BMC Complementary Medicine and Therapies
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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.
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Molecular Medicine Reports
·
2019
·Vol. 20
·期 6
·5084-5090
·
IF 1.851
Jinsan Yang,Cao Ma,Maoshu Zhang
Diabetes mellitus adversely affects human bones and increases the risk of developing osteoporosis. In the present study, treatment with 30 mmol/l glucose was used to establish a high glucose (HG) cell model in vitro. Plasmids were used to overexpress the P2X purinoceptor 7 (P2X7) gene. Brilliant blue G and (4‑benzoyl‑benzoyl)‑ATP were used as a P2X7 antagonist and agonist, respectively. Proliferation of osteogenic MC3T3‑E1 cells and alkaline phosphatase (ALP) activity were determined using MTT and colorimetric assays, respectively. Alizarin Red S was used to assess calcification of MC3T3‑E1 cells. Western blotting and reverse transcription‑quantitative PCR were performed to determine protein and mRNA expression levels. The results demonstrated that HG inhibited MC3T3‑E1 cell proliferation and P2X7 expression, reduced calcification, and downregulated the expression levels of ALP and osteocalcin (Ocn) in MC3T3‑E1 cells. Overexpression of P2X7 in HG conditions increased calcification and proliferation, and upregulated the levels of ALP and Ocn in MC3T3‑E1 cells. Inhibition of P2X7 downregulated the expressions of ALP and Ocn in MC3T3‑E1 cells under HG conditions. Therefore, the present results indicated that HG caused damage to osteogenic MC3T3‑E1 cells. Thus, P2X7 may be a regulatory factor that may be used to counteract the effects of HG on osteogenesis.
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IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL
·
2022
·Vol. 59
·期 1
·1-9
·
IF 2.723
Chen Sunyu,Xiao Zhanhao,Jiang Wenjin
Osteogenic differentiation is a crucial process of new bone formation. This study aimed to explore the roles and mechanism of SRY-Box Transcription Factor 2 (SOX2) on proliferation and osteogenic differentiation of MC3T3-E1 cells. Bone morphogenetic protein 2 (BMP2) was used to induce the osteogenic differentiation of MC3T3-E1 cells. The expression of SOX2 was determined by quantitative real-time PCR (RT-PCR) at different time points after induction. The SOX2 overexpression plasmids were constructed and transfected into MC3T3-E1 cells. Osteogenic differentiation was evaluated by Alizarin Red S staining and alkaline phosphatase (ALP) assay. The expressions of osteogenic differentiation markers including runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN) were detected by western blot assay. Luciferase reporter and CHIP assays were used to confirm that SOX2 regulated the transcriptional activation of leucine-rich repeat-containing G protein–coupled receptor 4 (LGR4). We found that SOX2 was down-regulated upon BMP2-induced osteogenic differentiation in MC3T3-E1 cells. Overexpression of SOX2 effectively inhibited osteogenic differentiation with decreased ALP activity, calcification, and osteogenic differentiation markers’ expression including Runx2, OPN, and OCN. LGR4 was identified as a target of SOX2, and the inhibitory effect of SOX2 on osteogenic differentiation was reversed by knockdown of LGR4. The present study confirmed that SOX2 suppressed osteogenic differentiation of MC3T3-E1 cells through targeting LGR4, which possesses a therapeutic strategy for bone formation and generation.
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Journal of Biomedical Nanotechnology
·
2019
·Vol. 15
·期 12
·2351-2362
·
IF 0
Xu Yingjie,Wu Xin,Wang Shuyi,Yang Changzhou,Li Ying,Cao Yi
Hydrogels have been widely used to mimic the biochemical and mechanical environments of native extracellular matrices for cell culture and tissue engineering. Among them, self-assembling peptide hydrogels are of special interest thanks to their great biocompatibility, designability
and convenient preparation procedures. In pioneering studies, self-assembling peptide hydrogels have been used for the culture of bone marrow cells. However, the low mechanical stability of peptide hydrogels seems to be a drawback for these applications, as bone marrow cells prefer hard substrates
for osteogenic differentiation. In this work, we explored the use of hydroxyapatite (HAP)-peptide hybrid hydrogels for three-dimensional (3D) culture and differentiation of osteogenic MC3T3-E1 cells. We used HAP nanoparticles as crosslinkers to increase the mechanical stability of peptide
hydrogels. Meanwhile, HAP provided unique chemical cues to promote the differentiation of osteoblasts. A phosphate group was introduced to the self-assembling peptide so that the peptide fibers could bind to HAP nanoparticles specifically and strongly. Rheological characterization indicated
that the hybrid hydrogels were mechanically more stable than the hydrogels containing only peptides and can be used for long term cell culture. Moreover, the hydrogels were biocompatible and showed very low cytotoxicity. The favorable mechanical properties of the hybrid hydrogels and the chemical
properties of HAP synergistically supported the differentiation of MC3T3-E1 cells. Based on these characterizations, we believe that these hybrid hydrogels can potentially be used as scaffolds for cartilage and bone regeneration in the future.
研究领域:
生物材料
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组织工程
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细胞生物学
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材料科学
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COLLOIDS AND SURFACES B-BIOINTERFACES
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2017
·
IF 3.887
Cuixia Yu,Junjun Zhuang,Lingqing Dong,Kui Cheng,Wenjian Weng
Hierarchical porous bioglass films on the tantalum were designed to enhance osteointegration of metallic implants. The films were prepared by a sol-gel method using P123 as the mesopore template and polystyrene microsphere as the nanopore template. The films with 5.4 nm mesopores and 100 nm nanopores (MBG-100) elicited an obviously elongated morphology of the cultured MC3T3-E1 cells, as a result, a higher alkaline phosphatase level was expressed. It is suggested that the nanopores play an important role in regulating cellular behavior by initial protein adsorption through nanopore curvatures. The mesopores were proven very effective for loading rhBMP-2, and the rhBMP-2 loaded on MBG-100 films showed a better function of enhancing osteogenic differentiation, which is attributed to that the nanopore structure could expedite rhBMP-2 release and provide a microenvironment for intensifying the interaction of rhBMP-2 with the cells. Hence, the cell osteogenic differentiation can be enhanced by hierarchical porous bioglass films through both the porous structure and rhBMP-2 induction.
研究领域:
生物材料
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细胞生物学
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组织工程
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Rong Rong,Hao Li,Xiangyang Dong,Liqun Hu,Xiaowen Shi,Yumin Du,Hongbing Deng,Yue Sa
Proper bone scaffolds should be biocompatible, mechanically robust and porous for cell migration. Here, pure silk fibroin (SF)- chitosan (CS) aerogel scaffolds reinforced with different amount of SF nanofibers (SF-CS/NF 1% , SF-CS/NF 2% and SF-CS/NF 3% ) are prepared for bone regeneration. Surface morphology and composition were analyzed to ensure successful integration of each component. Incorporating 3 % nanofibers endowed the aerogels with a resistance to 3.5 times the compressive stress of the pure SF-CS aerogels. The benefits of nanofibers were also confirmed by the high porosity of 72.3 ± 1.3 %, the regulated pore size and the high-water uptake ratio of 1770.4 ± 156.8 %. Enhanced cell viability of the aerogel scaffolds was verified with Cell Counting Kit-8 (CCK-8) assays, and confocal microscopy and scanning electron microscopy (SEM) images were taken to assess the cell migration and distribution. The cell differentiation on the aerogel scaffolds was evaluated with enzyme-linked immunosorbent assay (ELISA). Significantly higher level of Collagen type I (Col-I), osteocalcin (OCN), osteopontin (OPN), and alkaline phosphatase (ALP) expression was observed on SF-CS/NF 3% aerogels. This biocompatible nanofiber-reinforced aerogel scaffold facilitates osteogenic differentiation by rougher surface, enhanced mechanical strength and well-regulated pores. Thus, as-prepared scaffolds may be further applied in bone regeneration field.
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ACS Applied Materials & Interfaces
·
2017
·
IF 7.504
Xin Zhang,Junyu Chen,Xiang Pei,Jian Wang,Qianbing Wan,Shaokang Jiang,Chao Huang,Xibo Pei
Nanoscale zeolitic imidazolate framework-8 (ZIF-8)-modified titanium (ZIF-8@AHT) can enhance osteogenesis in vitro. In this study, we systematically and quantitatively examined the effects of ZIF-8@AHT on osteogenesis, and investigated its ability to form bone in vivo. First, we coated various quantities of nanoscale ZIF-8 crystals on alkali- and heat-treated titanium (AHT) by controlling the concentration of the synthesis solution. We then characterized the ZIF-8@AHT materials using scanning electron microscopy (SEM), powder X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), and determination of the sessile drop contact angle. To illustrate the combined effects of micro/nanotopography and ZIF-8@AHT composition on bone regeneration, we cultured MC3T3-E1 preosteoblast cells on various titanium substrates in vitro by setting pure titanium (Ti) and AHT as control groups. The ZIF-8@AHTs enhanced cell bioactivity compared with AHT and Ti, as evidenced by increased extracellular matrix (ECM) mineralization, collagen secretion and the upregulated expression of osteogenic genes (Alp, Col1, Opg, and Runx2) and osteogenesis-related proteins (ALP and OPG). ZIF-8@AHT-1/8 exhibited better osteogenic activity compared with the other ZIF-8@AHT groups investigated. We subsequently inserted Ti, AHT, and ZIF-8@AHT-1/8 implants into the healed first molars (M1s) of mice, and found that ZIF-8@AHT-1/8 also promoted osseointegration at the bone–implant interface. These results suggest that ZIF-8@AHT-1/8 has great potential for practical application in implant modification.
研究领域:
生物材料
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组织工程
·
材料科学
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COLLOIDS AND SURFACES B-BIOINTERFACES
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2016
·
IF 3.902
Kena Ma,Dan Huang,Jing Cai,Xinjie Cai,Lingling Gong,Pin Huang,Yining Wang,Tao Jiang
Metal orthopedic implants still face challenges in some compromised conditions, partly due to bio-inertness. The present study aimed to functionalize metallic implants with organic-inorganic nanocomposite (strontium-containing chitosan/gelatin) coatings through a simple single-step electrophoretic deposition under mild conditions. The surface characterization and in vitro cellular response were studied and compared with chitosan/gelatin (CS/G) coatings. SEM images suggested the inorganic nanoparticles may be encapsulated within or mixed with organic polymers. The XRD patterns showed that strontium carbonate was generated in the coatings. The TEM images revealed strontium-containing nanoparticles were released from the coatings in PBS. The continuous release after the initial burst release ensured the enduring effects of the functionalized surface. The tensile bond strength of the coatings to the substrates increased after the addition of strontium. In vitro cellular study confirmed that strontium-containing coatings supported the proliferation of MC3T3-E1 cells and exhibited excellent ability to enhance the differentiation of such pre-osteoblasts. Therefore, such organic-inorganic nanocomposite coatings are a promising candidate to functionalize orthopedic implant surfaces.
研究领域:
生物材料学
·
医学工程
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CARBOHYDRATE POLYMERS
·
2018
·
IF 5.158
Xueqian Wang,Mengliu Zhang,Dawei Zhang,Xinluan Wang,Huijuan Cao,Qian Zhang,Chunyan Yan
Curculigo orchioides , is a traditional Chinese medicine, is used in strengthening tendons and bones. We evaluated the anti-osteoporosis activity of the crude polysaccharide (CO70) isolated from the rhizomes of C. orchioides in ovariectomized rats. CO70 showed excellent anti-osteoporosis activity comparable to that of 17 β -estradiol. To explore the constituents responsible for the anti-osteoporosis activity of CO70, a novel homogeneous heteropolysaccharide, COP70-3, was isolated and purified from CO70. COP70-3 has a main backbone chain of (1→5)-linked α -L-Ara f , (1,3→5)-linked α -L-Ara f , (1→6)-linked β -D-Gal p , (1→4)-linked β -D-Man p , (1,2→5)-linked α -L-Ara f , (1→3)-linked β -L-Rha p , (1, 3→6)-linked β -D-Man p , (1→3)-linked α -D-Gal p A, (1,3→6)-linked β -D-Gal p and (1→6)-linked α -D-Glc p residues. Furthermore, 1.87 nM COP70-3 obviously promoted the differentiation of MC3T3-E1 cells, while 0.94 and 1.87 nM COP70-3 significantly improved the osteogenic mineralization rate. These data indicate that COP70-3 has favorable anti-osteoporosis activity in vitro .
研究领域:
药理学
·
生物化学
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Food Bioscience
·
2024
·
IF 4.8
Aonan Xia,Yang Jiang,Yuan-Kun Lee,Dong Liang,Bo Yang,Xiaoming Liu,Jianxin Zhao,Hao Zhang,Wei Chen
Fermented milk has abundant bioactive peptides, and there is a growing interest in seeking osteogenic peptides of dairy origin. This study employed computer-assisted screening to identify potential osteogenic peptides from fermented milk by Lactobacillus helveticus and investigated the effects of these osteogenic peptides on the proliferation, differentiation, and mineralization of MC3T3-E1 cells, along with their underlying mechanisms. Three osteogenic peptides were identified: AF10 (ARHPHPHLSF), AY17 (AAGGPGAPADPGRPTGY), and NE19 (NIPPLTQTPVVVPPFLQPE). Treatment with 0.2 μM of AF10, AY17, and NE19 promoted the proliferation of MC3T3-E1 cells by increasing the proportion of cells in the S phase and G2/M phase. Further molecular docking revealed significant differences in the interaction of the three osteogenic peptides with the critical domain (Asn12-Lys13-Leu14-Thr15-Gln16-Leu17-Gly18) of the epidermal growth factor receptor (EGFR). The three peptides exhibited different degrees of promotion on the differentiation and mineralization of MC3T3-E1 cells, as well as significant differences in the expression of genes and proteins related to ERK1/2, RUNX2, and BMP2. The AF10 peptide was selected for further validation of the effect of the mitogen-activated protein kinase (MAPK) signaling pathway on the proliferation and expression of differentiation and mineralization-associated genes in MC3T3-E1 cells by combining MAPK pathway inhibitors. Molecular dynamics simulation revealed the stability of AF10 when binding to EGFR. These findings provide valuable insights into the screening of osteogenic peptides of various origins and the development of functional dairy products accordingly.
研究领域:
食品科学
·
生物化学
·
分子生物学
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Food & Function
·
2019
·Vol. 11
·期 1
·1165-1175
·
IF 3.241
Xiaoxia Chen,Tianyu Li,Degang Qing,Jun Chen,Qian Zhang,Chunyan Yan
Humulus lupulus is a perennial climbing plant of the subfamily Cannabioideae native to the Northern Hemisphere. The primary use of H. lupulus is in the brewing industry, where it is an essential ingredient for imparting a unique flavor (bitterness and aroma) to beer. The female flowers of H. lupulus are also used in traditional Chinese medicine, but the biologically active ingredients underlying its benefits remain unclear. China is the largest producer and consumer of H. lupulus in Asia. Using the waste from the beer-brewing process of H. lupulus as raw materials, the biologically active polysaccharides can be screened. This is useful for the full utilization of H. lupulus, potentially leading to disease prevention and treatment. In this study, we isolated a homogeneous polysaccharide (HLP50-1) with a molecular weight of 49.13 kDa from female flowers of H. lupulus via a DEAE-Cellulose 52 anion exchange column and a Sephadex G-75 gel filtration column. Methylation, GC-MS, and NMR analyses revealed that the HLP50-1 comprised →4)-α-D-Glcp-(1→, →6)-α-D-Manp-(1→, →3)-α-L-Rhap-(1→, β-D-Glcp-(1→, α-L-Araf-(1→, →4,6)-2-OAc-β-D-Galp-(1→, β-D-Galp-(1→, →3,6)-β-D-Glcp-(1→, →2,3,4)-α-D-Xylp-(1→, →6)-α-D-Glcp-(1→, →3)-α-D-Galp-(1→, →4)-α-D-Galp-(1→. Advanced structural analysis showed that the HLP50-1 contained irregular fragments of different sizes and shapes with a smooth surface. The aggregates appeared be composed of accumulated crystals. Furthermore, the osteogenic activities of the HLP50-1 were evaluated via MC3T3-E1 cells in vitro. The results showed that 0.13 μM HLP50-1 led to outstanding proliferation, differentiation, and mineralization of the MC3T3-E1 cells. Therefore, HLP50-1 has osteogenic effects, and it may be a candidate for the treatment of osteoporosis. It has broad application prospects in functional foods, health-care products, and pharmaceuticals.
研究领域:
药理学
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分子生物学
·
生物技术
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Acta Biomaterialia
·
2026
·
IF 10.4
Zhongming Zhao,Liming Chen,Xiuqiang Li,Haoyang Hei,Hong Sun,Yi Gao,Mengmeng Yao,Chaojie Yu,Fanglian Yao,Hong Zhang,Junjie Li
Endoscopic submucosal dissection (ESD), while a gold-standard minimally invasive technique for treating early-stage intestinal cancers, poses significant challenges for postoperative mucosal repair, owing to the risk of complications such as bleeding, perforation, and infection. Existing strategies often fail to provide a bioactive healing microenvironment. This study presents a Resolvin E1 (RvE1)-loaded, mucin-based injectable hydrogel (MHSR) designed to address these limitations. Employing a component-based biomimetic approach, this hydrogel combines thiol-modified mucin (MUC-S) with zwitterionic methacrylated hyaluronic acid (HA-SB-MA) via thiol-ene click chemistry. The MHSR hydrogel demonstrates good injectability, self-healing properties, and adhesion to positively charged inflamed tissues via electrostatic interactions and disulfide bonds. It establishes a strong physical barrier that effectively resists biofouling from proteins, cells, and bacteria. Notably, MHSR orchestrates comprehensive mucosal regeneration by scavenging reactive oxygen species (ROS), modulating inflammatory responses, enhancing angiogenesis, restoring tight junction integrity, normalizing goblet cell function, and regulating gut microbiota. In the rat model of rectal ESD, MHSR significantly accelerated wound closure, reduced inflammation, and promoted functional restoration of the mucosal barrier. This multifunctional hydrogel system constitutes a highly promising biomimetic strategy for promoting mucosal repair and remodeling of the intestinal microenvironment after ESD. Statement of Significance The study reports a mucin-based injectable hydrogel (MHSR) loaded with Resolvin E1 for rectal mucosal repair after endoscopic submucosal dissection. It combines thiol-modified mucin and zwitterionic hyaluronic acid to form an adhesive, self-healing barrier with anti-biofouling properties. The hydrogel scavenges reactive oxygen species, modulates inflammation, promotes angiogenesis, restores mucosal integrity, and regulates gut microbiota. It significantly accelerates wound healing and reduces complications in a rat ESD model, offering a promising strategy for post-operative regenerative therapy.
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