Lentiviral vector mediated enhanced green fluorescent protein screening for stably transfected rabbit bone marrow mesenchymal stem cells / 西安交通大学学报(医学版)

Objective: To isolate, culture and identify rabbit bone mesenchymal stem cells (BMSCs) so as to explore the optimal conditions for lentiviral vector-mediated enhanced green fluorescent protein (eGFP) infection in rabbit BMSCs and screen stable transfected BMSCs in rabbits. Methods: BMSCs were obtained by whole bone marrow adherence method. The osteogenic, chondrogenic and adipogenic differentiation of BMSCs was made by alizarin red, toluidine blue and oil red O staining, respectively. The expressions of CD44 and CD90 were detected by immunofluorescence. The concentration of puromycin was used to screen the minimum lethal concentration of BMSCs; the lentiviral vector with multiplicity of infection (MOI) of 50, 100, 150 and 200 mediated eGFP BMSCs were infected; the fluorescence expression was observed under an inverted microscope, and the stable transformation system was screened with puromycin. Results: When MOI was 150, lentiviral vector-mediated eGFP infection of rabbit BMSCs was the most efficient. The optimum concentration of puromycin for stable transfection of rabbit BMSCs was 1.0 μg/mL. Conclusion: Rabbit BMSCs were successfully cultured in this experiment. The stem cells were labeled with lentivirus-mediated GFP and stable transfected rabbit BMSCs were screened. A simple and effective stem cell labeling method was established to label BMSCs in vivo.

Research Progress on Mechanisms of Vibration in Osteogenesis of Bone Mesenchymal Stem Cells / 医用生物力学

Vibration represents a micro reciprocating motion of a particle or object along a line or arc relative to a reference position, while the effect of low-magnitude high-frequency vibration (LMHFV) on skeletal system cells is similar to the mechanical stimulation of muscle movement. Bone mesenchymal stem cells (BMSCs), which have been identified as force-sensitive cells, exist in the bone marrows and have the potential of multi-lineage differentiation. Their biological characteristics can change functionally according to the appropriate stimulation in vitro, in order to reach the optimal demand of the stimulation. LMHFV can promote the osteogenic differentiation of BMSCs, therefore, the research on its mechanism can contribute to the application of vibration in the treatment of diseases such as osteoporosis, fracture, osteogenesis imperfecta, obesity as well as the promotion of orthodontic tooth movement. This paper summarizes the recent progress about the effects of vibration on BMSCs stem cells in osteogenesis and the possible mechanisms, so as to provide research ideas and methods for studying the mechanical as well as biological changes of BMSCs under vibration stimulation.

Effects of Microenvironment on Osteogenesis of Bone Marrow Mesenchymal Stem Cells Through Cytoskeleton Tension / 医用生物力学

Extracellular matrix is the main element to provide mechanical clues for cells. The response of stem cells to mechanical signals is mainly achieved through the cytoskeleton. After mechanical signal is transmitted, cytoskeleton can form contractile microfilaments that actively generate tension through reorganization induced by microenvironment changes. The mechanical signals can regulate gene expression through either coupling with the nuclear skeleton directly or being transformed by the second message. Recent studies have proven that cytoskeleton tension has a series of impact on lineage specification, proliferation, differentiation and apoptosis of bone mesenchymal stem cells (BMSCs). BMSCs are of great significance in bone reconstruction and clinical treatment. The possible mechanisms about mechanotransduction and its effects of cytoskeleton tension on osteogenesis of BMSCs after micro-environmental changes were summarized.

Research Progress on Mechanisms of Vibration in Osteogenesis of Bone Mesenchymal Stem Cells / 医用生物力学

Vibration represents a micro reciprocating motion of a particle or object along a line or arc relative to a reference position, while the effect of low-magnitude high-frequency vibration (LMHFV) on skeletal system cells is similar to the mechanical stimulation of muscle movement. Bone mesenchymal stem cells (BMSCs), which have been identified as force-sensitive cells, exist in the bone marrows and have the potential of multi-lineage differentiation. Their biological characteristics can change functionally according to the appropriate stimulation in vitro, in order to reach the optimal demand of the stimulation. LMHFV can promote the osteogenic differentiation of BMSCs, therefore, the research on its mechanism can contribute to the application of vibration in the treatment of diseases such as osteoporosis, fracture, osteogenesis imperfecta, obesity as well as the promotion of orthodontic tooth movement. This paper summarizes the recent progress about the effects of vibration on BMSCs stem cells in osteogenesis and the possible mechanisms, so as to provide research ideas and methods for studying the mechanical as well as biological changes of BMSCs under vibration stimulation.

Research progress on Mechanism of Fluid Shear Stress in Osteogenesis of Bone Mesenchymal Stem Cells / 医用生物力学

Cells are exposed to mechanical stress, such as fluid shear stress (FSS), mechanical strain, hydrostatic pressure in vivo. FSS is considered to be the most important stress during bone homeostasis and remodeling. At present, most studies are mainly about the FSS effect on osteocytes and osteoblasts. However, the effects of FSS on bone mesenchymal stem cell (BMSCs) are not fully understood. BMSCs are of great significance in bone reconstruction and clinical treatment, so researchers increasingly focus on the response of BMSCs to FSS. The response of BMSCs to FSS depends on the alteration of cytoskeleton, matrix stiffness and elasticity, osteogenic signaling pathways and so on. In this review paper, the recent researches about the mechanotransduction mechanism of FSS, and its effect on differentiation and function of BMSCs are summarized, so as to provide new insights for studying construction of tissue engineered bone and treatment of bone diseases.

Acupoint injection of BMSCs combined with Chinese herbs for capillary density in ischemic hind limb of diabetes mellitus rats / 中国针灸

OBJECTIVE@#To observe the effect of acupoint injection of bone mesenchymal stem cells (BMSCs) combined with Chinese herbs of benefiting for activating blood circulation for capillary density and arterioles density in skeletal muscle in ischemic hind limb of diabetes mellitus (DM) rats.@*METHODS@#A total of 80 rats were randomized into a normal sham operation group (10 rats) and a model group (70 rats). Disposable intraperitoneal injection of streptozotocin (STZ, 50.0 mg/kg) was used to establish DM model, and the rats in the model group were randomized into 7 subgroups, 10 rats in each one. The subgroups were the DM sham operation group, DM ischemic group, Chinese herb group (intragastric herbs of benefiting for activating blood circulation), local injection group (BMSCs local injection), local injection + Chinese herb group (BMSCs local injection combined with intragastric herbs of benefiting for activating blood circulation), acupoint injection group (BMSCs acupoint injection), acupoint injection + Chinese herb group (BMSCs acupoint injection combined with intragastric herbs of benefiting for activating blood circulation). The local injection was phosphate buffer (PBS) injection at the equidistant 5 points along the line between the ischemic tissue and the normal tissue a time. The acupoints were "Sanyinjiao" (SP 6), "Zhaohai" (KI 6), "Huantiao" (GB 30), "Housanli" (ST 36) and "Yanglingquan" (GB 34). 100 μL BMSCs with 1×10/mL was totally injected at the above acupoints for one rat, 20 μL an acupoint. 1.5 kg/L Chinese herbs were applied by intragastric administration, including 120 g Radix Astragali, 120 g Codonopsis, 48 g Radix Glycyrrhiza, 120 g Angelica sinensis, 120 g Blood Rattan, 48 g Achyranthes bidentata. Intragastric distilled water was used in the other non-Chinese herb groups. The expressions of α-smooth muscle actin (α-actin), latelet endothelial cell adhesion molecule (CD31) and von willebrand factor (vWF) in the skeletal muscle were detected with immunohistochemical SP two-step method.@*RESULTS@#Twenty-one days after intervention, the expressions of α-actin and CD31 on the operation hind limb were higher than those on the healthy hind limb in all the groups, except the Chinese herb group (<0.05<0.01). The vWF expressions on the operation side were lower than those on the healthy side in the Chinese herb group, the local injection group, the local injection + Chinese herb group and the acupoint injection + Chinese herb group (<0.05, <0.01). The α-actin expression on the operation side in the acupoint injection + Chinese herb group was higher than those in the normal sham operation group, DM sham operation group, the DM ischemic group and the local injection group (<0.05, <0.01). The CD31 expressions in the acupoint injection group, the acupoint injection + Chinese herb group, local injection + Chinese herb group were higher than those in the normal sham operation group, DM sham operation group and DM ischemic group (<0.05, <0.01). The CD31 expression in the acupoint injection + Chinese herb group was higher than those in the Chinese herb group and the local injection group (both <0.05). The vWF expressions in the local injection + Chinese herb group, the acupoint injection group and the acupoint injection + Chinese herb group lower than those in the DM sham operation group and the DM ischemic group (<0.05, <0.01).@*CONCLUSION@#schemia increases the expressions of the vascular density related factors of α-actin and CD31. It is more obvious for the increasing expressions of α-actin and CD31, and decreasing expression of vWF with the interventions of simple BMSCs injection and simple Chinese herbs of benefiting for activating blood circulation, especially with the combination of the above tow methods. It is indicated that acupoint injection of BMSCs combined with Chinese herbs of benefiting for activating blood circulation can improve the angiogenesis of ischemic tissue.

The roles of STAT3 in osteogenesis of bone mesenchymal stem cells / 医用生物力学

Orthodontic tooth movement is a dynamic process,which includes bone resorption on the pressure side and osteogenesis on the tension side.Bone mesenchymal stem cells (BMSCs),which are force-sensitive cells,have potentials for differentiation into cells with various types.Their biological characteristics can change functionally according to the appropriate stimulation in vitro,in order to reach the optimal demand of the stimulation.Many signal pathways are involved in osteogenesis.Signal transducers and activators of transcription 3 (STAT3) is a ubiquitously expressed transcription factor,mediating cell proliferation,differentiation,survival,apoptosis and cellular immunity.It has been reported that STAT3 can regulate the differentiation process of BMSCs into osteoblasts.This paper summarizes the recent progress about effect of STAT3 on bone differentiation of BMSCs and the possible mechanism.

The roles of STAT3 in osteogenesis of bone mesenchymal stem cells / 医用生物力学

Orthodontic tooth movement is a dynamic process, which includes bone resorption on the pressure side and osteogenesis on the tension side. Bone mesenchymal stem cells (BMSCs), which are force-sensitive cells, have potentials for differentiation into cells with various types. Their biological characteristics can change functionally according to the appropriate stimulation in vitro, in order to reach the optimal demand of the stimulation. Many signal pathways are involved in osteogenesis. Signal transducers and activators of transcription 3 (STAT3) is a ubiquitously expressed transcription factor, mediating cell proliferation, differentiation, survival, apoptosis and cellular immunity. It has been reported that STAT3 can regulate the differentiation process of BMSCs into osteoblasts. This paper summarizes the recent progress about effect of STAT3 on bone differentiation of BMSCs and the possible mechanism.

The roles of STAT3 in osteogenesis of bone mesenchymal stem cells / 医用生物力学

Orthodontic tooth movement is a dynamic process,which includes bone resorption on the pressure side and osteogenesis on the tension side.Bone mesenchymal stem cells (BMSCs),which are force-sensitive cells,have potentials for differentiation into cells with various types.Their biological characteristics can change functionally according to the appropriate stimulation in vitro,in order to reach the optimal demand of the stimulation.Many signal pathways are involved in osteogenesis.Signal transducers and activators of transcription 3 (STAT3) is a ubiquitously expressed transcription factor,mediating cell proliferation,differentiation,survival,apoptosis and cellular immunity.It has been reported that STAT3 can regulate the differentiation process of BMSCs into osteoblasts.This paper summarizes the recent progress about effect of STAT3 on bone differentiation of BMSCs and the possible mechanism.

The roles of STAT3 in osteogenesis of bone mesenchymal stem cells / 医用生物力学

Orthodontic tooth movement is a dynamic process,which includes bone resorption on the pressure side and osteogenesis on the tension side.Bone mesenchymal stem cells (BMSCs),which are force-sensitive cells,have potentials for differentiation into cells with various types.Their biological characteristics can change functionally according to the appropriate stimulation in vitro,in order to reach the optimal demand of the stimulation.Many signal pathways are involved in osteogenesis.Signal transducers and activators of transcription 3 (STAT3) is a ubiquitously expressed transcription factor,mediating cell proliferation,differentiation,survival,apoptosis and cellular immunity.It has been reported that STAT3 can regulate the differentiation process of BMSCs into osteoblasts.This paper summarizes the recent progress about effect of STAT3 on bone differentiation of BMSCs and the possible mechanism.

Impacts of ground-based microgravity simulation on biological responses of bone marrow mesenchymal stem cells and its underlying mechanisms: A mini-review / 医用生物力学

With the development of space life science, researches on ground-based microgravity simulation become more and more important for spaceflight to complement their limited missions. It is well known that bone marrow mesenchymal stem cells (BMSCs) are pluripotent, self-renewing cells with multi-lineage differentiation capacity on the ground, but their responses under microgravity and the underlying regulatory mechanisms are poorly understood. Ground-based microgravity simulation might affect cell proliferation, apoptosis and expression of surface molecules, and induce cytoskeletal reorganization, as well as alter the differentiation potential of BMSCs. In this review, how ground based microgravity simulation mediates BMSCs’ responses and its involved mechanisms are summarized to further understand the mechano-biological coupling in such process and provide theoretical references for space flight-induced pathophysiological alterations.

Culturing Rabbit Stem Cells in Vitro and Its Anabolism under Basic Fibroblast Growth Factor / 中国康复理论与实践

@#Objective To explore the changes of rabbit adipose stem cells(ASCs)and bone mesenchymal stem cells(BMSCs)cultured in vitro and the anabolism under basic fibroblast growth factor(bFGF).Methods BMSCs and ASCs were cultured with DMEM,DMEM/F12(2∶1)or α-MEM respectively.The 3rd generation ASCs and BMSCs were divided into 2 groups respectively:group A:ASCs cultured in chondrogenic medium(CM),group B:ASCs cultured in CM supplemented with bFGF 5 ng/ml,group C:BMSCs cultured in CM,group D:BMSCs cultured in CM supplemented with bFGF 5 ng/ml.Morphological changes were observed under inverted microscope.The 35SO42-incorporation and total hydroxyproline were measured.Results BMSCs and ASCs showed much higher growth rate when cultured in α-MEM medium comparison with that in DMEM or in DMEM/F12(2:1).Both stem cells attachment cultured in monolayer greatly increased and cell clones were abundant,while the cells attachment became rather difficult and cell clones were less after cutured in CM.All stem cells possessed a round-like morphology,and the cells in group B and D were more than that in the other 2 groups.The 35SO42-incorporation and total hydroxyproline synthesis of group B or D increased compared with that of group A or C,but there was no diference between group D and B.Conclusion The rabbit ASCs and BMSCs cultured in CM suppling with bFGF grow well and their metabolism increased.

Achievements and Prospects for Nonunion (review) / 中国康复理论与实践

@#Nonunion of longbones is a significant consequence in treating fractures, which is not easy to treat. Thes reviewed the basic knowledge and main progress regarding the cause, pathogenesis, pathophysiology of nonunion, and the methods that may be applied to the treatment of nonunion especially those non-operative one.