Nanofibrous Spongy Microspheres for the Delivery of Hypoxia-primed Human Dental Pulp Stem Cells to Regenerate Vascularized Dental Pulp

Dental pulp infection and necrosis are widespread diseases. Conventional endodontic treatments result in a devitalized and weakened tooth. In this work, we synthesized novel star-shaped polymer to self-assemble into unique nanofibrous spongy microspheres (NF-SMS), which were used to carry human dental pulp stem cells (hDPSCs) into the pulp cavity to regenerate living dental pulp tissues. It was found that NF-SMS significantly enhanced hDPSCs attachment, proliferation, odontogenic differentiation and angiogenesis, as compared to control cell carriers. Additionally, NF-SMS promoted vascular endothelial growth factor (VEGF) expression of hDPSCs in a 3D hypoxic culture. Hypoxia-primed hDPSCs/NF-SMS complexes were injected into the cleaned pulp cavities of rabbit molars for subcutaneous implantation in mice. After 4 weeks, the hypoxia group significantly enhanced angiogenesis inside the pulp chamber and promoted the formation of ondontoblast-like cells lining along the dentin-pulp interface, as compared to the control groups (hDPSCs alone group, NF-SMS alone group, and hDPSCs/NF-SMS group pre-cultured under normoxic conditions). Furthermore, in an in situ dental pulp repair model in rats, hypoxia-primed hDPSCs/NF-SMS were injected to fully fill the pulp cavity and regenerate pulp-like tissues with a rich vasculature and a histological structure similar to the native pulp

Melatonin Improved Waterlogging Tolerance in Alfalfa (Medicago sativa) by Reprogramming Polyamine and Ethylene Metabolism

Melatonin (MT), polyamines (PAs), and ethylene have been suggested to play key roles in plant growth and development in response to environmental abiotic stresses. However, the effect of melatonin on polyamine and ethylene metabolism under waterlogging stress has rarely been elucidated. The main purpose of this study was to investigate the effect of melatonin pretreatment on waterlogging stress in alfalfa. The experiment was arranged into four treatment groups control with water pretreatment (CK-MT), control with melatonin pretreatment (CK+MT), waterlogging pretreated with water (WL-MT) and waterlogging pretreated with melatonin (WL+MT), with three replications. Six-week-old alfalfa seedlings were pretreated with 100 μM melatonin and exposed to waterlogging stress for 10 days. Plant growth rate, different physiological characteristics, and gene expression level were measured. Results showed that waterlogging induced melatonin accumulation, and melatonin pretreatment increased endogenous MT levels for the control and water-logged plants. Waterlogging stress caused a significant reduction in plant growth, chlorophyll content, photochemical efficiency (Fv/Fm) and net photosynthetic rate (Pn), while also causing increased leaf electrolyte leakage (EL) and malondialdehyde (MDA) content. Pretreatment with melatonin alleviated the waterlogging-induced damage and reduction in plant growth, chlorophyll content, Fv/Fm and Pn. Waterlogging stress significantly increased leaf polyamines (Put, Spd, Spm) and ethylene levels, and the increased PAs and ethylene levels are coupled with higher metabolic enzymes and gene expressions. While pretreatment with melatonin further increased Put, Spd and Spm levels, it also decreased ethylene levels under waterlogging, and those increased PAs levels or decreased ethylene levels are regulated by the metabolic enzymes and gene expressions. The results in this study provide more comprehensive insight into the physiological and molecular mechanisms of melatonin-improved waterlogging tolerance in alfalfa. Furthermore, they suggested that melatonin improved waterlogging tolerance in alfalfa at least partially by reprogramming ethylene and PA biosynthesis, attributable to the increased PAs and decreased ethylene levels, which leads to more enhanced membrane stability and photosynthesis as well as less leaf senescence caused by ethylene

Nanofibrous Spongy Microspheres Enhance Odontogenic Differentiation of Human Dental Pulp Stem Cells

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/113700/1/adhm201500308.pd

Differential responses of CO2 assimilation, carbohydrate allocation and gene expression to NaCl stress in perennial ryegrass with different salt tolerance.

Little is known about the effects of NaCl stress on perennial ryegrass (Lolium perenne L.) photosynthesis and carbohydrate flux. The objective of this study was to understand the carbohydrate metabolism and identify the gene expression affected by salinity stress. Seventy-four days old seedlings of two perennial ryegrass accessions (salt-sensitive 'PI 538976' and salt-tolerant 'Overdrive') were subjected to three levels of salinity stress for 5 days. Turf quality in all tissues (leaves, stems and roots) of both grass accessions negatively and significantly correlated with GFS (Glu+Fru+Suc) content, except for 'Overdrive' stems. Relative growth rate (RGR) in leaves negatively and significantly correlated with GFS content in 'Overdrive' (P0.05 for turf quality). A greater up-regulation in the expression of SPS, SS, SI, 6-SFT gene was observed in 'Overdrive' than 'PI 538976'. A higher level of SPS and SS expression in leaves was found in 'PI 538976' relative to 'Overdrive'. Accumulation of hexoses in roots, stems and leaves can induce a feedback repression to photosynthesis in salt-stressed perennial ryegrass and the salt tolerance may be changed with the carbohydrate allocation in leaves and stems

Comparative Transcriptome Combined with Proteome Analyses Revealed Key Factors Involved in Alfalfa (Medicago sativa) Response to Waterlogging Stress

Alfalfa (Medicago sativa) is the most widely grown and most important forage crop in the world. However, alfalfa is susceptible to waterlogging stress, which is the major constraint for its cultivation area and crop production. So far, the molecular mechanism of alfalfa response to the waterlogging is largely unknown. Here, comparative transcriptome combined with proteomic analyses of two cultivars (M12, tolerant; M25, sensitive) of alfalfa showing contrasting tolerance to waterlogging were performed to understand the mechanism of alfalfa in response to waterlogging stress. Totally, 748 (581 up- and 167 down-regulated) genes were differentially expressed in leaves of waterlogging-stressed alfalfa compared with the control (M12_W vs. M12_CK), whereas 1193 (740 up- and 453 down-regulated) differentially abundant transcripts (DATs) were detected in the leaves of waterlogging-stressed plants in comparison with the control plants (M25_W vs. M25_CK). Furthermore, a total of 187 (122 up- and 65 down-regulated) and 190 (105 up- and 85 down-regulated) differentially abundant proteins (DAPs) were identified via isobaric tags for relative and absolute quantification (iTRAQ) method in M12_W vs. M12_CK and M25_W vs. M25_CK comparison, respectively. Compared dataset analysis of proteomics and transcriptomics revealed that 27 and eight genes displayed jointly up-regulated or down-regulated expression profiles at both mRNA and protein levels in M12_W vs. M12_CK comparison, whereas 30 and 27 genes were found to be co-up-regulated or co-down-regulated in M25_W vs. M25_CK comparison, respectively. The strongly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways for co-up-regulated genes at mRNA and protein levels in M12_W vs. M12_CK comparison were ‘Amino sugar and nucleotide sugar metabolism’, ‘Arginine and proline metabolism’ and ‘Starch and sucrose metabolism’, whereas co-up-regulated protein-related pathways including ‘Arginine and proline metabolism’ and ‘Valine, leucine and isoleucine degradation’ were largely enriched in M25_W vs. M25_CK comparison. Importantly, the identified genes related to beta-amylase, Ethylene response Factor (ERF), Calcineurin B-like (CBL) interacting protein kinases (CIPKs), Glutathione peroxidase (GPX), and Glutathione-S-transferase (GST) may play key roles in conferring alfalfa tolerance to waterlogging stress. The present study may contribute to our understanding the molecular mechanism underlying the responses of alfalfa to waterlogging stress, and also provide important clues for further study and in-depth characterization of waterlogging-resistance breeding candidate genes in alfalfa

Effects of fertilization and clipping on carbon, nitrogen storage, and soil microbial activity in a natural grassland in southern China.

Grassland managements can affect carbon (C) and nitrogen (N) storage in grassland ecosystems with consequent feedbacks to climate change. We investigated the impacts of compound fertilization and clipping on grass biomass, plant and soil (0-20 cm depth) C, N storage, plant and soil C: N ratios, soil microbial activity and diversity, and C, N sequestration rates in grassland in situ in the National Dalaoling Forest Park of China beginning July, 2011. In July, 2012, the fertilization increased total biomass by 30.1%, plant C by 34.5%, plant N by 79.8%, soil C by 18.8% and soil N by 23.8% compared with the control, respectively. Whereas the clipping decreased total biomass, plant C and N, soil C and N by 24.9%, 30.3%, 39.3%, 18.5%, and 19.4%, respectively, when compared to the control. The plant C: N ratio was lower for the fertilization than for the control and the clipping treatments. The soil microbial activity and diversity indices were higher for the fertilization than for the control. The clipping generally exhibited a lower level of soil microbial activity and diversity compared to the control. The principal component analysis indicated that the soil microbial communities of the control, fertilization and clipping treatments formed three distinct groups. The plant C and N sequestration rates of the fertilization were significantly higher than the clipping treatment. Our results suggest that fertilization is an efficient management practice in improving the C and N storage of the grassland ecosystem via increasing the grass biomass and soil microbial activity and diversity

Differential physiological and metabolic response to low temperature in two zoysiagrass genotypes native to high and low latitude.

Low temperature is one of the important limiting factors for growing season and geographical distribution of plants. Zoysiagrass (Zoysia Willd) is one of the widely used warm-season turfgrass that is distribute in many parts of the world. Zoysaigrass native to high latitude may have evolved higher cold tolerance than the ones native to low latitude. The objective of this study was to investigate the cold stress response in zoysiagrass native to diverse latitude at phenotypic, physiological and metabolic levels. Two zoysiagrass (Z. japonica) genotypes, Latitude-40 (higher latitude) and Latitude-22 (lower latitude) were subjected to four temperature treatments (optimum, 30/25°C, day/night; suboptimum, 18/12°C; chilling, 8/2°C; freezing, 2/-4°C) progressively in growth chambers. Low temperature (chilling and freezing) increased leaf electrolyte leakage (EL) and reduced plant growth, turf quality, chlorophyll (Chl) content, photochemical efficiency (Fv/Fm) and photosynthesis (Pn, net photosynthetic rate; gs, stomatal conductance; intercellular CO2; Tr, transpiration rate) in two genotypes, with more rapid changes in Latitude-22. Leaf carbohydrates content (glucose, fructose, sucrose, trehalose, fructan, starch) increased with the decreasing of temperature, to a great extend in Latitude-40. Leaf abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) content increased, while indole-3-acetic acid (IAA), gibberellic acid (GA3) and trans-zeatin ribside (t-ZR) content decreased with the reduction of temperature, with higher content in Latitude-40 than in Latitude-22. Chilling and freezing induced the up-regulation of C-repeat binding factor (ZjCBF), late embryogenesis abundant (ZjLEA3) and dehydration-responsive element binding (ZjDREB1) transcription factors in two genotypes, whereas those genes exhibited higher expression levels in Latitude-40, particularly under freezing temperature. These results suggested that zoysiagrass native to higher latitude exhibited higher freezing tolerance may attribute to the higher carbohydrates serving as energy reserves and stress protectants that stabilize cellular membranes. The phytohormones may serve signals in regulating plant growth, development and adaptation to low temperature as well as inducing the up-regulated ZjCBF, ZjLEA3 and ZjDREB1 expressions thus result in a higher cold tolerance

Effects of cadmium exposure on growth and metabolic profile of bermudagrass [Cynodon dactylon (L.) Pers].

Metabolic responses to cadmium (Cd) may be associated with variations in Cd tolerance in plants. The objectives of this study were to examine changes in metabolic profiles in bermudagrass in response to Cd stress and to identify predominant metabolites associated with differential Cd tolerance using gas chromatography-mass spectrometry. Two genotypes of bermudagrass with contrasting Cd tolerance were exposed to 0 and 1.5 mM CdSO4 for 14 days in hydroponics. Physiological responses to Cd were evaluated by determining turf quality, growth rate, chlorophyll content and normalized relative transpiration. All these parameters exhibited higher tolerance in WB242 than in WB144. Cd treated WB144 transported more Cd to the shoot than in WB242. The metabolite analysis of leaf polar extracts revealed 39 Cd responsive metabolites in both genotypes, mainly consisting of amino acids, organic acids, sugars, fatty acids and others. A difference in the metabolic profiles was observed between the two bermudagrass genotypes exposed to Cd stress. Seven amino acids (norvaline, glycine, proline, serine, threonine, glutamic acid and gulonic acid), four organic acids (glyceric acid, oxoglutaric acid, citric acid and malic acid,) and three sugars (xylulose, galactose and talose) accumulated more in WB242 than WB144. However, compared to the control, WB144 accumulated higher quantities of sugars than WB242 in the Cd regime. The differential accumulation of these metabolites could be associated with the differential Cd tolerance in bermudagrass