Isolation of a native osteoblast matrix with a specific affinity for BMP2

During their commitment and differentiation toward the osteoblast lineage, mesenchymal stem cells secrete a unique extracellular matrix (ECM) that contains large quantities of glycosaminoglycans (GAGs). Proteoglycans (PGs) are major structural and functional components of the ECM and are composed of a core protein to which one or more glycosaminoglycan sugar chains (GAGs) attach. The association of BMP2, a member of the TGF-β super-family of growth factors, and a known heparin-binding protein, with GAGs has been implicated as playing a significant role in modulating the growth factor’s in vitro bioactivity. Here we have characterised an osteoblast-derived matrix (MX) obtained from decellularised MC3T3-E1 cell monolayers for its structural attributes, using SEM and histology, and for its functional ability to maintain cell growth and viability. Using a combination of histology and anion exchange chromatography, we first confirmed the retention of GAGs within MX following the decellularisation process. Then the binding specificity of the retained GAG species within the MX for BMP2 was examined using a BMP2-HBP/EGFP (BMP2 Heparin-Binding Peptide/Enhanced Green Fluorescent Protein) fusion protein. The results of this study provide further evidence for a central role of the ECM in the regulation of BMP2 bioactivity, hence on mesenchymal stem cell commitment to the osteoblast lineage.     

干细胞与再生医学技术发展态势研究

20世纪以来,干细胞与再生医学技术一直是国际生物医学领域的热点前沿之一,它为保障人类生命健康、改善人类生存质量和延长人类寿命发挥不可替代的巨大作用。因此,美国、欧洲国家、日本和中国等科技大国均将该领域纳入了国家科研与产业发展的重点战略中,并通过专项扶持、政策补贴、立法保障等方式激励该领域的创新发展。通过对近年来国际科技战略和科技研发态势的梳理分析,发现该领域的国际战略布局规律,揭示我国在该领域的领先优势与弱点,为我国未来干细胞与再生医学技术发展提出相关参考建议。     

Patterns of daily stem growth in different tree species in a warm-temperate forest in northern China

Radial growth in trees responds to environmental changes in various ways ranging from immediate to hysteretic responses. However, species-specific tree radial growth patterns and their responses to short-term weather changes are not fully understood. Here, the daily stem radial changes (SRCs) in four common tree species, linden (Tilia mongolica), birch (Betula dahurica), oak (Quercus wutaishanica) and larch (Larix principis-rupprechtii), were monitored with high-resolution point dendrometers during the main growing seasons in 2017–2019 on Dongling Mountain, northern China. The SRC was differentiated into tree water deficit-induced stem shrinkage (TWD) and growth-induced irreversible stem expansion (GRO) to evaluate species-specific responses to weather variables and short-term drought events. We found that the TWD and GRO of the four species were significantly different. The TWD was influenced primarily by the vapor pressure deficit (VPD), whereas the GRO was influenced primarily by precipitation (P). In linden and birch, a larger proportion of the GRO occurred at higher air temperature (T_mean) and VPD values; in contrast, the range of these changes was lower in oak and larch. With the increased durations of drought periods, oak and larch experienced large and rapid increases in TWD, whereas birch and linden showed small and slow increases. These results indicate that oak and larch would be sensitive to warmer and drier weather conditions predicted for the future, while linden and birch would have a conservative growth strategy. Our results provide further insights into the physiology of these four tree species and allow us to better predict the growth response of forest dynamics under climate change.     

How do Araucaria angustifolia trees grow in overstocked stands?

The objective of this study was to evaluate growth along the stem of Araucaria angustifolia (Bertol.) Kuntze trees competing in overstocked stands, in order to identify periods when growth and trunk shape are differentiated during the trees' lifespan. The research was carried out in a planted forest of Araucaria angustifolia established in 1946 in the Açungui National Forest in Campo Largo, Paraná, Brazil, when these trees were 65 years old. One thinning was recorded, at some time between 1970 and 1980. Forty-six trees were selected and divided into three development classes (DC) at 65 years of age; these classes considered diameter at 1.30 m (breast height, dbh) with a range of 20 cm (from 10 cm to 70 cm). In addition to dbh, total tree height, and crown height and diameter were measured in the field. From each tree, 14 disks were removed to analyze growth rings and confirm the age of the stand. Some trees in the smallest DC (10 ≤ dbh < 30 cm) were the product of natural regeneration (younger trees that grew after the initial planting). In 63% of the trees, at least one growth ring was missing at breast height. Missing rings at breast height were more common in trees with smaller dbh and crown diameters. The need for more growing space was observed at different periods during the studied lifespan of the trees from three DCs. It resulted in changes in stems shape from conical to cylindrical. Different growth patterns could be observed during the lives of some trees as they outgrew their competition.     

Apoptosis in HepaRG and HL-7702 cells inducted by polyphyllin II through caspases activation and cell-cycle arrest

Rhizoma Paridis, a traditional Chinese medicine, has shown promise in cancer prevention and therapy. Polyphyllin II is one of the most significant saponins in Rhizoma Paridis and it has toxic effects on kinds of cancer cells. However, our results in this study proved that the polyphyllin II has hepatotoxicity in vitro through caspases activation and cell-cycle arrest. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide results indicated polyphyllin II inhibited proliferation, induced apoptosis in HepaRG cells and HL-7702 cells and showed a concentration and time-dependent. Then, we selected the innovative cell model-HepaRG cells to explore the mechanism of hepatotoxicity. Our data showed the reactive oxygen species (ROS) increased and the mitochondrial membrane potential decreased in HepaRG cells after administration of polyphyllin II. Besides, with the increase of concentration, the release of lactate dehydrogenase increased and the S phase of the cell cycle was arrested. Nevertheless, when pretreatment with antioxidant N-acetylcysteine, apoptotic cells decreased significantly, inhibited the production of ROS and improved the decrease of membrane potential in HepaRG cells. Moreover, polyphyllin II treatment increased levels of Fas, Bax, cytochrome c, activated caspase-3, -8, -9, cleaved poly(ADP-ribose) polymerase and decreased Bcl-2 expression levels. Finally, we identified two signal pathways of apoptosis induced by polyphyllin II including the death receptor pathway and the mitochondria pathway. This study confirmed the hepatotoxicity of the polyphyllin II in vitro, which has never been discovered and gave a wake-up call for the clinical application of Rhizoma Paridis.     

Maintenance of stem cell viability and differentiation potential following cryopreservation within 3-dimensional hyaluronic acid hydrogels

While significant progress has been made in directing the behavior of cells encapsulated within three-dimensional (3D) covalently crosslinked hydrogels, the capacity of these materials to support in situ cryopreservation of cells directly within the gels has not been assessed. Here, we demonstrate the retention of human mesenchymal stem cell (hMSC) viability within hyaluronic acid (HA) and polyethylene glycol based hydrogels via a facile gradual cooling and freezing protocol. Encapsulated cell viability was retained at similar rates in both materials systems regardless of initial duration in culture or adhesive ligand incorporation, indicating the versatility of the approach. Additionally, the cryopreservation protocol maintains stem cell differentiation potential; incubation in adipogenic differentiation media induced equal rates of hMSC adipogenesis in freeze-thawed and non-frozen HA based hydrogels on a per-cell basis. Collectively, these findings highlight the cryopreservation protocol as a platform technology that, in addition to contributing to an increased understanding of three-dimensional cell-matrix interactions, could enable the long-term preservation of tissue engineering constructs for clinical applications.     

Stem cell niche: Dynamic neighbor of stem cells

Stem cell niche is a specialized and dynamic microenvironment around the stem cells which plays a critical role in maintaining the stemness properties of stem cells. Over the years, advancement in the research activity has revealed the various important aspects of stem cell niche including cell-cell interaction, cell-extracellular matrix interaction, a large number of soluble signaling factors and various biochemical and biophysical cues (such as oxygen tension, flow, and shear and pore size). Stem cells have the potential to be a powerful tool in regenerative medicine due to their self-renewal property and immense differentiation potential. Recent progresses in in vitro culture conditions of embryonic stem cells, adult stem cells and induced pluripotent stem cells have enabled the researchers to investigate and understand the role of the microenvironment in stem cell properties. The engineered artificial stem cell niche has led to a better execution of stem cells in regenerative medicine. Here we elucidate the key components of stem cell niche and their role in niche engineering and stem cell therapeutics.     

膝关节内干细胞/祖细胞在软骨再生中作用研究进展

关节软骨(AC)由于缺乏血管、神经和淋巴,一旦损伤无法自我修复.虽然以外源性细胞为基础的治疗策略在一定程度上能够再生关节软骨,但仍然存在手术间隔长、供体有限、细胞体外培养易去分化和病原体传播等风险.成人膝关节存在许多类型干细胞/祖细胞(SCPCs),当软骨损伤时,就会被动员,迁移到损伤部位,参与再生修复.因此,基于趋化...     

Molecular mechanism of therapeutic approaches for human gastric cancer stem cells

Gastric cancer(GC)is a primary cause of cancer-related mortality worldwide,and even after therapeutic gastrectomy,survival rates remain poor.The presence of gastric cancer stem cells(GCSCs)is thought to be the major reason for resistance to anticancer treatment(chemotherapy or radiotherapy),and for the development of tumor recurrence,epithelial–mesenchymal transition,and metastases.Additionally,GCSCs have the capacity for self-renewal,differentiation,and tumor initiation.They also synthesize antiapoptotic factors,demonstrate higher performance of drug efflux pumps,and display cell plasticity abilities.Moreover,the tumor microenvironment(TME;tumor niche)that surrounds GCSCs contains secreted growth factors and supports angiogenesis and is thus responsible for the maintenance of the growing tumor.However,the genesis of GCSCs is unclear and exploration of the source of GCSCs is essential.In this review,we provide up-todate information about GCSC-surface/intracellular markers and GCSC-mediated pathways and their role in tumor development.This information will support improved diagnosis,novel therapeutic approaches,and better prognosis using GCSC-targeting agents as a potentially effective treatment choice following surgical resection or in combination with chemotherapy and radiotherapy.To date,most anti-GCSC blockers when used alone have been reported as unsatisfactory anticancer agents.However,when used in combination with adjuvant therapy,treatment can improve.By providing insights into the molecular mechanisms of GCSCs associated with tumors in GC,the aim is to optimize anti-GCSCs molecular approaches for GC therapy in combination with chemotherapy,radiotherapy,or other adjuvant treatment.