BACKGROUNDDendritic cells are professional antigen-presenting cells found in an immature state in epithelia and interstitial space, where they capture antigens such as pathogens or damaged tissue. Matrix metallopeptidase 13 (MMP-13), a member of the collagenase subfamily, is involved in many different cellular processes and is expressed in murine bone marrow-derived dendritic cells (DCs). The function of MMP-13 in DCs is not well understood. Here, we investigated the effect of MMP-13 on DC maturation, apoptosis, and phagocytosis.

METHODSBone marrow-derived dendritic cells were obtained from C57BL/6 mice. One short-interfering RNA specific for MMP-13 was used to transfect DCs. MMP-13-silenced DCs and control DCs were prepared, and apoptosis was measured using real-time polymerase chain reaction and Western blotting. MMP-13-silenced DCs and control DCs were analyzed for surface expression of CD80 and CD86 and phagocytosis capability using flow cytometry.

RESULTSCompared to the control DCs, MMP-13-silenced DCs increased expression of anti-apoptosis-related genes, BAG1 (control group vs. MMP-13-silenced group: 4.08 ± 0.60 vs. 6.11 ± 0.87, P = 0.008), BCL-2 (control group vs. MMP-13-silenced group: 7.54 ± 0.76 vs. 9.54 ± 1.29, P = 0.036), and TP73 (control group vs. MMP-13-silenced group: 4.33 ± 0.29 vs. 5.60 ± 0.32, P = 0.001) and decreased apoptosis-related genes, CASP1 (control group vs. MMP-13-silenced group: 3.79 ± 0.67 vs. 2.54 ± 0.39, P = 0.019), LTBR (control group vs. MMP-13-silenced group: 9.23 ± 1.25 vs. 6.24 ± 1.15, P = 0.012), and CASP4 (control group vs. MMP-13-silenced group: 2.07 ± 0.56 vs. 0.35 ± 0.35, P = 0.002). Protein levels confirmed the same expression pattern. MMP-13-silenced groups decreased expression of CD86 on DCs; however, there was no statistical difference in CD80 surface expression. Furthermore, MMP-13-silenced groups exhibited weaker phagocytosis capability.

CONCLUSIONThese results indicate that MMP-13 inhibition dampens DC maturation, apoptosis, and phagocytosis.

Animals ; Apoptosis ; drug effects ; physiology ; Bone Marrow Cells ; cytology ; Dendritic Cells ; cytology ; drug effects ; metabolism ; Female ; Lipopolysaccharides ; pharmacology ; Matrix Metalloproteinase 13 ; metabolism ; physiology ; Mice ; Mice, Inbred C57BL ; RNA, Small Interfering

BACKGROUNDThe functional improvement following bone marrow stromal cells (BMSCs) transplantation after stroke is directly related to the number of engrafted cells and neurogenesis in the injured brain. Here, we tried to evaluate whether 3-methyl-1-phenyl-2-pyrazolin-5-one (MCI-186), a free radical scavenger, might influence BMSCs migration to ischemic brain, which could promote neurogenesis and thereby enhance treatment effects after stroke.

METHODSRat transient middle cerebral artery occlusion (MCAO) model was established. Two separate MCAO groups were administered with either MCI-186 or phosphate-buffered saline (PBS) solution to evaluate the expression of stromal cell-derived factor-1 (SDF-1) in ischemic brain, and compared to that in sham group (n = 5/ group/time point[at 1, 3, and 7 days after operation]). The content of chemokine receptor-4 (CXCR4, a main receptor of SDF-1) at 7 days after operation was also observed on cultured BMSCs. Another four MCAO groups were intravenously administered with either PBS, MCI-186, BMSCs (2 × 106), or a combination of MCI-186 and BMSCs (n = 10/group). 5-bromo-2-deoxyuridine (BrdU) and Nestin double-immunofluorescence staining was performed to identify the engrafted BMSCs and neuronal differentiation. Adhesive-removal test and foot-fault evaluation were used to test the neurological outcome.

RESULTSMCI-186 upregulated the expression of SDF-1 in ischemic brain and CXCR4 content in BMSCs was enhanced after hypoxic stimulation. When MCAO rats were treated with either MCI-186, BMSCs, or a combination of MCI-186 and BMSCs, the neurologic function was obviously recovered as compared to PBS control group (P < 0.01 or 0.05, respectively). Combination therapy represented a further restoration, increased the number of BMSCs and Nestin+ cells in ischemic brain as compared with BMSCs monotherapy (P < 0.01). The number of engrafted-BMSCs was correlated with the density of neuronal cells in ischemic brain (r = 0.72 , P < 0.01) and the improvement of foot-fault (r = 0.70, P < 0.01).

CONCLUSIONMCI-186 might promote BMSCs migration to the ischemic brain, amplify the neurogenesis, and improve the effects of cell therapy.

Animals ; Antipyrine ; analogs & derivatives ; therapeutic use ; Bone Marrow Cells ; cytology ; physiology ; Brain Ischemia ; drug therapy ; metabolism ; therapy ; Chemokine CXCL12 ; metabolism ; Disease Models, Animal ; Infarction, Middle Cerebral Artery ; drug therapy ; metabolism ; therapy ; Male ; Mesenchymal Stromal Cells ; physiology ; Neurogenesis ; physiology ; Rats ; Rats, Sprague-Dawley ; Stroke ; drug therapy ; metabolism ; therapy

OBJECTIVETo explore the effects of bone mesenchymal stem cells (BMSCs) in restoring the functions of degenerative nucleus pulposus cells (dNPCs).

METHODSThe animal models of degenerative nucleus pulposus were established by means of acupuncture and aspiration. The BMSCs as well as the normal and degeneratived nucleus pulposus cells of SD rats were isolated and cultured. The BMSCs/alginate gel complex and dNPCs/alginate gel complex were used for indirect co-culture in vitro, which was set as experiment group. The NPCs and dNPCs cultured alone as positive and negative controls. The cell growth conditions were observed by light microscopy, and suitable cells were selected to combine alginate gel stents and cultured in transwell plate. Seven days later, nucleus pulposus cells of each group were recycled, and the mRNA expressions of Collagen2, SOX 9, and Aggrecan were detected by RT-PCR, and the Collagen1, 2, and Aggrecan were detected by Western blotting and immunofluorescence.

RESULTSAfter non-contact co-culture for 7 days, the mRNA levels of Collagen2, SOX 9, and Aggrecan increased apparently in BMSCs+dNPCs group, while it was significantly lower in dNPCs sample (all P<0.05). The content of Collagen2 and Aggrecan detected by Western blotting in BMSCs+dNPCs group got close to NPCs sample, but it was significantly higher than dNPCs samples (all P<0.05), while the content of Collagen1in BMSCs+dNPCs group got close to NPCs samples, but it was significantly lower than dNPCs sample (P<0.05). Immunofluorescence results showed that cytoplasm was dyed red and the color near the caryon became dark in BMSCs+dNPCs group and dNPCs group;however, the colored scope of cytoplasm and the dark colored scope surrounding the caryon in BMSCs+dNPCs group was obviously larger than dNPCs group.

CONCLUSIONUnder a 3D non-contact co-culture system, BMSCs can promote the expression of epimatrix of the dNPCs, which shows that BMSCs can restore the functions of dNPCs of intervertebral disc to certain extent.

Animals ; Bone Marrow Cells ; physiology ; Cells, Cultured ; Coculture Techniques ; Intervertebral Disc ; cytology ; pathology ; Mesenchymal Stromal Cells ; physiology ; Rats ; Rats, Sprague-Dawley

BACKGROUNDGallbladder carcinoma (GBC) has a high mortality rate, requiring synergistic anti-tumor management for effective treatment. The myxoma virus (MYXV) exhibits a modest clinical value through its oncolytic potential and narrow host tropism.

METHODSWe performed viral replication assays, cell viability assays, migration assays, and xenograft tumor models to demonstrate that bone marrow-derived stem cells (BMSCs) may enhance efficiency of intravenous MYXV delivery.

RESULTSWe examined the permissiveness of various GBC cell lines towards MYXV infection and found two supported single and multiple rounds of MYXV replication, leading to an oncolytic effect. Furthermore, we found that BMSCs exhibited tropism for GBC cells within a Matrigel migration system. BMSCs failed to affect the growth of GBC cells, in terms of tumor volume and survival time. Finally, we demonstrated in vivo that intravenous injection of MYXV-infected BMSCs significantly improves the oncolytic effect of MYXV alone, almost to the same extent as intratumoral injection of MYXV.

CONCLUSIONThis study indicates that BMSCs are a promising novel vehicle for MYXV to clinically address gallbladder tumors.

Animals ; Bone Marrow Cells ; cytology ; Cell Movement ; physiology ; Cell Survival ; physiology ; Female ; Gallbladder Neoplasms ; therapy ; virology ; Humans ; Immunohistochemistry ; Mice ; Myxoma virus ; pathogenicity ; Stem Cells ; cytology ; physiology ; Virus Replication ; physiology ; Xenograft Model Antitumor Assays

BACKGROUNDIn order to suggest an ideal source of adult stem cells for the treatment of nervous system diseases, MSCs from human adipose tissue and bone marrow were isolated and studied to explore the differences with regard to cell morphology, surface markers, neuronal differentiation capacity, especially the synapse structure formation and the secretion of neurotrophic factors.

METHODSThe neuronal differentiation capacity of human mesenchymal stem cells from adipose tissue (hADSCs) and bone marrow (hBMSCs) was determined based on nissl body and synapse structure formation, and neural factor secretion function. hADSCs and hBMSCs were isolated and differentiated into neuron-like cells with rat brain-conditioned medium, a potentially rich source of neuronal differentiation promoting signals. Specific neuronal proteins and neural factors were detected by immunohistochemistry and enzyme-linked immunosorbent assay analysis, respectively.

RESULTSFlow cytometric analysis showed that both cell types had similar phenotypes. Cell growth curves showed that hADSCs proliferated more quickly than hBMSCs. Both kinds of cells were capable of osteogenic and adipogenic differentiation. The morphology of hADSCs and hBMSCs changed during neuronal differentiation and displayed neuron-like cell appearance after 14 days' differentiation. Both hADSCs and hBMSCs were able to differentiate into neuron-like cells based on their production of neuron specific proteins including β-tubulin-III, neuron-specific enolase (NSE), nissl bodies, and their ability to secrete brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF). Assessment of synaptop hysin and growth-associated protein-43 (GAP-43) suggested synapse structure formation in differentiated hADSCs and hBMSCs.

CONCLUSIONSOur results demonstrate that hADSCs have neuronal differentiation potential similar to hBMSC, but with a higher proliferation capacity than hBMSC. Adipose tissue is abundant, easily available and would be a potential ideal source of adult stem cells for neural-related clinical research and application.

Adipose Tissue ; cytology ; Adult ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; cytology ; Middle Aged ; Nerve Regeneration ; physiology ; Neurons ; cytology ; Young Adult

Bone marrow mensenchymal stem cells (BM-MSCs) are capable of supporting the survival, differentiation and migration of hematopoietic stem cell, and have a profound application prospect in transplantation and treatment of graft-versus-host disease (GVHD). This review aims to illustrate the interaction between BM-MSCs and other intra-bone marrow cells, including hematopoietic stem cells, endothelial cells and osteoblasts. The investigation of their regulating mechanism will help better understanding of the BM-MSCs' role in hematopoiesis.
Bone Marrow Cells ; cytology ; Cell Communication ; physiology ; Endothelial Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology

BACKGROUNDBone marrow derived mesenchymal stem cells (BMdMSCs) can differentiate into cardiomyocyte-like cells induced by different inductors individually or collectively. In this study, by inducing BMdMSCs with p53 inhibitor (p-fifty three inhibitor-alpha, PFT-α), 5-azacytidine (5-AZA), angiotensin-II (Ang-II) and bone morphogenic protein-2 (BMP-2) we compared the influences of four inductors on the differentiation of rat BMdMSCs into caridomyocyte like-cells.

METHODSBMdMSCs were collected from the bone marrow of Sprague Dawley rats and after the fourth generation, the purified cells were divided into five groups: 5-AZA (10 µmol/L), Ang-II (0.1 µmol/L), PFT-α (20 µmol/L), BMP-2 (10 µg/L) and control. The purity of the BMdMSCs and the cardiac differentiation rates were obtained by flow cytometry. The expressions of cTnT in the BMdMSCs after four weeks of induction were detected by immunofluorescence and the expressions of cTnI and Cx43 detected by Western blotting. The green fluorescent levels reflecting intracellular calcium transient function were determined by laser scanning confocal microscopy. The total potassium current levels of cells were measured on patch clamp.

RESULTSAll inductors affected to a different degree the differentiation of BMdMSCs into cardiomyocyte-like cells and the expressions of cTnT, cTnI and Cx43 suggesting that the combination of inductors could be an improved method for cardiac regenerative medicine. In addition, the total potassium current level and calcium transient in PFT-α cardiomyocyte-like cells were higher than other groups.

CONCLUSIONSThe cardiac differentiation of BMdMSCs induced by PFT-α, 5-AZA, Ang-II and BMP-2 has been improved at different levels. PFT-α has an advantage of differentiation rate and electrophysiological function over other inductors.

Animals ; Blotting, Western ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Electrophysiology ; methods ; Male ; Mesenchymal Stromal Cells ; cytology ; Myocytes, Cardiac ; cytology ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To investigate whether miR-125b regulates the osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs) by modulating Smad4, a predicted target in silicon. METHODS: Smad4 3'-UTR-luciferase vector was constructed and dual-luciferase reporter gene assay was employed to examine the effect of miR-125b on luciferase activity. MSCs were isolated and cultured from human bone marrow, and then transfected with miR-125b mimics followed by induction of osteogenic differentiation. qRT-PCR and Western blot were used to detect the expressions of Smad4 mRNA and protein. MSCs were induced into the osteoblasts after transfecting with Smad4 siRNA, and the effect of Smad4 downregulation on osteogenic differentiation was observed by AKP activity and RUNX2 mRNA levels. RESULTS: miR-216b bound Smad4 3'-UTR and inhibited the luciferase activity (P<0.05). Smad4 mRNA and protein expressions were significantly down-regulated in the MSCs induced into osteogenic differentiation when miR-125b was overexpressed. Downregulation of Smad4 suppressed the AKP activity and RUNX2 mRNA expression, indicating that Smad4 siRNA simulated at least in part the function of miR-125b as the regulator of MSCs osteogenic differentiation. CONCLUSION miR-125b can suppress MSCs osteogenic differentiation by directly targeting Smad4.
Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Down-Regulation ; Female ; Humans ; Mesenchymal Stem Cells ; cytology ; MicroRNAs ; physiology ; Osteoblasts ; cytology ; Osteogenesis ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Smad4 Protein ; genetics ; metabolism ; Transfection ; Young Adult

The fungus Trichophyton schoenleinii (T. schoenleinii) is the causative agent of Trichophytosis and Tinea favosa of the scalp in certain regions of Eurasia and Africa. Human innate immune system plays an important role in combating with various pathogens including fungi. The inflammasome is one of the most critical arms of host innate immunity, which is a protein complex controlling maturation of IL-1β. To clarify whether T. schoenleinii is able to activate the inflammasome, we analyzed human monocytic cell line THP-1 for IL-1β production upon infection with T. schoenleinii strain isolated from Tinea favosa patients, and rapid IL-1β secretion from THP-1 cells was observed. Moreover, applying competitive inhibitors and gene specific silencing with shRNA, we found that T. schoenleinii induced IL-1β secretion, ASC pyroptosome formation as well as caspase-1 activation were all dependent on NLRP3. Cathepsin B activity, ROS production and K⁺ efflux were required for the inflammasome activation by T. schoenleinii. Our data thus reveal that the NLRP3 inflammasome plays an important role in host defense against T. schoenleinii, and suggest that manipulating NLRP3 signaling can be a novel approach for control of diseases caused by T. schoenleinii infection.
Animals ; Bone Marrow Cells ; cytology ; Carrier Proteins ; metabolism ; Caspase 1 ; metabolism ; Cell Line ; Dendritic Cells ; cytology ; metabolism ; microbiology ; Enzyme Activation ; Hot Temperature ; Humans ; Inflammasomes ; metabolism ; Interleukin-1beta ; biosynthesis ; metabolism ; Lysosomes ; metabolism ; Mice ; Monocytes ; cytology ; metabolism ; microbiology ; NLR Family, Pyrin Domain-Containing 3 Protein ; Potassium ; metabolism ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; Trichophyton ; physiology

Bone marrow derived mesenchymal stem cell (BMSC) is one of the crucial cell types which plays roles in wound healing of tissues. In the last decades, it was believed that BMSCs promoted wound healing by differentiating into multiple lineages and placing the wounded tissues. In recent years, a new viewpoint arose from evidences that the paracrine effect of BMSCs might play a more important role in the process of wound healing than differentiation. Understanding the role of BMSCs paracrine in wound healing would be vital to clarify the mechanism how BMSCs take part into the process of wound healing. In this paper, we review the new research processes of BMSCs paracrine in wound healing of tissues.
Animals ; Bone Marrow Cells ; cytology ; physiology ; Cell Differentiation ; physiology ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology ; Paracrine Communication ; physiology ; Wound Healing ; physiology