell is a kind of multi-potential stem cells and can differentiate into osteoblast. The differentiation has membrane bone formation and enchondral bone formation two channels, and is regulated by numerous signal transduction pathways. According to related literatures, we review the signal transduction pathways of BMP/Smads, Runx2/osterix, Hedgehog, Wnt/β-Catenin and MAPK.
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PURPOSE: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. MATERIAL & METHODS: Gel-HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel-HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. RESULTS: Gel-HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores (20-40 micrometer). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel-HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. CONCLUSION: The Gel-HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.
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Lapins , Phosphatase alcaline , Anthraquinones , Moelle osseuse , Colorimétrie , Durapatite , Test ELISA , Fémur , Cytométrie en flux , Microscopie électronique à balayage , Nanocomposites , Nanostructures , Ostéocalcine , Graines , Cellules souchesRésumé
PURPOSE: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. MATERIAL & METHODS: Gel-HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel-HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. RESULTS: Gel-HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores (20-40 micrometer). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel-HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. CONCLUSION: The Gel-HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.
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Lapins , Phosphatase alcaline , Anthraquinones , Moelle osseuse , Colorimétrie , Durapatite , Test ELISA , Fémur , Cytométrie en flux , Microscopie électronique à balayage , Nanocomposites , Nanostructures , Ostéocalcine , Graines , Cellules souchesRésumé
[Objective]To explore the potential of a novel collagen I /chitosan /Nano?-tricalcium phosphate(?-TCP)bilayered scaffold for being used in tissue engineering(TE)of osteochondral repair.[Method]Bilayered scaffolds were produced with collagen Ⅰ,chitosan and?-TCP,using a special cross-linking and freeze drying method.The pore size,porosity and interpores of the scaffold were observed by scanning electron microscopy(SEM).Rabbit bone mesenchymal stem cells(BMSC)were isolated and amplified,then inoculated onto the scaffold.By SEM scanning,the condition of the cells adhering onto the scaffold was observed.The proliferation of the cells on the scaffolds was examined using MTT method,and the growth curve was drawn.The cell-scaffold composite were then induced to differentiate towards cartilage by 3-D culturing,and then implanted into muscle pouches 2 weeks later.The result was observed 6 weeks later by HE staining,toluidine blue staining and type Ⅱ collagen immunohistochemistry.[Result]The scaffold possessed high porosity and proper pore size,the porosity was above 95%.BMSC could adhere onto the scaffold well,and the proliferation rate of the cells on the scaffolds was perfectly good.After in vitro induction,BMSC-scaffold composite can differentiate toward cartilage ectopicly.[Conclusion]The novel collagen I /chitosan /?-TCP bilayered scaffold possesses good pore structure and biocompatibility,and will possibly become a new biomaterial of TE used for osteochondral repair.
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@#ObjectiveTo investigate bone marrow mesenchymal stem cells(BMSC) express brain derived neurotrophic factor(BDNF) and nerve growth factor(NGF) and their protective effect for neural stem cells (NSCs).MethodsBMSC were obtained from rat tibiae and femurs and centrifuged with Ficoll. The passage 3 cells were chosen to make immunocytochemical stain for CD44, CD71 and CD45. The expression of BDNF and NGF was detected in BMSC with RT-PCR, as well as in the media with ELISA. The media that cultured BMSC were collected as BMSC condition media. NSCs were obtained from cerebral cortex of new-born rat and cultured in vitro. After different ratio of BMSC condition midia were added, NSCs were induced to apoptosis with heat-shock, then NSCs were dyed with Annexin V-FITC/PI apoptosis kit and apoptosis rates were tested with flow cytometry. ResultsBMSC were CD44(+), CD45(-), CD71(+) and expressed BDNF and NGF mRNA. BDNF and NGF could be tested in the media of cultured BMSC and increase with cultured time. BMSC condition media could reduce the ratio of heat-induced apoptosis of NSCs, and more BMSC condition media showed better effect. ConclusionBMSC can express neurotrophic factores and protect neural stem cells from heat-induced apoptosis.
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Animaux , Enfant , Humains , Souris , Rats , Phosphatase alcaline , Moelle osseuse , Collagène , Dexaméthasone , Sialoprotéine liant les intégrines , Ostéoblastes , Ostéocalcine , Ostéogenèse , Ostéopontine , Rodentia , Cellules souches , Cellules stromales , Donneurs de tissus , Vitamine DRésumé
@#ObjectiveTo explore the feasibility of bone marrow stromal cells (BMSC) of rats differentiating into endothelial cells.MethodsBone marrow cells were obtained by an aseptic technique. Afterwards, the obtained cells were divided into two groups: cells in test group were propagated in 1640 medium with recombinant human GM-CSF (rhGM-CSF) (400ng/ml), and that in control group were propagated in medium with 1640 medium only. The differentiated cells were detected by specific immunology marker by immunocytochemistry and immunofluorescence at day 8.ResultsThe differentiated cells demonstrated the characters of endothelial cells under phase contrast microscopy. Cells of the test groups demonstrated specific characters by immunocytochemistry stain.ConclusionBone marrow stromal cells can multiply vigorously and differentiate into cells with endothelial cells characters in the medium with high concentration rhGM-CSF.
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Objective To further determine their possible synergistic effect on accelerating wound healing, adenovirus vector containing recombinant human hPDGF-A and hBD2 genes was constructed and the expression of exogenous genes in transformed mesenchymal stem cells derived from rat bone marrow was observed. Methods By putting IRES in the middle of hPDGF-A and hBD2, these two genes were expected to be expressed individually. The shuttle vector was named as pAdTrack-hPDGF-A-IRES2-hBD2, which homologously recombinated with Adeasy-1 in BJ5183 cells and formed the mammalian expression vector pAdeasy-hPDGF-A-IRES2-hBD2. Furthermore, the recombinant vector was packaged in 293 cells into infectious recombinant adenovirus, which were used to infect BMSCs. The expression of hPDGF-A and hBD2 in BMSCs was detected by RT-PCR. Results We successfully constructed recombinant adenovirus vector that simultaneously expressed hPDGF-A and hBD2. The expressions of hPDGF-A and hBD2 were confirmed by RT-PCR on transformed BMSCs. Conclusion The established BMSCs that overexpressed hPDGF-A and hBD2 provide a new strategy of combining cell therapy and gene therapy to promote wound healing, especially the chronic one.
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Objective To investigate the affect of body positions on biochemical indexes. Methods By autogenous contrast and cross matched survey, 107 volunteers divided into 3 season patches of winter, spring and summer, blood samples were drawn from the same part in both standing and lying positions。From19 persons, blood samples were collected respectively after standing and sitting for 15 min, lying for 15 min and 30 min and then sitting for another 15 min。 The blood samples were analyzed for 32 biochemical indexes on analyzer。Results 25 biochemical indexes in sitting position were significantly different from those in lying position (P0。05)。Conclusions Changing body position can result in obvious physiological variation of 28 biochemical indexes, particularly of those related to protein. Such result may lead to abnormality in some marginal values. It suggests body position should not be neglected in analyzing laboratory data.
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Objective:To evaluate the periodontal tissue regeneration effect of implantation of bone marrow stromal cells(BMSCs)-root-coralline hydroxyapatite(CHA) complex.Methods:BMSCs of beagle dogs were in vitro cultured on CHA scaffold and on tooth root surface respectively.Then BMSCs-root samples were inserted into BMSCs-CHA scaffolds to form BMSCs-CHA-root complexes.5 complexes were implanted on one side of mandible in each of the dogs 1 month after the 4th,5th and 6th teeth had been extracted.Root-CHA without BMSCs was implanted on another side as the control.All dogs were killed 16 weeks after implantation,and the periodontal tissue regeneration were observed histologically.Results:More new bone formation and more mature alveolar bone were observed in the test group than that in the control,and there was a clearly periodontal ligament-like tissue formation only in the test group.Conclusion:The complex of BMSC-root-CHA can improve the periodontal tissue regeneration.
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Objective:To study the effect of bone marrow stroma cell( BM SC) transplantation combined with multi-pore mineralization ossein(Bio-Oss) in the plerosis of mandibular defect.Methods:A round whole-layer defect in the diameter of 1.5 cm was created on each side of mandible in 16 whit e New Zealand rabbits.The rabbits were randomly divided into two groups.In group 1 BMSCs were collected and cultured with mineralization induce medium for 2 wee ks. Then cell suspension at the density of 5?10 7 /ml and gelatum were mixed a nd dripped into Bio-Oss bones, incubated in DMEM in 50 ml/L CO 2 incuba tor at 37 ℃ for 4-6 h.The BMSC-Bio-Oss was applied to make plerosis of the m andibular defects.In group 2 the defects were repaired with the Bio-Oss ossein only.Samples were collected after 8 and 12 weeks after opreation respectively an d observed with radiology and histology.Results:12 weeks after o peration, (1)in group 1 previous defects were basically consisted of new bone tissue;(2)in group 2 the previous defects were mainly consisted of fibrous tis sue. Conculsions:BMSC-Bio-Oss transplantation may effectively repair bone defect.