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BACKGROUND:How to avoid denervated muscular atrophy is a key factor to improve the therapeutic efficacy on peripheral nerve injuries. OBJECTIVE:To study the effect of basic fibroblast growth factor (bFGF) gene-transfected bone marrow mesenchymal stem cels against denervated muscle atrophy. METHODS: bFGF genes were transfected into rat bone marrow mesenchymal stem cels using viral transfection method, and then MTT, immunohistochemical staining, hematoxylin-eosin staining, RT-PCR, western blot, and ELISA methods were used to detect the transfection efficiency and product expression. Thirty-two Sprague-Dawley rats were selected to make animal models of sciatic nerve injury, and subjected to multi-point intramuscular injection of bFGF-transfected bone marrow mesenchymal stem cels (experimental group) or cel culture fluid (control group). At 2, 4, 6, 8 weeks after transfection, the gastrocnemius muscle tissues were harvested to detect action potential, residual wet weight, and cross-sectional area of muscle fibers. RESULTS AND CONCLUSION:The bFGF gene was successfuly transfected into bone marrow mesenchymal stem cels using the viral transfection method. The residual wet weight, cross-sectional area and residual action potential of the gastrocnemius muscle were significantly better in the experimental group than the control group (P < 0.05). These findings indicate that bFGF gene-transfected bone marrow mesenchymal stem cels transplanted into the denervated muscle can retard the development of muscle atrophy. Cite this article:Yu N, Wang YS, Qi CP.Application of basic fibroblast growth factor gene-transfected bone marrow mesenchymal stem cels in denervated muscle atrophy. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):89-94.
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BACKGROUND:Choosing an effective means to label and trace the distribution, differentiation and migration of celsin vivo help to further explore the specific mechanism of cels that exert a therapeutic effect. OBJECTIVE:To understand the migration and localization of BrdU-labeled human umbilical cord mesenchymal stem cels in brain injury model rats. METHODS:Human umbilical cord blood samples were obtained, and the isolation of human umbilical cord mesenchymal stem cels was carried out. The primary and passage culture were performed. The phenotype of cels was detected by flow cytometry. Passage 3 human umbilical cord mesenchymal stem cels were labeled using BrdU, and the cel proliferation was detected using MTT method. BrdU-labeled cels were injected into brain injury ratsvia the tail vein. At 14 days after transplantation, brain tissues in the injury region were cut into sections and the migration and location of the umbilical cord mesenchymal stem cels were observed under inverted fluorescence microscope. RESULTS AND CONCLUSION: Cel surface specific markers CD45 and CD34 were detected by flow cytometry, but the cels could not express CD44, CD105 and CD29. Based on the cel growth curve, the cels came into a conditioning period at 1-3 days of seeding and came into a logarithmic phase at 3-5 days. BrdU-positive cels were visible at the injury region after 14 days, indicating that in the rats, transplanted human umbilical cord mesenchymal stem cels migrated from the peripheral blood to the site of brain injury to achieve the effective repair of injured parts. Cite this article:Liu HL, Liu ZJ, Chen XB, Hu WZ, Ding BQ. Migration and localization of umbilical cord mesenchymal stem cels implanted into brain injury model rats. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):31-35.
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BACKGROUND:Adipose-derived mesenchymal stem cels are a kind of pluripotent stem cels that have the potential of self-renewal and proliferation, and have low immunogenicity and immunomodulatory role. OBJECTIVE:To study the effects of adipose-derived mesenchymal stem cels on T cel immune status of alergic rhinitis mouse models. METHODS:Sixty mice were randomly assigned into six groups (sensitized/chalenged/treatment): experimental group 1 was given ovalbumin/ovalbumin/high-dose adipose-derived mesenchymal stem cels, experimental group 2 given ovalbumin/ovalbumin/low-dose adipose-derived mesenchymal stem cels, experimental group 3 given ovalbumin/ovalbumin/PBS, experimental group 4 given ovalbumin/ovalbumin/0, and experimental group 5 given PBS/PBS/0, and normal control group given no treatment. In the former five groups, intraperitoneal injection of 200 μL ovalbumin sensitizing solution or PBS was conducted for basic sensitization at days 0, 7, 14; 20 μL ovalbumin chalenging solution or PBS was given for chalenging at days 15-19. In the former three groups, 0.1 mL of high-dose, low-dose adipose-derived mesenchymal stem cels or PBS was givenviathe tail vein, respectively, at days 20-22 after sensitization and chalenge. At 48 hours after final treatment, ELISA was used to detect serum levels of interleukin-4, interleukin-6, interleukin-10 and interferon-γ, and fluorogenic quantitative PCR used to detect the mRNA expressions of these cytokines in the spleen. Migration of fluorescent-labeled adipose-derived mesenchymal stem cels in the nasal mucosa was observed under fluorescence microscope, and pathological changes of the nasal mucosa were observed through hematoxylin-eosin staining. RESULTS AND CONCLUSION:Compared with the experimental group 4, the levels of interleukin-4 and interleukin-6 in the serum and spleen were significantly lower in the experimental group 1 (P 0.05). Fluorescent-labeled adipose-derived mesenchymal stem cels could migrate into the nasal mucosa, and the number of migrated cels was notably higher in the experimental group 1 than experimental group 2. Eosinophil infiltration in the nasal mucosa was remarkably aleviated in the experimental groups 1 and 2. These findings suggest that adipose-derived mesenchymal stem cels play a non-specific immunomodulatory effect dose-dependently by regulating Th1/Th2 immune imbalances and deficiencies of Treg cels.
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BACKGROUND:The non-specific immune suppression method is generaly used for treatment of systemic lupus erythematosus, but poor prognosis, such as infection and high recurrence rate, exists. OBJECTIVE:To evaluate the therapeutic effect of bone marrow mesenchymal stem cel transplantation on systemic lupus erythematosus in mice. METHODS:Sixteen mice with systemic lupus erythematosus were equivalently randomized into control and experimental groups, or then subjected to passage 3 bone marrow mesenchymal stem cel transplantation or the equal volume of normal saline via the tail vein, respectively. Mouse urine samples were colected to detect urine protein levels by Bradford method. Blood samples from the tip of the mouse tail were extracted to detect serum anti-ds-DNS antibody concentration by radioimmunoassay. Mouse kidney tissues were taken and observed pathohistologicaly through hematoxylin-eosin staining and immunohistochemistry staining under microscope. Flow cytometry was used to detect the expression of CD4+CD25+T cels in the inner canthus blood, fresh spleen and thymus. RESULTS AND CONCLUSION:Within 10 weeks after cel transplantation, the urine protein levels in the two groups were gradualy increased, and the rising velocity was higher in the control group than in the experimental group. From the 4th to 10th week, the urine protein levels in the experimental group were significantly lower than those in the control group (P 0.05). The serum anti-ds-DNA antibody concentration in the experimental group was significantly lower than that in the control group (P < 0.05). Taken together, bone marrow mesenchymal stem cel transplantation can improve the pathological damage in systemic lupus erythematosus mice, and has a certain therapeutic effect on systemic lupus erythematosus.
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BACKGROUND:Studies have shown that bone marrow mesenchymal stem cel transplantation can improve disease conditions by reducing inflammation. OBJECTIVE:To explore the therapeutic efficacy of bone marrow mesenchymal stem cels on chronic asthma rats. METHODS: A rat model of chronic asthma was established by intraperitonealy injected and aerosolized ovalbumin. After modeling, rats were given 4×105 and 8×105 bone marrow mesenchymal stem celsvia the tail vein, respectively. Thirty days later, the lung tissues were observed pathologicaly using hematoxylin-eosin staining; RT-qPCR and ELISA methods were employed to test the changes in interleukin-10, tumor necrosis factor-α and interferon-γ levels in lung tissue and peripheral blood, respectively. RESULTS AND CONCLUSION:Rat models of chronic asthma were successfuly established after intraperitoneal injection of ovalbumin combined with aerosolized ovalbumin. After 30 days of cel treatment, the structure of lung tissues were obviously recovered, and the levels of interleukin-10, tumor necrosis factor-α and interferon-γ showed some improvement in lung tissue and peripheral blood, but there were no differences between the two groups. In conclusion, bone marrow mesenchymal stem cels show some potential role in the treatment of chronic asthma.
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BACKGROUND:There is a high morbidity after spinal cord injury, and the therapeutic strategy is limited to early surgical intervention, medication and post-treatment exercise that only can improve the motor function slightly. However, there is no effective cure method. OBJECTIVE:To study the effect of partition-type spinal cord catheter combined with bone marrow stromal stem cels on T8 spinal cord transection damage in rats. METHODS:Fifty rats were randomized into five groups (n=10 per group): group I, T8 spinal cord transection (5 mm) was made in rats with no treatment; group II, the partition-type tube was inserted into the injured site after modeling; group III, partition-type tube combined with bone marrow stromal stem cels was implanted into the injured site after modeling; group IV, partition-type tube combined with polyglycolic acid fibers was implanted into the injured site after modeling; group V, partition-type tube combined with bone marrow stromal stem cels and polyglycolic acid fibers was implanted into the injured site after modeling. RESULTS AND CONCLUSION:At 2 and 12 weeks postoperatively, Basso, Beattie and Bresnahan scores were significantly higher in the groups III and IV than the groups I, II, IV (P < 0.05). At 12 weeks postoperatively, the latency of motor evoked potential below the injury plane was significantly decreased in group V compared with groups I, II, III, IV (P < 0.05). Immunohistochemical results displayed that in the groups III and V, regenerated nerve fibers grew positively and arranged orderly among the tubes, and there was no obvious winding phenomenon. Under transmission electron microscopy, a certain number of myelinated nerve fibers were found as bridges among groups. These findings indicate that the partition-type chitosan tube combined with bone marrow stromal stem cels has a good connection with the injured spinal cord a good connection to restore part of electrophysiological properties, accelerate the axon regeneration, recover the motor function, thereby providing a new direction for the treatment of spinal cord injury. Cite this article:Zhao XW, Liu X, Yu DP, Rong H, Yu XS, Yang CS, Liu T, Zhao TB. Partition-type spinal cord catheter combined with bone marrow stromal stem cels in the repair of spinal cord transection injury in rats. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):42-48.
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BACKGROUND:Bone marrow mesenchymal stem cels have low immunogenicity and can induce immune tolerance. At present, the mechanism of immune regulation of bone marrow mesenchymal stem cels is not completely understood. It has been rarely reported whether the bone marrow mesenchymal stem cels can migrate to the thymus after transplantation. OBJECTIVE:To observe the distribution and survival of bone marrow mesenchymal stem cels in the thymus of aging rats after transplantation. METHODS: Bone marrow mesenchymal stem cels cultured in vitrowere transfected by adenovirus vectors expressing green fluorescent protein. Transfected bone marrow mesenchymal stem cels were injected into the portal vein of aging rats. At days 3, 7, 14, 21 after transplantation, the survival of bone marrow mesenchymal stem cels homing to the thymus was observed under fluorescence microscope. At day 3 after transplantation, thymus tissues were taken and stained with hematoxylin-eosin for pathological observation. RESULTS AND CONCLUSION:Green fluorescent protein-labeled bone marrow mesenchymal stem cels had a strong green fluorescence at days 3 and 7 after transplantation, and the cel contour was clear. There was no significant difference in the mean absorbance values at days 3 and 7 (P> 0.05). Expression of green fluorescent protein was weakened significantly at days 14 and 21 compared with that at day 3 (P < 0.05). At 3 days after transplantation, the transplanted bone marrow mesenchymal stem cels were clearly visible in the thymus, and acute rejection was not observed. The results show that bone marrow mesenchymal stem cels can migrate to the damaged thymus tissue through the blood circulation, and can survive at least 1 week.
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BACKGROUND:Conditioned medium from mesenchymal stem cels (MSC-CM) may represent a promising alternative to MSCs transplantation. Previous studies have shown that inflammatory activation can strengthen the multiple biological potencies of MSCs; however, normal MSCs with insufficiency of immunocompetence and migration ability are not effective for tissue damage repair. OBJECTIVE:To investigate differential effects of MSC-CM with and without inflammatory activation on radiation-induced intestinal injury.METHODS:MSCs from the bone marrow of SD rats were separated, cultured and identified, and then co-cultured with non-irradiated IEC-6 or irradiated IEC-6 in a transwel system for 24 hours. Then, MSCs with inflammatory activation were cultured alone for another 48 hours. After that, the supernatant was colected as non-activated MSC-CM (MSC-CMNOR) and MSC-CM under radiation-induced inflammatory condition (MSC-CMIR). Rats were exposed to 14 Gy whole abdominal irradiation and randomly divided into four groups: control group, radiation injury group (DMEM/F12), MSC-CMNOR group and MSC-CMIR group. Continuous administration was givenvia tail vein and intraperitoneal implantation of Alzet microosmotic pumps. Intestinal samples were colected at 1, 3, 7 days after radiation for analysis of short circuit variation, at 3 days after radiation for analysis of intestinal epithelium ultrastructure, and at 1, 3, 5, 7, 14 days after radiation for histological observation of the intestinal epithelium using hematoxylin-eosin staining. Blood samples were colected at 1, 3, 7 days after radiation for analysis of serum xylose levels. In addition, the survival state and survival time of rats were observed and recorded. RESULTS AND CONCLUSION: The short circuit variation responding to electrical field stimulation was significantly reduced at al frequencies, but it was significantly improved in the MSC-CMIR group. Similarly, the intestinal absorption (serum xylose levels) was also significantly impaired by irradiation, but improved by delivery of MSC-CMIR (P < 0.05). At 3 days after MSC-CMIR infusion, the intestinal epithelium exhibited an increase in crypt size and vilous length (P < 0.05). Under the electron microscope, a reduction in intestinal microvili and open tight junctions in irradiated intestinal epithelium was found, and the intestine from rats treated with MSC-CMIR had more obvious tight junctions. In addition, treatment with MSC-CMIR dramaticaly improved the survival rate and mean survival time of irradiated rats as compared to those treated with DMEM/F12 or MSC-CMNOR (P < 0.05). Taken together, the present study demonstrated that MSC-CMIR , but not non-activated MSC-CM, improves the structural and functional restoration of the smal intestine after radiation-induced intestinal injury.
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BACKGROUND:It has been demonstrated to be effective for the improvement of heart function after acute myocardial infarction with intravenous or intramyocardial administration of bone marrow mesenchymal stromal cels. However, little is known regarding the effect of the combination of intravenous and intramyocardial administration of mesenchymal stromal cels on the heart function of a chronic myocardial infarction model. OBJECTIVE:To study the effect of intravenous and intramyocardial administration of bone marrow mesenchymal stromal cels on the heart function of a rat chronic myocardial infarction model and the relevant mechanism. METHODS:Bone marrow mesenchymal stromal cels isolated from Lewis rats were expandedex vivo. BrdU-labeled bone marrow mesenchymal stromal cels (3×106) were administeredvia the femoral vein and the myocardial surface respectively into rat models of chronic myocardial infarction in cel transplantation group. The equal volume of PBS was injected into the same place in control group. Four weeks after injection, echocardiography was performed to evaluate the heart function, and then the heart tissues were harvested for immunohistochemistry examination. The total blood vessel density in the scar area was evaluated. RESULTS AND CONCLUSION:At 4 weeks after cel implantation, the left ventricular function was not improved in the two groups. The immunohistochemistry staining showed that (1) the mesenchymal stromal cels in the myocardium did not differentiate to myocardial cels; (2) there was no significant difference in the total blood vessel density in the scar area between the cel transplantation and control groups. Taken together, the combined intravenous and intramyocardial administration of bone marrow mesenchymal stromal cels cannot improve heart function in a rat chronic myocardial infarction model.
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BACKGROUND:Chemotherapy drugs can damage the ovarian function in women of childbearing age, and even lead to premature ovarian failure. Therefore, to improve and restore the ovarian function in patients has become an important issue. OBJECTIVE:To explore the therapeutic effect and feasibility of bone marrow mesenchymal stem cel therapy against chemotherapy-induced ovarian damage. METHODS:Rat models of chemotherapy-induced premature ovarian failure were established, and injected with PKH26-labeled bone marrow mesenchymal stem cels. At 15, 30, 45, 60 days after cel transplantation, five rats were selected respectively to detect folicle-stimulating hormone and estradiol levels, and then, the rats were kiled to take the right ovary for pathological examination. The number of ovarian folicles was detected under light microscope. At 30 days after cel transplantation, another two rats were selected to mate with male rats to observe the difference in the reproductive activity. RESULTS AND CONCLUSION:Four of 22 rats (18%) gradualy recovered their estrous cycle after cel transplantation, with the decreased folicle-stimulating hormone level and increased estradiol level. Moreover, the number of folicles was reduced. Al of these indicated that the ability to have children in rats was not damaged.These experimental findings suggest that bone marrow mesenchymal stem cels can partialy improve the ovarian function of rats under chemotherapy.
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BACKGROUND:Due to the different transplantation time after myocardial infarction, the homing ability of bone marrow mesenchymal stem cels to damaged tissues as wel as the repairing role wil be very different. OBJECTIVE:To explore the optimal window time for the homing of bone marrow mesenchymal stem cels to the myocardial tissue after myocardial infarction. METHODS: Eighteen Chinese miniature pigs were modeled by the ligation of left anterior descending coronary artery. BrdU-labeled bone marrow mesenchymal stem cels were injectedvia the coronary artery at 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks after modeling, respectively. Then, the animals were kiled at 3 days after cel transplantation to detect the home of bone marrow mesenchymal stem cels in the infarct zone. RESULTS AND CONCLUSION:BrdU-labeled positive cels with brown nuclei were visible at 1 day, 3 days, 1 week, 2 weeks, 3 weeks, and 4 weeks after myocardial infarction, especialy at 1 week after myocardial infarction (P < 0.05). It indicates that the best homing window for bone marrow mesenchymal stem cels was at week after myocardial infarction, when the stem cel transplantation is given to be able to promote myocardial repair.
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BACKGROUND:Bone marrow mesenchymal stem cels have a therapeutic effect on acute lung injury, but the mechanism is unclear. If the mechanism is understood, the majority of patients with acute lung injury can obtain a benefit. OBJECTIVE:To explore the possible mechanism underlying bone marrow mesenchymal stem cels in the treatment of acute lung injury with sepsis in rats. METHODS: (1) Thirty-six adult Wistar rats were randomly divided into three groups, sham operation group (sham group), sepsis group and bone marrow mesenchymal stem cels group (cel treatment group). In the sepsis and cel treatment groups, animal models of sepsis with acute lung injury were established by cecal ligation and puncture, while in the sham group, the cecum was not ligated and punctured. Then, 1 mL normal saline was injected via the femoral vein in the sepsis and sham groups, and 1 mL bone marrow mesenchymal stem cel suspension (1×109/L) was injected into the cel treatment group. After 6 hours, interleukin 10 and macrophage inflammatory protein-2 levels in serum were measured in the three groups. Lung tissues were taken for pathological observation using hematoxylin-eosin staining. (2) Rat alveolar macrophages were obtained by bronchoalveolar lavage, seeded into 24-wel culture plates, and divided into three groups: control group (group A), sepsis model group (group B) and intervention group of bone marrow mesenchymal stem cels (group C). Normal saline, septic plasma, and co-intervention of septic plasma and mesenchymal stem cels were used in the groups A, B, C, respectively. Then, cels in the three groups were cultured in a 5% CO2 incubator at 37℃ for 1 hour. After that, alveolar macrophages were taken to detect whether nuclear factor-κB (P65) protein entered into the nucleus using laser scanning confocal microscopy. RESULTS AND CONCLUSION: (1) The results of animal experiments showed that compared with the sham group, the macrophage inflammatory protein-2 levels in the sepsis group and cel treatment group were significantly increased (P 0.05); inflammatory cel infiltration, interstitial pulmonary edema and pulmonary hemorrhage existed in the sepsis and cel treatment groups, but these symptoms were significantly reduced in the cel treatment group compared with the sepsis group. (2) Results from cel experiments showed that compared with the group A, in group B and group C, the number of nuclear factor-κB (P65) proteins into the nucleus was significantly higher (P < 0.05), but it was lower in the group C than the group B (P < 0.05). These findings indicate that bone marrow mesenchymal stem cels in acute lung injury with sepsis can regulate nuclear factor-κB (P65) protein of alveolar macrophages into the nucleus, reduce expression of macrophage inflammatory protein-2, and thereby play a protective role in the lungvia reducing neutrophil infiltration. Temporarily, this study cannot explain whether bone marrow mesenchymal stem cels have an effect on interleukin 10.
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BACKGROUND:There are most single-center studies about bone marrow stem cels applied to treat decompensated cirrhosis, but the therapeutic results are not ideal. It is possibly related to aging, physical weakness, poor bone marrow hematopoietic function, less available number of stem cels and feeble ability of regeneration and proliferation in liver cirrhosis patients. Umbilical cord mesenchymal stem cels are characterized of easy to obtain, wide source and weak immunogenicity. Co-transplantation of bone marrow stem cels and umbilical cord mesenchymal stem cels may improve the therapeutic effects on decompensated cirrhosis patients. OBJECTIVE:To investigate the efficacy and safety of co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels on decompensated cirrhosis.METHODS:Thirty-two decompensated cirrhosis patients were randomly divided into two groups: in stem cel group, 13 patients received co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels based on regular medical treatment; in control group, 19 patients only underwent the regular medical treatment. Al the patients were folow-up for 1 year. Alanine aminotransferase, albumin, total bilirubin, prothrombin time, Child-Pugh score and Model for End-Stage Liver Disease score, 1-year survival rate, Quality of Life score and adverse reactions related to stem cel therapy were observed and recorded in the two groups at 4, 12, 52 weeks after treatment. RESULTS AND CONCLUSION:At 4, 12, 52 weeks after treatment, improvements in the liver function, prothrombin time, Child-Pugh score and Model for End-Stage Liver Disease score were found in the two groups, but there was no difference between the two groups (P > 0.05). At 4 weeks after transplantation, the clinical symptoms and Quality of Life score in the stem cel group were significantly improved, which were better than those in the control group (P 0.05). In addition, the 1-year survival rate showed no difference between the two groups, and no severe adverse reactions related to stem cel therapy occurred during the folow-up. Co-transplantation of umbilical cord mesenchymal stem cels and bone marrow stem cels is safe and effective to improve the clinical symptoms of decompensated cirrhosis patients. However, further studies with larger samples are warranted to better clarify the co-transplantation effects.
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BACKGROUND:Stromal cel-derived factor-1 has a strong chemotaxis to bone marrow mesenchymal stem cels, and both of them can promote wound healing. However, there are less studies on their correlation with skin wound healing. OBJECTIVE:To investigate the effects of stromal cel-derived factor-1 on bone marrow mesenchymal stem cels migration and skin wound repair. METHODS: Thirty SD rats were divided into five groups at random. Bone marrow mesenchymal stem cels labeled with PKH-26 were injected into the rat caudal vein. After 1 week, skin wound models were established. Then, different concentrations (1, 2, 10, 50 μg/L) of stromal cel-derived factor-1 were injected via multi-points on the skin wound. The skin wound healing was observed and recorded at 14 days after injection. The number and distribution of bone marrow mesenchymal stem cels were observed by the fluorescent staining at different time points. The pathological changes of wound tissue were observed by hematoxylin-eosin staining. The expression of colagen I and colagen III were detected by western blot assay. RESULTS AND CONCLUSION:Stromal cel-derived factor-1 at 10 μg/L could induce the largest number of bone marrow mesenchymal stem cels to the skin wound and achieve the best repair results. Stromal cel-derived factor-1 could also regulate the expression of colagen I and colagen III in the wound, and when the concentration of stromal cel-derived factor-1 was 10 μg/L, the expressions of colagen I and colagen II reached the peak. These findings indicate that the appropriate concentration of stromal cel-derived factor-1 is better to promote the migration of bone marrow mesenchymal stem cels, thereby contributing to skin wound repair.
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BACKGROUND: Core decompression alone for osteonecrosis of femoral head easily causes fovea of femoral head and colapse of inner microstructure. Therefore, autologous bone is needed for filing and supporting. Moreover, bone marrow stem cel transplantation can decrease the incidence of femoral head colapse. OBJECTIVE:To discuss the clinical effects of core decompression and bone grafting combined with autotransplantation of bone marrow mesenchymal stem cels for osteonecrosis of femoral head. METHODS: A total of 33 patients were treated by core decompression and bone grafting combined with autotransplantation of bone marrow mesenchymal stem cels in the Fourth Department of Bone Surgery, Central Hospital Affiliated to Shenyang Medical Colege in China from December 2012 to May 2013. RESULTS AND CONCLUSION:After the treatment by core decompression and bone grafting combined with autotransplantation of bone marrow mesenchymal stem cels, Harris hip function score increased and pain disappeared in patients with osteonecrosis of femoral head. They could do various labors. Radiographs or CT examination displayed normal femoral head in 30 hips, accounting for 79%. Pain significantly reduced. Normal or slight limp walking was found in 15 hips, accounting for 40%. There were 35 hips in patients, whose walking distance was extended, accounting for 92%. 24 hips dysfunction was improved markedly, accounting for 63%. Al results suggested that core decompression and bone grafting combined with autotransplantation of bone marrow mesenchymal stem cels improved the local blood supply of femoral head, and played a positive role in promoting the necrotic bone absorption and bone repairing.
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BACKGROUND:Human telomerase reverse transcriptase (hTERT) is the first choice for regulating the proliferation and directional differentiation, with multiple biological effects. OBJECTIVE:To observe the therapeutic effect of hTERT-transfected bone marrow mesenchymal stem cels transplantation in diabetic rats. METHODS: Bone marrow mesenchymal stem cels from Sprague-Dawley rats were culturedin vitro and transfected with retrovirus PLXSN carrying hTERT. RT-PCR was used to detect the hTERT expression in the bone marrow mesenchymal stem cels before and after transfection. Sixty male Sprague-Dawley rats were selected and equaly randomized into four groups: normal control group, transfection group, cel transplantation group, and model group. In the latter three groups, rats were injected with 45 mg/kg chain urea to establish diabetes models, and then injectedvia the tail vein with 0.2 mL hTERT-transfected bone marrow mesenchymal stem cels, 0.2 mL bone marrow mesenchymal stem cels, and 0.2 mL normal saline, respectively. RESULTS AND CONCLUSION:At 48 hours after hTERT transfection, the expression of hTERT mRNA was detected in the bone marrow mesenchymal stem cels, and mainly concentrated in the nuclei. At 14 days after transfection, the fasting glucose level in the model group was higher than that in the normal control group (P 0.05). These findings indicate that the transplantation of hTERT-transfected bone marrow mesenchymal stem cels is effective in the treatment of diabetic rats.
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BACKGROUND:Platelet-derived endothelial cel growth factor (PD-ECGF) can promote revascularization in fat transplantation. OBJECTIVE: To explore the dual effects of PD-ECGF and adipose-derived mesenchymal stem cels on the survival rate of fat grafts. METHODS:(1) Adipose-derived mesenchymal stem cels were isolated from the inguinal subcutaneous fat of New Zealand white rabbits, and then cultured. Passage 3 adipose-derived mesenchymal stem cels were divided into experimental group (Lenti-PD-ECGF-EGFP transfected adipose-derived mesenchymal stem cels), control group (Lenti-EGFP transfected adipose-derived mesenchymal stem cels) and blank group (adipose-derived mesenchymal stem cels with no transfection). (2) Lenti-PD-ECGF-EGFP transfected adipose-derived mesenchymal stem cels were cultured in DMEM complete medium, and then mixed with fat tissues as group A; adipose-derived mesenchymal stem cels with no transfection were cultured in DMEM complete medium and then mixed with fat tissues as group B; DMEM complete medium with no cels served as group C. Then, the grafts in groups A, B, C were respectively injected subcutaneously into the upper left, lower left and upper right parts of the rabbits’ black. RESULTS AND CONCLUSION:(1) In the experimental group, PD-ECGF mRNA and protein expressions were significantly higher than those in the control and blank groups (P < 0.05), and cel proliferation was also the fastest. (2) Graft weight and the number of capilaries were greater in group A than groups B and C. These findings indicate that PD-ECGF transfection of adipose-derived mesenchymal stem cels not only can continuously express the PD-ECGF protein, but also can promote the proliferation of adipose-derived mesenchymal stem cels.
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BACKGROUND:Bone marrow mesenchymal stem cels can be used to repair neurons, but have no ideal outcomes on nervous system injuries. OBJECTIVE:To investigate the effects of bone marrow mesenchymal stem cel transplantation combined with propofol on the hind limb function and electrophysiological changes of rats with spinal cord injury. METHODS:Eighty adult Wistar rats were selected to make animal models of spinal cord injury, and then randomized into four groups (n=20): bone marrow mesenchymal stem cel group, control group, combination group, propofol group. At 6 hours after modeling, rats in these four groups were injectedvia the tail vein with bone marrow mesenchymal stem cel suspension, cel culture medium, bone marrow mesenchymal stem cel suspension+propofol solution, and propofol solution using a 1 mL syringe, respectively. Rat motor function was assessed by Basso Beattie Bresnahan score, modified Tarlov score and inclined plane test before and at 1 day, 3 days, 1-4 weeks after modeling. Under fluorescence microscope, the survival and distribution of PKH-26-labeled bone marrow mesenchymal stem cels were observed at 4 weeks after modeling, and meanwhile, hematoxylin-eosin staining was used for pathological observation. Horseradish peroxidase tracer analysis was performed to analyze regeneration of nerve fibers, and motor and somatosensory evoked potentials were used to analyze the neurophysiological recovery of rats. RESULTS AND CONCLUSION:(1) The motor function of the rat hind limb recovered best in the combination group, better in the bone marrow mesenchymal stem cel group and propofol group, but worse in the control group. (2) There were a smal amount of nerve axon-like structures and smal syringomyelia in the bone marrow mesenchymal stem cel group and propofol group, but the combination group had more axon-like structures and no syringomyelia. In the control group, no axons but spinal cord defects and syringomyelia formed. (3) The amount of horseradish peroxidase-positive nerve fibers and the number of PKH-26 positive cels were ranked as folows: control group propofol group and bone marrow mesenchymal stem cel group > combination group, and there were significant differences between the groups (P < 0.05). (5) Amplitudes of motor and somatosensory evoked potentials were arranged as folows: control group < propofol group and bone marrow mesenchymal stem cel group < combination group, and the differences were statisticaly significant (P < 0.05). These findings indicate that both propofol and bone marrow mesenchymal stem cels can promote synaptic regeneration and improve the electrophysiological function and motor function of rats with spinal cord injury. Their combination has a better role than propofol and bone marrow mesenchymal stem cels used alone.
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BACKGROUND:Bone marrow stem cels combined with traditional surgery regimen can significantly improve the therapeutic effects on bone nonunion, which are considered to have an important application value. OBJECTIVE:To explore therapeutic effect of bone marrow mesenchymal stem cels on bone nonunion under micro-damage environment. METHODS:Forty New Zealand white rabbits were selected and randomized into experimental and control groups, 20 rabbits in each group. Bone marrow of the tibia was extracted to isolate and culture bone marrow mesenchymal stem cels. Passage 3 cels with the order of magnitudes of 107 were labeled by superparamagnetic iron oxide nanoparticles. A 15-mm bone defect was made at the middle of the radius of the rabbit forelimb. Bone nonunion appeared at 6 weeks after bone defects. Bone marrow mesenchymal stem cels combined with iliac particles were implanted into the bone defect of rabbits in the experimental group, and only iliac particles were implanted into the bone defect of rabbits in the control group. Within 12 weeks after implantation, the bone nonunion was observed through gross morphology, X-ray observation, and pathological observation. RESULTS AND CONCLUSION:After implantation, a remarkable calus was found in the experimental group, and the bone defect recovered gradualy until it was completely healed; in the control group, there was no calus, and the bone marrow cavity was closed and ful of granulation tissues. In the experimental group, there were actively proliferated cartilage tissues, bone particles were fused, osteoid structures appeared, and osteoblasts proliferated progressively; in the control group, poor cartilage hyperplasia was found, and there were a large amount of dead bone tissues but no fused bone particles and osteoblasts. In the experimental group, X-ray films on the defected radium showed cloudiness-like shadow, the bone marrow cavity was recanalized, and the skeleton was shaped wel; in the control group, few bone particles were absorbed, the bone marrow cavity was partly recanalized, and the injured bone was not healed with osteosclerosis. These findings indicate that under the micro-damage environment, bone marrow mesenchymal stem cels can differentiate into osteoblasts to repair bone defects-induced bone nonunion.
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BACKGROUND:Studies have shown that human telomerase reverse transcriptase (hTERT) has the ability to enhance cel proliferation, maintain telomere length, prolonged cel life cultured in vitro. OBJECTIVE:To observe the effect of hTERT gene-modified bone marrow mesechymal stem cel transplantation on neural function recovery of rats with cerebral infarction. METHODS:Rat models of middle cerebral artery occlusion were established and randomized into model group, cel transplantation group and hTERT-modified cel transplantation group, with 20 rats in each group. Rats in the three groups were respectively injected via tail vein with 1 mL PBS, passage 9 bone marrow mesenchymal stem cel suspension (2.5×107/L) and hTERT-modified passage 9 bone marrow mesenchymal stem cel suspension (2.5×107/L), respectively. Modified neurological severity scores were determined before and after transplantation; RT-PCR and western blot assay were used to measure hTERT expression at gene and protein levels; TUNEL method was adopted to detect cel apoptosis in the brain. RESULTS AND CONCLUSION:hTERT-modified bone marrow mesenchymal stem cels had prolonged cel cycle, and with the increase in passage number, the cels showed good growth with no changes in morphology. The expressions of hTERT mRNA and protein were superior in the hTERT-modified cel transplantation group than the cel transplantation group, and there was a significant difference (P < 0.05). Modified neurological severity scores and number of apoptotic cels were decreased significantly in the hTERT-modified cel transplantation group compared with the other two groups (P < 0.05). These findings indicate that hTERT-modified bone marrow mesenchymal stem cels can promote neural functional recovery of rats with cerebral infarction.