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Abstract Objectives Osteoblasts are derived from Bone Marrow-derived Mesenchymal Stem Cells (BM-MSCs), which play an indispensable role in bone formation. In this study, the authors aim to investigate the role of IRF4 in the osteogenic differentiation of BM-MSCs and its potential molecular mechanism. Methods The authors used lentivirus infection to overexpress IRF4 in BM-MSCs. The expression of IRF4 and osteogenesis-related genes were detected by qRT-PCR and western blot analysis. The osteogenic differentiation of BM-MSCs was evaluated by Alkaline Phosphatase (ALP) activity, Alizarin red staining, and Alkaline Phosphatase (ALP) staining. Chromatin Immunoprecipitation (ChIP), Dual-Luciferase reporter assay and RNA Immunoprecipitation Assay were applied to confirm the regulatory mechanism between IRF4, miR-636 and DOCK9. Results The authors found IRF4 was down-regulated during the osteogenic differentiation of BM-MSCs, and IRF4 overexpression could decrease the osteogenic differentiation of BM-MSCs by specifically promoting the reduction of Alkaline Phosphatase (ALP) activity and down-regulating osteogenic indicators, including OCN, OPN, Runx2 and CollA1. Mechanistically, IRF4 activated microRNA-636 (miR-636) expression via binding to its promoter region, and Dedicator of Cytokinesis 9 (DOCK9) was identified as the target of miR-636 in BM-MSCs. Moreover, the damage in the capacity of osteogenic differentiation of BM-MSCs induced by IRF4 overexpression could be rescued by miR-636 inhibition. Conclusions In summary, this paper proposed that IRF4/miR-636/DOCK9 may be considered as targets for the treatment of osteoporosis (OP).
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Cancer stem cells (CSCs) are a subpopulation of cancer cells with functions similar to those of normal stem cells. Although few in number, they are capable of self-renewal, unlimited proliferation, and multi-directional differentiation potential. In addition, CSCs have the ability to escape immune surveillance. Thus, they play an important role in the occurrence and development of tumors, and they are closely related to tumor invasion, metastasis, drug resistance, and recurrence after treatment. Therefore, specific targeting of CSCs may improve the efficiency of cancer therapy. A series of corresponding promising therapeutic strategies based on CSC targeting, such as the targeting of CSC niche, CSC signaling pathways, and CSC mitochondria, are currently under development. Given the rapid progression in this field and nanotechnology, drug delivery systems (DDSs) for CSC targeting are increasingly being developed. In this review, we summarize the advances in CSC-targeted DDSs. Furthermore, we highlight the latest developmental trends through the main line of CSC occurrence and development process; some considerations about the rationale, advantages, and limitations of different DDSs for CSC-targeted therapies were discussed.
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Icariin, a prenylated flavonol glycoside isolated from Epimedium, has been considered as a potential alternative therapy for osteoporosis. The present study aimed to clarify the detailed molecular mechanisms of action of icariin on osteoblast function, using bone marrow-derived mesenchymal stem cells (BM-MSCs). BM-MSCs were first stimulated by icariin. Then, gene and protein expression of cAMP/PKA/CREB signaling molecules were analyzed by RT-PCR and western blotting (WB), and alkaline phosphatase (ALP) was analyzed in cell lysates by ELISA. MTT assays indicated that icariin did not have significant effects on cell viability up to 1 µM. Icariin showed a dose-dependent effect on the alkaline phosphatase activity of BM-MSCs. WB analysis showed that icariin treatment of BM-MSCs significantly enhanced the protein expression of protein kinase A (PKA) and cAMP-responsive element binding protein (CREB), while RT-PCR results showed that icariin dose-dependently increased the mRNA levels of PKA and CREB. Icariin induced BM-MSC differentiation by BMP2, Smad1, and Runx2. RT-PCR and WB results indicated that icariin significantly increased the expression of BMP2, Smad1, and Runx2 in BM-MSCs. These results suggest that icariin is an agonist of the cAMP/PKA/CREB pathway in BM-MSC differentiation, raising the possibility that it could be used in the treatment of osteoporosis.
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BACKGROUND: Mesenchymal stem cells (MSCs) and/or biological scaffolds have been used to regenerate articular cartilage with variable success. In the present study we evaluated cartilage regeneration using a combination of bone marrow (BM)-MSCs, Hyalofast™ and/or native cartilage tissue following full thickness surgical cartilage defect in rabbits. METHODS: Full-thickness surgical ablation of the medial-tibial cartilage was performed in New Zealand white (NZW) rabbits. Control rabbits (Group-I) received no treatment; Animals in other groups were treated as follows. Group-II: BMMSCs (1 × 10⁶ cells) + Hyalofast™; Group-III: BMMSCs (1 × 10⁶ cells) + cartilage pellet (CP); and Group-IV: BMMSCs (1 × 10⁶ cells) + Hyalofast™+ CP. Animals were sacrificed at 12 weeks and cartilage regeneration analyzed using histopathology, International Cartilage Repair Society (ICRS-II) score, magnetic resonance observation of cartilage repair tissue (MOCART) score and biomechanical studies. RESULTS: Gross images showed good tissue repair (Groups IV>III>Group II) and histology demonstrated intact superficial layer, normal chondrocyte arrangement, tidemark and cartilage matrix staining (Groups III and IV) compared to the untreated control (Group I) respectively. ICRS-II score was 52.5, 65.0, 66 and 75% (Groups I–IV) and the MOCART score was 50.0, 73.75 and 76.25 (Groups II–IV) respectively. Biomechanical properties of the regenerated cartilage tissue in Group IV closed resembled that of a normal cartilage. CONCLUSION: Hyalofast™ together with BM-MSCs and CP led to efficient cartilage regeneration following full thickness surgical ablation of tibial articular cartilage in vivo in rabbits. Presence of hyaluronic acid in the scaffold and native microenvironment cues probably facilitated differentiation and integration of BM-MSCs.
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Animals , Rabbits , Bone Marrow , Cartilage , Cartilage, Articular , Chondrocytes , Cues , Hyaluronic Acid , Mesenchymal Stem Cells , New Zealand , Osteoarthritis , RegenerationABSTRACT
Objective To compare the hepatic differentiation potential of human umbilical cord mesenchymal stem cells (UC-MSCs) with bone marrow mesenchymal stem cells (BM-MSCs).Methods UC-MSCs and BM-MSCs derived from passage 3 were induced by IMDM supplemented with 20 μg/L HGF and 20 mg/L α-FGF.The medium was changed twice a week.The concentrations of albumin and urea nitrogen from cultural medium were measured to compare the differentiation ability of the two cells.We also examined the expression of hepatic related genes by real-time RT-PCR.Results UC-MSCs manifested significandy stronger proliferation potential than BM-MSCs.Both UC-MSCs and BM-MSCs could be induced into hepatocyte-like cells.The morphology of UC-MSCs tended to be more mature than BM-MSCs and they had more cytoplasmic granules.After 4 weeks,the levels of albumin and urea nitrogen from the cultural medium of the UC-MSCs group were significantly higher than the BM-MSCs group (P ＜ 0.05).Real-time PCR showed the expressions of four liver related genes CK18,G6P,HGF and ALB in the UC-MSCs group were significantly higher than the BM-MSCs group (P ＜ 0.05).Conclusion UC-MSCs had higher hepatic differentiation potential than BM-MSCs.Thus,UC-MSCs are more suitable than BM-MSCs for tissue engineering in the treatment of end-stage liver diseases.
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Aim To investigate the effects of berberine combined with bone marrow mesenchymal stem cells ( BM-MSCs) on the energy metabolism of human um-bilical vein endothelial cells ( HUVECs ) under the condition of high glucose. Methods ①The state of cell reproduction and cell proliferating activity were de-termined by MTT assay. ②The cell cycle was detected by flow cytometry ( FCM ) . ③DNA damage of cells was measured by comet tail assay. ④The contents of ATP, ADP and AMP were determined by high perform-ance liquid chromatography ( HPLC ) and the level of energy charge ( EC) was calculated. ⑤The expression of CCR and COX mRNA was detected by reverse tran-scription-polymerase chain reaction ( RT-PCR) . ⑥The expressions of COX and CCR were detected by Western blot. Results ①The proliferating activity of HUVECs declined apparently and the proliferation decreased af-ter high glucose intervention. Meanwhile, the quantity of cells during S +G2 dropped dramatically and there was certain degree of damage to DNA. The berberine and BM-MSCs respectively improved the proliferating activity and the proliferation in different degrees, in-creased the quantity of cells during S+G2 and promo-ted the repair of DNA ( P <0 . 01 ) , and so did the combination of the two, with a better effect than each of them alone. ②After high glucose intervention and the damage caused, the content of both ATP and ADP of HUVECs was reduced, and EC level also declined significantly, while the content of AMP increased. The berberine and BM-MSCs respectively up-regulated the content of ATP and ADP ( P<0. 01 ) , and so did the combination of the two, with a better effect than each of them alone. ③After high glucose intervention and the damage caused, the expression of COX, CCR mR-NA and protein decreased obviously. Yet, all of the three gained a dramatic increase when the berberine, BM-MSCs or the combination of the two were added ( P<0. 01 or P <0. 05 ) , among which the combination worked more effectively. Conclusions The berber-ine, BM-MSCs and the joint use of the two could im-prove the energy metabolism of HUVECs, which had been damaged by high glucose, probably because the berberine and BM-MSCs could up-regulate the expres-sion of COX, CCR mRNA and protein, which leads to the hydrolyzation of glucose oxide and thus the im-provement of blood environment and the enhancement of glucose's supply and intake of HUVECs. Then, here comes the final result: the improvement of the energy metabolism of damaged vascular endothelial cells by high glucose.
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BACKGROUND AND OBJECTIVES: Irradiated wound healing is a highly complex and dynamic process. The latest technology making a huge difference in this process is stem cell therapy. The goal of this study was to evaluate the use of bone marrow-derived mesenchymal stem cells (BM-MSCs) or human amniotic epithelial cells (HAECs) in the healing of irradiated wounds. METHODS AND RESULTS: Forty five male albino rats were subjected to whole body 6 gray gamma radiations. One day post irradiation, full-thickness incisional wound was created in the tibial skin. The rats were randomly equally divided into three groups. The incisions of the first group (gp I) were injected intra-dermally with saline before stitching and those of both the second (gp II) and the third groups (gp III) were intradermally injected with BM-MSCs and HAECs before stitching respectively. Animals were sacrificed after the third, seventh and fourteenth days postoperative. The healing process was assessed histopathologically. CXCL-5, SDF-1 and Transforming growth factor-beta 1 (TGF-beta1) expression were also detected in biopsies from all wounds. Expression of TGF-beta1 in gp I was more than the other groups leading to severe inflammation, deficient healed dermis and delayed reepithelialization. SDF-1 expression was high in gp II while CXCL-5 expression was high in gp III causing accelerated wound healing. BM-MSCs showed a great effect on the quality of the dermis, while superiority of the epithelium and its appendages were achieved in HAECs group. CONCLUSIONS: Using BM-MSCs and HAECs could be used safely in case of irradiated wounds.
Subject(s)
Animals , Humans , Male , Rats , Biopsy , Dermis , Epithelial Cells , Epithelium , Gamma Rays , Inflammation , Mesenchymal Stem Cells , Skin , Stem Cells , Transforming Growth Factor beta1 , Wound HealingABSTRACT
Objective To investigate the feasibility of transplantation of mesenchymal stem cell (aisc) into expanion skin. Methods MSCs were isolated from porker's bone marrow and cultured in vitro. Pigs were randomly divided into four groups: Group A was injected with MSCs on the local expansion; Group B was injected with MSCs from ear vein; Group C was planted expander only; Group D was the contronl group. Each side of pig's spinal column was implanted with three expander in groups A, B and C. The same volume of NS was injected at the fixed time, the marked area measured after 7, 14, 28 days of expansion, the difference of those area was compared between groups. The differentiation of BM-MSC was detected by immunofluorescence. Results Flow-cytometric analysis showed that these BMSCs expressed CD90 and CD29 highly but did not express CD34 or CD45. The expansion area (cm2) of groups A, B, C and D was 34.05±0. 92, 31.83±l. 07,30. 10±0.79, and 18. 27±0.25, respectively (P＜0. 01). Immunofluorescence showed that the positive expression rate of CD31, PCNA in groups A and B was higher than that in groups C and D, in which the expression was the highest in group A. Conclusions Allogeneic transplantation of BM-MSC can promote skin expansion and the effect of local transplantation group is most significant.
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Diabetic neuropathy is one of the most frequent and troublesome complications of diabetes. Although there has been a continuous increase in the incidence of diabetic neuropathy, treatments have yet to be found that effectively treat diabetic neuropathy. Neurotrophic factors are proteins that promote the survival of specific neuronal populations. They also play key roles in the regeneration of peripheral nervous system. Recent evidence from diabetic animal models and human diabetic subjects suggest that reduced availability of neurotrophic factors may contribute to the pathogenesis of diabetic neuropathy. One way to reverse this effect is to take advantage of the finding that bone marrow derived mesenchymal stem cells (BM-MSCs) promote peripheral nerve repair and the functioning of neurotrophic factors. Therefore, we speculated that treatment with BM-MSCs could be a viable therapeutic strategy for diabetic neuropathy. The present study was designed to examine the possible beneficial effect of BM-MSCs on functions of neurotrophic factors in diabetic neuropathy. To assess this possibility, we used an in vivo streptozotocin-induced diabetic neuropathy mouse model. Quantitative real-time polymerase-chain reacion showed that BM-MSCs significantly increase expression levels of neurotrophic factors. Also, BM-MSCs ameliorated nerve conduction velocity in streptozotocin-treated mice. These results may help to elucidate the mechanism by which BM-MSCs function as a cell therapy agent in diabetic neuropathy.
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Animals , Humans , Mice , Bone Marrow , Diabetic Neuropathies , Imidazoles , Incidence , Mesenchymal Stem Cells , Models, Animal , Nerve Growth Factors , Neural Conduction , Neurons , Nitro Compounds , Peripheral Nerves , Peripheral Nervous System , Proteins , Regeneration , Cell- and Tissue-Based TherapyABSTRACT
Nicotinamide and Exendin-4 could promote transdifferentiation of rat BM-MSCs into insulin-producing cells in vitro, which expressed insulin, C-peptide and related genes.