Metabolic utilization of human osteoblast cell line hFOB 1.19 under normoxic and hypoxic conditions: A phenotypic microarray analysis

2021 ◽  
pp. 153537022098546
Author(s):  
Yan Chao Cui ◽  
Yu Sheng Qiu ◽  
Qiong Wu ◽  
Gang Bu ◽  
Amira Peli ◽  
...  
Keyword(s):  
Cell Growth ◽  
Bone Regeneration ◽  
Bone Repair ◽  
Hypoxic Condition ◽  
Human Osteoblast ◽  
Osteoblast Cells ◽  
Hypoxic Conditions ◽  
New Findings ◽  
Phenotype Microarrays ◽  
Oxygen Concentrations

Osteoblasts play an important role in bone regeneration and repair. The hypoxia condition in bone occurs when bone undergoes fracture, and this will trigger a series of biochemical and mechanical changes to enable bone repair. Hence, it is interesting to observe the metabolites and metabolism changes when osteoblasts are exposed to hypoxic condition. This study has looked into the response of human osteoblast hFOB 1.19 under normoxic and hypoxic conditions by observing the cell growth and utilization of metabolites via Phenotype MicroArrays™ under these two different oxygen concentrations. The cell growth of hFOB 1.19 under hypoxic condition showed better growth compared to hFOB 1.19 under normal condition. In this study, osteoblast used glycolysis as the main pathway to produce energy as hFOB 1.19 in both hypoxic and normoxic conditions showed cell growth in well containing dextrin, glycogen, maltotriose, D-maltose, D-glucose-6-phospate, D-glucose, D-mannose, D-Turanose, D-fructose-6-phosphate, D-galactose, uridine, adenosine, inosine and α-keto-glutaric acid. In hypoxia, the cells have utilized additional metabolites such as α-D-glucose-1-phosphate and D-fructose, indicating possible activation of glycogen synthesis and glycogenolysis to metabolize α-D-glucose-1-phosphate. Meanwhile, during normoxia, D-L-α-glycerol phosphate was used, and this implies that the osteoblast may use glycerol-3-phosphate shuttle and oxidative phosphorylation to metabolize glycerol-3-phosphate. Impact statement Currently, researchers understand that bone cells experience hypoxia during bone injury or fracture. Such stress condition exerts effect on bone regeneration and repair. However, there is limited knowledge on the metabolites and metabolism changes that occur in osteoblast cells when they undergo inherent regeneration and repair under hypoxia. This manuscript describes the use of Phenotype MicroArrays to observe the response of human osteoblast cells under normoxic and hypoxic conditions in terms of cell growth and utilization of metabolites. The human osteoblast cultured under these two different oxygen concentrations showed different growth curve and utilization of metabolites, suggesting oxygen levels play a role in bone repair and healing. We have deduced the main metabolites for osteoblast cells to produce energy under normoxic and hypoxic conditions. The new findings in this research help researchers to understand how hypoxia can influence utilization of metabolites in osteoblast cells, which serve as important knowledge to improve methods for bone regeneration.

scholarly journals Bone defect reconstruction via endochondral ossification: A developmental engineering strategy

2021 ◽  
Vol 12 ◽  
pp. 204173142110042
Author(s):  
Rao Fu ◽  
Chuanqi Liu ◽  
Yuxin Yan ◽  
Qingfeng Li ◽  
Ru-Lin Huang
Keyword(s):  
Bone Regeneration ◽  
Bone Defect ◽  
Bone Repair ◽  
Endochondral Ossification ◽  
Current Knowledge ◽  
Endochondral Bone ◽  
Defect Reconstruction ◽  
Hypoxic Conditions ◽  
Bone Defect Reconstruction

Traditional bone tissue engineering (BTE) strategies induce direct bone-like matrix formation by mimicking the embryological process of intramembranous ossification. However, the clinical translation of these clinical strategies for bone repair is hampered by limited vascularization and poor bone regeneration after implantation in vivo. An alternative strategy for overcoming these drawbacks is engineering cartilaginous constructs by recapitulating the embryonic processes of endochondral ossification (ECO); these constructs have shown a unique ability to survive under hypoxic conditions as well as induce neovascularization and ossification. Such developmentally engineered constructs can act as transient biomimetic templates to facilitate bone regeneration in critical-sized defects. This review introduces the concept and mechanism of developmental BTE, explores the routes of endochondral bone graft engineering, highlights the current state of the art in large bone defect reconstruction via ECO-based strategies, and offers perspectives on the challenges and future directions of translating current knowledge from the bench to the bedside.

Icariin Stimulates hFOB 1.19 Osteoblast Proliferation and Differentiation via OPG/RANKL Mediated by the Estrogen Receptor

2020 ◽  
Vol 22 (1) ◽  
pp. 168-175 ◽  
Author(s):  
Lin-Jun Sun ◽  
Chong Li ◽  
Xiang-hao Wen ◽  
Lu Guo ◽  
Zi-Fen Guo ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Estrogen Receptor ◽  
Protein Expression ◽  
Chinese Herb ◽  
Human Osteoblast ◽  
Osteoblast Cells ◽  
Expression Ratio ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Mrna And Protein Expression

Background:: Icariin (ICA), one of the main effective components isolated from the traditional Chinese herb Epimedium brevicornu Maxim., has been reported to possess extensive pharmacological actions, including enhanced sexual function, immune regulation, anti-inflammation, and antiosteoporosis. Methods:: Our study was designed to investigate the effect of ICA on cell proliferation and differentiation and the molecular mechanism of OPG/RANKL mediated by the Estrogen Receptor (ER) in hFOB1.19 human osteoblast cells. Results:: The experimental results show that ICA can stimulate cell proliferation and increase the activity of Alkaline Phosphatase (ALP), Osteocalcin (BGP) and I Collagen (Col I) and a number of calcified nodules. Furthermore, the mRNA and protein expression of OPG and RANKL and the OPG/ RANKL mRNA and protein expression ratios were upregulated by ICA. The above-mentioned results indicated that the optimal concentration of ICA for stimulating osteogenesis was 50ng/mL. Subsequent mechanistic studies comparing 50ng/mL ICA with an estrogen receptor antagonist demonstrated that the effect of the upregulated expression is connected with the estrogen receptor. In conclusion, ICA can regulate bone formation by promoting cell proliferation and differentiation and upregulating the OPG/RANKL expression ratio by the ER in hFOB1.19 human osteoblast cells.

scholarly journals Bioresorbable Magnesium-Based Alloys as Novel Biomaterials in Oral Bone Regeneration: General Review and Clinical Perspectives

2021 ◽  
Vol 10 (9) ◽  
pp. 1842
Author(s):  
Valentin Herber ◽  
Begüm Okutan ◽  
Georgios Antonoglou ◽  
Nicole G. Sommer ◽  
Michael Payer
Keyword(s):  
Additive Manufacturing ◽  
Bone Regeneration ◽  
Modulus Of Elasticity ◽  
Bone Repair ◽  
Clinical Results ◽  
Porous Scaffolds ◽  
General Review ◽  
Synthetic Materials ◽  
Bone Preservation

Bone preservation and primary regeneration is a daily challenge in the field of dental medicine. In recent years, bioresorbable metals based on magnesium (Mg) have been widely investigated due to their bone-like modulus of elasticity, their high biocompatibility, antimicrobial, and osteoconductive properties. Synthetic Mg-based biomaterials are promising candidates for bone regeneration in comparison with other currently available pure synthetic materials. Different alloys based on Mg were developed to fit clinical requirements. In parallel, advances in additive manufacturing offer the possibility to fabricate experimentally bioresorbable metallic porous scaffolds. This review describes the promising clinical results of resorbable Mg-based biomaterials for bone repair in osteosynthetic application and discusses the perspectives of use in oral bone regeneration.

scholarly journals Photocuring Hyaluronic Acid/Silk Fibroin Hydrogel Containing Curcumin Loaded CHITOSAN Nanoparticles for the Treatment of MG-63 Cells and ME3T3-E1 Cells

Polymers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 2302
Author(s):  
Qingwen Yu ◽  
Zhiyuan Meng ◽  
Yichao Liu ◽  
Zehao Li ◽  
Xing Sun ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Bone Regeneration ◽  
Water Uptake ◽  
Silk Fibroin ◽  
Bone Repair ◽  
Ph Value ◽  
Chitosan Nanoparticles ◽  
Vitro Assay ◽  
Long Term Treatment

After an osteosarcoma excision, recurrence and bone defects are significant challenges for clinicians. In this study, the curcumin (Cur) loaded chitosan (CS) nanoparticles (CCNP) encapsulated silk fibroin (SF)/hyaluronic acid esterified by methacrylate (HAMA) (CCNPs-SF/HAMA) hydrogel for the osteosarcoma therapy and bone regeneration was developed by photocuring and ethanol treatment. The micro or nanofibers networks were observed in the CCNPs-SF/HAMA hydrogel. The FTIR results demonstrated that alcohol vapor treatment caused an increase in β-sheets of SF, resulting in the high compression stress and Young’s modulus of CCNPs-SF/HAMA hydrogel. According to the water uptake analysis, SF caused a slight decrease in water uptake of CCNPs-SF/HAMA hydrogel while CCNPs could enhance the water uptake of it. The swelling kinetic results showed that both the CCNPs and the SF increased the swelling ratio of CCNPs-SF/HAMA hydrogel. The accumulative release profile of CCNPs-SF/HAMA hydrogel showed that the release of Cur from CCNPs-SF/HAMA hydrogel was accelerated when pH value was decreased from 7.4 to 5.5. Besides, compared with CCNPs, the CCNPs-SF/HAMA hydrogel had a more sustainable drug release, which was beneficial for the long-term treatment of osteosarcoma. In vitro assay results indicated that CCNPs-SF/HAMA hydrogel with equivalent Cur concentration of 150 μg/mL possessed both the effect of anti-cancer and promoting the proliferation of osteoblasts. These results suggest that CCNPs-SF/HAMA hydrogel with superior physical properties and the bifunctional osteosarcoma therapy and bone repair may be an excellent candidate for local cancer therapy and bone regeneration.

scholarly journals Hypoxia Transcriptomic Modifications Induced by Proton Irradiation in U87 Glioblastoma Multiforme Cell Line

2021 ◽  
Vol 11 (4) ◽  
pp. 308
Author(s):  
Valentina Bravatà ◽  
Walter Tinganelli ◽  
Francesco P. Cammarata ◽  
Luigi Minafra ◽  
Marco Calvaruso ◽  
...  
Keyword(s):  
Glioblastoma Multiforme ◽  
Cell Line ◽  
Acute Hypoxia ◽  
Hypoxic Condition ◽  
Biological Processes ◽  
Microarray Gene Expression ◽  
Hypoxic Conditions ◽  
Gene Expression Analyses ◽  
Shed Light ◽  
New Strategies

In Glioblastoma Multiforme (GBM), hypoxia is associated with radioresistance and poor prognosis. Since standard GBM treatments are not always effective, new strategies are needed to overcome resistance to therapeutic treatments, including radiotherapy (RT). Our study aims to shed light on the biomarker network involved in a hypoxic (0.2% oxygen) GBM cell line that is radioresistant after proton therapy (PT). For cultivating cells in acute hypoxia, GSI’s hypoxic chambers were used. Cells were irradiated in the middle of a spread-out Bragg peak with increasing PT doses to verify the greater radioresistance in hypoxic conditions. Whole-genome cDNA microarray gene expression analyses were performed for samples treated with 2 and 10 Gy to highlight biological processes activated in GBM following PT in the hypoxic condition. We describe cell survival response and significant deregulated pathways responsible for the cell death/survival balance and gene signatures linked to the PT/hypoxia configurations assayed. Highlighting the molecular pathways involved in GBM resistance following hypoxia and ionizing radiation (IR), this work could suggest new molecular targets, allowing the development of targeted drugs to be suggested in association with PT.

scholarly journals Overexpression of CRABP2 inhibits dexamethasone-induced apoptosis in human osteoblast cells

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Haiping Zhang ◽  
Ziliang Yu ◽  
Farui Sun ◽  
Jin Jin
Keyword(s):  
Protein Expression ◽  
Underlying Mechanism ◽  
R Package ◽  
Treated Group ◽  
Gene Expression Omnibus ◽  
Human Osteoblast ◽  
Osteoblast Cells ◽  
Gene And Protein Expression ◽  
Osteoblast Apoptosis ◽  
Induced Apoptosis

Abstract Background The purpose of the current study was to explore the role and underlying mechanism of cellular retinoic acid binding protein 2 (CRABP2) in dexamethasone (DEX)-induced apoptosis in human osteoblast cells. Methods GSE10311 was downloaded from the Gene Expression Omnibus (GEO) database to identify the differentially expressed genes (DEGs) by the limma/R package. Primary human osteoblast was isolated and treated with different concentration of DEX (0, 10-8, 10-7, 10-6, 10-5, and 10-4 mol/L), and cell viability and flow cytometry were used to detect cell proliferation and apoptosis. A CRABP2 overexpression plasmid (oe-CRABP2) was used to overexpress CRABP2, and western blotting was conducted to detect protein expression. Results We found that CRABP2 was downregulated in the DEX-treated group. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated that DEGs were associated with PI3K/Akt signaling pathway. DEX downregulated CRABP2 gene and protein expression, inhibited viability, and induced human osteoblast apoptosis. Overexpression of CRABP2 reversed DEX-induced apoptosis in human osteoblast. Moreover, overexpression of CRABP2 delayed the progression of DEX-induced osteonecrosis of the femoral head (ONFH) animal model. Conclusion In conclusion, CRABP2 is effective at inhibiting DEX-induced human osteoblast apoptosis and delayed ONFH progression.

scholarly journals Zn-Containing Membranes for Guided Bone Regeneration in Dentistry

Polymers ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1797
Author(s):  
Manuel Toledano ◽  
Marta Vallecillo-Rivas ◽  
María T. Osorio ◽  
Esther Muñoz-Soto ◽  
Manuel Toledano-Osorio ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Bone Regeneration ◽  
Bone Healing ◽  
Bone Repair ◽  
Complex Modulus ◽  
Bacterial Colonization ◽  
Guided Bone Regeneration ◽  
M2 Macrophage

Barrier membranes are employed in guided bone regeneration (GBR) to facilitate bone in-growth. A bioactive and biomimetic Zn-doped membrane with the ability to participate in bone healing and regeneration is necessary. The aim of the present study is to state the effect of doping the membranes for GBR with zinc compounds in the improvement of bone regeneration. A literature search was conducted using electronic databases, such as PubMed, MEDLINE, DIMDI, Embase, Scopus and Web of Science. A narrative exploratory review was undertaken, focusing on the antibacterial effects, physicochemical and biological properties of Zn-loaded membranes. Bioactivity, bone formation and cytotoxicity were analyzed. Microstructure and mechanical properties of these membranes were also determined. Zn-doped membranes have inhibited in vivo and in vitro bacterial colonization. Zn-alloy and Zn-doped membranes attained good biocompatibility and were found to be non-toxic to cells. The Zn-doped matrices showed feasible mechanical properties, such as flexibility, strength, complex modulus and tan delta. Zn incorporation in polymeric membranes provided the highest regenerative efficiency for bone healing in experimental animals, potentiating osteogenesis, angiogenesis, biological activity and a balanced remodeling. Zn-loaded membranes doped with SiO2 nanoparticles have performed as bioactive modulators provoking an M2 macrophage increase and are a potential biomaterial for promoting bone repair. Zn-doped membranes have promoted pro-healing phenotypes.

Hypoxic tolerance of Chinese black sleeper Bostrichthys sinensis embryos at heartbeat stage

2006 ◽  
Vol 86 (6) ◽  
pp. 1473-1476 ◽  
Author(s):  
Shixi Chen ◽  
Wanshu Hong ◽  
Wei Zhou ◽  
Qiyong Zhang
Keyword(s):  
Dissolved Oxygen ◽  
Exposure Time ◽  
Hypoxic Conditions ◽  
Hypoxic Tolerance ◽  
Heartbeat Rate ◽  
Bostrichthys Sinensis ◽  
Oxygen Concentrations

Tolerance of hypoxia in Chinese black sleeper (Bostrichthys sinensis) embryos at heartbeat stage was examined at different oxygen concentrations. Embryonic response to hypoxic conditions was expressed in terms of the intensity of variation in heartbeat rate (V). Exposure of the embryos at 25°C to 0.5, 1.0 and 1.5 mg/l dissolved oxygen (DO), caused bradycardia, which was developed within the first 10 min of hypoxia, followed by a plateau, and lasted until termination of the hypoxia. The V values were significantly affected by DO concentrations (P<0.01). Exposure of the embryos to 0.2 mg/l DO at 25°C caused a periodic heartbeat (including a period of heartbeat and a period of silence). This phenomenon was first recorded in the present study. During the period of heartbeat, the heartbeat rates were faster at first (147±5 beats per min), and then decreased gradually until the period of silence. As the exposure time increased, the duration of heartbeat was prolonged significantly from 43.4±2.4 second to 126.2±8.2 second (P<0.01), and the duration of silence was also prolonged significantly from 68.0±5.5 second to 247.9±11.5 second (P<0.01). At the beginning of exposure, the primary heartbeat rates displayed tachycardia, and their V values were significantly lower than the V values of average heartbeat rates (P<0.05). However, the V values were not significantly different between primary heartbeat rate and average heartbeat rate after 90 min exposure (P>0.05).

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