BACKGROUND: Our previous studies have demonstrated that bone marrow mesenchymal stem cells can ameliorate airway inflammation and regulate levels of inflammatory factors. Moreover, galectin-1 secreted by bone marrow mesenchymal stem cells can inhibit the immune function of dendritic cells in vitro. However, the effect of galectin-1 secreted by bone marrow mesenchymal stem cells on asthma is unknown.
OBJECTIVE: To investigate the effect of galectin-1 secreted by bone marrow mesenchymal stem cells on airway inflammation of asthmatic mice.
METHODS: A recombinant lentiviral vector, pLVX-gal-1-shRNA, was constructed for RNA interference of galectin-1 gene and then was transferred into bone marrow mesenchymal stem cells. Forty female BALB/c mice were equally randomized into normal control group, asthmatic group, bone marrow mesenchymal stem cells treatment group (cell treatment group), galectin-1 treatment group and galectin-1 interference group. The number of total inflammatory cells and differential cells in the mouse bronchoalveolar lavage fluid was determined. Furthermore, hematoxylin-eosin staining was used to compare airway inflammation among five groups.
RESULTS AND CONCLUSION: Four short-hairpin RNA sequences targeting mouse galectin-1 mRNA were designed. A real time-quantitative polymerase chain reaction demonstrated that bone marrow mesenchymal stem cells were inhibited most by interference site 249 RNA sequences which were later used in vivo study. Accumulation of inflammation cells, particularly eosinophils, around the airway and in the bronchoalveolar lavage fluid was evidence in asthmatic mice compared to the normal control group. However, bone marrow mesenchymal stem cells engraftment or protein of galectin-1 infusion significantly reduced inflammatory infiltration both in the airway and in the bronchoalveolar lavage fluid. Moreover, eosinophils in the bronchoalveolar lavage fluid and in the airway attenuated dramatically in the cell treatment group and galectin-1 treatment group. However, there was no effect on inflammation accumulation in the bronchoalveolar lavage fluid and airway by infusion of galectin-1-transfected bone marrow mesenchymal stem cells to asthmatic mice. These results indicate that galectin-1 secreted by bone marrow mesenchymal stem cells could alleviate airway inflammation in the asthmatic mouse.
BACKGROUND: Seeking proper methods to promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells can provide therapeutic ideas for the treatment of osteoporosis.
OBJECTIVE: To study the mechanism of lithium chloride on the proliferation and differentiation of rat bone marrow mesenchymal stem cells.
METHODS: Rat bone marrow mesenchymal stem cells were cultured with the whole bone marrow cell inoculation and adherent purification, followed by intervention with 0, 2, 5 nmol/L lithium chloride. Cell counting kit-8 was used to detect the effect of lithium chloride on the proliferation of bone marrow mesenchymal stem cells. Alizarin red staining was used to determine calcified nodules. PNPP method was applied to measure alkaline phosphatase activity. The contents of glycogen synthase kinase 3β, phosphorylated glycogen synthase kinase 3β and β-catenin in rat bone marrow mesenchymal stem cells were determined by western blot.
RESULTS AND CONCLUSION: Lithium chloride, 2 and 5 nmol/L, promoted the proliferation of bone marrow mesenchymal stem cells and
5 nmol/L lithium chloride had the strongest proliferative effect. Compared with 0 nmol/L group, more calcified nodules and higher alkaline phosphatase activity were observed in the 2 and 5 nmol/L groups, especially in the 5 nmol/L group. Both 2 and 5 nmol/L lithium chloride upregulated the expression of phosphorylated glycogen synthase kinase 3β and β-catenin proteins, but showed no difference in the expression of glycogen synthase kinase 3β. In conclusion, lithium chloride can promote the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells, and the possible mechanism is through the regulation of Wnt/β-catenin signaling pathway-related protein expression.
Simvastatin promotes osteogenic differentiation of bone marrow mesenchymal stem cells
Simvastatin promotes osteogenic differentiation of bone marrow mesenchymal stem cells [J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 3961-.
BACKGROUND: Cholinergic receptor agonists can promote the secretion of residual salivary cells in patients with radiation-induced salivary gland injury. However, there are serious adverse reactions associated with its long-term use and limited therapeutic effects on severely radioactive salivary gland damage. Bone marrow mesenchymal stem cells are a kind of cells with multipotential differentiation potential and almost unlimited proliferative capacity, which have the potential to treat radiation-induced salivary gland injury.
OBJECTIVE: To explore the mechanism of bone marrow mesenchymal stem cells repairing radiation-induced salivary gland injury.
METHODS: Bone marrow mesenchymal stem cells were extracted from mouse bone marrow, and co-cultured with the second-generation salivary gland acinar cells for 24 hours by a 3D co-culture system. One hundred and fifty C57 mice were randomly divided into normal control group, radiation-induced salivary gland injury group and bone marrow mesenchymal stem cells+radiation-induced salivary gland injury group (stem cell treatment group). A mouse model of radiation-induced salivary gland injury was made by an electron linear accelerator (15 Gy). One week after irradiation, the mice in the stem cell treatment group were subcutaneously injected with 2×109/L bone marrow mesenchymal stem cell suspension at multiple points of the salivary gland, and those in the other two groups were injected with the same amount of saline.
RESULTS AND CONCLUSION: After 24 hours of co-culture, bone marrow mesenchymal stem cells differentiated into salivary gland acinar cells, with a polygon-like shape and expressed α-amylase. Compared with the normal control group, the salivary flow, salivary gland mass, and salivary amylase level were significantly decreased, acinar cell structure was markedly damaged, and the Notch expression level in the salivary gland was decreased in the radiation-induced salivary gland injury group. Compared with the radiation-induced salivary gland injury group, the above indexes in the bone marrow mesenchymal stem cells+radiation-induced salivary gland injury group were significantly restored. These findings indicate that bone marrow mesenchymal stem cell transplantation can effectively treat radiation-induced salivary gland injury, and its effect may be achieved by regulating the Notch expression level in the salivary gland.
BACKGROUND: Previous studies have focused on the effects of inflammatory factors-activated nuclear factor-kappa B signaling pathway on bone cells. Little has been reported on the effects of micro-inflammatory environment on osteogenic differentiation and nuclear factor-kappa B signaling pathways of human periodontal ligament stem cells.
OBJECTIVE: To investigate the effect of tumor necrosis factor-alpha on osteogenic differentiation and nuclear factor-kappa B signaling pathway of periodontal ligament stem cells.
METHODS: Human periodontal ligament stem cells were cultured in vitro by enzymatic digestion combined with tissue explant method, and the cell source was identified by immunohistochemical staining. The study protocol was approved by the Ethics Committee of the Affiliated Hospital of Qinghai University, and informed consent was obtained from each patient prior to the enrollment in the study. The cells were cultured in simple osteogenic induction medium (control group) or osteogenic induction medium containing 10 μg/L tumor necrosis factor-alpha (tumor necrosis factor-alpha group). Alizarin red staining and alkaline phosphatase activity were used to detect the effect of tumor necrosis factor-alpha on osteogenic differentiation of periodontal ligament stem cells. Real-time quantitative RT-PCR was used to detect the effect of tumor necrosis factor-alpha on the expression of osteogenic differentiation genes, Osterix and Runx2. Western blot was used to detect the effect of tumor necrosis factor-alpha on nuclear factor-kappa B signaling pathway.
RESULTS AND CONCLUSION: Immunohistochemical staining showed that human periodontal ligament stem cells were positive for anti-vimentin and negative for anti-keratin. After alizarin red staining, mineralized nodules formed in the tumor necrosis factor-alpha group were fewer in number, smaller in scope and lighter in staining than those in the control group. Alkaline phosphatase activity in the tumor necrosis factor-alpha group was significantly lower than that in the control group (P < 0.05). Real-time quantitative RT-PCR results showed that the expression levels of Osterix and Runx2 mRNA in tumor necrosis factor-alpha group were significantly lower than those in the control group (P < 0.05). Western blot assay showed that, compared with the control group, the expression of p-IκBα protein increased significantly in the tumor necrosis factor-alpha group (P < 0.05), while the expression of p65 and IκBα proteins decreased (P < 0.05). These findings indicate that tumor necrosis factor-alpha can inhibit the osteogenic differentiation of human periodontal ligament stem cells by activating the nuclear factor-kappa B signaling pathway.
BACKGROUND: Numerous studies addressing cells in osteoarthritis still focus on the in vitro culture of articular chondrocytes in animal models. Degeneration in the animal models is not completely consistent with that in human osteoarthritis. How to construct an in vitro cell model using chondrocytes from human osteoarthritis and to study changing trend of its characteristic proteins is the key to simulating the cartilage degeneration in human osteoarthritis in vivo.
OBJECTIVE: To investigate the in vitro isolation and culture of human chondrocytes from osteoarthritis patients, to observe the morphological characteristics of human osteoarthritis chondrocytes from primary to third generation, and to study the changes in biological characteristics of chondrocyte-related proteins in different generation.
METHODS: The study protocol was in line with the ethic requirements of Chinese PLA General Hospital with the approval No. S2017-23-7. Six cases undergoing arthroplasty for severe osteoarthritis (2 males and 4 females, age 62-72 years old with a mean age of (68.3±3.39) years) were enrolled. The chondrocytes from abandoned cartilage tissue in these patients were isolated and cultured by one-step enzymatic digestion (type II collagenase), and then subcultured, so as to construct an in vitro chondrocyte culture system for osteoarthritis. The morphology of cells at different generations was observed by inverted phase contrast microscope. Hematoxylin-eosin staining, toluidine blue staining and type II collagen immunofluorescence staining were used for cell identification. Western blot was used to detect the expressions of Col2a, Aggrecan and matrix metalloproteinase-13 in each generation of chondrocytes at the fusion rate of 70%.
RESULTS AND CONCLUSION: After digestion using type II collagenase, the cells scattered around the tissue mass could be observed at about 1 week of culture, and these cells could be subcultured for further study after about 3 weeks. Morphological observation, hematoxylin-eosin staining, toluidine blue staining and type II collagen immunofluorescence staining proved that human chondrocytes were successfully cultured. The relative expression of Col2a and Aggrecan in chondrocytes at the third generation was significantly decreased as compared with that in primary cells (P < 0.01), and the expression gradually decreased with the subculture times. Matrix metalloproteinase-13 expression gradually increased with the increase of subculture times (P < 0.01). In conclusion, chondrocytes can be successfully isolated from the osteoarthritis specimens by one-step digestion of type II collagenase and then subcultured. The dedifferentiation of osteoarthritis chondrocytes in vitro increases with the increase of subculture times and the expression of functional proteins decreases as a whole. The chondrocytes within three generations present with cartilage degeneration in osteoarthritis and may be the best choice for experimental studies.
BACKGROUND: Since mandibular distraction osteogenesis has achieved significant repercussions in clinical practice, many scholars have been working on studies that enhancing osteogenesis and shortening the course of treatment. Among them, stem cell transplantation has attracted much attention as a hotspot.
OBJECTIVE: To summarize the current progress and deficiency of stem cell-based tissue engineering technology assisting mandibular distraction osteogenesis.
METHODS: Following the PRISMA guidelines, a computer-based search of PubMed, WOS, CNKI and WanFang databases was performed by the first author for relevant articles with the keywords of “distraction osteogenesis,mandibular, mandible, stem cells” in English and Chinese, respectively. Animal models, types of stem cells and routes of transplantation in the selected literature were extracted and counted up. The results were tabulated and introduced briefly.
RESULTS AND CONCLUSION: Fifty-one articles were strictly selected as per the eligibility criteria, analyzed and summarized. In the selected literature, the timing, methods and types of stem cell transplantation vary greatly. The majority of animal experimental studies have confirmed that stem cell-based tissue engineering technology can enhance bone and blood vessel regeneration in mandibular distraction osteogenesis. Explorations on the selection and application of stem cells in mandibular distraction osteogenesis assisted by stem cell-based tissue engineering technology cannot only provide ideas for future treatment, but also lay the foundation for clinical trials.
BACKGROUND: In recent years, with the development of stem cell and regeneration technologies, bone tissue engineering has been rapidly developed. Despite there are a variety of seed cells for bone tissue engineering, researchers need to decide which types of stem cells are the best suitable for their specific purposes. To date, there is no ideal treatment for oral bone tissue repair, although a plenty of therapeutic methods have been proposed. Therefore, it will provide a new idea for repairing oral bone defects by fully utilizing the potential of stem cells during bone regeneration.
OBJECTIVE: To review the osteogenic potential of stem cells from different sources on oral bone regeneration, thereby providing a basis for selecting seed cells for tissue engineering.
METHODS: Databases of CBM, CNKI, PubMed, Elsevier, and Web of Science were retrieved with the keywords of “stem cell, bone repair, alveolar bone regenerate, osteogenesis, bone tissue” in English and Chinese, respectively. After initial screening of titles and abstracts, irrelevant articles were excluded and 81 eligible articles were included in final analysis.
RESULTS AND CONCLUSION: Tissue engineering technology provides a new approach to oral bone regeneration, but the choice of stem cells has not been standardized. Many types of stem cells have been used for bone tissue regeneration, but their roles in the body vary due to different origins of stem cells. Therefore, choosing a suitable type of stem cells is important for the repair of specific tissue damage. In this review, we compared the advantages and disadvantages of embryonic stem cells, mesenchymal stem cells and induced pluripotent stem cells in the oral bone regeneration, and attempted to provide a reference for the selection of seed cells for the regeneration and repair of oral bone tissue.
BACKGROUND: Bone marrow mesenchymal stem cells are important seed cells in bone tissue engineering repair. However, it is difficult to control the cell proliferation and differentiation. Although previous studies have focused on biochemical regulation, little is reported on the mechanical microenvironment for controlling the proliferation and differentiation of bone marrow mesenchymal stem cells.
OBJECTIVE: To summarize the research status and recent progress in the effect of mechanical microenvironment on the proliferation and differentiation of bone marrow mesenchymal stem cells.
METHODS: A search with the keywords of “mechanical microenvironment; mesenchymal stem cells; stiffness; proliferation and differentiation” in Chinese and English was conducted in the CNKI, WanFang and PubMed, respectively. Initially 117 articles were retrieved, and 65 eligible articles were finally summarized.
RESULTS AND CONCLUSION: Different mechanical forces cause different effects on the proliferation and differentiation of bone marrow mesenchymal stem cells. Under mechanical tension and fluid shear forces, the cells mostly differentiate into osteoblasts. Under compressive and hydrostatic pressure, the majority of the cells differentiate into chondrocytes, while a small amount of the cells are induced to differentiate into osteoblasts. Mechanical signals from different basal mechanics have an effect on neurogenesis adipogenesis, myogenesis and osteogenesis in the mesenchymal stem cells. There are different optimal conditions for the cells differentiating into different tissues. Different self-components of the cells also play an important role in their response to mechanical signals. To explore the relationship between mechanical dynamics and proliferation and differentiation of mesenchymal stem cells, we establish a simulated matrix and scaffold system similar to the mechanical microenvironment of bone marrow mesenchymal stem cells.
BACKGROUND: Since human embryonic stem cells were successfully obtained in 1998, clinicians have been trying to use stem cells in wound healing. Importantly, stem cells can regulate wound inflammation.
OBJECTIVE: To investigate the application and prospect of adipose-derived stem cells in wound inflammation.
METHODS: The first author searched CNKI and PubMed databases (from 1999 to October 2018) with the retrieval words of “adipose-derived stem cells, wound, inflammation” in Chinese and English, respectively. Literatures regarding adipose-derived stem cells, wound and inflammation were selected. The articles published lately or authoritatively in the same field were preferred.
RESULTS AND CONCLUSION: Wound healing is a complex and orderly process, and closely related to wound inflammation. With the development of regenerative medicine, the application of stem cells brings a new approach to wound healing. Adipose-derived stem cells have a promising prospect as characterized by the strong abilities of proliferation and multidirectional differentiation, easy to obtain, and abundant sources. To date, adipose-derived stem cells are still one of the most popular candidates for regulating wound inflammation and promoting wound healing.
BACKGROUND: Human umbilical cord blood contains a large number of stem cells that are capable of differentiating into hematopoietic, epithelial, endothelial and neural tissues both in vitro and in vivo. Thus, human umbilical cord blood has a therapeutic potential to a wide variety of diseases.
OBJECTIVE: To summarize the clinical application of human umbilical cord blood cells in the treatment of non-hematopoietic diseases.
METHODS: A computer-based retrieval of PubMed and CNKI databases was performed in order to search relevant articles published from 2001 till now, using the keywords of “umbilical cord stem cells, umbilical cord blood mesenchymal stem cells, umbilical cord blood” in English and Chinese, respectively. After removal of repetitive or irrelevant articles, 64 articles were finally reviewed.
RESULTS AND CONCLUSION: The clinical research of umbilical cord blood stem cells involves many fields such as the circulatory system, endocrine system and nervous system, in most of which good experimental results have been achieved. Findings from in-depth basic and clinical studies on biological and immunological characteristics of cord blood indicate an extensive application of umbilical cord blood stem cells that will be a new therapeutic choice for patients.