Osteogenic differentiation of rat bone marrow stem cells induced by acanthopanax senticosus glycoside / 解放军医学杂志

Objective To investigate the effect of acanthopanax on inducing osteogenesis of rat bone marrow stem cells (BMSCs). Methods BMSCs were extracted from SD rats, and the surface antigens CD45, CD29 and CD90 were identified by flow cytometry at the third generation. Different concentrations of acanthopanax were added to the classical osteogenic medium to make it being 9 groups: A(1×10–4mol/L), B(1×10–5mol/L), C(1×10–6mol/L), D(1×10–7mol/L), E(1×10–8mol/L), F(1×10–9mol/L), G(1×10–10mol/L), H(classical osteogenic group), and I(negative control group). The cell counting kit CCK-8 was used to detect cell proliferation, RT-qPCR was performed to detect the mRNA expression of bone morphogenetic protein (BMP), and then the optimal concentration of acanthopanax was selected and used to the later experiments. On the 12th day of culturing with optimal concentration, RT-qPCR was performed to detect osteogenic differentiation-related gene expression: RUNX, OSX, BSP and OCN. Western blotting was used to detect the levels of transmembrane receptor protein 1 (Notch1) and hairy enhancer of split 1 (Hes1). On the 21th day of culturing, the mineralized calcium nodules were stained with alizarin red. Results The surface antigens of the third generation BMSCs were consistent with the stem cell identification criteria. CCK-8 results indicated that the proliferation of BMSCs was inhibited in group A and group B 120h after cultivation, so the two groups were discarded in the later culture. RT-qPCR results showed that among groups C-G with acanthopanax, the expression of BMP in group E (1×10–8mol/L) was the highest (4.91±0.46), so 1×10–8mol/L was selected as the optimal concentration of acanthopanax to finish the later experiments. The results of RT-qPCR showed that the expression of OSX was significantly higher in group E (30.72±1.96) than in group I (1.02±0.27) and group H (9.99±0.59, P<0.05); the expression of BSP (8.15±0.47) was higher than in group I (1.09±0.31) and group H (6.03±0.8, P<0.05); and the expression of OCN (5.91±0.68) was higher than in group I (1.18±2.91) and group H (3.05±0.53, P<0.05). However, the expression of RUNX was higher in group E (1.99±0.09) than in group I (1.02±0.19, P<0.05), but was lower than in group H (2.51±0.06, P<0.05). Western blotting suggested that the Notch1 in group E (4608±103) was higher than in group I (2638±308), but lower than in group H (5218±182, P<0.05); Hes1 expression in group E (8885±17) was higher than in group I ( 6241±461), but lower than in H group (12289±629, P<0.05). The alizarin red staining indicated that the number of mineralized calcium nodules was higher in group E than in group H, suggesting that the osteogenic effect in group E (with acanthopanax concentration of 10–8mol/L) is better than in group H. Conclusion Acanthopanax may cooperate with dexamethasone to promote the osteogenesis of BMSCs, which may be related to the Notch signaling pathway.

Hypoxia-stressed cardiomyocytes promote early cardiac differentiation of cardiac stem cells through HIF-1/Jagged1/Notch1 signaling

Hypoxia is beneficial for the differentiation of stem cells transplanted for myocardial injury, but mechanisms underlying this benefit remain unsolved. Here, we report the impact of hypoxia-induced Jagged1 expression in cardiomyocytes (CMs) for driving the differentiation of cardiac stem cells (CSCs). Forced hypoxia-inducible factor 1 (HIF-1) expression and physical hypoxia (5% O) treatment could induce Jagged1 expression in neonatal rat CMs. Pharmacological inhibition of HIF-1 by YC-1 attenuated hypoxia-promoted Jagged1 expression in CMs. An ERK inhibitor (PD98059), but not inhibitors of JNK (SP600125), Notch (DAPT), NF-B (PTDC), JAK (AG490), or STAT3 (Stattic) suppressed hypoxia-induced Jagged1 protein expression in CMs. c-Kit CSCs isolated from neonatal rat hearts using a magnetic-activated cell sorting method expressed GATA4, SM22 or vWF, but not Nkx2.5 and cTnI. Moreover, 87.3% of freshly isolated CSCs displayed Notch1 receptor expression. Direct co-culture of CMs with BrdU-labeled CSCs enhanced CSCs differentiation, as evidenced by an increased number of BrdU/Nkx2.5 cells, while intermittent hypoxia for 21 days promoted co-culture-triggered differentiation of CSCs into CM-like cells. Notably, YC-1 and DAPT attenuated hypoxia-induced differentiation. Our results suggest that hypoxia induces Jagged1 expression in CMs primarily through ERK signaling, and facilitates early cardiac lineage differentiation of CSCs in CM/CSC co-cultures HIF-1/Jagged1/Notch signaling.

The efficacy and long-term safety of autologous bone marrow stem cells transplantation in patients with hepatitis B virus-associated decompensated liver cirrhosis / 中华传染病杂志

Objective To evaluate the efficacy and long-term safety of autologous bone marrow stem cells(ABMSC)transplantation in patients with hepatitis B virus(HBV)-associated decompensated liver cirrhosis.Methods This was an open-label,prospective matched case-control study.Thirty patients with HBV-associated decompensated liver cirrhosis hospitalized at the Third Affiliated Hospital of Sun Yat-Sen University from January 2005 to June 2010 were collected and infused with stem cells(stem cell group). Another thirty patients in control group were matched according to baseline characteristics and treated with standard medicine therapy.The patients in stem cell group were treated with stem cells infusion by hepatic artery or portal vein based on standard medicine therapy.All the patients were followed up for 5 to 10 years after surgery. Biochemical indicators were evaluated within the first 48 weeks after transplantation.The complications of cirrhosis and the cumulative incidence rate of hepatocellular carcinoma(HCC)were observed.Measurement data with normal distribution were analyzed by t test. Measurement data with non-normal distribution were compared by Mann-Whitney test.Count data were compared by χ2 test.The cumulative incidence rate of HCC development was compared by Kaplan-Meier analysis.Results The bone marrow aspiration and transplantation surgery were well tolerated in all patients in stem cell group.No complication related to stem cell transplantation therapy was observed. The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil) and prothrombin time(PT)decreased,albumin level increased,while model for end-stage liver disease (MELD)scores decreased in both groups after treatment.Serum albumin level in stem cell group increased and ALT level decreased markedly at week 4,compared with that in control group at week 4(Z=2.188,P=0.029,Z=3.296,P=0.001,respectively).In stem cell group,21 patients received stem cells transplantation by hepatic artery and 9 patients by portal vein.Biochemical indicators were improved in all patients compared to baseline.However,there was no statistically significant differences between hepatic artery group and portal vein group.The median follow-up time was 6 years.Two patients in stem cell group and 1 patient in control group died(χ2=0.351,P=0.554).Six patients in stem cell group (20.0%)and 11 patients(36.7%)in control group developed HCC.There was no significant differences in the cumulative incidence rate of HCC between two groups(χ2= 0.148,P= 0.701).Hepatorenal syndrome did not development in either group.There were no statistically significant differences in the rates of complications including spontaneous peritonitis,hepatic encephalopathy and gastrointestinal hemorrhage between two groups after 5 to 10 years of follow-up(χ2=0.162,P=0.688,χ2=1.071,P=0.301,χ2=1.071,P=0.301,respectively).Conclusion ABMSC transplantation in patients with HBV-associated decompensated liver cirrhosis improves liver function transiently and has long-term safety.

Autologous bone marrow stem cells implantation to bone inducing active material combined with core decompression in the treatment of early femoral head osteonecrosis / 中华显微外科杂志

Objective To observe the curative effect of autologous bone marrow stem cells implantation to bone inducing active material combined with core decompression in the treatment of early femoral head osteonecrosis (FHON).Methods From April,2010 to March,2012,in Department of Orthopaedics,the First Affiliated Hospital of Zhengzhou University,a total of 79 adult patients with 108 hips suffered from the early stage FHON were treated with autologous bone marrow stem cells implantation to bone inducing active material combined with core decompression through the core of the femoral canal,male of 65 cases,female of 14 cases,the mean age was 29.5 (20-50) years old.According to the etiology classification:the alcohol-induced FHON was in 54 patients with 66 hips,steroid-induced FHON in 14 patients with 20 hips,steroid and alcohol-induced ONFH was in 11patients with 22 hips.According to association research circulation osseous (ARCO)classifying,Ⅰ-A,Ⅰ-B,Ⅱ-A,Ⅱ-B phases were 6,16,8,and 78 hips,respectively.There were 43 hips in left side and 65 hips in the right side.Results All patients were followed up from 4 to 6 (4.8 ± 0.6) years.Compared with before operation,the scores of all patients were significantly increased (P ＜ 0.05).All patients with hip pain symptoms were relieved or disappeared.The healing tine of the patients in all age groups was statistically significant (P ＜ 0.05),and with the increase of age,the healing time was prolonged.The excellent and good rates of Ⅰ-A,Ⅰ-B,Ⅱ-Aand Ⅱ-B were 100％ (6 / 6),100％ (16/16),100％ (8/8),and 98.7％ (77/78).The X-ray showed that coarse channel osteogenic phenomenon is obvious,there is 1 case collapse of femoral head of stage Ⅱ-B,the rest were not collapse.Conclusion The treatment of early osteonecrosis of the femoral head with autologous bone marrow stem cells implantation to bone inducing active material combined with core decompressionis definitely effective,especially in patients with ARCO:Ⅰ-A,Ⅰ-B and Ⅱ-A phase,and the effect of ARCO:Ⅰ-A and Ⅱ-A is the best.

Isolation, expansion and differentiation of mesenchymal stromal cells from rabbits' bone marrow / Isolamento, expansão e diferenciação de celulas tronco mesenquimais oriundas da medula óssea de coelhos

Tissue engineering has been a fundamental technique in the regenerative medicine field, once it permits to build tri-dimensional tissue constructs associating undifferentiated mesenchymal cells (or mesenchymal stromal cells - MSCs) and scaffolds in vitro. Therefore, many studies have been carried out using these cells from different animal species, and rabbits are often used as animal model for in vivo tissue repair studies. However, most of the information available about MSCs harvesting and characterization is about human and murine cells, which brings some doubts to researchers who desire to work with a rabbit model in tissue repair studies based on MSCs. In this context, this study aimed to add and improve the information available in the scientific literature providing a complete technique for isolation, expansion and differentiation of MSCs from rabbits. Bone marrow mononuclear cells (BMMCs) from humerus and femur of rabbits were obtained and to evaluate their proliferation rate, three different culture media were tested, here referred as DMEM-P, DMEM´S and α-MEM. The BMMCs were also cultured in osteogenic, chondrogenic and adipogenic induction media to prove their multipotentiality. It was concluded that the techniques suggested in this study can provide a guideline to harvest and isolate MSCs from bone marrow of rabbits in enough amount to allow their expansion and, based on the laboratory experience where the study was developed, it is also suggested a culture media formulation to provide a better cell proliferation rate with multipotentiality preservation.

A engenharia de tecidos tem sido uma técnica fundamental no campo da medicina regenerativa, uma vez que permite a criação de peças teciduais tri-dimensionais por meio da associação de células mesenquimais indiferenciadas (ou células estromais mesenquimais - CEMs) e moldes de biomateriais in vitro. Assim, muitos estudos têm sido realizados utilizando estas células oriundas de diferentes espécies animais, e os coelhos são frequentemente utilizados como um modelo animal para estudos in vivo de reparação tecidual. No entanto, a maioria das informações disponíveis sobre a coleta e caracterização de CEMs referem-se às células humanas e murinas, o que traz algumas dúvidas para pesquisadores que desejam trabalhar com coelhos em estudos de reparação de tecidos baseados em CEMs. Neste contexto, o presente estudo objetivou contribuir e aprimorar as informações disponíveis na literatura científica fornecendo uma técnica completa para o isolamento, expansão e diferenciação das MSCs de coelhos. Células mononucleares da medula óssea (CMMOs) do úmero e fêmur de coelhos foram obtidas e, para avaliar sua taxa de proliferação, três meios de cultura diferentes foram testadas, aqui referidos como DMEM-P, DMEM'S e α-MEM. As CMMOs também foram cultivadas em meios de indução osteogênico, condrogênico, e linhagens adipogênico para provar a sua multipotencialidade. Concluiu-se que as técnicas sugeridas neste estudo podem fornecer um guia para a coleta e isolamento de CEMs da medula óssea de coelhos em quantidade suficiente para permitir a sua expansão e, com base na experiência de laboratório onde o estudo foi desenvolvido, é também sugerida uma formulação de meio de cultivo para proporcionar uma melhor taxa de proliferação celular com preservação da multipotencialidade.

Clinical research about liver function of hepatolenticular degeneration combined liver fibrosis by transplantation of bone marrow stem cells / 中国免疫学杂志

Objective:To evaluate the influence of bone marrow stem cells ( BMSCs ) in the serological indicators of hepatolenticular degeneration combined liver fibrosis.Methods:60 cases were randomly divided into 3 groups: penicillamine group, BMSCs group and BMSCs+penicillamine group.BMSCs(2 ml)were injected into vein with normal saline(100 ml) every 10 days ( 3 times for a period of treatment).Liver tissue pathology biopsy was inspected and TBIL, ALT, ALB, CHE, PDGF-BB, TGF-β1, IL-6 and TNF-αwere detected at 0,4 ,8 and 12 weeks.Results: The level of serological indicators about liver functions were reduced in every group, while the changes in BMSCs+penicillamine group were especially obvious ( P<0.05 ).Conclusion: Penicillamine combined with BMSCs was effective in the improvement of liver functions of hepatolenticular degeneration combined liver fibrosis.

Effect of Activating Blood to Resolve Stagnation on Cells Expressing CD34 and Vascular Endothelial Growth Factor in Rats with Acute Myocardial Infarction / 中国康复理论与实践

@#Objective To explore the effect of Activating Blood to Resolve Stagnation on the expression of CD34 and vascular endothelial growth factor (VEGF) in rats with acute myocardial infarction. Methods 32 Sprague-Dawley rats were randomly divided into sham operation group (A, n=8), model group (B, n=8), Xuesaitong Injection + granulocyte colony- stimulating factor (G- CSF) group (C, n=8) and G-CSF group (D, n=8). Corresponding medicine was given to each group 3 hours after modeling, for 6 days. Pathomorphological changes were observed through HE staining, and the expression of CD34, VEGF and Ki-67 were observed through immunohistochemical staining. Results The expressions of CD34, VEGF and Ki-67 were higher in groups B, C and D than in group A (P<0.05), and were higher in group groups C and D than in group B (P<0.05). The expressions of CD34 and VEGF were higher in group C than in group D (P<0.05). However, there was no significant difference in the expression of Ki-67 between 2 groups (P>0.05). Conclusion The expression of CD34 and VEGF increases with Activating Blood to Resolve Stagnation method, which is superior to using G-CSF only. Activating Blood to Resolve Stagnation may play an important role in the treatment of acute myocardial infarction.

Effect of Activating Blood to Resolve Stagnation on Cells Expressing CD34 and Vascular Endothelial Growth Fac-tor in Rats with Acute Myocardial Infarction / 中国康复理论与实践

Objective To explore the effect of Activating Blood to Resolve Stagnation on the expression of CD34 and vascular endotheli-al growth factor (VEGF) in rats with acute myocardial infarction. Methods 32 Sprague-Dawley rats were randomly divided into sham opera-tion group (A, n=8), model group (B, n=8), Xuesaitong Injection + granulocyte colony-stimulating factor (G-CSF) group (C, n=8) and G-CSF group (D, n=8). Corresponding medicine was given to each group 3 hours after modeling, for 6 days. Pathomorphological changes were observed through HE staining, and the expression of CD34, VEGF and Ki-67 were observed through immunohistochemical staining. Re-sults The expressions of CD34, VEGF and Ki-67 were higher in groups B, C and D than in group A (P0.05). Conclusion The expression of CD34 and VEGF in-creases with Activating Blood to Resolve Stagnation method, which is superior to using G-CSF only. Activating Blood to Resolve Stagna-tion may play an important role in the treatment of acute myocardial infarction.

Acupuncture-moxibustion with Tong Du Method on the Bone Marrow Stem Cells in Patients with Ischemic Stroke / 上海针灸杂志

ObjectiveTo observe the effect ofTong Du acupuncture-moxibustion, conventional acupuncture, and medication on the bone marrow stem cells (BMSCs) mobilization markers CD34+and CD133+, neurological deficit score in patients with ischemic stroke.Method Totally 120 eligible subjects with ischemic stroke were randomized into aTong Du acupuncture-moxibustion group (group A), a conventional acupuncture group (group B), and a medication group (group C), 40 in each group. In addition to the conventional medication given to all three groups, patients in group A also received moxibustion at Baihui (GV20), bloodletting at Dazhui (GV14), and acupuncture at Fengchi (GB20), Quchi (LI11), Hegu (LI4), Zusanli (ST36), and Sanyinjiao (SP6); patients in group B also received acupuncture at Fengchi, Quchi, Hegu, Zusanli, and Sanyinjiao; group C received patent Chinese medicine Tong Xin Luo capsules. The intervention lasted 14 d in all three groups. The CD34+and CD133+and neurological deficit score were evaluated and compared before and after intervention.Result The three treatment methods all significantly increased the level of CD34 + and CD133+in peripheral blood (P0.05); the three treatment methods all improved the neurological deficit score to various extent, and the result in group A was significantly different from that in group B and C (P<0.05,P<0.01), and there was also a significant difference between group B and C (P<0.05).ConclusionsTong Du acupuncture-moxibustion, conventional acupuncture, and medication can influence the BMSCs and neurological deficit to different extent; the effect ofTong Du acupuncture-moxibustion on the BMSCs and neurological deficit is more significant than that of the conventional acupuncture and medication, suggesting thatTong Du acupuncture-moxibustion method should have advantage in treating cerebral infarction.

Advances in research of the reversal of hepatolenticular degeneration with liver fibrosis by transplantation of bone marrow mesenchymal stem cells / 实用医学杂志

Objective To evaluate the effect of bone marrow stem cells (BMSCs) transplantation in the treatment of hepatolenticular degeneration of liver fibrosis. Methods Sixty cases with confirmed hepatolenticular degeneration of liver fibrosis were randomly divided into 3 groups: penicillamine group [40 mg/(kg·d)], BMSCs group and BMSCs + penicillamine group. Autologous BMSCs (2 mL) were injected into vein with normal saline (100 mL). Liver tissue pathology biopsy was inspected and the changes in HA, PCⅢ, LN, CⅣ, TIMP-1 and MMP-1 were observed at 0, 4th, 8th and 12th week during the therapeutic process. Results The level of serum fibrotic markers were reduced in every group , while the changes in BMSCs + penicillamine group were especially obvious (P < 0.05). Conclusion Penicillamine combined with BMSCs was effective in the therapy of hepatolenticular degeneration of liver fibrosis.

Study on the concentration of 17β-estradiol in BMSCs and different tissues of rats / 实用医学杂志

Objective To investigate the concentration of 17β-estradiol in bone marrow stem cells (BMSCs) and different tissues of rats, and study the function of estradiol produced in extragonadal sites prelimi-narily. Methods (1) The concentration of 17β-estradiol in lysate of BMSCs and in supernatant cell culture media were detected by Elisa method after rats BMSCs were serum-free cultured in 0 h , 24 h and 48 h respective ly . (2) The tissues of organs were grinded and broken under ultrasonic wave , then washed and weighed. The con-centration of 17β-estradiol in different tissues was detected by Elisa method. Results Compared with the concen-tration of 17β-estradiol at 24 h, the concentration of 17β-estradiol at 48h significantly increased (P 0.05). Conclusions (1) 17β-estradiol can be secreted by BMSCs , and the concentration is proportional to the time to some degree. But whether the extragonadal estrogen can function locally is still unclear. (2) The concentration of 17β-estradiol is non- gonad dependent but whether it is secreted locally remains to be elucidated. The non- gonad estrogen could be the estrogen source of bone metabolism in order to sustain bone health after menopause.

In vitro differentiation of islet-like cells from mouse bone-marrow stem cells on a PLGA scaffold / 国际生物医学工程杂志

Objective To investigated the potential influences of poly(lactic-co-glycolicacid)(PLGA)scaffold as a platform on the differentiation of mouse bone-marrow stem cells to islet-like cells.Methods Mouse bonemarrow stem cells were grown and differentiated in culture with or without PLGA scaffold,and cell morphology and functions were compared within these groups.Results The PLGA scaffold showed fine biological compatibility.Differentiated islet-like cells were dithizone (DTZ) positive,insulin and C-peptide double positive,glucagon positive and somatostatin positive in both groups.Under electron microscope there were ultrastructures similar to that of islet β cells in cells of both groups.Cells with PLGA scaffold secreted more insulin under high level glucose stimulation (P＜0.01).Conclusions PLGA scaffold was biologically compatible and improved function of the differentiated islet-like cells.

Xinfukang drug-containing serum on bone marrow stem cell transcription and secretion of SDF-1α / 国际生物医学工程杂志

Objective To clarify the effects of Xinfukang containing-serum on stromal cell-derived factor-1α (SDF-1α) translation and protein secretion of bone marrow stem cells (BMSCs).Methods BMSCs were isolated and amplified using bone marrow culture method,and were identified by flow cytometry.mRNA and protein secretion of SDF-1α were detected by quantitative PCR (q-PCR) and enzyme linked immunosorbent assay (ELISA),respectively.Results The expression of SDF-1α mRNA were significantly increased after 72 h in drug-containing serum,and SDF-1α mRNA in the experimental group was approximately 200 times as that in the control group (P＜0.05).Secretion of SDF-1 α in the experimental group (277.561 1 ± 15.651 8) pg/ml was nearly doubled compared with that in the control group (153.107 1±14.765 1) pg/ml (P＜0.05).Conclusions BMSCs from whole bone marrow adherent culture have high purity,and drug-containing serum can promote BMSCs to express SDF-1 α mRNA and secretion of SDF-1 α.

Orthogonal Compatibility of Icariin, Psoralen, Oleanolic Acid, Stilbene Glucoside on Regulation of Bmp2, Smad1, and Smad 4 Induced Osteogenic Differentiation of BMSCs / 世界科学技术-中医药现代化

This study was aimed to observe four kinds of kidney-tonification medicine, which were Epimedium, pso-ralen, Ligustrum lucidum, Polygonum with the active ingredient of icariin, psoralen, oleanolic acid, stilbene glucoside and their orthogonal compatibility. There were two kinds of non-kidney tonification medicine, which were Chuanx-iong and astragalus with the active ingredient of TMP and astragaloside. The observation was made on the regulatory role of rat bone marrow stem cells (BMSCs). A total of 65 SD rats were randomly divided into the normal control group, positive transformed control group, kidney-tonification compatibility group (including Group 1, Group 2, Group 3, Group 4, Group 5, Group 6, Group 7, Group 8, and Group 9), non-kidney tonification medicine control group (in-cluding TMP group and astragaloside group). Intragastric administration of medication was given to the kidney-tonifi-cation compatibility group and the non-kidney tonification medicine control group, once a day for 3 consecutive days. Intragastric administration of equal amount of normal saline was given to the normal control group and the posi-tive transformed control group. On the third day of intragastric administration, rats in each group were sacrificed. Serum containing medication was used in the culture of BMSCs for 6, 12, or 18 days. ELISA method was used to quantitatively detect the expression activity and content of BMP2 on the 6th, 12th, or 18th day, in order to evaluate the degree of bone cell differentiation degree. Real-time quantitative PCR method was used for detection of expression of Bmp2, Smad1, Smad4 mRNA in serum containing medication in the culture of BMSCs on the 18th day. The results showed that the kidney-tonification compatibility can improve the expression activity and content of BMP2 culture in vitro, with the peak on the 12th day. The kidney-tonification compatibility groups can upregulate expressions of Bmp2, Smad1, Smad4 mRNA. It was concluded that the active ingredient compatibility of kidney-tonification medicine can promote BMSCs. Its mechanism may be related to the upregulation of expression of Bmp2, Smad1, Smad4 mRNA, and the activity and content of Bmp2.

Impact of nano-structured porous polycaprolactone combined with bone marrow stromal cells on CTGF expression in rabbit impaired articular cartilage / 实用医学杂志

Objective To study nano-structured porous polycaprolactone combined with bone marrow stromal cells on CTGF expression in rabbit impaired cartilage. Methods 50 rabbits were randomly divided into normal group, model group, NSP-PCL group, BMSCs group, or NSP-PCL + BMSCs group. A model of rabbit impaired cartilage was surgically established. Then NSP-PCL, BMSCs, and NSP-PCL + BMSCs were separately administered to the latter three groups once a week from the 2nd week to the 5th week after the procedure. The impaired cartilage tissues were collected 24 h after the last administration. The cartilage tissues were pathologically examined by H & E staining. Number of surviving BMSCs was detected by Hoeehst33342 Markers 1 week and 3 weeks after transplantation. Levels of CTGF protein in cartilage were determined by immunohistochemistry and Western blotting. Results In the model group, cartilage layer became thinner, with proliferation of fibroblast cells and a obvious cartilage surface hollow; New cartilage cells appeared in the surface hollow of the impaired cartilage in each treatemnt group, with a thicker layer. The number of transplanted BMSCs after 3 weeks was significantly increased in BMSCs group and NSP-PCL + BMSCs group. As compared with the model group, levels of CTGF protein were increased in each treatment group, with significant differences (P < 0.05). Conclusions Nano-porous polycaprolactone combined with bone marrow mesenchymal stem cells can repair cartilage damage by enhancing the expression of CTGF protein.

Effect of transplantation of bone marrow stem cells on myocardial infarction size in a rabbit model / 世界急诊医学杂志（英文）

@#BACKGROUND: Intravenous transplantation has been regarded as a most safe method in stem cell therapies. There is evidence showing the homing of bone marrow stem cells (BMSCs) into the injured sites, and thus these cells can be used in the treatment of acute myocardial infarction (MI). This study aimed to investigate the effect of intravenous and epicardial transplantion of BMSCs on myocardial infarction size in a rabbit model. METHODS: A total of 60 New Zealand rabbits were randomly divided into three groups: control group, epicardium group (group I) and ear vein group (group II). The BMSCs were collected from the tibial plateau in group I and group II, cultured and labeled. In the three groups, rabbits underwent thoracotomy and ligation of the middle left anterior descending artery. The elevation of ST segment>0.2 mV lasting for 30 minutes on the lead II and III of electrocardiogram suggested successful introduction of myocardial infarction. Two weeks after myocardial infarction, rabbits in group I were treated with autogenous BMSCs at the infarct region and those in group II received intravenous transplantation of BMSCs. In the control group, rabbits were treated with PBS following thoracotomy. Four weeks after myocardial infarction, the heart was collected from all rabbits and the infarct size was calculated. The heart was cut into sections followed by HE staining and calculation of infarct size with an image system. RESULTS: In groups I and II, the infarct size was significantly reduced after transplantation with BMSCs when compared with the control group (P<0.05). However, there was no significant difference in the infarct size between groups I and II (P>0.05). CONCLUSION: Transplantation of BMSCs has therapeutic effect on MI. Moreover, epicardial and intravenous transplantation of BMSCs has comparable therapeutic efficacy on myocardial infarction.

Allogeneic bone combined with autologous bone marrow stem cell transplantation for the treatment of benign bone tumors and tumor-like lesions / 中国组织工程研究

BACKGROUND:Al ogenic bone is a clinical commonly used bone graft material, but the osteoinductive capacity is the biggest problem. OBJECTIVE:To evaluate the effect of al ogeneic bone combined with autologous bone marrow stem cells on the repair of bone defects after scraping or resection of benign bone tumors and tumor-like lesions. METHODS:Sixty-five cases of benign bone tumors (including patients with tumor-like lesions) were divided into two groups according to bone graft. There were 35 cases in the composite bone marrow stem cells for bone graft group, and 20-40 mL red bone marrow were extracted from anterosuperior iliac spine or iliac spine on both sides according to the expected amount of bone graft, then the bone marrow stem cells were isolated, purified, cultured and amplified for standby, and the bone marrow stromal stem cells and al ogeneic bone particles were ful y blended before bong graft. After tumor scraping or resection, the blended bone marrow stromal stem cells and al ogeneic bone particles were implanted into the bone defect region. In the bone graft group, the bone defect was implanted with al ogeneic bone soaked with saline for half an hour. X-ray examination was performed at 1, 3, 6 and 12 months after treatment to compare the fuzzy boundary and the time for disappear, and the postoperative complications were observed. RESULTS AND CONCLUSION:Al the 62 patients were fol owed-up for more than 12 months. The fuzzy boundary time and disappear time in the composite bone marrow stem cells for bone graft group were shorter than those in the bone graft group (P<0.05). In the composite bone marrow stem cells for bone graft group, one case appeared rejection and healed after treated with immunosuppressive agents for 2 weeks, and no complication observed in two groups. The results indicate that al ogeneic bone composite autologous bone marrow stem cells for bone graft can promote bone fusion and bone defect healing.

La terapia celular en la cardiopatía isquémica / Cell therapy for ischemic heart disease

La cardiopatía isquémica es la principal causa de muerte e insuficiencia cardiaca a nivel mundial. Esto hace de vital importancia el desarrollo de nuevas modalidades terapéuticas, que disminuyan la mortalidad y complicaciones a largo plazo en estos pacientes. Una de las principales líneas de investigación a nivel mundial es la regeneración miocárdica a partir de células progenitoras, con el fin de mejorar la función sistólica y diastólica de los pacientes con cardiopatía isquémica, además de incrementar su sobrevida. Con bases teóricas y fisiológicas sobre la función de estas células, se han llevado a cabo con gran entusiasmo a nivel mundial, estudios en animales y humanos para tratar de definir la utilidad del empleo de las células madre, en el manejo de los pacientes con cardiopatía isquémica. En la actualidad, la terapia regenerativa en la cardiopatía isquémica es considerada una herramienta terapéutica novedosa, de beneficios teóricos considerables y pocos efectos adversos. En esta revisión presentamos los fundamentos científicos básicos que apoyan el empleo de esta terapia, la evidencia clínica actual sobre su beneficio. Señalamos los puntos controversiales y las perspectivas sobre su empleo y utilidad a corto y largo plazo.

Ischemic heart disease is the leading cause of death and heart failure worldwide. That is why it is important to develop new therapeutic modalities to decrease mortality and long-term complications in these patients. One of the main lines of research worldwide is myocardial regeneration, using progenitor cells in order to improve systolic and diastolic function in patients with ischemic heart disease, as well as to increase their survival. There have been carried out, with great enthusiasm worldwide, human and animal studies to define the usefulness of stem cells in the management of patients with ischemic heart disease. Today, regenerative therapy in ischemic heart disease is considered a novel therapeutic tool, with substantial theoretical benefits and few side effects. Here we present the scientific principles that support the use of this therapy, discuss the current clinical evidence available; and point out the controversial issues still not clarified on its use and usefulness in the short and long term.

Autologous Bone Marrow Stem Cell Therapy Shows Functional Improvement in Hemorrhagic Stroke.

In hemorrhagic stroke, damage to the brain tissue is inevitable and no effective treatment for functional improvement is currently available except neurorehabilitation. Stem cell therapy is a rapidly growing field and has recently opened new avenues for brain repair strategies. We present a case study of a 69-year-old female treated with stem cell therapy for right-sided hemiplegia caused due to left thalamic hemorrhagic stroke. Inspite of regular physiotherapy, the patient had constant residual neurodeficit, one year after the stroke, which was severely incapacitating. In view of the same, the patient was given intrathecal autologous bone marrow derived stem cell therapy as part of the neuroregeneration and rehabilitation therapy (NRRT) along with rehabilitation. After the therapy, patient showed functional as well as neurological improvements (cognition and motor strength) without any side effects. There is accumulating experimental data showing the benefits of cell transplantation on functional recovery after hemorrhagic stroke. This case study supports the concept of neuroregeneration with bone marrow stem cells as a novel strategy having great therapeutic potential. However, large clinical studies are needed to further investigate autologous bone marrow stem cell therapy in addition to neurorehabilitation for treating the disability in hemorrhagic stroke.

CD34+ bone marrow stem cells homing to embolized rabbit lung tissue / 第二军医大学学报

Objective To observe the homing of bone marrow stem cells (BMSCs) to the embolized lung tissues after mobilization and to investigate the potential mechanisms. Methods Thirty healthy Chinese big-ear rabbits were randomized into two groups regardless of gender: pulmonary thromboembolism (PTE) group (model group), PTE+granulocyte colony-stimulating factor (G-CSF) mobilization group (experimental group), with each group having 15 animals. Stable PTE models were established by injecting autologous thrombus into the femoral veins of the animals (model group). The animals in the experimental group received daily hypodermic injection of G-CSF (10 μg/[kg��d]) for five days, which was started 4 days before the establishing PTE models and ended at one day after the models were established. The animals were sacrificed 24 hours after modeling in both groups for general sample observation. Hematoxylin-eosin (HE) staining was done for the samples. Immunohistochemical analysis was done to detect the expressions of CD34 and SDF-1 in the embolized area, edge area, and normal area. A medical image processing software was used to analyze the results of immunohistochemical results to calculate the relative contents of CD34 and SDF-1. Results (1)General sample observation: the animals exhibited the damage of lung tissues in both groups, including splinter hemorrhage sites, local pale region and atelectasis. (2)Microscopic pathological observation: the embolized areas of all animals exhibited interstitial edema and hyperemia, and many red blood cells entered the pulmonary alveoli. Increased monocyte infiltration was detected in the experimental group. (3)Results of immunohistochemical analyses and image analyses of CD34: Most CD34 expression was found in the embolized area, very weak expression was found in the edge area, and hardly any was detected in the normal area. CD34 expression in embolized area was significantly higher in the experimental group than that in the model group (P<0.01). More monocytes expressing CD34 were seen in the experimental group and only a small number of them were found in the model group. (4) Results of immunohistochemical analyses and image analyses of SDF-1: Most SDF-1 expression was found in the embolized area, and no expression was found in the edge area and normal area. SDF-1 expression in embolized area was significantly higher in experimental group than in the model group (P<0.01). Conclusion BMSCs expressing CD34 can home to the embolized lung tissues after PTE. The increased SDF-1 expression may be one of the mechanisms of BMSCs homing to embolized lung tissues after PTE. Mobilization with G-CSF can increase SDF-1 expression in embolized area, which can attract more BMSCs to home to embolized lung tissues.