Effect of gamma irradiation on nuclear factor-kappa B in cultured bone marrow stromal cells

Objective: To explore the effect of gamma irradiation on nuclear factor-kappa B in cultured bone marrow stromal cells. Methods: Immunocytochemistry, Western blot and electrophoretic mobility shift assay (EMSA) were used. Results: The expression of NF-kB in cultured mouse bone marrow stromal cells (BM-SCs) on the level of protein was elevated after exposure to 60Co in the dosage of 8. 0 Gy with the use of im-munocytochemistry and Western blot. The activity of nuclear factor-kappa B in cultured BMSCs was significantly increased after exposure to gamma irradiation by using EMSA. The activity peak was at the 4th h after irradiation. Conclusion: Our results suggest that the activation of nuclear factor-kappa B in the BMSCs after irradiation may be involved in the protection of BMSCs against apoptosis and in the recovery of hematopoiesis after radiation.

Optimization of adipose tissue-derived mesenchymal stromal cells transplantation for bone marrow repopulation following irradiation

BACKGROUND Bone marrow(BM)suppression is one of the most common side effects of radiotherapy and the primary cause of death following exposure to irradiation.Despite concerted efforts,there is no definitive treatment method available.Recent studies have reported using mesenchymal stromal cells(MSCs),but their therapeutic effects are contested.AIM We administered and examined the effects of various amounts of adipose-derived MSCs(ADSCs)in mice with radiation-induced BM suppression.METHODS Mice were divided into three groups:Normal control group,irradiated(RT)group,and stem cell-treated group following whole-body irradiation(WBI).Mouse ADSCs(mADSCs)were transplanted into the peritoneal cavity either once or three times at 5×10^(5) cells/200μL.The white blood cell count and the levels of,plasma cytokines,BM mRNA,and BM surface markers were compared between the three groups.Human BM-derived CD34+hematopoietic progenitor cells were co-cultured with human ADSCs(hADSCs)or incubated in the presence of hADSCs conditioned media to investigate the effect on human cells in vitro.RESULTS The survival rate of mice that received one transplant of mADSCs was higher than that of mice that received three transplants.Multiple transplantations of ADSCs delayed the repopulation of BM hematopoietic stem cells.Anti-inflammatory effects and M2 polarization by intraperitoneal ADSCs might suppress erythropoiesis and induce myelopoiesis in sub-lethally RT mice.CONCLUSION The results suggested that an optimal amount of MSCs could improve survival rates post-WBI.

Synergistic effects of brain-derived neurotrophic factor and retinoic acid on inducing the differentiation of bone marrow stromal cells into neuron-like cells in adult rats in vitro

BACKGROUND： Under induction of retinoic acid （RA）, bone marrow stromal cells （BMSCs） can differentiate into nerve cells or neuron-like cells, which do not survive for a long time, so those are restricted to an application. Other neurotrophic factors can also differentiate into neuronal cells through inducing BMSCs; especially, brain-derived neurotrophic factor （BDNF） can delay natural death of neurons and play a key role in survival and growth of neurons. The combination of them is beneficial for differentiation of BMSCs. OBJECTIVE： To investigate the effects of BDNF combining with RA on inducing differentiation of BMSCs to nerve cells of adult rats and compare the results between common medium group and single BDNF group. DESIGN： Randomized controlled animal study SETTING： Department of Neurology, Affiliated Hospital of Xuzhou Medical College MATERIALS： The experiment was carried out in the Clinical Neurological Laboratory of Xuzhou Medical College from September 2003 to April 2005. A total of 24 SD rats, of either gender, 2 months old, weighing 130-150 g, were provided by Experimental Animal Center of Xuzhou Medical College [certification： SYXK （su） 2002-0038]. Materials and reagents： low-glucose DMEM medium, bovine serum, BDNF, RA, trypsin, separating medium of lymphocyte, monoclonal antibody of mouse-anti-nestin, neuro-specific enolase, glial fibrillary acidic protein （GFAP） antibody, SABC kit, and diaminobenzidine （DAB） color agent. All these mentioned above were mainly provided by SIGMA Company, GIBCO Company and Boshide Company. METHODS： Bone marrow of SD rats was selected for density gradient centrifugation. BMSCs were undertaken primary culture and subculture; and then, those cells were induced respectively in various mediums in total of 3 groups, including control group （primary culture）, BDNF group （20 μg/L BDNF） and BDNF＋RA group （20 μg/L BDNF plus 20 μg/L RA）. On the 3^rd and the 7^th days after induction, BMSCs were stained immunocytochemically with nestin （sign of nerve stem cells）, neuron-specific enolase （NSE, sign of diagnosing neurons） and GFAP （diagnosing astrocyte）, and evaluated cellular property. MAIN OUTCOME MEASURES ： Induction and differentiation in vitro of BMSCs in 3 groups RESULTS： （1） Induction and differentiation of BMSCs： Seven days after induction, cells having 2 or more apophyses were observed. Soma shaped like angle or erose form, which were similar to neurons and glial cells having strong refraction. （2） Results of immunocytochemical detection： Three days after induction, rate of positive cells in BDNF＋RA group was higher than that in BDNF group and control group [（86.15±4.58）%, （65.43±4.23）%, （4.18±1.09）%, P 〈 0.01]. Seven days after induction, rate of positive cells was lower in BDNF group and BDNF＋RA group than that in both groups at 3 days after induction [（31.12±3.18）%, （29.35±2.69）%, P 〈 0.01]; however, amounts of positive cells of NSE and GFAP were higher than those at 3 days after induction （P 〈 0.01）; meanwhile, the amount in BDNF＋RA group was remarkably higher than that in BDNF group （P 〈 0.01）. CONCLUSION： Combination of BDNF and RA can cooperate differentiation of BMSCs into neurons and astrocyte, and the effect is superior to single usage of BDNF.

Effects of granulocyte colony-stimulating factor and stem cell factor, alone and in combination, on the biological behaviours of bone marrow mesenchymal stem cells

Aim: The effects of granulocyte colony- stimu-lating factor (G-CSF) and stem cell factor (SCF) on the proliferation and osteogenic differentia-tion capacity of bone marrow mesenchymal stem cells (MSCs) were studied in the experi-ment. Methods: Bone marrow MSCs were col-lected from rabbits successfully, and treated with various concentrations of G-CSF, SCF or a combination of the two. Flow cytometric ana-lyse, MTT test, CFU-F assay, and alkaline phosphatase (ALP) activity measurement were employed. Results: The results of flow cytome-try showed that immunophenotype of the cells were CD29+/CD45-, CD105+/ CD34–, CD90+/ HLADR–. MSCs were shown to constitutively express low levels of c-kit which could be en-hanced by SCF. G-CSF and SCF had an obvious facilitative effect on the proliferation of MSCs in a dose-dependent fashion. In addition, G-CSF and SCF would be effective in reversibly pre-venting their differentiation, as showed by the decrease of ALP activity, leading to self-renewal rather than differentiative cell divisions. The effects of G-CSF were superior to SCF. And cells in the group treated with combination of G-CSF and SCF showed more powerful effects than the groups treated with G-CS, SCF, or none of the two. Conclusion: On the whole, these studies demonstrated that MSCs responsed to G-CSF, SCF, and to G-CSF plus SCF in a manner that suppressed differentiation, and promotes proliferation and self-renewal, and support the view that these factors could act synergistically.

The Effect of Sheng Bai Solution on Irradiated Mice Bone Marrow Cell Division Index and DNA Content

Before and after general irradiation with 60Co-γ, mice were orally given Sheng Bai Solution (SBS) for one week. SBS alleviated the irradiation-induced reduction of bone marrow cell chromosome division index. The irradiation-induced decrease of marrow DNA amount, thymic and splenic fractions, and total leukocyte number were restored to some extent. SBS also helped to ameliorate general condition of patients.

Sustained mutagenic effect in bone marrow cells induced by signal nuclide ^(134)Cs retention in skeleton

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Effect of Ligustrazine on Expressions of Basic Fibroblast Growth Factor and Its Receptor in Bone Marrow of Mice with Acute Radiation Injury

Objective: To study the expressions of basic fibroblast growth factor (bFGF) and its receptor (bFGFR) in bone marrow of mice with acute radiation injury, and to evaluate the effect of Ligustrazine (Lt) on them. Methods: Fifty-six Kunming mice of clean grade were randomly divided into 3 groups, the normal group, the control group and the Lt group. Mice in the latter two groups were once homogeneously systemic irradiated with 6.0 Gy of 60 Co, with the absorption dose rate of 0. 56 Gy/min, then treated with saline (0.2 ml/ mice) or Lt (2 mg/mice) respectively, twice a day through gastrogavage for successive 13 days. Mice were sacrificed in batch on the 3rd, 7th and 14th day by cervical dislocation to collect the bilateral femoral bone marrow for preparing bone marrow mono-nuclear cell (BMMNC) suspension. The bFGFR expression on surface of BMMNC was determined by flow cytometry; and the bFGF expres-sion level in one side of femoral bone marrow tissue was detected by immunohistochemistry with SABC-AP assay. Results: The bFGF expression in bone marrow of mice on the 3rd, 7th and 14th day after acute radiation injury all were significantly lower than that of the normal mice (P<0.05 or P<0.01). The expressions of bFGF and bFGFR in the Lt group detected were significantly higher than that in the control group detected at the corresponding time points (P<0.05 or P < 0.01). Conclusion:By way of enhancing bFGF expression in bone marrow and bFGFR expression on surface of BMMNC to accelerate the repairing of hemopoietic micro-environment in bone marrow might be one of the mechanisms of Lt in promoting hemopoietic function reconstitution after acute radiation injury.Original article on CJITWM (Chin) 2004;24(5):439

The effect of neuropeptides on proliferation of rat bone marrow mesenchymal stem cells

Objective To investigate the effects and mechanism of calcitonin gene-related peptide(CGRP)and substance P (SP) on proliferation of rat bone marrow mesenchymal stem cells.Methods The rBMSCs were isolated using whole bone marrow

Updates in the pathophysiological mechanisms of Parkinson's disease: Emerging role of bone marrow mesenchymal stem cells

AIM: To explore the approaches exerted by mesenchymal stem cells (MSCs) to improve Parkinson&#x02019;s disease (PD) pathophysiology.METHODS: MSCs were harvested from bone marrow of femoral bones of male rats, grown and propagated in culture. Twenty four ovariectomized animals were classified into 3 groups: Group (1) was control, Groups (2) and (3) were subcutaneously administered with rotenone for 14 d after one month of ovariectomy for induction of PD. Then, Group (2) was left untreated, while Group (3) was treated with single intravenous dose of bone marrow derived MSCs (BM-MSCs). SRY gene was assessed by PCR in brain tissue of the female rats. Serum transforming growth factor beta-1 (TGF-&#x003b2;1), monocyte chemoattractant protein-1 (MCP-1) and brain derived neurotrophic factor (BDNF) levels were assayed by ELISA. Brain dopamine DA level was assayed fluorometrically, while brain tyrosine hydroxylase (TH) and nestin gene expression were detected by semi-quantitative real time PCR. Brain survivin expression was determined by immunohistochemical procedure. Histopathological investigation of brain tissues was also done.RESULTS: BM-MSCs were able to home at the injured brains and elicited significant decrease in serum TGF-&#x003b2;1 (489.7 &#x000b1; 13.0 vs 691.2 &#x000b1; 8.0, P &#x0003c; 0.05) and MCP-1 (89.6 &#x000b1; 2.0 vs 112.1 &#x000b1; 1.9, P &#x0003c; 0.05) levels associated with significant increase in serum BDNF (3663 &#x000b1; 17.8 vs 2905 &#x000b1; 72.9, P &#x0003c; 0.05) and brain DA (874 &#x000b1; 15.0 vs 599 &#x000b1; 9.8, P &#x0003c; 0.05) levels as well as brain TH (1.18 &#x000b1; 0.004 vs 0.54 &#x000b1; 0.009, P &#x0003c; 0.05) and nestin (1.29 &#x000b1; 0.005 vs 0.67 &#x000b1; 0.006, P &#x0003c; 0.05) genes expression levels. In addition to, producing insignificant increase in the number of positive cells for survivin (293.2 &#x000b1; 15.9 vs 271.5 &#x000b1; 15.9, P &#x0003e; 0.05) expression. Finally, the brain sections showed intact histological structure of the striatum as a result of treatment with BM-MSCs.CONCLUSION: The current study sheds light on the therapeutic potential of BM-MSCs against PD pathophysiology via multi-mechanistic actions.

Local low-dose X-ray radiation promotes homing of mesenchymal stem cells to the injured mouse spinal cord

BACKGROUND： Bone marrow-derived mesenchymal stem cells （BMSCs） are a potentially useful source for cell replacement therapy following spinal cord injury. However, the homing characteristics of BMSCs in vivo remain unclear. Low-dose radiation has been shown to promote homing of BMSCs to exposed sites. OBJECTIVE： To investigate the effects of low-dose local radiation to non-injured areas on the ability of human BMSCs to home to the injured mouse spinal cord, as well as recovery of spinal cord injury. DESIGN, TIME AND SE＇I-FING： A randomized, controlled, animal experiment was performed at the Central Laboratory, Second Affiliated Hospital of Soochow University between October 2007 and October 2008. MATERIALS： BMSCs were isolated from four adult, human donors. METHODS： Fifty adult, female, Balb/c mice were subjected to adjusted weight-drop impact resulting in complete paraplegia. Three days later, mice were randomly assigned to a radiation ＋ transplantation group （n = 23） and a transplantation group （n = 20）. In total, 2 x 106 carboxyfluorescein diacetate succinimidyl ester-labeled BMSCs were injected into each mouse via the caudal vein. Mice in the radiation ＋ transplantation group received 2.5 Gy local X-ray irradiation 2 hours before BMSCs injection. MAIN OUTCOME MEASURES： The homing of BMSCs to injured cord and irradiated skin after transplantation was observed by fluorescence microscope; the structure recovery of injured cord was assessed by magnetic resonance imaging. RESULTS： Compared with the transplantation group, at 24 hours after transplantation, the number of BMSCs was significantly increased in the injured area and the exposed site （P 〈 0.05）, and inflammation and edema were significantly alleviated in the injured cord in the radiation ＋ transplantation group. CONCLUSION： Local low-dose radiation has the potential to promote homing of BMSCs and recovery of spinal cord injury, although the radiated region was not injured area.

Radiogenotoxicological effect of signal nuclide ^(134)Cs on somatic and germ cells

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p53-dependent and independent apoptosis induced by ionizing radiation in murine bone marrow cells

APOPTOSIS or programmed cell death is a model of cell death, along with typical morphological and biochemical changes. The cell death, occurring during embryonic development, T and B cell maturation and endocrine-induced atrophy, is a physiological apoptosis which controls the amount of cells in the body. Apoptosis can also be induced by various factors, such as ionizing radiation and anti-tumour agents which damage DNA, and has been proposed to be a consequence of the induced expression of some tumour-related genes or increase of the proteins sta-

Laser Device for the Protection of Biological Objects from the Damaging Action of Ionizing Radiation

Laser irradiation device for the protection of biological objects from the action of ionizing radiation to be used in practice has been manufactured （invention patent RU 2 428 228 C2）. Research of the action of y-radiation itself as well as of the combined action of laser devices on survival, weight, skin and the general mitotic index of the bone marrow cells （mitotic index of all nucleus-containing cells of the bone marrow） of C57BL/6 experimental young mice was carried out. The mice were irradiated with ionizing （whole body irradiation） and laser radiation, separately one by one in a special frame device. Laser radiation in the dose 1 mJ/cm^2 irradiated only the back of a mouse, or both the back and the abdomen of mice. In case of combined irradiation of mice, the time interval between two types of irradiation did not exceed 30 min. First, the mice were exposed to y-radiation then to laser radiation. The method of the laser radiation-protection of biological subjects contributes to an increase in the viability of mice, prevents the damages of skin and also increases the mitotic activity of mice bone marrow cells.

Protective Effect of Renshen Yangrong Decoction(人参养荣汤) on Bone Marrow against Radiation Injury in Mouse

Objective：To explore the effect of Renshen Yangrong Decoction（人参养荣汤,RYD） in protecting bone marrow from radiation injury.Methods：One hundred and eighty Kuming mice were subjected to the three tests for anti-radiation injury effect evaluation,i.e.the test of peripheral white blood cell（WBC） count, the test of bone marrow nucleated cell count,and the bone marrow micronucleus test,using 60 mice for each test.The mice in each test were divided into 6 groups：the blank control group,the model control group,the positive control group treated by Shiyiwei Shenqi Tablet（十一味参芪片,1.0 g/kg）,and three RYD groups treated with high（42.0 g/kg）,moderate（21.0 g/kg）,and low（10.5 g/kg） doses of crude drugs of RYD,with 10 mice in each group.The treatment was given by gastrogavage perfusion continuously for 7-14 days before mice received ^（60）Co-γray radiation and continued until the end of the experiment.The body weights of the mice were monitored,the changes in peripheral WBC and bone marrow nucleated cells were counted,and the variation in bone marrow micronucleated cells was observed on the respective appointed days.Results：A significant decrease in body weight,peripheral WBC count,and bone marrow nucleated cell count,as well as marked changes in bone marrow micronucleated cells were observed in the mice after radiation,indicating that the radiation injury model was successfully established.As compared with the model control group,the decrease in body weight,peripheral WBC count,and bone marrow nucleated cell count,as well as the increase in bone marrow micronucleus cell count in the high dosage RYD treated group were obviously inhibited or lessened （P0.05 or P0.01）.Conclusion：RYD showed obvious protective effect in mice with bone marrow injury induced by radiation.

Effect of Ligustrazine on CD_(34) Antigen Expression of Bone MarrowCells in Immune-Induced Aplastic Anemia Mice

Objective:To explore the effect of ligustrazine on CD34 antigen expression of bone marrow cells in immune-induced aplastic anemia（AA）mice.Methods:The model of immune aplastic anemia mice was in duced by means of 6.0GY60γ-rayirr adi at ion and lym phocyte infusion through tail vein.Immuneinduce dAA micewere divided into 3 groups:thenormal group,the AA control group and the ligustrazine group.Miceof the ligustrazine group were fed by 4 mg of ligustrazine in jecti on twicea day bygastro gavage.On the 10 thday,CD34 an tigen expression intensity of bone marrow cell membrane was measured by flowcy tometer an alysis system.Results:CD34 antigen expressionin tensity of ligustrazine group was 77.6±6.5,with no statistic difference from that in normal group（80.0±2.6),while that of the control group was muchhigher（68.6±4.5,P＜0.05).Conclusion:Ligustrazine could promote proliferation of stem and progenitor cell of AA mice through in fluencing on bone marrowmicro-environment so as to increase the CD34antigen expression of bone marrow cells.

Effect of moxibustion at “Guanyuan”( CV4) and “Sanyinjiao”( SP 6) on bone morphology,metabolism and ERα of bone marrow mesenchymal stem cells in the ovariectomized rats

Objective To observe the effects of moxibustion at'Guanyuan'(CV 4) and'Sanyinjiao'(SP 6) on bone morphology,metabolism and ERαof bone marrow mesenchymal stem cells (MSCs) in the ovariectomized rats and explore the underlying mechanism of moxibustion at Guanyuan (CV 4) and Sanyinjiao (SP 6) on the regulationof bone metabolism. Methods A total of 60 SD ratswere randomized into a normal group (20 rats) and anovariectomy group (40 rats).

Effect of intracoronary autologous bone marrow mononuclear cells transplantation on arrhythmia in canines

Objective To observe the survival and the differentiation of grafted bone marrow cells(BM-MNCs)in host myocardium.To observe whether BM-MNCs transplantation can potentially cause arrhythmia and whether the BM-MNCs transplantation can alter the spatial distribution of connexins,important mediator for arrhythmia gen-

Injectable bone marrow microniches by co-culture of HSPCs with MSCs in 3D microscaffolds promote hematopoietic reconstitution from acute lethal radiation

Hematopoietic syndrome of acute radiation syndrome(h-ARS)is an acute illness resulted from the damage of bone marrow(BM)microenvironment after exposure to radiation.Currently,the clinical management of h-ARS is limited to medication-assisted treatment,while there is still no specific therapy for the hematopoietic injury from high-dose radiation exposure.Our study aimed to assemble biomimetic three-dimensional(3D)BM microniches by co-culture of hematopoietic stem and progenitor cells(HSPCs)and mesenchymal stem cells(MSCs)in porous,injectable and viscoelastic microscaffolds in vitro.The biodegradable BM microniches were then transplanted in vivo into the BM cavity for the treatment of h-ARS.We demonstrated that the maintenance of HSPCs was prolonged by co-culture with MSCs in the porous 3D microscaffolds with 84μm in pore diameter and 11.2 kPa in Young’s modulus compared with 2D co-culture system.Besides,the minimal effective dose and therapeutic effects of the BM microniches were investigated on a murine model of h-ARS,which showed that the intramedullary cavity-injected BM microniches could adequately promote hematopoietic reconstitution and mitigate death from acute lethal radiation with a dose as low as 1000 HSPCs.Furthermore,the mRNA expression of Notch1 and its downstream target gene Hes1 of HSPCs were increased when co-cultured with MSCs,while the Jagged1 expression of the co-cultured MSCs was upregulated,indicating the significance of Notch signaling pathway in maintenance of HSPCs.Collectively,our findings provide evidence that biomimetic and injectable 3D BM microniches could maintain HSPCs,promote hematopoiesis regeneration and alleviate post-radiation injury,which provides a promising approach to renovate conventional HSPCs transplantation for clinical treatment of blood and immune disorders.

Adipocyte Derived Mesenchymal Stromal Cell and Platelet Lysate: Ideal Cell and Supplement for the Treatment of Immune-Inflammatory Diseases?

Background: Mesenchymal stromal cells (MSC) are being tested for the treatment of immune diseases. MSC are present in several adult tissues which milieu may influence MSC behavior particularly under inflammatory conditions. Additionally, culture conditions also can modify cell function or state of activation. Methods: To address the influence of the MSC source on its characteristics, we studied a xenofree, platelet lysate supplemented MSC from dental pulp, adipose tissue and bone marrow, co-cultured with isolated T cells and PBMC subset, and studied the effect of culture animal or human supplements immunomodulatory effect. Results: All three sources were efficient in inhibiting T cells. Among all MSC sources, as also described by others, adipose MSC was capable to significantly induce Treg phenotype and decrease T CD8+. Furthermore, comparing fetal bovine serum and platelet lysate, results demonstrate that platelet lysate alone is capable to induce immunomodulatory phenotype. Additional studies have to be made to elucidate the PL immunomodulatory effect.

全反式视黄酸调控颌骨骨髓间充质干细胞成骨分化双向效应的体外研究

目的·探究不同浓度全反式视黄酸(all-trans retinoic acid,ATRA)对大鼠颌骨骨髓间充质干细胞(jaw bone marrow mesenchymal stem cells,jBMSCs)成骨分化的影响。方法·通过全骨髓贴壁法分离培养4周龄Sprague-Dawley(SD)大鼠jBMSCs。运用流式细胞术鉴定jBMSCs表面抗原。利用碱性磷酸酶(alkaline phosphatase,ALP)染色/茜素红染色、油红O染色及阿尔辛蓝染色分别对成骨诱导、成脂诱导及成软骨诱导后的jBMSCs进行多向分化潜能检测。分别使用ATRA浓度为0.01、0.1、1、5、10、20μmol/L的成骨诱导液对jBMSCs进行体外成骨诱导,并使用二甲基亚砜(dimethyl sulfoxide,DMSO)作为对照组,利用CCK8进行细胞活力检测。采用ALP染色和茜素红染色对各浓度组jBMSCs的成骨分化能力进行检测,并筛选后续实验浓度。利用实时荧光定量聚合酶链反应(quantitative real-time polymerase chain reaction,qPCR)、免疫荧光染色分析不同浓度ATRA下jBMSCs成骨相关基因mRNA与蛋白表达水平。结果·流式细胞术分析显示,98%以上的P1代jBMSCs表现为CD29+CD90+CD31−CD45−,与骨髓间充质干细胞表面抗原特点相符。ALP染色/茜素红染色、油红O染色及阿尔辛蓝染色结果证明P1代jBMSCs具有成骨、成脂、成软骨多向分化能力。ALP染色/茜素红染色结果显示,0.01、0.1和1μmol/L ATRA组jBMSCs成骨活性和矿化能力较对照组增强,而继续提升ATRA浓度则使成骨活性与矿化能力减弱,浓度高于5μmol/L时开始低于对照组水平(均P<0.05)。qPCR分析发现,0.1、1μmol/L ATRA组成骨相关基因如Alp、骨唾液酸蛋白(bone sialoprotein,Bsp)、Ⅰ型胶原蛋白α1(collagen typeⅠα1,Col1a1)、骨钙素(osteocalcin,Ocn)表达水平较对照组上升,而继续提升ATRA浓度则使基因表达水平下降,ATRA浓度高于5μmol/L时开始低于对照组水平(均P<0.05)。免疫荧光染色结果显示,0.1、1μmol/L ATRA组较对照组成骨相关蛋白成骨细胞特异性转录因子SP7、ALP及OCN表达增强,而继续提升ATRA浓度则使蛋白表达下降,浓度高于5μmol/L时开始低于对照组水平(均P<0.05)。结论·较低浓度(0.1、1μmol/L)ATRA可促进大鼠jBMSCs成骨分化能力,且该促进效应在0.1μmol/L浓度时达到峰值,进一步提升浓度可使该促进效应减弱。较高浓度(5、10、20μmol/L)ATRA对大鼠jBMSCs成骨分化呈现抑制效应。研究在体外证明ATRA对大鼠jBMSCs成骨分化具双向效应,并鉴定了0.1μmol/L ATRA为大鼠jBMSCs成骨分化的最适浓度,为体内研究的开展与全反式视黄酸的临床应用提供了参考依据。