ABSTRACT
Once damaged, cartilage has poor intrinsic capacity to repair itself. Current cartilage repair strategies cannot restore the damaged tissue sufficiently. It is hypothesized that biomimetic scaffolds, which can recapitulate important properties of the cartilage extracellular matrix, play a beneficial role in supporting cell behaviors such as growth, cartilage differentiation, and integration with native cartilage, ultimately facilitating tissue recovery. Adipose-derived stem cells regenerated cartilage upon the sequential release of transforming growth factor ß1(TGFß1) and fibroblast growth factor 2(FGF2) using a nanofibrous scaffold, in order to get the recovery of functional cartilage. Experiments in vitro have demonstrated that the release sequence of growth factors FGF2 to TGFß1 is the most essential to promote adipose-derived stem cells into chondrocytes that then synthesize collagen II. Mouse subcutaneous implantation indicated that the treatment sequence of FGF2 to TGFß1 was able to significantly induce multiple increase in cartilage regeneration in vivo. This result demonstrates that the group treated with FGF2 to TGFß1 released from a nanofibrous scaffold provides a good strategy for cartilage regeneration by making a favorable microenvironment for cell growth and cartilage regeneration.
Subject(s)
Cell Differentiation , Fibroblast Growth Factor 2 , Nanofibers , Stem Cells , Tissue Scaffolds , Transforming Growth Factor beta1 , Animals , Fibroblast Growth Factor 2/pharmacology , Transforming Growth Factor beta1/metabolism , Transforming Growth Factor beta1/pharmacology , Mice , Nanofibers/chemistry , Cell Differentiation/drug effects , Tissue Scaffolds/chemistry , Stem Cells/cytology , Stem Cells/drug effects , Stem Cells/physiology , Chondrogenesis/drug effects , Cartilage/drug effects , Cartilage/cytology , Cartilage/physiology , Adipose Tissue/cytology , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/physiology , Cells, Cultured , Tissue Engineering/methodsABSTRACT
Diffuse Intrinsic Pontine Gliomas (DIPGs) are deadly brain cancers in children for which there is no effective treatment. This can partly be attributed to preclinical models that lack essential elements of the in vivo tissue environment, resulting in treatments that appear promising preclinically, but fail to result in effective cures. Recently developed co-culture models combining stem cell-derived brain organoids with brain cancer cells provide tissue dimensionality and a human-relevant tissue-like microenvironment. As these models are technically challenging, we aimed to establish whether interaction with the organoid influences DIPG biology and thus warrants their use. To address this question DIPG24 cells were cultured with pluripotent stem cell-derived cortical organoids. We created "mosaic" co-cultures enriched for tumour cell-neuronal cell interactions versus "assembloid" co-cultures enriched for tumour cell-tumour cell interactions. Sequential window acquisition of all theoretical mass spectra (SWATH-MS) was used to analyse the proteomes of DIPG fractions isolated by flow-assisted cell sorting. Control proteomes from DIPG spheroids were compared with DIPG cells isolated from mosaic and assembloid co-cultures. This suggested changes in cell interaction with the external environment reflected by decreased gene ontology terms associated with adhesion and extracellular matrix, and increased DNA synthesis and replication, in DIPG24 cells under either co-culture condition. By contrast, the mosaic co-culture was associated with neuron-specific brahma-associated factor (nBAF) complex signalling, a process associated with neuronal maturation. We propose that co-culture with brain organoids is a valuable tool to parse the contribution of the brain microenvironment to DIPG tumour biology.
ABSTRACT
Phosphate tailings (PT) was used to reduce the release of heavy metals (HMs) during pyrolysis and the leachable rate of residual HMs, and simultaneously improve the bioavailability of phosphorus in the sludge-based biochar. The concentration of heavy metals and the fractions determined by BCR method was used to investigate the release and the transformation of Zn, Pb, Mn, Ni and Cu during pyrolysis involved with the effects of temperature and the addition of PT. The respective pyrolysis experiments shows that the release of Zn and Pb increases with temperature for both sewage sludge (SS) and PT, and the bioavailable fractions (F1 + F2) of Mn, Ni, and Cu increases with temperature for PT. During co-pyrolysis, blended samples released lower quantities of Zn and Pb and presented lower bioavailability of HMs than the individual SS or PT. A synergistic effect of co-pyrolysis was evident for volatile Zn and Pb. The decomposition of CaMg (CO3)2 from PT produced CaO, by which the volatile ZnCl2 and PbCl2 were transformed into ZnO and PbO with less volatility and higher reactivity with SiO2 and Al2O3 than the chlorides. Then SiO2 and Al2O3 from SS acted as the final stabilizer to immobilize the oxides. The final product combined with SiO2 and Al2O3, such as ZnSiO4 and ZnAl2O4, were detected. The addition of PT also introduced more Ca and P into sludge to produce biochar with higher concentration of apatite phosphorus with higher bioavailability.
Subject(s)
Metals, Heavy , Phosphates , Phosphorus , Pyrolysis , Sewage , Sewage/chemistry , Metals, Heavy/chemistry , Metals, Heavy/analysis , Phosphorus/chemistry , Phosphates/chemistry , Charcoal/chemistryABSTRACT
PURPOSE: To compare the clinical efficacy of two surgical interventions in treating advanced stages TB of the pubis and pubic symphysis. METHODS: Between June 2010 and January 2020, 33 cases of the advanced pubis and pubic symphysis TB were treated with a one-stage debridement procedure (debridement only group, n = 15) or a one-stage debridement with bone grafting and plate fixation procedure (debridement + plating group, n = 18). The visual analog scale (VAS) score, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), operation time, intraoperative blood loss, complications, time of bone graft fusion, and improvement in the mental component summary (MCS) and physical component summary (PCS) of Short Form-36 (SF-36) were compared and analyzed. RESULTS: All patients were followed for 24.9 (SD 1.6) months. All patients were completely cured of the pubis and pubic symphysis TB with no recurrence. There were no significant differences (P >0.05) between the two groups in terms of age, follow-up period and intraoperative blood loss. The post-operative VAS scores, ESR and CRP levels, PCS and MCS scores of two groups significantly improved compared to pre-therapy. The mean operation time in debridement + plating group was 140.9 (43.2) min, which was significantly longer than in debridement only group [94.9(21.8) min, P < 0.01]. The final follow-up (FFU) indices of the VAS score in debridement only group were higher than those in debridement + plating group [1.9 (0.8) vs 1.3 (0.5), P=0.012]. A satisfactory average bony fusion time of 12.2 (3.3) months was achieved in debridement + plating group . CONCLUSIONS: A one-stage debridement, bone grafting, and reconstruction plate fixation procedure achieved reconstruction of the integrity and stability of the pelvic ring, pain relief, and rapid cure of bone TB. This procedure is a safe and effective treatment option for advanced pubis and pubic symphysis TB.
Subject(s)
Pubic Symphysis , Spinal Fusion , Tuberculosis, Spinal , Humans , Adult , Retrospective Studies , Blood Loss, Surgical , Tuberculosis, Spinal/surgery , Pubic Bone , Pubic Symphysis/surgery , Thoracic Vertebrae/surgery , Spinal Fusion/methods , Debridement/methods , Treatment Outcome , Lumbar Vertebrae/surgeryABSTRACT
This retrospective study aimed to compare the clinical efficacy of the posterior procedure with the combined anterior and posterior procedure in the surgical management of lumbar Brucella spondylitis. From January 2015 to June 2020, a total of 62 patients presenting with lumbar Brucella spondylitis underwent either one-stage posterior pedicle fixation, debridement, and interbody fusion (Group A, n = 33) or anterior debridement, bone grafting, and posterior instrumentation (Group B, n = 29). All patients were followed up for an average of 25.4 ± 1.5 months and achieved complete resolution of lumbar Brucella spondylitis. No significant differences between the groups were observed in terms of age or pre-operative, three-month postoperative and final follow-up indices of the VAS, ESR, CRP, lordosis angle, ODI scores, fusion time, and time of serum agglutination test conversion to negative (P > 0.05). Each patient exhibited notable improvements in neurological function, as assessed by the JOA score rating system. Group A demonstrated significantly shorter operative duration, intraoperative blood loss, and hospital stay compared to Group B (P < 0.05). Superficial wound infection was observed in one case in Group A, whereas Group B experienced one case each of intraoperative peritoneal rupture, postoperative ileus, iliac vein injury, and superficial wound infection. This study supports the efficacy of both surgical interventions in the treatment of lumbar Brucella spondylitis, with satisfactory outcomes. However, the posterior approach demonstrated advantages, including reduced surgical time, diminished blood loss, shorter hospital stays, and fewer perioperative complications. Consequently, the one-stage posterior pedicle fixation, debridement, and interbody fusion represent a superior treatment option.
Subject(s)
Brucella , Brucellosis , Spinal Fusion , Spondylitis , Wound Infection , Humans , Adult , Retrospective Studies , Lumbar Vertebrae/surgery , Spinal Fusion/methods , Debridement/methods , Spondylitis/surgery , Brucellosis/surgery , Treatment OutcomeABSTRACT
Advanced oxidation process (AOP) based on peroxymonosulfate (PMS) has aroused extensive discussion in the degradation of organic pollutants due to the strong oxidative ability of SO4â¢-. Great attention has been paid to developing transition metal catalysts for PMS activation. Still, few studies focused on the co-catalysis effect of non-redox metals. To study the co-catalysis of Mg and develop a more efficient metal catalyst, the CoMg2Mn-LDO was prepared by a co-precipitation method accompanied by calcination. The material showed an excellent ability for PMS activation. 97.1% of Orange â ¡ was degraded within 15 min with the reaction rate constant (kobs) of 0.539 min-1 when pH equals 6.7, the dosages of CoMg2Mn-LDO and PMS were 90 mg L-1 and 100 mg L-1, respectively. By contrast, the value of kobs was 0.375 min-1 for the system of Co3Mn-LDO/PMS at the same experimental conditions. The electron paramagnetic resonance (EPR) and quenching experiments results proved the existence of O2â¢-, SO4â¢- and HO⢠in the CoMg2Mn-LDO/PMS system and the dominant role of SO4â¢- in Orange â ¡ degradation. The synergistic effects among Co, Mn, and Mg were found to be responsible for the outstanding catalytic ability of CoMg2Mn-LDO. The presence of Mg could not only promote the formation of Mg-HSO5- and CoOH+ complexes but also reduce the leaching of Co and Mn, which accelerated the generation of free radicals and decreased secondary pollution risk. Based on the overall analysis, reasonable activation mechanisms of PMS and possible degradation pathways of Orange â ¡ in this reaction system were proposed. This work proves that Mg could be applied as an effective co-catalytic element and provides new insight into developing transition metal catalysts for PMS-based AOPs.
Subject(s)
Azo Compounds , Peroxides , Benzenesulfonates , CobaltABSTRACT
Multiple abiotic stress is known as a type of environmental unfavourable condition maximizing the yield and growth gap of crops compared with the optimal condition in both natural and cultivated environments. Rice is the world's most important staple food, and its production is limited the most by environmental unfavourable conditions. In this study, we investigated the pre-treatment of abscisic acid (ABA) on the tolerance of the IAC1131 rice genotype to multiple abiotic stress after a 4-day exposure to combined drought, salt and extreme temperature treatments. A total of 3285 proteins were identified and quantified across the four treatment groups, consisting of control and stressed plants with and without pre-treatment with ABA, with 1633 of those proteins found to be differentially abundant between groups. Compared with the control condition, pre-treatment with the ABA hormone significantly mitigated the leaf damage against combined abiotic stress at the proteome level. Furthermore, the application of exogenous ABA did not affect the proteome profile of the control plants remarkably, while the results were different in stress-exposed plants by a greater number of proteins changed in abundance, especially those which were increased. Taken together, these results suggest that exogenous ABA has a potential priming effect for enhancing the rice seedlings' tolerance against combined abiotic stress, mainly by affecting stress-responsive mechanisms dependent on ABA signalling pathways in plants.
Subject(s)
Abscisic Acid , Oryza , Abscisic Acid/pharmacology , Abscisic Acid/metabolism , Oryza/genetics , Proteome/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Stress, Physiological/genetics , Gene Expression Regulation, Plant , Plants, Genetically Modified/genetics , DroughtsABSTRACT
Photoreceptor cells are highly susceptible to oxidative-stress-induced damage due to their high metabolic rate. Oxidative stress plays a key role in driving pathological events in several different ocular diseases, which lead to retinal degeneration and ultimately blindness. A growing number of studies have been performed to understand downstream events caused by ROS induced oxidative stress in photoreceptor cells; however, the underlying mechanisms of ROS toxicity are not fully understood. To shed light on ROS induced downstream pathological events, we employed a tandem mass tag (TMT) labelling-based quantitative mass-spectrometric approach to determine proteome changes in 661W photoreceptor cells following oxidative stress induction via the application of different concentrations of H2O2 at different time points. Overall, 5920 proteins were identified and quantified, and 450 differentially expressed proteins (DEPs) were identified, which were altered in a dose and time dependent manner in all treatment groups compared to the control group. These proteins were involved in several biological pathways, including spliceosome and ribosome response, activated glutathione metabolism, decreased ECM-receptor interaction, oxidative phosphorylation, abnormally regulated lysosome, apoptosis, and ribosome biogenesis. Our results highlighted ECM receptor interaction, oxidative phosphorylation and spliceosome pathways as the major targets of oxidative stress that might mediate vascular dysfunction and cellular senescence.
ABSTRACT
Polydatin is an active polyphenol displaying multifaceted benefits. Recently, growing studies have noticed its potential therapeutic effects on bone and joint disorders (BJDs). Therefore, this article reviews recent in vivo and in vitro progress on the protective role of polydatin against BJDs. An insight into the underlying mechanisms is also presented. It was found that polydatin could promote osteogenesis in vitro, and symptom improvements have been disclosed with animal models of osteoporosis, osteosarcoma, osteoarthritis and rheumatic arthritis. These beneficial effects obtained in laboratory could be mainly attributed to the bone metabolism-regulating, anti-inflammatory, antioxidative, apoptosis-regulating and autophagy-regulating functions of polydatin. However, studies on human subjects with BJDs that can lead to early identification of the clinical efficacy and adverse effects of polydatin have not been reported yet. Accordingly, this review serves as a starting point for pursuing clinical trials. Additionally, future emphasis should also be devoted to the low bioavailability and prompt metabolism nature of polydatin. In summary, well-designed clinical trials of polydatin in patients with BJD are in demand, and its pharmacokinetic nature must be taken into account.
ABSTRACT
INTRODUCTION: Previous research has suggested that vanishing white matter disease (VWMD) astrocytes fail to fully differentiate and respond differently to cellular stresses compared to healthy astrocytes. However, few studies have investigated potential VWMD therapeutics in monoculture patient-derived cell-based models. METHODS: To investigate the impact of alterations in astrocyte expression and function in VWMD, astrocytes were differentiated from patient and control induced pluripotent stem cells and analyzed by proteomics, pathway analysis, and functional assays, in the absence and presence of stressors or potential therapeutics. RESULTS: Vanishing white matter disease astrocytes demonstrated significantly reduced expression of astrocyte markers and markers of inflammatory activation or cellular stress relative to control astrocytes. These alterations were identified both in the presence and absence of polyinosinic:polycytidylic acid stimuli, which is used to simulate viral infections. Pathway analysis highlighted differential signaling in multiple pathways in VWMD astrocytes, including eukaryotic initiation factor 2 (EIF2) signaling, oxidative stress, oxidative phosphorylation (OXPHOS), mitochondrial function, the unfolded protein response (UPR), phagosome regulation, autophagy, ER stress, tricarboxylic acid cycle (TCA) cycle, glycolysis, tRNA signaling, and senescence pathways. Since oxidative stress and mitochondrial function were two of the key pathways affected, we investigated whether two independent therapeutic strategies could ameliorate astrocyte dysfunction: edaravone treatment and mitochondrial transfer. Edaravone treatment reduced differential VWMD protein expression of the UPR, phagosome regulation, ubiquitination, autophagy, ER stress, senescence, and TCA cycle pathways. Meanwhile, mitochondrial transfer decreased VWMD differential expression of the UPR, glycolysis, calcium transport, phagosome formation, and ER stress pathways, while further modulating EIF2 signaling, tRNA signaling, TCA cycle, and OXPHOS pathways. Mitochondrial transfer also increased the gene and protein expression of the astrocyte marker, glial fibrillary acidic protein (GFAP) in VWMD astrocytes. CONCLUSION: This study provides further insight into the etiology of VWMD astrocytic failure and suggests edaravone and mitochondrial transfer as potential candidate VWMD therapeutics that can ameliorate disease pathways in astrocytes related to oxidative stress, mitochondrial dysfunction, and proteostasis.
Subject(s)
Leukoencephalopathies , White Matter , Humans , Astrocytes/metabolism , Edaravone/metabolism , Eukaryotic Initiation Factor-2/metabolism , Leukoencephalopathies/genetics , Mitochondria/metabolism , White Matter/metabolismABSTRACT
In this protocol we describe our workflow for analyzing complex, multi-condition quantitative proteomic experiments, with the aim to extract biological insights. The tool we use is an R package, PloGO2, contributed to Bioconductor, which we can optionally precede by running correlation network analysis with WGCNA. We describe the data required and the steps we take, including detailed code examples and outputs explanation. The package was designed to generate gene ontology or pathway summaries for many data subsets at the same time, visualize protein abundance summaries for each biological category examined, help determine enriched protein subsets by comparing them all to a reference set, and suggest key highly correlated hub proteins, if the optional network analysis is employed.
Subject(s)
Proteins , Proteomics , Proteomics/methods , Gene Ontology , WorkflowABSTRACT
Bone defect repair remains a challenge in orthopedics. This study describes the development and potential effectiveness of vascular endothelial growth factor (VEGF)/bone morphogenetic protein-2 (BMP-2) shell-core microspheres for promoting bone regeneration. Poly(L-lactic acid)/polylactic-co-glycolic acid (PLLA/PLGA) core-shell microspheres loaded with VEGF and BMP-2 were prepared by a coaxial electrospray technique, and their surface morphology, core-shell distribution, and particle size were examined. Different groups of microspheres were prepared with different placement of the growth factors, and the encapsulation efficiency and in vitro release curves were measured. Additionally, the effects of the different groups of microspheres on the proliferation and differentiation of osteoblasts and vascular endothelial cells were investigated. The prepared microspheres had a core-shell structure with good homogeneity and dispersion, a clear boundary, and a smooth surface. On scanning electron microscopy, the mean diameter of the microspheres was similar for all six preparations (P > 0.05). During in vitro release, growth factor was initially released via a brief burst release from the outer shell of the microsphere followed by a slower sustained release. The release of growth factors from the inner core remained relatively slow and sustained. Sequential release of different growth factors was achieved through the inconsistent release rates from the microsphere shell and inner core. All groups of microspheres showed no cytotoxicity, good biocompatibility, and the ability to promote osteoblast proliferation. The microspheres loaded with BMP-2 also promoted osteoblast differentiation, and VEGF-loaded microspheres promoted the proliferation and differentiation of vascular endothelial cells. The BMP-2 (core)/VEGF (shell) microsphere group best promoted osteoblast differentiation. The microspheres prepared in this study exhibited slow sequential release of BMP-2 and VEGF and showed good biocompatibility along with the ability to promote osteoblast differentiation and vascular endothelial cell proliferation.
Subject(s)
Endothelial Cells , Vascular Endothelial Growth Factor A , Microspheres , Delayed-Action Preparations/pharmacology , Vascular Endothelial Growth FactorsABSTRACT
The repair and reconstruction of bone defects remain a challenge in orthopedics. The present study offers a solution to this problem by developing a vascular endothelial growth factor (VEGF)/bone morphogenetic protein 2 (BMP-2) shell-core microspheres loaded on 3D-printed porous titanium alloy via gelatin coating to prepare a titanium-alloy microsphere scaffold release system. The composite scaffold was characterized via scanning electron microscope (SEM) and energy disperse spectroscopy (EDS), and the effect of the composite scaffold on the adhesion, proliferation, and differentiation of osteoblasts were determined in vitro. Furthermore, a rabbit femoral defect model was established to verify the effect of the composite scaffold on osteogenesis and bone formation in vivo. The results demonstrated that the composite scaffold could release VEGF and BMP-2 sequentially. Meanwhile, the composite scaffold significantly promoted osteoblast adhesion, proliferation, and differentiation (p < 0.05) compared to pure titanium alloy scaffolds in vitro. Furthermore, the composite scaffold can exhibit significant osteogenic differentiation (p < 0.05) than gelatin-coated titanium alloy scaffolds. The in vivo X-rays demonstrated that the implanted scaffolds were in a good position, without inflammation and infection. Micro-CT and quantitative results of new bone growth illustrated that the amount of new bone in the composite scaffold is significantly higher than that of the gelatin-coated and pure titanium alloy scaffolds (p < 0.05). Similarly, the fluorescence labeling and V-G staining of hard tissue sections indicated that the bone integration capacity of the composite scaffold was significantly higher than the other two groups (p < 0.05). This research suggests that VEGF/BMP-2 shell-core microspheres loaded on 3D-printed titanium alloy porous scaffold through gelatin hydrogel coating achieved the sequential release of VEGF and BMP-2. Most importantly, the in vitro and in vivo study findings have proven that the system could effectively promote osteogenic differentiation and osseointegration.
ABSTRACT
This retrospective study aimed to evaluate midterm outcomes of surgical management of multi-segment thoracic spinal tuberculosis by single-stage posterior transforaminal debridement and interbody fusion with instrumentation. From January 2007 to October 2015, 42 adult patients with thoracic spinal tuberculosis involving three or more levels underwent single-stage posterior transforaminal debridement, interbody fusion and instrumentation At a mean follow-up of 73.5 ± 9.6 months, all patients were eligible for final evaluation. All displayed improved biochemical markers and pain scores at 3 months and improved physiologic levels at the end of treatment. Visual analogue and 36-Item Short-Form Health Survey scores were significantly improved compared with preoperative values. All 30 patients with preoperative neurological deficits experienced neurologic improvement. Thoracic kyphosis angle decreased significantly from 34.4° ± 4.5° to 22.0° ± 2.6°. A mean kyphotic angle loss of 1.7° ± 1.1° was recorded at the final follow-up, and bone fusion was observed at a mean of 10.6 ± 2.1 months, with no instrumentation failures. One patient experienced delayed incisional healing and five patients suffered postoperative intercostal neuralgia that were cured by conservative treatment. There were no graft failures or implant breakages. This study showed the utility of a single-staged procedure combining posterior transforaminal debridement and interbody fusion with instrumentation, and demonstrated promising results.
Subject(s)
Kyphosis , Spinal Fusion , Tuberculosis, Spinal , Adult , Humans , Tuberculosis, Spinal/surgery , Follow-Up Studies , Debridement/methods , Retrospective Studies , Thoracic Vertebrae/surgery , Spinal Fusion/methods , Kyphosis/surgery , Treatment Outcome , Lumbar Vertebrae/surgeryABSTRACT
The effect of titanium scaffold geometry on the bone regeneration ability of the scaffold remains unclear. Here, selective laser melting as a 3D printing technology was used to create porous titanium alloy scaffolds with two unit structures: a hollow hexagonal prism (group A) and a hollow triangular prism (group B). The structures and morphologies of the scaffolds were characterized before mechanical properties were simulated. Cell adhesion behaviors, osteoblast activity and proliferation, and alkaline phosphatase (ALP) activity were evaluated, in addition to in vivo testing in an animal model. The results showed that the two scaffolds made of Ti6Al4V had compression moduli similar to that of human cortical bone (116.91 ± 0.01 and 174.29 ± 2.21 MPa vs. 89-164 MPa). The two scaffolds were nontoxic to cells and had good biocompatibility, while group A scaffolds facilitated cell adhesion. The number of cells increased gradually in culture. The ALP activity of cells on group A scaffolds demonstrated higher osteogenic ability than that of group B scaffolds. The in vivo tests showed that all scaffolds retained their shape well after implantation, and no obvious inflammatory reaction or infection in surrounding tissues was found. Based on fluorescence staining, mature new bone formation was found at week 12. Group A scaffolds showed better bone integration ability compared with group B scaffolds. The percentage of new bone area in group A (7.5%) was higher than that in group B (6.5%). This research suggests that the hollow hexagonal prism structure of porous scaffolds can promote osteogenic differentiation and osseointegration better than the triangular prism structure.
Subject(s)
Osteogenesis , Titanium , Alloys/pharmacology , Animals , Humans , Porosity , Printing, Three-Dimensional , Tissue Scaffolds/chemistry , Titanium/chemistry , Titanium/pharmacologyABSTRACT
Drought is responsible for major losses in rice production. Root tips contain meristematic and elongation zones that play major roles in determination of root traits and adaptive strategies to drought. In this study we analysed two contrasting genotypes of rice: IR64, a lowland, drought-susceptible, and shallow-rooting genotype; and Azucena, an upland, drought-tolerant, and deep-rooting genotype. Samples were collected of root tips of plants grown under control and water deficit stress conditions. Quantitative proteomics analysis resulted in the identification of 7294 proteins from the root tips of IR64 and 6307 proteins from Azucena. Data are available via ProteomeXchange with identifier PXD033343. Using a Partial Least Square Discriminant Analysis on 4170 differentially abundant proteins, 1138 statistically significant proteins across genotypes and conditions were detected. Twenty two enriched biological processes showing contrasting patterns between two genotypes in response to stress were detected through gene ontology enrichment analysis. This included identification of novel proteins involved in root elongation with specific expression patterns in Azucena, including four Expansins and seven Class III Peroxidases. We also detected an antioxidant network and a metallo-sulfur cluster assembly machinery in Azucena, with roles in reactive oxygen species and iron homeostasis, and positive effects on root cell cycle, growth and elongation.
Subject(s)
Oryza , Oryza/metabolism , Droughts , Proteomics , Meristem/genetics , Gene Expression Regulation, Plant , Genotype , Plant Roots/genetics , Plant Roots/metabolismABSTRACT
OBJECTIVE: This study investigated the molecular mechanism of whether hUC-MSCs-EVs repressed PTEN expression and activated the PI3K/AKT pathway through miR-29b-3p, thus inhibiting LPS-induced neuronal injury. METHODS: hUC-MSCs were cultured and then identified. Cell morphology was observed. Alizarin red, oil red O, and alcian blue staining were used for inducing osteogenesis, adipogenesis, and chondrogenesis. EVs were extracted from hUC-MSCs and identified by transmission electron microscope observation and Western blot. SCI neuron model was established by 24h lipopolysaccharide (LPS) induction. After the cells were cultured with EVs without any treatment, uptake of EVs by SCI neurons, miR-29b-3p expression, cell viability, apoptosis, caspase-3, cleaved caspase-3, caspase 9, Bcl-2, PTEN, PI3K, AKT, and p-Akt protein levels, caspase 3 and caspase 9 activities, and inflammatory factors IL-6 and IL-1ß levels were detected by immunofluorescence labeling, RT-qPCR, MTT, flow cytometry, Western blot, caspase 3 and caspase 9 activity detection kits, and ELISA. The binding sites between PTEN and miR-29b-3p were predicted by the database and verified by dual-luciferase assay. RESULTS: LPS-induced SCI cell model was successfully established, and hUC-MSCs-EVs inhibited LPS-induced apoptosis of injured spinal cord neurons. EVs transferred miR-29b-3p into LPS-induced injured neurons. miR-29b-3p silencing reversed EV effects on reducing LPS-induced neuronal apoptosis. miR-29b-3p reduced LPS-induced neuronal apoptosis by targeting PTEN. After EVs-miR-inhi and si-PTEN treatment, inhibition of the PI3K/AKT pathway reversed hUC-MSCs-EVs effects on reducing LPS-induced neuronal apoptosis. CONCLUSION: hUC-MSCs-EVs activated the PI3K/AKT pathway by carrying miR-29b-3p into SCI neurons and silencing PTEN, thus reducing neuronal apoptosis.
Subject(s)
Extracellular Vesicles , Mesenchymal Stem Cells , MicroRNAs , Spinal Cord Injuries , Alcian Blue/metabolism , Alcian Blue/pharmacology , Apoptosis , Caspase 3/metabolism , Caspase 9/metabolism , Caspase 9/pharmacology , Extracellular Vesicles/metabolism , Humans , Interleukin-6/metabolism , Lipopolysaccharides/pharmacology , MicroRNAs/genetics , MicroRNAs/metabolism , Neurons/metabolism , PTEN Phosphohydrolase , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Spinal Cord Injuries/genetics , Spinal Cord Injuries/metabolism , Spinal Cord Injuries/therapy , Umbilical Cord/metabolismABSTRACT
Rice crops are often subject to multiple abiotic stresses simultaneously in both natural and cultivated environments, resulting in yield reductions beyond those expected from single stress. We report physiological changes after a 4 day exposure to combined drought, salt and extreme temperature treatments, following a 2 day salinity pre-treatment in two rice genotypes-Nipponbare (a paddy rice) and IAC1131 (an upland landrace). Stomata closed after two days of combined stresses, causing intercellular CO2 concentrations and assimilation rates to diminish rapidly. Abscisic acid (ABA) levels increased at least five-fold but did not differ significantly between the genotypes. Tandem Mass Tag isotopic labelling quantitative proteomics revealed 6215 reproducibly identified proteins in mature leaves across the two genotypes and three time points (0, 2 and 4 days of stress). Of these, 987 were differentially expressed due to stress (cf. control plants), including 41 proteins that changed significantly in abundance in all stressed plants. Heat shock proteins, late embryogenesis abundant proteins and photosynthesis-related proteins were consistently responsive to stress in both Nipponbare and IAC1131. Remarkably, even after 2 days of stress there were almost six times fewer proteins differentially expressed in IAC1131 than Nipponbare. This contrast in the translational response to multiple stresses is consistent with the known tolerance of IAC1131 to dryland conditions.
Subject(s)
Oryza/physiology , Stress, Physiological/physiology , Abscisic Acid/metabolism , Gases/metabolism , Gene Expression Regulation, Plant , Genotype , Oryza/genetics , Photosynthesis , Plant Leaves/metabolism , Plant Proteins/metabolism , Proteome/metabolism , Proteomics , Stress, Physiological/geneticsABSTRACT
Following recent advances in osteoimmunology, there is growing recognition of the vital role of immune cells in the osteogenesis process. The 3D-printed scaffold, as a substitute for injured and/or diseased bone tissues, has demonstrated satisfactory pro-osteogenetic performance. However, whether immune cells prompt the above pro-osteogenetic performance has not been elucidated in detail. In the present study, highly controllable Ti-6Al-4V scaffolds with different pore geometries were fabricated using a selective laser-melting technique, to reveal their osteoimmunological functions with macrophages. The results showed that macrophages displayed characteristics of M2 phenotype in response to scaffolds. As a result, an anti-inflammatory microenvironment was generated. When the pore geometry was considered, such observations were more apparent with the hexagonal pore scaffold than with the triangular one. In addition, inhibition of the toll-like receptor signaling pathway in macrophages has been proposed to cause the above phenomenon. Upon applying conditioned media derived from macrophages on pre-osteoblasts, the hexagonal pore scaffold group was found to significantly enhance osteoblastic differentiation, via macrophage-to-implant interactions. However, the effect of triangular pore scaffold was not statistically significant compared to that of hexagonal pore scaffolds or nonporous samples. In an attempt to quantify scaffold pore geometries, it was suggested that pores with higher circularity values tended to induce M2 polarization of macrophages, promote an anti-inflammatory milieu, and therefore, achieve better osteogenetic performance via immunomodulation.
Subject(s)
Osteogenesis , Titanium , Alloys , Lasers , Macrophages , Porosity , Tissue Scaffolds , Titanium/pharmacologyABSTRACT
The preconditioning of human sperm with sublethal nitrosative stress before cryopreservation can potentially improve the thawed sperm quality. However, the underlying mechanisms behind this protective strategy are not entirely understood. We compared the cryosurvival of human sperm exposed to 0.01 µM nitric oxide (NO) throughout the cryopreservation and used multiplexed quantitative proteomics approach to identify changes in the proteome profile of preconditioned sperm cells. Semen samples were obtained from 30 normospermia donors and then each sample was divided into three equal parts: fresh (F), frozen-control (C), and frozen exposed to nitric oxide (NO). The sperm undergoing mild sublethal stress showed higher values for motility and viability compared to the frozen control sperm. Moreover, out of 2912 identified proteins, 248 proteins were detected as differentially abundant proteins (DAPs) between cryopreserved groups and fresh group (F) (p < 0.05). Gene ontology (GO) analysis of differentially abundant proteins indicated that the abundance of proteins associated with glycolysis, gluconeogenesis, and fertilization processes was reduced while oxidative phosphorylation pathway was increased in abundance in cryopreserved sperm compared to the fresh sperm. Moreover, redox protein such as thioredoxin 17 was increased in abundance in the NO group compared to the control freezing group. Therefore, the pre-conditioning of sperm prior to cryopreservation may play an important role in maintaining the redox balance in mitochondria of sperm after freezing. Overall, our results indicate that arylsulfatase A (ARSA), serine protease 37 (PRSS37), and sperm surface protein (SP17) may potentially serve as protein biomarkers associated with screening the fertilization potential of the thawed sperm.