INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Hua Zhang,Zhao-Yuan Feng,Zhen-Zhou Li,Yi-Sen Wang,Zhe-Xuan Mu,Xiao-Hong Lv,Zhen-Yu Wang,Zi-Luan Fan
In this study, a polysaccharide component was extracted from Pinus pumila using biomimetic-microwave assisted alkali tandem extraction and then purified by DEAE-52 and Sephadex G-200. A new purified polysaccharide (PSP c-a) was determined through in vitro tracking. The structural features were characterized using a HPGP, FTIR, IC and NMR spectroscopy. The analysis showed that there may be →3, 6)-α-Galp (1 → 3, 4)-α-Glcp (1 → 5)-α-Manp (1→ in the backbone of PSP c-a, →3, 4)-α-Glcp (1→ and →3, 4)-α-Glcp (1 → α-Arap (1→ in the branch of PSP c-a. To improve the effects of PSP c-a on osteoblast MC3T3-E1, PSP c-a was biologically and chemically modified by α-glucosidase together with Sr. Finally, a new thermostable complex Sr-PSP c-a-1 was synthesized. The effects of PSP c-a and Sr-PSP c-a-1 on osteoblasts MC3T3-E1 were compared. It was found that both were found to have a positive effect on MC3T3-E1, and Sr-PSP c-a-1 was more effective than PSP c-a. Therefore, Sr-PSP c-a-1 should be a potential candidate for promoting skeletal tissue regeneration.
研究领域:
生物化学
·
材料科学
·
医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Orthopaedic Surgery and Research
·
2020
·Vol. 15
·期 1
·1-8
·
IF 1.777
Zhang Wei,Cui Sheng-Yu,Yi Hong,Zhu Xin-Hui,Liu Wei,Xu You-Jia
Background The dysregulation of proliferation and apoptosis plays a significant role in the pathogenesis of postmenopausal osteoporosis (PO). MicroRNAs play an important role in regulating apoptosis of MC3T3-E1 cells. However, the role and potential mechanism of miR-708 for regulating H2O2-induced apoptosis is unknown. This study aimed to investigate the protective function of miR-708 in H2O2-induced apoptosis of MC3T3-E1 osteoblasts.Methods MC3T3-E1 was co-cultured with H2O2 for 8 h, then, flow cytometry, malondialdehyde (MDA), and glutathione peroxidase (Gpx) levels were measured to establish the oxidative model. MiRNA microarray was performed to assess differentially expressed miRNAs between control and H2O2-treated MC3T3-E1 cells. We then performed RT-PCR to identify the relative expression of miR-708 and PTEN. After transfected MC3T3-E1 with miR-708 mimics, flow cytometry, MDA, and Gpx level were performed to identify the apoptosis rate and oxidative stress in these groups. Furthermore, we small interfering RNA of PTEN to identify the role of PTEN in H2O2-induced apoptosis of MC3T3-E1 cells.Results H2O2 (100 nM) could significantly induce the apoptosis of MC3T3-E1 cells. Moreover, H2O2 could significantly increase the MDA level and downregulated Gpx level. RT-PCR found that H2O2 significantly decrease the level of miR-708. Compared with H2O2 group, H2O2 + miR-708 mimic significantly decreased the apoptosis rate.Conclusion smiR-708 plays a protective role in H2O2-induced MC3T3-E1 osteoblasts apoptosis and its protective effect is proceeded by regulating ROS level and PTEN expression level.
研究领域:
分子生物学
·
细胞生物学
·
骨质疏松研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Experimental and Therapeutic Medicine
·
2022
·Vol. 23
·期 3
·1-10
·
IF 2.751
Furong Wang,Fucai Zhang,Feng Zheng
Long non‑coding (lnc)RNA KCNQ1 opposite strand/antisense transcript 1 (Kcnq1ot1) has been shown to regulate multiple biological processes. However, the functional role of Kcnq1ot1 in osteoporosis and the underlying mechanism are still unclear. The present study aimed to investigate the function of lncRNA Kcnq1ot1 in osteogenic differentiation. Alkaline phosphatase (ALP) activity was measured using an ALP assay kit. Western blotting was performed to assess the expression levels of osteogenic differentiation‑associated proteins. Reverse transcription‑quantitative PCR was performed to detect Kcnq1ot1, microRNA (miR)‑98‑5p and T‑box transcription factor 5 (Tbx5) expression levels. The binding of Kcnq1ot1 with miR‑98‑5p and that of miR‑98‑5p with Tbx5 were predicted by starBase and TargetScan databases, respectively, and verified using dual luciferase reporter assays. The mineralization of MC3T3‑E1 cells was observed using an Alizarin red S staining assay. The results revealed that expression of Kcnq1ot1 was increased and that of miR‑98‑5p was decreased during osteogenic differentiation. Additionally, Kcnq1ot1 was shown to target miR‑98‑5p and inhibit its expression. Inhibiting miR‑98‑5p reversed the inhibitory effect of Kcnq1ot1 knockdown on osteogenic differentiation and mineralization of MC3T3‑E1 cells. Furthermore, Kcnq1ot1 regulated Tbx5 expression via miR‑98‑5p. Overexpressing miR‑98‑5p or downregulating Tbx5 expression reversed the promotive effect of Kcnq1ot1 overexpression on osteogenic differentiation and mineralization of MC3T3‑E1 cells. In conclusion, these findings suggested that Kcnq1ot1 may promote bone formation by inhibiting miR‑98‑5p and upregulating Tbx5. Kcnq1ot1, miR‑98‑5p and Tbx5 may therefore serve as promising targets for the treatment of osteoporosis.
研究领域:
分子生物学
·
细胞生物学
·
骨骼生物学
·
遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biomedical Materials
·
2024
·Vol. 19
·期 2
·025002
·
IF 3.9
Liu Wenxiu,Han Guojiang,Qin Liying,Dong Wenli,Han Baoqin,Jin Liming,Yang Yan
This study aimed to synthesize bioactive glass (BG) and phosphorylated chitosan (PCS), and fabricate a BG/PCS composite scaffold. The physical properties (mechanical strength, swelling degree, and degradation rate) of the BG/PCS scaffold were tested. The in vitro mineralization properties of composite scaffolds in simulated body fluid were investigated. MC3T3-E1 cell responses with the BG/PCS scaffold were investigated using live/dead cell staining, actin staining, alkaline phosphatase (ALP) activity, and Alizarin red staining. Our results showed that the scaffold had an inner porous structure, good swelling properties, and good degradation rate. After immersion in SBF, the scaffolds demonstrated high properties in inducing mineralization. Leaching solutions of the composite scaffolds exhibited good cytocompatibility. MC3T3-E1 cells adhered, spread, and proliferated on the scaffold. The BG/PCS composite scaffold showed osteo-inductive activity by increasing ALP activity and calcium deposition. Our results indicated that the BG/PCS scaffold had potential applications as a bone-defect repair biomaterial.
研究领域:
生物材料学
·
组织工程学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
COMPOSITES PART B-ENGINEERING
·
2019
·
IF 6.864
Yin Zhang,Jiapan Luan,Shengxiang Jiang,Xiang Zhou,Mingming Li
The 3-aminopropyltriethoxysilane (APTES) was grafted on mesoporous bioactive glass (MBG) to prepare amino-functionalized mesoporous bioglass (N-MBG) by reflux agitation method. The corresponding cell experiments were performed to evaluate the influences of bioactive glass on MC3T3-E1 cells in vitro stimulation. The results displayed that the initial zeta-potential on the surface of MBG is −0.06 mV, after grafting APTES, changing to a positive charge about +5.30 mV. The surface of N-MBG was distributed positive charge, which promoted cell adhesion and protein adsorption . SEM images and fluorescence micrographs of MC3T3-E1 cells showed that N-MBG had well biocompatibility in vitro compared with that of BG and MBG. In addition to surface properties, N-MBG with lower degradation rate and pH value also had played a crucial role in promoting bone cell proliferation and differentiation. These results suggested that N-MBG would be a promising material for bone tissue regeneration .
研究领域:
生物材料学
·
细胞生物学
✓ 已验证
产品关联
DOI
高影响因子
Materials Science & Engineering C-Materials for Biological Applications
·
2015
·
IF 3.088
Qingqing Yao,Wei Li,Shanshan Yu,Liwei Ma,Dayong Jin,Aldo R. Boccaccini,Yong Liu
Novel chitosan–polyvinyl pyrrolidone/45S5 Bioglass® (CS-PVP/BG) scaffolds were prepared via foam replication and chemical cross-linking techniques. The pristine BG, CS-PVP coated BG and genipin cross-linked CS-PVP/BG (G-CS-PVP/BG) scaffolds were synthesized and characterized in terms of chemical composition, physical structure and morphology respectively. Resistance to enzymatic degradation of the scaffold is improved significantly with the use of genipin cross-linked CS-PVP. The bio-effects of scaffolds on MC3T3-E1 osteoblast-like cells were evaluated by studying cell viability, adhesion and proliferation. The CCK-8 assay shows that cell viability on the resulting G-CS-PVP/BG scaffold is improved obviously after cross-linking of genipin. Cell skeleton images exhibit that well-stretched F-actin bundles are obtained on the G-CS-PVP/BG scaffold. SEM results present significant improvement on the cell adhesion and proliferation for cells cultured on the G-CS-PVP/BG scaffold. The drug release performance on the as-synthesized scaffold was studied in a phosphate buffered saline (PBS) solution. Vancomycin is found to be released in burst fashion within 24 h from the pristine BG scaffold, however, the release period from the G-CS-PVP/BG scaffold is enhanced to 7 days, indicating improved drug release properties of the G-CS-PVP/BG scaffold. Our results suggest that the G-CS-PVP/BG scaffolds possess promising physicochemical properties, sustained drug release capability and good biocompatibility for MC3T3-E1 cells' proliferation and adhesion, suggesting their potential applications in areas such as MC3T3-E1 cell stimulation and bone tissue engineering.
研究领域:
生物材料学
·
组织工程学
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Molecular Biosciences
·
2025
·
IF 4
Lian Zhu,Liu Xie,Ziming Wang,Kai-Lin Li,Wei Cai
Introduction The bioactive compound 3,5-DiCQA, derived from Duhaldea nervosa, has been traditionally utilized in folk remedies for bone fractures and osteoporosis. However, its therapeutic mechanisms remain unclear. Methods We employed UHPLC-Q Exactive Orbitrap MS-based cell metabolomics to investigate the molecular mechanisms of 3,5-DiCQA in MC3T3-E1 cells. Cell proliferation was assessed via MTT assay, differentiation by alkaline phosphatase (ALP) activity, and mineralization through alizarin red staining and cetylpyridinium chloride quantification. Metabolomic profiling compared drug-treated and control groups. Results Results from MTT assays demonstrated that 3,5-DiCQA significantly promoted cell proliferation at 100 μM. Alkaline phosphatase (ALP) assays and alizarin red staining revealed enhanced osteoblast differentiation and mineralization, respectively. Calcification deposition was significantly increased in the calcified stained cells by cetylpyridinium chloride quantization, indicating that 3,5-DiCQA can promote the mineralization of MC3T3-E1 cells. Metabolomic analysis identified key metabolic changes, including the downregulation of phytosphingosine and upregulation of sphinganine and citric acid. Discussion These findings suggest that 3,5-DiCQA promotes osteoblast proliferation, differentiation and mineralization through pathways such as sphingolipid metabolism, arginine and proline metabolism, mucin type O-glycan biosynthesis and the citrate cycle (TCA cycle). This study provides insights into the therapeutic potential of 3,5-DiCQA for osteoporosis and highlights the utility of metabolomics in elucidating traditional Chinese medicine (TCM).
研究领域:
药理学
·
代谢组学
·
骨生物学
✓ 已验证
DOI
PubMed
高影响因子
CURRENT PHARMACEUTICAL BIOTECHNOLOGY
·
2021
·Vol. 22
·期 5
·609-621
·
IF 2.837
Zheng Jing,Changyuan Wang,Shijie Wen,Yue Jin,Qiang Meng,Qi Liu,Jingjing Wu,Huijun Sun,Mozhen Liu
Background: Osteoporosis, characterized by bone loss, usually occurs with the increased bone resorption and decreased bone formation. H2O2-induced MC3T3-E1 cells are commonly used for the study of osteoblastic activities, which play a crucial role in bone formation. Objective: This study aimed to investigate the effects of Phosphocreatine (PCr) on the osteoblastic activities in H2O2-induced MC3T3-E1 cells and elaborate on the possible molecular mechanism. Methods: The Osteoprotegerin (OPG)/Receptor Activator of NF-ΚB Ligand (RANKL) ratio and osteogenic markers were detected to investigate the effects of PCr on osteoblastic activities, and the osteoblastic apoptosis was detected using Hochest staining. Moreover, oxidative stress, Adenosine Triphosphate (ATP) generation and the expression of Sirtuin 1 (SIRT1), Forkhead Box O 1 (FOXO1) and Peroxisome Proliferator-Activated Receptor Γ Coactivator-1α (PGC-1α) were also examined to uncover the possible molecular mechanism in H2O2-induced MC3T3-E1 cells. Result: The results showed that PCr promoted the osteoblastic differentiation by increasing the expression levels of osteogenic markers of Alkaline Phosphatase (ALP) and Runt-related transcription factor 2 (Runx2), as well as increased the OPG/RANKL ratio and suppressed the osteoblastic apoptosis in H2O2-induced MC3T3-E1 cells. Moreover, treatment with PCr suppressed reactive oxygen species (ROS) over-generation and promoted the ATP production as well as increased the PGC-1α, FOXO1 and SIRT1 protein expression levels in H2O2-induced MC3T3-E1 cells. Conclusion: PCr treatment could promote osteoblastic activities via suppressing oxidative stress and increasing the ATP generation in H2O2-induced MC3T3-E1 cells. In addition, the positive effects of PCr on osteoblasts might be regulated by SIRT1/FOXO1/ PGC-1α signaling pathway.
研究领域:
医学
·
分子生物学
✓ 已验证
DOI
高影响因子
BIOLOGICAL TRACE ELEMENT RESEARCH
·
2019
·Vol. 196
·期 1
·214-222
·
IF 2.431
Cao Zheng,Geng Xue,Jiang Xinpeng,Gao Xiang,Liu Kexiang,Li Yanfei
Aluminum (Al) inhibits osteoblast-mediated bone formation by oxidative stress, resulting in Al-induced bone disease. Melatonin (MT) has received extensive attention due to its antioxidant and maintenance of bone health effect. To evaluate the protective effect and mechanism of MT on AlCl 3 -induced osteoblast dysfunction, MC3T3-E1 cells were treated with MT (100 μM) and/or AlCl 3 (8 μM). First, MT alleviated AlCl 3 -induced osteoblast dysfunction, presenting as the reduced apoptosis rate as well as increased cell viability, alkaline phosphatase (ALP) activity, and type I collagen (COL-1) level. Then, MT significantly attenuated AlCl 3 -induced oxidative stress, presenting as the reduced reactive oxygen species and 8-hydroxy-2′-deoxyguanosine levels as well as increased glutathione level and superoxide dismutase activity. Finally, MT protected MC3T3-E1 cells against p53-dependent apoptosis and differentiation suppression, as assessed by Caspase-3 activity, protein levels of p53, Bcl-2-associated X protein (Bax), B cell lymphoma gene 2 (Bcl-2), cytosolic Cytochrome c, Runt-related transcription factor 2 (Runx2), and Osterix, as well as the mRNA levels of Bax, Bcl-2, Runx2, Osterix, ALP, and COL-1. Overall, our findings demonstrate MT attenuates AlCl 3 -induced apoptosis and osteoblastic differentiation suppression by inhibiting oxidative stress in MC3T3-E1 cells.
研究领域:
药理学
·
细胞生物学
·
毒理学
·
骨骼健康
✓ 已验证
DOI
PubMed
高影响因子
Regenerative Biomaterials
·
2020
·Vol. 8
·期 1
·
IF 4.882
Sun Yingxiao,Liu Xingdan,Tan Ji,Lv Dan,Song Wengang,Su Rui,Li Ling,Liu Xuanyong,Ouyang Liping,Liao Yun
Polyetheretherketone (PEEK) has been used as an implant material because it has similar mechanical properties to natural bone. However, inferior osseointegration and bioinertness hamper the clinical application of PEEK. In this study, the surfaces of sulfonated three-dimensional (3D) PEEK porous structures were loaded with different concentrations of strontium ranelate, a compound commonly used in the treatment or prevention of osteoporosis by promoting bone formation and inhibiting bone resorption. Field-emission scanning electron microscopy was used to characterize the topography of the structures, elemental carbon, oxygen and strontium contents were measured by X-ray photoelectron spectroscopy, and surface zeta potentials and water-contact angle were also measured. The results indicated that strontium ranelate was successfully loaded onto the 3D porous structures. In vitro cellular results showed that strontium ranelate-treated sulfonated PEEK (SP-SR) strengthened the adhesion of MC3T3-E1 cells. The activity of alkaline phosphatase, collagen secretion and extracellular matrix mineralization deposition of MC3T3-E1 cells were also improved on the surface of SP-SR. These results indicate that SP-SR could serve a new implant candidate for surgical treatment.
研究领域:
生物材料
·
骨科
·
细胞生物学
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Experimental and Therapeutic Medicine
·
2021
·Vol. 23
·期 2
·1-8
·
IF 2.447
Yao Duan,Hejia Li,Xiaohong Dong,Zhaoli Geng,Xinyi Xu,Yi Liu
The present study aimed to investigate whether VEGF was involved in bisphosphonate (BP)‑induced apoptosis and differentiation of osteoblasts. Murine MC3T3‑E1 osteoblasts were stimulated with zoledronic acid (ZA) for 7 days. VEGF mRNA and protein expression levels were determined via reverse transcription‑quantitative PCR and western blot analysis, respectively. Cell viability was evaluated using Cell Counting Kit‑8 assay. In addition, the cell apoptotic rate and the expression levels of apoptosis‑related proteins were measured using a TUNEL staining kit and western blot analysis, respectively. To evaluate mineralization, cells were stained with alizarin red, while the secretion levels of alkaline phosphatase (ALP) were measured using the corresponding assay kit. Finally, the expression levels of differentiation‑related proteins and proteins of the Nod‑like receptor family pyrin domain‑containing 3 (NLRP3)/caspase 1/gasdermin D (GSDMD) pyroptosis pathway were measured by western blot analysis. VEGF expression level was notably decreased in ZA‑stimulated MC3T3‑E1 cells. However, the viability of these cells was enhanced following VEGF addition. Furthermore, VEGF attenuated apoptosis, promoted mineralization and increased ALP activity in ZA‑stimulated MC3T3‑E1 cells. The ZA‑mediated decrease in the protein expression of the osteogenic genes osteopontin, osteocalcin and runt‑related transcription factor 2 was restored after MC3T3‑E1 cell treatment with 10 ng/ml VEGF. The present study demonstrated that VEGF could attenuate BP‑induced apoptosis and differentiation of MC3T3 cells by regulating the NLRP3/caspase 1/GSDMD pathway.
研究领域:
细胞生物学
·
分子生物学
·
药理学
·
骨质疏松研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BMC Complementary Medicine and Therapies
·
2023
·Vol. 23
·期 1
·1-11
·
IF 3.9
Cai Weiye,Sun Bin,Song Chao,Liu Fei,Wu Zhengliang,Liu Zongchao
Background In mouse, it was discovered that resveratrol (Res) enhanced osteoporosis (OP) by boosting osteogenesis. Besides, Res can also have an impact on MC3T3-E1 cells, which are crucial for the control of osteogenesis and thus increase osteogenesis. Although some articles have discovered that Res enhanced autophagy to promote the value-added differentiation of MC3T3, it is unclear exactly how this affects the process of osteogenesis in mouse. Therefore, we will show that Res encourages MC3T3-E1 proliferation and differentiation in mouse pre-osteoblasts and further investigate the autophagy-related mechanism for this impact.Methods (1) MC3T3-E1 cells were separated into blank control group and various concentrations (0.01, 0.1, 1, 10, 100µmol/L) of group in order to determine the ideal Res concentration. In the Res group, Cell Counting Kit-8 (CCK-8) was used to measure the proliferation activity of pre-osteoblasts in mice in each group after resveratrol intervention. Alkaline Phosphatase (ALP) and alizarin red staining were used to gauge the degree of osteogenic differentiation, and RT-qPCR was used to measure the expression levels of Runx2 and OCN in the osteogenic differentiation ability of the cells. (2) In the experiment, four groups were set up: the control group, 3MA group, Res group, and Res + 3MA group. To examine cell mineralization, ALP and alizarin red staining were utilized. RT-qPCR and Western blot detection of cell autophagy activity levels and osteogenic differentiation capacity in each group following intervention.Results (1) Resveratrol might increase the number of mice pre-osteoblast, with the impact being most pronounced at 10µmol/L (P < 0.05). The nodules developed substantially more often than in the blank control group, and Runx2 and OCN expressions significantly increased (P < 0.05). (2) In contrast to the Res group, after 3MA purine blocked autophagy, the Res + 3MA group’s alkaline phosphatase staining and the development of mineralized nodules were reduced. Runx2, OCN, LC3II / LC3I expression decreased, p62 expression increased (P < 0.05).Conclusion The present study partially or indirectly demonstrated that Res may, through increased autophagy, induce osteogenic differentiation of MC3T3-E1 cells.
研究领域:
药理学
·
细胞生物学
·
分子生物学
·
骨骼研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）