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Objective@#To determine the effect of transplanting bone marrow mononuclear cells (BMMCs) on the expression of glial fibrillary acidic protein (GFAP) and Nogo-A around the ischemic foci after focal cerebral ischemia and reperfusion, and to study any role of BMMCs in nerve function recovery.@*Methods@#BMMCs were isolated from the bone marrow of Sprague-Dawley rats. Cerebral ischemia and reperfusion was performed using a nylon thread to occlude the right middle cerebral artery for 2h followed by 24h of reperfusion. The qualified models were selected according to the Longa scale. The 48 models selected were randomly divided into a model group and an observation group, each of 24. Each group was further divided into 7d, 14d and 21d subgroups. 100μl of BMMCs (5×106 /ml) were slowly injected into the ischemic lateral striata of the observation group. The rats in the model group were similarly injected, but with buffered saline solution. The rats were evaluated using the Longa scale after 7d, 14d and 21d. The rats were then sacrificed and the brain was resected. Immunohistochemical assays quantified the expression of GFAP and Nogo-A around the ischemic foci.@*Results@#Compared with the model group, the rats in the observation group showed less neurological deficit on the 21st day, significantly greater expression of GFAP and significantly less Nogo-A expression on days 14 and 21. Nogo-A expression on the 21st day was also significantly lower than on the 14th day in the observation group.@*Conclusion@#BMMC transplantation can promote recovery from nerve damage after focal cerebral ischemia, which is probably related to enhanced expression of GFAP and restrained expression of Nogo-A in the brain tissues surrounding ischemic lesions.
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Cellular therapy in acute ischemic stroke (AIS) is a research hotspot in the field of neu-roscience in recent years. Compared with other cells,bone marrow mononuclear cells (BMMNCs) are one of the most attractive therapeutic cells because BMMNCs can be rapidly isolated from bone marrow,are enriched with stem cells and permit autologous applications. Numerous basic researches showed that BMMNCs trans-plantation can decrease infarct volume and promote neurological outcomes in animal stroke model,indicating BMMNCs transplantation may has therapeutic values in acute ischemic stroke,and the transformation from basic research to clinical applications is on the key phase. In this paper,the progress of BMMNCs transplan-tation is reviewed in acute ischemic stroke on the aspects of BMMNCs component,methods of purification, route of transplantation,therapeutic mechanisms and problems from basic research to clinical application.
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Objective To explore the neuroprotection and mechanisms of bone marrow mononuclear cells (BMMNCs),and evaluate whether ERK1/2 signaling pathway was involved in it.Methods384 healthy male SD rats,which were 6-8 week old,weighting 250-280 g,were selected.The middle cerebral artery occlusion (MCAO) model was established in SD rats using the suture method.The rats were randomly divided into sham operation group,model group,BMMNCs group and ERK1/2 inhibitor group,with 96 rats in each group.At the time of 24 h after the successful modeling,200 μl PBS solution was injected into the caudal vein of the rats in the model group,200 μl PBS solution containing 5×106 BMMNCs was injected into the rats in the BMMNCs group and the ERK1/2 inhibitor group.meanwhile,5 μl PD98059 was injected into the lateral ventricle of the brain of rats in the ERK1/2 inhibitor group.At the time points of 3 d,7 d and 14 d,the modified neurological severity scores (mNSS) was used to evaluate the neurological function,the volume of cerebral infarction was assessed by TTC staining,the pERK1/2,Bax,Bcl-2 and caspase-3 levels were detected by Western blot,and the effect of BMMNCs on activation of microglia was detected by immunofluorescence assay.Results(1)At each time point,the mNSS and the volume of cerebral infarction of the model group were significantly higher than those of the sham operation group (P0.05).(2)At each time point,the pERK1/2,Bcl-2,Bax and caspase-3 protein levels of the model group were significantly higher than those of the sham operation group (P0.05).(3) At each time point,microglia (Iba1 positive) in ischemic penumbra of the BMMNCs group was significantly more than those of the model group,and it was increased with the time extension (P0.05).ConclusionBMMNCs can reduce the apoptosis through ERK1/2 signaling pathway,thus improving the neurological function and reducing the infarct scope.
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Cerebral Palsy (CP) is a disabling condition that affects a child's life and his/her family irreversibly. It is usually a non-progressive condition but improvement over time is rarely seen. The condition can be due to prenatal hypoxia, metabolic, genetic, infectious, traumatic or other causes. It is therefore a heterogeneous group that results in functional motor disability associated with different degrees of cognitive abnormalities. There are no treatments that can cure or even improve CP and the best available approach aims at functional, social and nutritional supportive care and counseling. In this paper, we report 17 sequential patients with CP treated with intrathecal administration of Bone Marrow Mononuclear Cells (BMMC). All patients had an uneventful post-injection course with 73% of the evaluable patients treated having a good response using the Gross Motor Function Classification System (GMFCS). The average improvement was 1.3 levels on the GMFCS with cognitive improvements as well.
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Humans , Hypoxia , Bone Marrow , Cerebral Palsy , Classification , Counseling , Injections, Spinal , Nutritional Support , Stem CellsABSTRACT
Objective: To explore the effects and the possible mechanism of combined injection of Angelica sinensis polysaccharide (ASP) and cytarabine (Ara-C) on the bone marrow mononuclear cells (BMMCs) of the transplanted human leukemia mouse model. Methods: K562 cells (2×107)were transplanted into the tail vein of mice to establish the transplanted human leukemia NOD/SCID mouse model. Then the leukemia mice were randomly divided into model, ASP, Ara-C, and ASP + Ara-C groups. After the 30 d transplantation, the mice were ip injected with ASP (200 mg/kg/d), Ara-C (2.5 mg/kg/d), and ASP (200 mg/kg/d) + Ara-C (2.5 mg/kg/d), respectively for 14 d, and the mice in the model group were injected with saline (equal volume and time). The next day after the treatment, the eyeball blood was collected to detect the amount and classification of white blood cells (WBC). The femurs were taken to count BMMCs of each femur. The proliferation of BMMCs was detected by CCK-8; The distribution of cell cycles was analyzed by flow cytometry (FCM); The capability of colony forming was examined by CFU-Mix cultivation; The ratio of the SA-β-Gal staining positive BMMCs was counted; The aging related proteins of P16, Rb, CDK4, and CyclinD1 were detected by Western blotting. Results: Compared with the model group, ASP or Ara-C injected alone and their combined injection can obviously reduce the amount of the peripheral blood WBC, the percentage of neutrophiles, and the number of femur BMMCs; effectively inhibit the proliferation of BMMCs, CFU-Mix forming, and the ratio of S stages; markedly raise the percentage of lymphocytes, ratio of G1 stages, and the percentage of SA-β-Gal positive cells; down-regulate the expression of the aging related proteins of CDK4 and CyclinD1; and up-regulate the expression of P16 and Rb protein. The effects of ASP + Ara-C group were much better than those in the other groups. Conclusion: The aging mechanism of BMMCs for the transplanted human leukemia mice induced by ASP and Ara-C may be ascribed to the regulation of the expression of the aging related proteins of P16, Rb, CDK4, and CyclinD1. Our research provides a new idea to treat leukemia in clinic.
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Objective To detect the expression of autophagy-related gene Atg3 and Atg5 in bone marrow mononuclear cells (BMMNCs) from children with acute leukemia(AL),so as to explore the relationship between autophagy and the pathogenesis of AL in children.Methods Seventy-four bone marrow specimens were obtained from children with AL in the First Affiliated Hospital of Zhengzhou University Pediatrics Hematology Ward,including 37 cases of initially diagnosed AL without any treatment,28 cases of AL in complete remission,9 cases of refractory or relapse AL and 28 bone marrow specimens from children without tumor were also collected as the control group.BMMNCs were separated by Lydroxypropylmethyl Cellulose.After BMMNCs were stained by Monodansylcadaverine,the autophagy phenomenon was observed by using fluorescence microscope,and the ratio of autophagy was detected by using flow cytometry.Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of Atg3 mRNA and Atg5 mRNA in each group.Results It was found that autophagy phenomenon was more common in the initially diagnosed group and the refractory/relapse group,and the autophagy ratio in both groups was respectively (17.07 ±2.31) ％,(15.37 ± 1.59) ％,respectively,which were obviously higher than that of the control group (2.71 ± 1.57) ％ and that of the complete remission group.The differences were statistically significant (t =28.29,20.96,all P ＜ 0.01).The autophagy ratio in complete remission group was (3.48 ± 1.94) ％,and compared with the control group,the difference was of no statistical significance(t =1.634,P ＞ 0.05).The autophagy ratio in the refractory/relapse group higher than that in the complete remission group (t =16.61,P ＜ 0.05).The expressions of Atg3 mRNA and Atg5 mRNA in initially diagnosed group and refractory/relapse group were higher than those of the complete remission group and control group,and the differences were statistically significant (F =67.592,106.160,all P ＜ 0.008) ; the difference between complete remission group and control group was of no statistical significance (P ＞ 0.008).Conclusions The autophagy ratio and the expressions of Atg3 mRNA and Atg5 mRNA in initially diagnosed group and the refractory/ relapse group were both obviously higher.It was revealed that higher autophagy activity,which was caused by upregulated expressions of Atg3 mRNA and Atg5 mRNA,had a closely connection with the mechanism of occurrence,development and resistance of AL in children.
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Objective To investigate the effect of intravenously transplanted bone marrow mononuclear cells (BMMNCs) on brain injury and inflammation after intracerebral hemorrhage(ICH) in rats.Methods Experimnental ICH models were performed by stereotaxic injection collagenase Ⅳ into caudate putamen,rats that underwent ICH were randomly divided into four groups:sham operation group,ICH group,PBS group,BMMNC-treated group.The BMMNCs were injected intravenously into rats after ICH.The neurobehavioral function was evaluated on days 1,3,7,14 by the modified neurological severity score,and the brain edema was examined by wet-dry weighting method on day 3 after cell transplantation.Immumofluorecence staining was used to identify the number of activation of microglia and infiltration of neutrophils in the brain after ICH.Results The neurological score in BMMNCtreated group on days 7,14 was significantly improved compared with those in ICH group and PBS group(P＜O.05).Compared to the ICH group ((81.09 ± 0.83) ％) and PBS group ((80.99 ± 0.79) ％),there was a significant decreasc in thc brain water content in BMMNC-treated group((78.62±0.97) ％) (P＜0.05).The number of activation of microglia and infiltration of neutrophils were both significantly lower in BMMNC-treated group ((55.8±22.1)/mm2,(49.6± 12.9)/mm2) compared to ICH group and PBS group (respectively (125.0 ± 20.7) /mm2,(86.8±13.6/mm2))(P＜0.01).Conclusion Administration of BMMNCs can significantly reduce edema and improve neurologic function by inhibiting the activation of microglia and infiltration of neutrophils.
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Objective To investigate the effect of treatment with transplanted bone marrow mononuclear cells under different transplant paths on neovascularization in ischemic cerebral infarction rats. Methods The MCAO models were established by intraluminal vascular suture method. All the successful models were randomly divided into control group, intravenous transplantation group and subarachnoid transplantation group according to the random number table. The Extraction,isolation and culture of bone marrow mononuclear cells were extracted,i?solated and cultured. The models in the control group were injected 200μl PBS through the femoral vein. The mod?els in the intravenous transplantation group were injected 200μl containing 2×107 bone marrow mononuclear cells through the femoral vein. The models in the subarachnoid transplantation group were injected 200μl containing 2× 107 bone marrow mononuclear cells through the subarachnoid. The VEGF levels in cerebrospinal fluid of rats were detected using quantitative ELISA method. The vascular regenerations in the infarct area were observed by using quantitative immunofluorescence. Results After transplantation of bone marrow mononuclear cells 7 d,14 d and 28 d,the VEGF levels of rat cerebrospinal fluid in subarachnoid transplantation group were (207.4±8.9)pg/ml, (171.2±10.3)pg/ml and (143.8±13.8)pg/ml,respectively,which were all higher than the VEGF levels in intra?venous transplantation group,the differences were all statistically significant (P<0.05). After transplantation 14 d and 28 d,the BrdU?positive cells in subarachnoid transplantation group were ( 2043. 8 ± 514. 2) and ( 1834. 8 ± 307.4) ,respectively,which were higher than the intravenous transplantation group,the differences were all statisti?cally significant (P<0.05). The microvessel counts in subarachnoid transplantation group were (384.6±45.1) and (514.8±51.3),respectively,which were higher than that in the intravenous transplantation group,and the differ?ences were all statistically significant (P<0.05) . Conclusions Bone marrow mononuclear cells transplantation can increase VEGF secretion,and promote angiogenesis infarct. The effect of subarachnoid transplantation is better.
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Objective To comparethe reprogramming efficiencies of mouse bone marrow mesenchymal stem cells (BMSCs), mouse bone marrow mononuclear cells (BM-MNCs) and mouse embryonic fibroblasts (MEFs) into induced pluripotent stem cells (iPS cells). Methods BMSCs, BM-MNCs and MEFs were infected with lentivirus (LV-efla-mOrt4- IRES-EGFP, LV-efla-mklf4-IRES-EGFP, LV-efla-mK/4-IRES-EGFP and LV-efla-mc-Mcc-IRES-EGFP) at a multiplicity of infection. Reprogramming efficiencies of BMSCs, BM-MNCs and MEFs were compared by counting the number of alkaline phosphatase (AP)-positive clones. Pluripotency of the clones was evaluated by detecting the expression of embryonic stem cells markers and assessing their ability to formembryoid bodies and teratomas. Results iPS cells derived from BMSCs, BM-MNCs and MEFs were all able to grow into clones with clear borders, to express enbryonic stem cell-specific cell surface markers (Nanog, Rex-1 and SSEA-1), and to express characteristic genes of all three germ layers both in vitro and vivo. The AP- positive clones derived from BMSCs were notably less than those from BM-MNCs and MEFs. Conclusion BMSCs, BM-MNCs and MEFs can all reprogram into iPS cells, but the reprogramming efficiency of BMSCs in adherent culture is lower than those of BM-MNCs and MEFs.
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Objective Observe the influence on the Gilial cell-line derived neurotrophic factor (GDNF) after treatment stroke rats with Bone marrow mesenchymal stem cells (BMSCs) or Bone marrow mononuclear cells (BMNCs)combined with traditional Chinese medicine (TCM).Methods Separating and cultivating BMSCs and BMNCs were.140 Wistar rats were divided into 7 groups randomly,normal group,pretended surgery group,model group,BMNC group,BMNC+TCM group,BMSC group,BMSC+TCM group.The other five groups were performed for 2 hours middle cerebral arterial occlusion (MCAO) except normal group and pretended surgery group.Intervention methods in each group after 24 hours of MCAO:model group:Subarachnoid injection of 100 μl 0.01M PBS,BMNC group and BMNC + TCM group,Subarachnoid injection of 2 × 107 BMNCS,BMSC group and BMSC+TCM group,Subarachnoid injection of 2 × 107 BMSCS,BMNC +TCM group and BMSC+TCM group were treated united with TCM on the transplantation day (po.Qd.).GDNF level in all groups' rat brain were analyzed by Enzyme-linked immuno sorbent assay (ELISA) method at the 4th day and the 28th day after transplantation.Results The GDNF level of model group [(62.60±4.05) pg/ml] is higher than normal group's [(53.46 ± 3.91)pg/ml] at the 28th day (P＜ 0.05).The GDNF levels of BMNC group [(194.21 ±39.56)pg/ml,(67.70±4.73)pg/ml] and BMSC group [(169.83±28.84)pg/ml,(82.66±32.23)pg/ml] are higher than model group's at the 4th and 28th day(P＜0.05).The GDNF level of BMSC group is higher than BMNC group's at the 28th day(P＜0.05).The GDNF levels of group BMNC+TCM group[(560.61 ± 194.84) pg/ml,(265.83 ±93.58) pg/ml and BMSC+TCM group[(370.93 ±46.19) pg/ml,(247.34±98.02)pg/ml] are higher significantly than BMNC group's or BMSC group's at the 4th and 28th day(P＜0.05).At the 4th day the GDNF level of the BMNC+TCM group is higher than BMSC+TCM group's(P＜0.05).Conclusion Subarachnoid transplantation of BMNCs or BMSCs will increase the GDNF level in brain of MCAO rats.The transplantantion combined with TCM can inprove the capability of the enhance.That reflect the advantage of transplantantion bone marrow origin stem cell united with TCM.
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ObjectiveTo investigate the expression of CCL2 and its effects on the proliferation and adhesiveness on leukemia cells in patients with acute lymphoblastic leukemia(ALL). MethodsThe bone marrow mesenchymal cells (BMMSC) and bone marrow mononuclear cells (BMMNC) of 30 ALL patients and 30 healthy controls were studied, and CCL2 level was evaluated by ELISA. CCL2 gene mRNA level in ALL was determined by RT-PCR. The cell proliferation and adhesiveness were detected by MTT assays. The cell counts were measured by flow cytometry. ResultsThe BM plasma levels of CCL2 in patients at diagnosis were significantly higher than that in healthy controls [(780.12±129.61) pg/ml vs (120.49±25.21) pg/ml,t =4.96, P =0.00]. Supernatant levels of CCL2 in BMMSC were significantly higher than that of BMMNC [(572.38±35.39) pg/ml vs (193.85±15.45) pg/ml,t =5.37,P =0.00]in vitro.CCL2 cannot induce leukemia proliferation alone,but could induce leukemia proliferation in BMMNC and BMMSC co-culture in a dose- and time-dependent manner.CCL2 could increase the leukemia adhesive to the BMMSC compared with control (r =0.824,P =0.02).ConclusionPatients with B type ALL had higher levels of CCL2 which was secreted by BMMSC. The leukemia could induce the BMMSC to secrete CCL2. CCL2 could promote the survival and proliferation of leukemia in the presence of BMMSC and BMMNC, and enhance ALL cells adhesion toBMMSC in a dose-dependent manner.
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Introdução e objetivos: a insuficiência hepática aguda (IHA), apesar de rara, permanece como uma condição rapidamente progressiva e frequentemente fatal. A intoxicação por acetaminofen (APAP) induz necrose hepática maciça e frequentemente leva à morte por edema cerebral. Terapias celulares são de grande interesse como potenciais tratamentos para IHA. Neste projeto foi avaliado o potencial terapêutico das células mononucleares da medula óssea (CMMO) em um modelo experimental de IHA induzida por APAP em camundongos. Métodos: A IHA foi induzida em camundongos C57Bl/6, previamente submetidos à dieta alcoólica por três semanas, através da administração de APAP na dose de 300 mg/kg por via intraperitoneal. Após a indução da IHA, os camundongos foram transplantados, por via endovenosa, com 107 CMMO...
Introduction and objectives: a cute liver failure (IHA), although rare, remains a rapidly progressive and often fatal condition. Poisoning by acetaminophen (APAP) induces a massive hepatic necrosis and often leads to death by cerebral edema. Cell therapies are of great interest as potential treatments for IHA. In this project we evaluated the therapeutic potential of bone marrow mononuclear cells (BMC) in an experimental model of IHA induced by APAP in mice. Methods: The IHA was induced in C57BL/6 mice previously submitted to the alcohol diet for three weeks by the administration of APAP at a dose of 300 mg / kg, intraperitoneally. After induction of IHA, the mice were transplanted intravenously with 107 BMC...
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Animals , Cytokines/analysis , Cytokines/immunology , Liver Failure, Acute/complications , Liver Failure, Acute/diagnosis , Liver Failure, Acute/mortality , Liver Failure, Acute/pathology , Bone Marrow/immunology , Bone Marrow/innervation , Cell- and Tissue-Based Therapy/methods , Cell- and Tissue-Based Therapy/mortalityABSTRACT
Objective To investigate the effect and safety of autologons bone margow-mononuclear cell (BM-MNC)transplantation after the bone marrow stimulation in patients with thromboangiitis obliterans (TAO).Methods The bone marrows of 12 patients were stimulated by an injection of the recombinant human granuloeyte-macmphage colony-stimulatory factor(GCSF)for 3-5 days.150-200 ml bone marrow was drown from the iliac spine and the autologous BM-MNC were obtained in each patients.Fifteen lower limbs of 12 patients received implantation of the autologous BM-MNC by an intramuscular iniecdon.A series of subjective indexes(including improvement of pain and cold sensation)and objeetive indexes [including increase of ankle braehial index(ABI),transcutaneons oxygen pressure(TcPO2)and improvement of foot skin ulcer] were used to evaluate the effects.Results The outcomes were evaluated after 2 months of transplantation.The pain-relief rate and the cold feeling improvement rate were 86.7%(13/15)and 93.3%(14/15)respectively.The ABI were 0.38 ±0.05 vs.0.61 ±0.14(P<0.05)before transplantation and 2 months after transplantation respectively.increased in 66.7%(10/15)limbs.The TcPO2 of the ischemic legs increased from(27.47±2.85)mm Hg to(43.53 ±8.38)mm Hg(t=-7.03,P<0.05)after the transplantation,and the improvement rate of TcPO2 was 93.3%(14/15).Skin ulcers in improved in 8/9 limbs.Twelve patients were followed up for all average period of 10 months.The patients'symptoms improved in 80.0%(12/15)limbs,as to the objective index the ABI was0.57±0.13,TcPO2 was(42.07 ±7.81)mm Hg,which improved significandy compared to before treatment(t=-5.33,-7.80,Ps<0.05).skin ulcer healing rate was 66.7%(6/9).The ischemic symptoms in 2 patients were not relieved.There WBS no mortality and high level amputation in all subjects.The complications,such as proliferative retinopathy,malitpmnt tumor,myocardial infarction,stroke or hemangioma were not found in any patients.Conclusion In patients with TAO,intramuscular transplantation of autologous BM-MNC after the bone marrow stimulation has advantages of less bone marrow aspiration,more mononuclear cell content and relatively high safety.It may be a new and effective method to alleviate symptoms and accelerate the healing of skin ulcer.
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Objective: To assess the effect of auto-bone marrow mononuclear cells (BMMNCs) transplantation on reendothelialization and neointima formation in vein grafts. Methods: BMMNCs were extracted from the bone marrows of adult rabbit under sterile environment and were labelled with DAPI before transplanted into vein grafts. Twenty adult rabbits were randomly divided into 2 groups: BMMNCs transplantation group(group I, n 10) and PBS transplantation group (group II, n=10). The left external jugular vein of 20 rabbits were harvested and transplanted between ipsolateral common carotid artery (AVG). Three days later, animals in group I were transplanted with BMMNCs(6x108 cells)/100 μl via periotic veins and those in group II were injected with 100 μl PBS. Animals were killed 4 weeks later and graft veins were harvested to observe the reendothelialization and the thickness of vein grafts. Results: We found that the transplanted cells survived and were incorporated into the endothelium of vein grafts in group I. The endothelium integrity of the vein grafts in group I was significantly better than that of group II. The intima thickness of vein grafts in group I was significantly thicker than that of group II. Conclusion: BMMCs transplantation therapy may improve re-endothelialization of the vein graft and inhibits intimal hyperplasi the vein graft.
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A terapia celular para doenças cardiovasculares representa uma nova e promissora opção terapêutica, principalmente para a cardiopatia isquêmica. A injeção de células mononucleares de medula óssea (CMMO) pela via intracoronariana (ICO) é a mais estudada em ensaios clínicos. Embora já se tenha documentado efeitos benéficos com essa terapia, dados relativos aos mecanismos de interação entre as células transplantadas e o ambiente microvascular cardíaco são escassos. A avaliação das CMMO na microcirculação miocárdica tem o potencial de esclarecer seus mecanismos de ação, abrindo novas possibilidades terapêuticas. O objetivo do estudo foi descrever e avaliar uma inovadora técnica de injeção ICO direta e visualização in vivo das CMMO na microcirculação miocárdica de pequenos roedores, após um período de isquemia miocárdica global (IMG) seguida de reperfusão, na presença e na ausência da aterosclerose. Materiais e Métodos: A técnica de transplante cardíaco heterotópico cervical (TCHC) em murinos foi modificada com a inserção e fixação de um microcateter conectado a uma seringa, no tronco braquiocefálico do animal doador. IMG foi induzida ocluindo-se a artéria nutridora do enxerto com um microclamp vascular, por 1 hora. As CMMO foram isoladas e marcadas antes da injeção ICO. Microscopia intravital de fluorescência (MIF) foi usada para análise da microcirculação coronariana do ventrículo direito (VD), incluindo arteríolas, capilares e vênulas pós-capilares. Parâmetros de perfusão e permeabilidade microvasculares e as interações entre as CMMO e células endoteliais foram estudados. O impacto da aterosclerose na recuperação, fenótipo e função celular também foi avaliado. Resultado: A MIF permitiu análise detalhada da microcirculação coronariana e da cinética das CMMO injetadas pela via ICO. A IMG afetou a microcirculação, reduzindo a densidade capilar funcional (DCF). Tal redução foi maior na presença de aterosclerose...
Cell therapy for cardiovascular disease represents a promising new therapeutic option, especially for ischemic heart disease. The intracoronary (ICo) injection of bone marow mononuclear cells (BMMC) is the most commonly studied rote in clinical trials. Although it has been documented beneficial effects with this therapy, data pertaining the mechanisms of interaction between transplanted cells and cardiac microvascular environment are lacking. Assessement of BMMC in myocardial microcirculation has the potential to clarify its mechanisms of action, opening new therapeutic possibilities. The objective of the study was to describe and to evaluate an innovative technique of direct ICo injection and in vivo visualization of BMMC in myocardial small rodents, after a period of global cardiac ischemia (GMI) and reperfusion, in the presence and absence of atherosclerosis. Materials and Methods: The syngeneic murine cervical heterotopic heart transplantation (CHHT) technique was modified by inserting and fixing a catheter connected to a syringe into the donor animal brachiocephalic trunk. GMI was induced occluding the nutritive artery of the graft for 1 hour. Bone marrow mononuclear cells were isolated and labeled prior to ICo injection. Intravital fluorescence microscopy (IFM) was used for analysis of the coronary microcirculation of the right ventricle (RV), including arterioles, capillaries and venules. Parameters of microvascular perfusion, microvascular permeability and the interactions between the BMMC and endothelial cells were studied. The impact of atherosclerosis on the recovery, phenotype and cellular function was also evaluated. Results: The IFM has allowed detailed analysis of both the coronary microcirculation, as the kinetics of BMMC. The GMI affected microcirculation, reducing the functional capillary density (FCD). This reduction was higher in the presence of atherosclerosis. The main area of cell retention was the capillary network...
Subject(s)
Animals , Atherosclerosis , Bone Marrow Cells , Coronary Circulation , Injections, Intra-Arterial/trends , Myocardial Ischemia/therapy , Leukocytes, Mononuclear/transplantation , Microcirculation , Microscopy, Fluorescence/methods , Rodentia , Bone Marrow Transplantation/methodsABSTRACT
Objective To track the magnetically labeled bone marrow mononuclear stem cells (BM-MNCs) in canine myocardial infarction (MI) model with MR. Methods BM-MNCs were labeled with Feridex effectively in vitro and then injected intramyocardially in 8 MI model dogs. Serial MR was performed with 1.5T MR scanner to show the location of the labeled cells compared with histology. Results The injection sites of labeled BM-MNCs could be located on the 1st and 2nd week, but disappeared on the 4th week. Corresponding to these sites, Prussian blue staining consistently showed that large clusters of cells were labeled by dense intracellular iron at the scar tissue. Conclusion Feridex labeling BM-MNCs enables ready detection in the beating heart on a conventional MR scanner after transplantation into canine infarcted myocardium.
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@#Objective To investigate whether uninduced autologous bone-marrow mononuclear cell (ABM-MNC) could survive and differentiate into myocardial cells and endothelial cells in the infarcted heart. Methods 40 male big-ear Japanese rabbits were divided into two groups randomly: the transplanted group (n=20) and the control group (n=20). The model of acute myocardial infarction was made by left anterior descending artery ligation, which was confirmed by ECG. The cardiac function was evaluated by the echocardiography. 7 days later, BrdU labeled ABM-MNCs were injected into infarcted and marginal area myocardium in the transplanted group, while the control rabbits were injected with saline. 6 weeks later, the hearts were harvested for histology and immunohistochemistry evaluation. Results In the transplanted group, viable cells labeled with BrdU could be identified in the infarcted area, and myocytes and endothelial cells labeled with BrdU can also be found in the border area, these cells demonstrate myogenic differentiation with the expression of α-Actin by immunostaining. Moreover, the vessel density of the transplanted group in the borders of the infarction was higher than the control group (P<0.05), but there was no difference in the infarcted areas between two groups (P>0.05). At the 6 weeks after experiment, the cardiac function was improved in both groups, but the transplanted group improved more than that in the control group (P<0.05). Conclusion Autologous bone-marrow mononuclear cells injected into the infarcted myocardium could survive in both the infarcted and the border areas, differentiated into endothelial cells and other cells which have obtained the characters of myocytes, and increase the vessel density in border area, improved the cardiac function.
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Objective To investigate whether uninduced autologous bone-marrow mononuclear cell (ABM-MNC) could survive and differentiate into myocardial cells and endothelial cells in the infarcted heart. Methods 40 male big-ear Japanese rabbits were divided into two groups randomly: the transplanted group (n=20) and the control group (n=20). The model of acute myocardial infarction was made by left anterior descending artery ligation, which was confirmed by ECG. The cardiac function was evaluated by the echocardiography. 7 days later, BrdU labeled ABM-MNCs were injected into infarcted and marginal area myocardium in the transplanted group, while the control rabbits were injected with saline. 6 weeks later, the hearts were harvested for histology and immunohistochemistry evaluation. Results In the transplanted group, viable cells labeled with BrdU could be identified in the infarcted area, and myocytes and endothelial cells labeled with BrdU can also be found in the border area, these cells demonstrate myogenic differentiation with the expression of α-Actin by immunostaining. Moreover, the vessel density of the transplanted group in the borders of the infarction was higher than the control group (P＜0. 05), but there was no difference in the infarcted areas between two groups (P＞0.05). At the 6 weeks after experiment, the cardiac function was improved in both groups, but the transplanted group improved more than that in the control group (P＜0.05). Conclusion Autologous bone-marrow mononuclear cells injected into the infarcted myocardium could survive in both the infarcted and the border areas, differentiated into endothelial cells and other cells which have obtained the characters of myocytes, and increase the vessel density in border area, improved the cardiac function.
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Objective To observe the effect of allogenic bone marrow mononuclear cells(BM-MNCs) transplantation on myocardial apoptosis after acute myocardial infarction(AMI) in rats.Methods 40 Wistar rats were randomly divided into control group(n=20) and transplantation group(n=20).Myocardium around the infarcted left ventricular area of the rats in transplantation group were injected with BM-MNCs suspension beneath the epicardium.Myocardium the area of control group was injected with culture solution.Results After 4 weeks of the operation,the myocardial apoptosis index,the TNF-? content and the PDCD5 mRNA of transplantation group were all notably less than those of control group(P