1.Astragalus Promotes Osteogenic Differentiation of hBMSCs and Alleviates Osteoporosis by Targeting SOX11 Via miR-181d-5p.
Yuan XIAO ; Yong Li SITU ; Ting Ting WANG ; Shang KONG ; Jiang Qi LIU ; Hong NIE
Biomedical and Environmental Sciences 2025;38(10):1287-1301
OBJECTIVE:
This study aimed to investigate the effect of Astragalus (AST) on osteoporosis (OP) and the downstream mechanisms.
METHODS:
Human bone marrow-derived mesenchymal stem cells (hBMSCs) were induced to differentiate into osteogenic cells. After transfection with relevant plasmids, cell proliferation, cell cycle progression, and apoptosis were assessed. Alizarin red staining was used to detect calcium nodules in the cells, alkaline phosphatase (ALP) staining was used to detect ALP activity in the cells, and quantitative reverse transcription-polymerase chain reaction and western blotting were used to determine RUNX2 and Osterix expression levels. An OP rat model was established using ovariectomy and micro-computed tomography scanning. Hematoxylin and eosin staining and Masson's trichrome staining were used to evaluate the pathological conditions of bone tissues, while immunohistochemistry was conducted to detect RUNX2 in bone tissues.
RESULTS:
AST promoted the osteogenic differentiation of BMSCs, reduced miR-181d-5p expression levels, and increased SOX11 expression levels. Restoring miR-181d-5p expression or reducing SOX11 expression levels reversed the effects of AST on the osteogenic differentiation of hBMSCs. miR-181d-5p was found to target SOX11 in hBMSCs. AST improved OP in rats, and miR-181d-5p overexpression or SOX11 inhibition reversed the therapeutic effects of AST on OP in rats.
CONCLUSION
AST promoted the osteogenic differentiation of hBMSCs and alleviated OP by targeting SOX11 via miR-181d-5p.
Osteogenesis/drug effects*
;
Animals
;
MicroRNAs/genetics*
;
Mesenchymal Stem Cells/drug effects*
;
Osteoporosis/drug therapy*
;
Humans
;
Cell Differentiation/drug effects*
;
Astragalus Plant/chemistry*
;
Rats
;
Rats, Sprague-Dawley
;
Female
;
SOXC Transcription Factors/genetics*
;
Plant Extracts/pharmacology*
;
Cells, Cultured
;
Drugs, Chinese Herbal/pharmacology*
2.Ginsenoside Rb3 regulates the phosphorrylated extracellular signal-regulated kinase signaling pathway to alleviate inflammatory responses and promote osteogenesis in rats with periodontitis.
Xueying ZHANG ; Xin MENG ; Zhizhen LIU ; Kang ZHANG ; Honghai JI ; Minmin SUN
West China Journal of Stomatology 2025;43(2):236-248
OBJECTIVES:
To explore the promoting effect of ginsenoside Rb3 (Rb3) on osteogenesis in periodontitis environment, and to explain its mechanism.
METHODS:
Human periodontal ligament stem cells (hPDLSCs) were cultured by tissue block method and identified by flow cytometry. Cell counting kit-8 (CCK8) method and calcein acetoxymethyl ester/propidium iodide staining were used to detect the effect of Rb3 on the viability of hPDLSCs cells. In vitro cell experiments were divided into control group, 10 μg/mL lipopolysaccharides (LPS) group, 10 μg/mL LPS+100 μmol/L Rb3 group and 10 μg/mL LPS+200 μmol/L Rb3 group. Alkaline phosphatase (ALP) staining was used to detect the ALP activity of hPDLSCs in each group after osteogenesis induction. The expression of hPDLSCs interleukin-6 (IL-6), interleukin-8 (IL-8), runt-related transcription factor 2 (RUNX2) and transforming growth factor-β (TGF-β)genes in each group after osteogenesis was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) method. Western blot was used to detect the protein expression of hPDLSCs phosphorrylated extracellular signal-regulated kinase (p-ERK) in each group. Sprague-Dawley rats were randomly divided into the control group, ligation group and ligation+Rb3 group. The left molar-maxillary tissue was subjected to micro-computed tomography (micro-CT) scanning. After the scanning, the left molar-maxilla was made into periodontal tissue sections. Hematoxylin-eosin (HE) staining was used to detect the infiltration and loss of adhesion of inflammatory cells. Masson staining was used to detect the destruction of gingival collagen fibers. Immunofluorescence staining was used to detect the protein expression of RUNX2 and p-ERK. The expression of TGF-β in rat gingival tissue was detected by qRT-PCR. The protein expression of IL-6 in peripheral serum of rats was detected by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was used to detect the proportion of Treg cells in rat heart blood. The experimental data were statistically analyzed by Graph Pad Prism10.1.2 software.
RESULTS:
Rb3 had no effect on the cell activity of hPDLSCs. The results of qRT-PCR and ALP staining showed that Rb3 could inhibit the gene expression of IL-6 and IL-8 in inflammatory hPDLSCs, promote TGF-β gene and promote the osteogenic differentiation of inflammatory hPDLSCs. Western blot showed that Rb3 inhibited the protein expression of inflammatory hPDLSCs p-ERK. The results from micro-CT, Masson staining, and HE staining demonstrated that Rb3 promotes alveolar bone formation in rats with periodontitis, while simultaneously inhibiting the destruction of periodontal fibrous tissue, reducing attachment loss, and suppressing inflammatory cell infiltration. The results of flow cytometry showed that Rb3 could promote the differentiation of Treg cells in peripheral blood of periodontitis rats. The results of ELISA and qRT-PCR showed that Rb3 could inhibit the protein expression of IL-6 and promote the gene expression of TGF-β in periodontitis rats. Immunofluorescence results showed that Rb3 could promote the protein expression of RUNX2 and inhibit the protein expression of p-ERK in periodontitis rats.
CONCLUSIONS
Rb3 can reduce the inflammatory reaction of periodontal tissues in periodontitis rats, and promote the osteogenic differentiation of hPDLSCs by regulating p-ERK pathways.
Animals
;
Ginsenosides/pharmacology*
;
Osteogenesis/drug effects*
;
Periodontitis/metabolism*
;
Rats
;
Periodontal Ligament/cytology*
;
Humans
;
Core Binding Factor Alpha 1 Subunit/metabolism*
;
Stem Cells/drug effects*
;
Interleukin-6/metabolism*
;
Rats, Sprague-Dawley
;
Interleukin-8/metabolism*
;
Cells, Cultured
;
MAP Kinase Signaling System/drug effects*
;
Transforming Growth Factor beta/metabolism*
;
Signal Transduction
;
Male
;
Phosphorylation
;
Lipopolysaccharides
;
Extracellular Signal-Regulated MAP Kinases/metabolism*
;
Alkaline Phosphatase/metabolism*
3.In vitro osteogenic performance study of graphene oxide-coated titanium surfaces modified with dopamine or silane.
Qinglin WU ; Yingzhen LAI ; Yanling HUANG ; Zeyu XIE ; Yanyin LIN
West China Journal of Stomatology 2025;43(3):336-345
OBJECTIVES:
This study aimed to compare the osteogenic performance differences of titanium surface coatings modified by dopamine or silanized graphene oxide, and to provide a more suitable modification scheme for titanium surface graphene oxide coatings.
METHODS:
Titanium was subjected to alkali-heat treatment and then modified with dopamine and silanization, respectively, followed by coating with graphene oxide. Control and experimental groups were designed as follows: pure titanium (Ti) group; titanium after alkali-heat treatment (Ti-NaOH) group; titanium after alkali-heat treatment and silanization modification (Ti-APTES) group; titanium after alkali-heat treatment and dopamine modification (Ti-DOPA) group; titanium with silanization-modified surface decorated with graphene oxide (Ti-APTES/GO) group; titanium with dopamine-modified surface decorated with graphene oxide (Ti-DOPA/GO) group. The physical and chemical properties of the material surfaces were analyzed using scanning electron microscopy (SEM), contact angle goniometer, X-ray photoelectron spectroscopy (XPS), and Raman spectrometer. The proliferation and adhesion morphology of mouse embryonic osteoblast precursor cells MC3T3-E1 on the material surfaces were observed by cell viability detection and immunofluorescence staining followed by laser confocal microscopy. The effects on the osteogenic differentiation of MC3T3-E1 cells were studied by alkaline phosphatase (ALP) staining, alizarin red staining and quantification, and real-time quantitative polymerase chain reaction.
RESULTS:
After modification with graphene oxide coating, a thin-film-like structure was observed on the surface under SEM. The hydrophilicity of all experimental groups was improved, among which the Ti-DOPA/GO group had the best hydrophilicity. XPS and Raman spectroscopy analysis showed that the modified materials exhibited typical D and G peaks, and XPS revealed the presence of a large number of oxygen-containing functional groups on the surface. CCK8 assay showed that all groups of materials had no cytotoxicity, and the proliferation level of the Ti-APTES/GO group was higher than that of the Ti-DOPA/GO group. Under the laser confocal microscope, the cells in the Ti-DOPA/GO and Ti-APTES/GO groups spread more fully. The Ti-DOPA/GO and Ti-APTES/GO groups had the deepest ALP staining, and the Ti-APTES/GO group had the most alizarin red-stained mineralized nodules and the highest quantitative result of alizarin red staining. In the Ti-DOPA/GO and Ti-APTES/GO groups, the expression of the early osteogenic-related gene RUNX2 reached a relatively high level, while in the expression of the late osteogenic-related genes OPN and OCN, the Ti-APTES/GO group performed better than the Ti-DOPA/GO group.
CONCLUSIONS
Ti-APTES/GO significantly outperformed Ti-DOPA/GO in promoting the adhesion, proliferation, and in vitro osteogenic differentiation of MC3T3-E1 cells.
Titanium/chemistry*
;
Graphite/chemistry*
;
Dopamine/chemistry*
;
Animals
;
Mice
;
Osteogenesis
;
Osteoblasts/cytology*
;
Surface Properties
;
Cell Proliferation
;
Silanes/chemistry*
;
Cell Adhesion
;
Coated Materials, Biocompatible/chemistry*
;
Cell Differentiation
;
Alkaline Phosphatase/metabolism*
;
Microscopy, Electron, Scanning
4.Investigating the protective effect of naringenin on hydrogen peroxide induced oxidative damage of human periodontal ligament stem cells by regulating the forkhead box protein O-1/β-catenin pathway.
Li ZHANG ; Shiyuan PENG ; Feiyang TANG ; Jingwei JIAN ; Shuosheng YUAN ; Xiaomei XU
West China Journal of Stomatology 2025;43(4):559-569
OBJECTIVES:
Investigating the protective effect of naringenin (NAR) on the osteogenic potential of human periodontal ligament stem cells (hPDLSCs) under oxidative stress and its related mechanisms.
METHODS:
The oxidative damage model of hPDLSCs was established using hydrogen peroxide (H2O2) andthe hPDLSCs were treated with different concentrations of NAR and 0.5 μmol/L forkhead box protein O-1 (FOXO1) inhibitor AS1842856. After that, the cell counting kit-8 (CCK8) was used to determine the optimal concentrations of H2O2 and NAR. The alkaline phosphatase (ALP) staining and real time fluorescent quantitative reverse transcription polymerase chain reaction (qRT-PCR) were employed to assess the expression of ALP, runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN) in hPDLSCs of each group. The enzyme-linked immunosorbent assay (ELISA) and 2',7'-dichlorofluorescin diacetate (DCFH-DA) staining were utilized to evaluate the expression of reactive oxygen species (ROS), malondialdehyde (MDA) and lactate dehydrogenase (LDH) in hPDLSCs. Meanwhile, qRT-PCR and western blot were used to detect the expression levels of FOXO1 and β-catenin, both are pathway related genes and proteins.
RESULTS:
H2O2 exposure led to an increase in oxidative damage in hPDLSCs, characterized by a rise in intracellular ROS levels and increased expression of MDA and LDH (P<0.05). At the same time, the osteogenic differentiation ability of hPDLSCs decreased, as evidenced by lighter ALP staining and reduced expression levels of osteogenic differentiation-related genes ALP, RUNX2 and OCN (P<0.05). Co-treatment with NAR alleviated the oxidative damage in hPDLSCs, enhanced their antioxidant capacity, and restored their osteogenic ability. The FOXO1 inhibitor AS1842856 downregulated the expression of β-catenin (P<0.05) and significantly diminished both the antioxidant effect of NAR and its ability to restore osteogenesis (P<0.05).
CONCLUSIONS
NAR can enhance the antioxidant capacity of hPDLSCs by activating the FOXO1/β-catenin signaling pathway within hPDLSCs, thereby mitigating oxidative stress damage and alleviating the loss of osteogenic capacity.
Humans
;
Oxidative Stress/drug effects*
;
Periodontal Ligament/cytology*
;
Hydrogen Peroxide
;
Forkhead Box Protein O1/metabolism*
;
Stem Cells/cytology*
;
Flavanones/pharmacology*
;
beta Catenin/metabolism*
;
Osteogenesis/drug effects*
;
Signal Transduction
;
Core Binding Factor Alpha 1 Subunit/metabolism*
;
Alkaline Phosphatase/metabolism*
;
Osteocalcin/metabolism*
;
Cells, Cultured
;
Cell Differentiation/drug effects*
5.Clinical and histological evaluation of three-dimensional printing individualized titanium mesh for alveolar bone defect repair.
Pengyu ZHAO ; Gang CHEN ; Yi CHENG ; Chao WANG ; Dan CHEN ; Haitao HUANG
West China Journal of Stomatology 2025;43(4):592-602
OBJECTIVES:
To evaluate the osteogenic efficacy of three-dimensional printing individualized titanium mesh (3D-PITM) as a scaffold material in guided bone regeneration (GBR).
METHODS:
1) Patients undergoing GBR for alveolar bone defects were enrolled as study subjects, and postoperative healing complications were recorded. 2) Postoperative cone beam computed tomography (CBCT) scans acquired at least 6 months post-surgery were used to calculate the percentage of actual bone formation volume. 3) Alveolar bone specimens were collected during the first-stage implant surgery for histomorphometric analysis. This analysis quantitatively measured the proportions of newly formed bone and newly formed unmineralized bone within the specimens. Specimens were categorized into three groups based on healing complications (good healing group, wound dehiscence group, 3D-PITM exposure group) to compare differences in the proportions of newly formed bone and newly formed unmineralized bone.
RESULTS:
1) Twelve patients were included. Guided bone regeneration failed in one patient, and 3D-PITM exposure occurred in three patients (exposure rate: 25%). 2) The mean percentage of actual bone formation volume in the 11 successful guided bone regeneration cases was 95.23%±28.85%. 3) Histomorphometric analysis revealed that newly formed bone constituted 40.35% of the alveolar bone specimens, with newly formed unmineralized bone accounting for 13.84% of the newly formed bone. Intergroup comparisons showed no statistically significant differences (P>0.05) in the proportions of newly formed bone or newly formed unmineralized bone between the good healing group and the wound dehiscence group or the 3D-PITM exposure group.
CONCLUSIONS
3D-PITM enables effective bone augmentation. Radiographic assessment demonstrated favorable bone formation volume, while histological analysis confirmed substantial formation of newly formed mineralized bone within the surgical site.
Humans
;
Printing, Three-Dimensional
;
Titanium
;
Cone-Beam Computed Tomography
;
Bone Regeneration
;
Osteogenesis
;
Surgical Mesh
;
Tissue Scaffolds
;
Alveolar Process/surgery*
;
Adult
;
Male
;
Middle Aged
;
Female
;
Wound Healing
;
Guided Tissue Regeneration, Periodontal/methods*
;
Alveolar Bone Loss/surgery*
6.Effect of trapezoidal and modified triangular flaps on mucosal blood supply and osteogenesis after guided bone regeneration.
Shuangzhen CHEN ; Xianyue ZHANG ; Xiaofeng JIA ; Rong XIA ; Fan JIANG
West China Journal of Stomatology 2025;43(5):679-688
OBJECTIVES:
Color doppler flow imaging (CDFI) and cone-beam computed tomography (CBCT) were utilized to evaluate changes in mucosal vascular parameters and the osteogenic effects following guided bone regeneration (GBR) in the maxillary anterior region using trapezoidal or modified triangular flaps.
METHODS:
Patients undergoing single maxillary anterior dental implant surgery with GBR were randomly allocated into two groups: a trapezoidal flap group and a modified triangular flap group. After GBR surgery, the mucosal vascular parameters at the surgical site were assessed at various time intervals (preoperative, 2 h, 1 and 3 days, and 1, 2, and 4 weeks postoperative) using CDFI. In addition, the effects of bone augmentation were evaluated through the analysis of CBCT images obtained preoperatively, 2 h, and 6 months postoperative.
RESULTS:
The buccal mucosa in the edentulous area had a lower blood flow rate than the corresponding tooth in the same jaw, and the difference was statistically significant (P<0.001). The mucosal blood flow rate in the surgical area increased compared with that in the preoperative period. The peak flow rate was recorded at 2 weeks postoperatively and then decreased to levels comparable to those of the reference tooth. A statistically significant difference was observed between the two groups (P<0.05). The buccal alveolar ridge width of the implant platform was reduced by (1.3±0.9) mm in the trapezoidal flap group and (0.9±0.7) mm in the modified triangular flap group, respectively, at 6 months postoperatively, compared with 2 h postoperative. The buccal alveolar ridge width of the 5 mm from the implant platform was reduced by (0.9±0.6) mm and (0.3±0.6) mm, respectively. The buccal alveolar ridge width of the 10 mm from the implant platform was reduced by (0.6±0.8) mm and (0.2±0.6) mm, respectively. The height of the alveolar ridge was reduced by (1.9±1.4 ) mm and (1.4±1.3) mm. The change in graft volume was (136±78 ) mm3 and (114±85) mm3. However, the differences between the two groups were not statistically significant (P>0.05).
CONCLUSIONS
When a tooth is missing, blood flow to the buccal mucosa on the side of the missing tooth is reduced. The modified triangular flap group demonstrated superior microcirculation of blood flow in the operative area after GBR of the maxillary anterior teeth. Trapezoidal and modified triangular flaps achieved the anticipated bone augmentation during bone augmentation surgery in the maxillary anterior region, with no considerable effect on the changes in alveolar bone size parameters.
Humans
;
Surgical Flaps/blood supply*
;
Bone Regeneration
;
Mouth Mucosa/blood supply*
;
Cone-Beam Computed Tomography
;
Osteogenesis
;
Maxilla/surgery*
;
Male
;
Female
;
Guided Tissue Regeneration, Periodontal/methods*
7.Preparation of polycaprolactone-polyethylene glycol-concentrated growth factor composite scaffolds and the effects on the biological properties of human periodontal ligament stem cells.
Li GAO ; Mingyue ZHAO ; Shun YANG ; Runan WANG ; Jiajia CHENG ; Guangsheng CHEN
West China Journal of Stomatology 2025;43(6):819-828
OBJECTIVES:
This study investigated the effects of a polycaprolactone (PCL)-polyethylene glycol (PEG) scaffold incorporated with concentrated growth factor (CGF) on the adhesion, proliferation, and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs).
METHODS:
The PCL-PEG-CGF composite scaffold was fabricated using an immersion and freeze-drying technique. Its microstructure, mechanical properties, and biocompatibility were systematically characterized. The hPDLSCs were isolated through enzymatic digestion, and the hPDLSCs were identified through flow cytometry. Third-passage hPDLSCs were seeded onto the composite scaffolds, and their adhesion, proliferation and osteogenic differentiation were assessed using CCK-8 assays, 4',6-diamidino-2-phenylindole (DAPI) staining, alkaline phosphatase (ALP) staining, alizarin red staining, and Western blot analysis of osteogenesis-related proteins [Runt-related transcription factor 2 (Runx2), ALP, and morphogenetic protein 2 (BMP2)].
RESULTS:
Scanning electron microscopy revealed that the PCL-PEG-CGF composite scaffold exhibited a honeycomb-like structure with heterogeneous pore sizes. The composite scaffold exhibited excellent hydrophilicity, as evidenced by a contact angle (θ) approaching 0° within 6 s. Its elastic modulus was measured at (4.590 0±0.149 3) MPa, with comparable hydrophilicity, fracture tensile strength, and fracture elongation to PCL-PEG scaffold. The hPDLSCs exhibited significantly improved adhesion to the PCL-PEG-CGF composite scaffold compared with the PCL-PEG scaffold (P<0.01). Additionally, cell proliferation was markedly improved in all the experimental groups on days 3, 5, and 7 (P<0.01), and statistically significant differences were found between the PCL-PEG-CGF group and other groups (P<0.01). The PCL-PEG-CGF group showed significantly elevated ALP activity (P<0.05), increased mineralization nodule formation, and upregulated expression of osteogenic-related proteins (Runx2, BMP2 and ALP; P<0.05).
CONCLUSIONS
The PCL-PEG-CGF composite scaffold exhibited excellent mechanical properties and biocompatibility, enhancing the adhesion and proliferation of hPDLSCs and promoting their osteogenic differentiation by upregulating osteogenic-related proteins.
Humans
;
Polyesters/chemistry*
;
Periodontal Ligament/cytology*
;
Polyethylene Glycols/chemistry*
;
Stem Cells/cytology*
;
Tissue Scaffolds
;
Cell Proliferation
;
Osteogenesis
;
Cell Differentiation
;
Cell Adhesion
;
Bone Morphogenetic Protein 2/metabolism*
;
Cells, Cultured
;
Alkaline Phosphatase/metabolism*
;
Core Binding Factor Alpha 1 Subunit/metabolism*
;
Intercellular Signaling Peptides and Proteins/pharmacology*
;
Tissue Engineering/methods*
8.Effects of Gly mutations N-terminal to the integrin-binding sequence on the structure and function of recombinant collagen.
Fei LI ; Yuxi HOU ; Ben RAO ; Xiaoyan LIU ; Yaping WANG ; Yimin QIU
Chinese Journal of Biotechnology 2025;41(4):1573-1587
Collagen, a vital matrix protein for various tissue and functions in animals, is widely applied in biomaterials. In type Ⅰ collagen, missense mutations of glycine (Gly) in the Gly-Xaa-Yaa triplet of the triple helix are a major cause of osteogenesis imperfecta (OI). Clinical manifestations exhibit marked heterogeneity, spanning a broad disease spectrum from mild skeletal fragility (Type Ⅰ) to severe limb deformities (Type Ⅲ) and perinatal lethal forms (Type Ⅱ). This study utilized recombinant collagen as a model to further elucidate whether Gly→Ala/Val mutations at the N-terminus of the integrin-binding sequence GFPGER affect collagen structure and function, and to explore the underlying mechanisms by which missense mutations impact the biological function of collagen. By introducing Ala and Val substitutions at seven Gly positions N-terminal to the GFPGER sequence, we systematically assessed the effects of these amino acid replacements on the triple-helical structure, thermal stability, integrin-binding ability, and cell adhesion of recombinant collagen. All constructs formed a stable triple-helix structure, with slightly compromised thermal stability. Gly→Val substitutions increased the susceptibility of recombinant collagen to trypsin, which suggested local conformational perturbations in the triple helix. In addition, Gly→Val substitutions significantly reduced the integrin-binding affinity and decreased HT1080 cell adhesion, with the effects stronger than Gly→Ala substitutions. Compared with Gly→Ala substitutions, substitution of Gly with the larger residue Val had enhanced negative effects on the structure and function of recombinant collagen. These findings provide new insights into the molecular mechanisms of osteogenesis imperfecta and offer theoretical references and experimental foundations for the design of collagen sequences and the development of collagen-based biomaterials.
Recombinant Proteins/biosynthesis*
;
Glycine/genetics*
;
Humans
;
Osteogenesis Imperfecta/genetics*
;
Integrins/metabolism*
;
Collagen/metabolism*
;
Collagen Type I/metabolism*
;
Amino Acid Substitution
;
Mutation
;
Mutation, Missense
9.Mechanism analysis of ω-3 polyunsaturated fatty acids in alleviating oxidative stress and promoting osteogenic differentiation of MC3T3-E1 cells through activating Nrf2/NQO1 pathway.
Jiahui HUANG ; Long CHEN ; Chen XU ; Haojie YU ; Shishuai ZHOU ; Jianzhong GUAN
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(11):1459-1467
OBJECTIVE:
To explore the mechanism by which ω-3 polyunsaturated fatty acids (hereinafter referred to as "ω-3") exert antioxidant stress protection and promote osteogenic differentiation in MC3T3-E1 cells, and to reveal the relationship between ω-3 and the key antioxidant stress pathway involving nuclear factor E2-related factor 2 (Nrf2) and NAD (P) H quinone oxidoreductase 1 (NQO1) in MC3T3-E1 cells.
METHODS:
The optimal concentration of H 2O 2 (used to establish the oxidative stress model of MC3T3-E1 cells in vitro) and the optimal intervention concentrations of ω-3 were screened by cell counting kit 8. MC3T3-E1 cells were divided into blank control group, oxidative stress group (H 2O 2), low-dose ω-3 group (H 2O 2+low-dose ω-3), and high-dose ω-3 group (H 2O 2+high-dose ω-3). After osteoblastic differentiation for 7 or 14 days, the intracellular reactive oxygen species (ROS) level was measured by fluorescence staining and flow cytometry, and the mitochondrial morphological changes were observed by biological transmission electron microscope; the expression levels of Nrf2, NQO1, heme oxygenase 1 (HO-1), Mitofusin 1 (Mfn1), and Mfn2 were detected by Western blot to evaluate the cells' antioxidant stress capacity; the expression levels of Runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN) were detected by immunofluorescence staining and Western blot; osteogenic potential of MC3T3-E1 cells was evaluated by alkaline phosphatase (ALP) staining and alizarin red staining.
RESULTS:
Compared with the oxidative stress group, the content of ROS in the low and high dose ω-3 groups significantly decreased, and the protein expressions of Nrf2, NQO1, and HO-1 significantly increased ( P<0.05). At the same time, the mitochondrial morphology of MC3T3-E1 cells improved, and the expressions of mitochondrial morphology-related proteins Mfn1 and Mfn2 significantly increased ( P<0.05). ALP staining and alizarin red staining showed that the low-dose and high-dose ω-3 groups showed stronger osteogenic ability, and the expressions of osteogenesis-related proteins RUNX2 and OCN significantly increased ( P<0.05). And the above results showed a dose-dependence in the two ω-3 treatment groups ( P<0.05).
CONCLUSION
ω-3 can enhance the antioxidant capacity of MC3T3-E1 cells under oxidative stress conditions and upregulate their osteogenic activity, possibly through the Nrf2/NQO1 signaling pathway.
Oxidative Stress/drug effects*
;
NF-E2-Related Factor 2/metabolism*
;
NAD(P)H Dehydrogenase (Quinone)/metabolism*
;
Animals
;
Mice
;
Osteogenesis/drug effects*
;
Cell Differentiation/drug effects*
;
Fatty Acids, Omega-3/pharmacology*
;
Signal Transduction/drug effects*
;
Osteoblasts/drug effects*
;
Reactive Oxygen Species/metabolism*
;
Cell Line
;
Hydrogen Peroxide/pharmacology*
;
Core Binding Factor Alpha 1 Subunit/metabolism*
;
Antioxidants/pharmacology*
;
Heme Oxygenase-1/metabolism*
10.Comparison of therapeutic effects of tibial transverse transport microcirculation reconstruction and periosteal distraction in the treatment of early diabetic foot.
Bi-Hui SONG ; Kang-Quan SHOU ; Tong-Zhu BAO ; Hua-Rui YANG ; Ya-Dong TAN
China Journal of Orthopaedics and Traumatology 2025;38(9):910-916
OBJECTIVE:
To compare clinical efficacy of tibial transverse transport (TTT) microcirculation reconstruction and periosteal distraction in treating patients with early diabetic foot(DF).
METHODS:
From June 2021 to June 2024, 60 patients with DF were admitted and divided into bone transport group and stretch group according to different treatment methods. There were 30 patients in bone transport group, including 16 males and 14 females;aged from 48 to 65 years old with an average of (55.59±3.78) years old;the course of disease ranged from 2 to 9 months with an average of(5.95±1.32) months;TTT microcirculation reconstruction surgery was performed. There were 30 patients in distraction group, including 17 males and 13 females;aged from 47 to 67 years old with an average of (55.24±3.81) years old;the course of disease ranged from 2 to 10 months with an average of (5.68±1.54) months;periosteal distraction surgery was performed. The skin temperature of the affected feet, the time of getting out of bed and walking after operation, the time of full weight-bearing, the wound healing time and complications were compared between two groups;the pain was evaluated by visual analogue scale (VAS) before operation and one month after operation respectively;the changes of blood flow velocity of dorsal foot arteries, ankle brachial index(ABI), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF) before and after operation at 3 months were compared between two groups.
RESULTS:
All patients were followed up for 3 to 4 months with an average of (3.52±0.12) months. There were no statistically significant differences in comparison of foot skin temperature, postoperative walking time, full weight-bearing time and complications between two groups (P>0.05). The wound healing time of bone transport group (61.26±7.31) days was shorter than that of distraction group (70.17±7.15) days, and the difference was statistically significant (P<0.05). Postoperative VAS at 1 month of bone transport group (2.19±0.21) was lower than that of distraction group (2.55±0.20), and the difference was statistically significant (P<0.05). At 3 months after operation, the blood flow velocity of dorsal foot artery, ankle-brachial index, EGF and bFGF in bone transport group were(34.73±4.18) cm·s-1, (0.95±0.13), (716.61±71.13) pg·ml-1 and (175.69±31.28) pg·ml-1, respectively;which were higher than that of distraction group (31.86±3.23) cm·s-1, (0.84±0.11), (677.37±70.21) pg·ml-1, (149.26±30.13) pg·ml-1, and the differences were statistically significant (P<0.05). There was no recurrence of ulcers in situ or at other sites in both groups during follow-up.
CONCLUSION
Compared with periosteal distraction, TTT microcirculation reconstruction surgery has a definite effect in the treatment of early DF. It could effectively reduce pain level, improve blood flow indicators and vascular endothelial function of the foot, and has a relatively high safety.
Humans
;
Male
;
Female
;
Middle Aged
;
Aged
;
Tibia/blood supply*
;
Diabetic Foot/physiopathology*
;
Microcirculation
;
Periosteum/surgery*
;
Plastic Surgery Procedures/methods*
;
Osteogenesis, Distraction

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