1.Subtalar arthroereisis for treatment of pediatric flexible flatfoot:relationship between radiographic indicators and clinical efficacy
Guangtao LIAO ; Ziyu FENG ; Xiaoyong FU ; Qinglan ZHAO ; Chao CHEN ; Jinsong HONG
Chinese Journal of Tissue Engineering Research 2026;30(3):661-670
BACKGROUND:Pediatric flexible flatfoot is a common foot deformity that often leads to foot pain and reduced quality of life.OBJECTIVE:To explore the relationship between radiographic parameters and clinical efficacy of subtalar arthroereisis in the treatment of pediatric flexible flatfoot.METHODS:A retrospective study was conducted on 56 pediatric patients(mean age of 11.8 years)who underwent subtalar arthroereisis at Guangzhou Orthopedic Hospital between January 2022 and May 2023.All patients underwent detailed radiographic examinations and clinical evaluations before and after surgery,including the American Orthopaedic Foot & Ankle Society Ankle-Hindfoot score and Visual Analog Scale score.Paired t-tests and independent t-tests were used to compare changes in radiographic parameters and clinical scores before and after surgery.Correlation analyses were conducted to evaluate the relationship between radiographic parameters and clinical outcomes.RESULTS AND CONCLUSION:(1)All radiographic parameters significantly improved during the 8 to 12-month follow-up after surgery(P<0.001).(2)Clinical evaluation results indicated that the American Orthopaedic Foot & Ankle Society Ankle-Hindfoot score significantly improved from 66.2±6.0 preoperatively to 91.3±5.8 postoperatively,and the Visual Analog Scale score significantly decreased from 3.1±0.8 preoperatively to 1.3±0.8 postoperatively(P<0.001).(3)Independent t-tests showed a significant difference in postoperative the first metatarsal angle and Visual Analog Scale score grades(P=0.043),with a smaller the first metatarsal angle associated with less postoperative pain;preoperative lateral arch angle showed a significant difference between the"excellent"and"good"groups in postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot scores(P=0.033),suggesting that a smaller preoperative posterior arch angle might predict better postoperative foot function recovery.(4)Correlation analysis showed that preoperative posterior arch angle(r=-0.486,P<0.01)and heel pitch angle(r=-0.344,P<0.01)were significantly negatively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot,while preoperative medial longitudinal arch angle(r=0.293,P<0.05)was significantly positively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot.Postoperative medial longitudinal arch angle(r=0.331,P<0.05)and lateral arch angle(r=0.387,P<0.01)were significantly positively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot,whereas postoperative Bohler's angle(r=-0.272,P<0.05),posterior arch angle(r=-0.461,P<0.01),and heel pitch angle(r=-0.318,P<0.01)were significantly negatively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot.(5)It is concluded that subtalar arthroereisis is significantly effective in correcting pediatric flexible flatfoot,and improvements in radiographic parameters are closely related to clinical efficacy.Preoperative and postoperative radiographic evaluations can serve as important reference indicators for predicting postoperative clinical outcomes,guiding clinicians to optimize treatment plansand rehabilitation programs.
2.Subtalar arthroereisis for treatment of pediatric flexible flatfoot:relationship between radiographic indicators and clinical efficacy
Guangtao LIAO ; Ziyu FENG ; Xiaoyong FU ; Qinglan ZHAO ; Chao CHEN ; Jinsong HONG
Chinese Journal of Tissue Engineering Research 2026;30(3):661-670
BACKGROUND:Pediatric flexible flatfoot is a common foot deformity that often leads to foot pain and reduced quality of life.OBJECTIVE:To explore the relationship between radiographic parameters and clinical efficacy of subtalar arthroereisis in the treatment of pediatric flexible flatfoot.METHODS:A retrospective study was conducted on 56 pediatric patients(mean age of 11.8 years)who underwent subtalar arthroereisis at Guangzhou Orthopedic Hospital between January 2022 and May 2023.All patients underwent detailed radiographic examinations and clinical evaluations before and after surgery,including the American Orthopaedic Foot & Ankle Society Ankle-Hindfoot score and Visual Analog Scale score.Paired t-tests and independent t-tests were used to compare changes in radiographic parameters and clinical scores before and after surgery.Correlation analyses were conducted to evaluate the relationship between radiographic parameters and clinical outcomes.RESULTS AND CONCLUSION:(1)All radiographic parameters significantly improved during the 8 to 12-month follow-up after surgery(P<0.001).(2)Clinical evaluation results indicated that the American Orthopaedic Foot & Ankle Society Ankle-Hindfoot score significantly improved from 66.2±6.0 preoperatively to 91.3±5.8 postoperatively,and the Visual Analog Scale score significantly decreased from 3.1±0.8 preoperatively to 1.3±0.8 postoperatively(P<0.001).(3)Independent t-tests showed a significant difference in postoperative the first metatarsal angle and Visual Analog Scale score grades(P=0.043),with a smaller the first metatarsal angle associated with less postoperative pain;preoperative lateral arch angle showed a significant difference between the"excellent"and"good"groups in postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot scores(P=0.033),suggesting that a smaller preoperative posterior arch angle might predict better postoperative foot function recovery.(4)Correlation analysis showed that preoperative posterior arch angle(r=-0.486,P<0.01)and heel pitch angle(r=-0.344,P<0.01)were significantly negatively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot,while preoperative medial longitudinal arch angle(r=0.293,P<0.05)was significantly positively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot.Postoperative medial longitudinal arch angle(r=0.331,P<0.05)and lateral arch angle(r=0.387,P<0.01)were significantly positively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot,whereas postoperative Bohler's angle(r=-0.272,P<0.05),posterior arch angle(r=-0.461,P<0.01),and heel pitch angle(r=-0.318,P<0.01)were significantly negatively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot.(5)It is concluded that subtalar arthroereisis is significantly effective in correcting pediatric flexible flatfoot,and improvements in radiographic parameters are closely related to clinical efficacy.Preoperative and postoperative radiographic evaluations can serve as important reference indicators for predicting postoperative clinical outcomes,guiding clinicians to optimize treatment plansand rehabilitation programs.
3.Multi-dimensional Holographic Characterization of Zhejiang Characteristic Atractylodis Macrocephalae Rhizoma with Nine-time Repeating Steaming and Processing
Xin WU ; Cuiwei CHEN ; Qiao YU ; Chao FENG ; Hongyan ZHANG ; Yan CHEN ; Caihua SUN ; Gang CAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):197-205
ObjectiveHistorically documented Zhejiang Atractylodis Macrocephalae Rhizoma(Baizhu) possesses premium characteristics such as phoenix-like head and crane-like neck, pronounced sweetness, and fragrant aroma. However, its current market circulation is low, and the processed products with Zhejiang-style characteristics are at the risk of being lost. This study aims to preserve the ancient Zhejiang-style processing techniques and evaluate them using modern scientific methods. MethodsMultidimensional intelligent sensory evaluation was used to digitally characterize the "quality-structure" of the external appearance of nine-steamed and nine-processed Baizhu medicinal materials(intermediate processed products) and the "odor-taste" of the internal quality of its decoction pieces(slices), and the appearance parameters were digitally characterized by colorimeter, texture analyzer, electronic nose and electronic tongue, the chemical composition was analyzed via ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS). Then, cluster analysis on the differences in odor between the medicinal materials(intermediate processed products) and decoction pieces(slices) of nine-steamed and nine-processed Baizhu was conducted, as well as the differences in taste between water-soluble and alcohol-soluble extracts of the decoction pieces(slices), and the correlation analysis of chroma value-alcohol-soluble extract content-component response value. ResultsThe nine-steamed and nine-processed Baizhu had a dark brown to black epidermis, a brownish-yellow to brownish-gray cross-section, a slightly tough texture, a faint odor, and a slightly sweet, bitter and pungent taste. Texture analyzer measurements revealed minimal adhesion and maximum recovery in the middle section of the characteristic processed Baizhu, consistent with the processing endpoint of thorough steaming and cooking. The head section showed the highest internal hardness, elasticity and chewiness, indicating a denser texture in this area. The electronic nose sensor could clearly distinguish the difference between the medicinal materials and its decoction pieces, with a more significant clustering effect at 60 ℃ for 30 minutes compared to ambient temperature headspace for 2 hours, highlighting the significant impact of the baking degree before slicing on the quality. The electronic tongue taste signal map clearly distinguished the differences between water-soluble and alcohol-soluble extracts of nine-steamed and nine-processed Baizhu decoction pieces, and the addition of auxiliary materials during processing could enhance its alcohol-soluble extract content. A total of 82 chemical components were identified in the characteristic processed Baizhu. After processing, the contents of 58 components increased, while 24 components decreased. Correlation analysis revealed significant negative correlations(P<0.01) between ethanol-soluble extract content and colorimetric values of brightness(L*), yellow-bule value(b*), and total color difference(E*ab). E*ab showed marked negative correlations(P<0.05) with the response values of isochlorogenic acid A and C. ConclusionThis study establishes a modern intelligent sensory evaluation model for multidimensional holographic characterization of nine-steamed and nine-processed Baizhu, clarifying the correlation between increased isochlorogenic acid content and the visual color appearance after different steaming cycles, as well as its intrinsic alcohol-soluble extracts. This provides a reference for quality evaluation and processing standards of the Zhejiang-style characteristic processed products.
4.Evaluation of the public health governance capacity in Jiangsu Province
Peiyu FENG ; Anning MA ; Peiwu SHI ; Qunhong SHEN ; Chaoyang ZHANG ; Zheng CHEN ; Chuan PU ; Lingzhong XU ; Zhaohui GONG ; Tianqiang XU ; Panshi WANG ; Chao HAO ; Zhi HU ; Mo HAO ; Hua WANG ; Chengyue LI
Shanghai Journal of Preventive Medicine 2026;38(2):146-152
ObjectiveTo evaluate the public health governance capacity in Jiangsu Province and provide an optimized pathway for the construction of a “strong, rich, beautiful, and high-quality” new Jiangsu. MethodsA total of 806 policy documents, 658 public information reports, and 148 research literatures related to public health governance capacity in Jiangsu Province from January 1995 to December 2023 were collected. The status of current public health goverance was assessed based on the evaluation criteria suitable for public health systems, and the strengths and the weaknesses of the system were identified. ResultsThe public health governance capability of Jiangsu Province was scored at 738.3 points, ranking 3rd nationally. Maternal health care and emergency response capacities achieved leading positions nationwide, both ranking 2nd. Jiangsu had exhibited a standardized guidance in the strategic level, a well-established management mechanism, an extensive coverage in information collection, and a scientifically established health targets setting. However, bottlenecks remained, including an unclear division of responsibilities across organizational departments, an insufficient public-health workforce, the absence of a stable growth mechanism for government funding investment, and difficulties in promptly identifying public needs. ConclusionJiangsu’s public-health system demonstrates leading nationally, yet several components remain underdeveloped. Future efforts should consolidate advantages while addressing weaknesses, further diversify content and forms, establish a stable funding increase mechanism, and clarify departmental functions, thereby providing solid health support for realizing the developmental goals of a “strong, rich, beautiful and high-quality” new Jiangsu.
5.Clinical Importance of BAIAP2L1 Expression in Cervical Cancer and Its Effect on Malignant Phenotype of Cervical Cancer Cells
Jueying ZHAO ; Zhuoying HAN ; Lulu FENG ; Chenlong WANG ; Li ZHANG ; Chao LUO ; Qilong WANG
Cancer Research on Prevention and Treatment 2025;52(6):481-490
Objective To explore the expression characteristics of BAIAP2L1 in cervical cancer (CC) and its regulatory role in tumor cell metastasis. Methods The correlation between BAIAP2L1 expression and clinical prognosis was analyzed by using a public database. GO pathway enrichment and clinicopathological correlation analyses were conducted by employing R language. The effect of BAIAP2L1 knockdown on CC cell proliferation, invasion, migration, and epithelial-mesenchymal transition (EMT) were further investigated through gene silencing approaches. Results BAIAP2L1 expression was significantly upregulated in CC tissues (Padj <0.001) and it was identified as an independent risk factor for patient mortality (HR=2.808, P=0.03). Elevated BAIAP2L1 levels showed significant correlations with poor overall survival, advanced T/N stage, recurrence, and metastasis (all P<0.05). Functional enrichment analysis revealed its involvement in tumor metastasis-related pathways. The knockdown of BAIAP2L1 significantly attenuated CC cell proliferation, invasion, and migration and suppressed key EMT processes (all P<0.05). Conclusion BAIAP2L1 is overexpressed in CC tissues and associated with patient prognosis and metastasis. The targeted inhibition of BAIAP2L1 can effectively curb tumor progression.
6.Identification of unknown pollutants in drinking water based on solid-phase extraction and supramolecular solvent extraction
Zixin QIAN ; Yuhang CHEN ; Chao FENG ; Yuanjie LIN ; Qian XU ; Ziwei LIANG ; Xinyu WANG ; Dasheng LU ; Ping XIAO ; Zhijun ZHOU
Journal of Environmental and Occupational Medicine 2025;42(7):854-861
Background With the progression of industrialization, an increasing number of emerging contaminants are entering aquatic environments, posing significant threats to the safety of drinking water. Therefore, establishing a system for identifying unknown hazardous factors and implementing safety warning mechanisms for drinking water is of paramount importance. Among these efforts, non-target screening plays a critical role, but its effectiveness is largely constrained by the scope of coverage of sample pre-treatment methods. Objective To integrate modern chromatography/mass spectrometry techniques with advanced data mining methods to develop a non-discriminatory sample pre-treatment method for comprehensive enrichment of unknown contaminants in drinking water, laying a technical foundation for the discovery and identification of unknown organic hazardous factors in drinking water. Methods A non-discriminatory pre-treatment method based on supramolecular and solid-phase extraction was developed. The final target compounds including 333 pesticides, 194 pharmaceuticals and personal care products (PPCPs), and 59 per- and polyfluoroalkyl substances (PFASs) were used for optimizing the pre-treatment method, confirming its coverage. The impacts of different eluents on the absolute recovery rates of target compounds were compared to select the conditions with the highest recovery for sample pre-treatment. The effects of different supramolecular solvents and salt concentrations on target compound recovery were also evaluated to determine the most suitable solvent and salt concentration. Results The solid-phase extraction elution solvents, supramolecular extraction solvents, and salt concentrations were optimized based on the target compound recovery rates. The optimal recovery conditions were achieved using 2 mL methanol, 2 mL methanol (containing 1% formic acid), 2 mL ethyl acetate, 2 mL dichloromethane, hexanediol supramolecular solvent, and 426 mg salt. The detection method developed based on these conditions showed a good linear relationship for all target compounds in the range of 0.1-100.0 ng·mL−1, with R² > 0.99. The method’s limit of detection ranged from 0.01 ng−1 to 0.95 ng−1, and 95% of target compounds were recovered in the range of 20%-120%, with relative standard deviation (RSD) less than 30%, indicating good precision. Conclusion The combined pre-treatment method of solid-phase extraction and supramolecular solvent extraction can effectively enrich contaminants in drinking water across low, medium, and high polarities, enabling broad-spectrum enrichment of diverse trace contaminants in drinking water. It provides technical support for broad-spectrum, high-throughput screening and identification of organic pollutants in drinking water, and also serves as a reference for establishing urban drinking water public safety warning systems.
7.Prevalence of Schistosoma japonicum infections in wild rodents in key areas during the elimination phase
Chao LÜ ; Xiaojuan XU ; Jiajia LI ; Ting FENG ; Hai ZHU ; Yifeng LI ; Ling XU ; Zhihong FENG ; Huiwen JIANG ; Xiaoqing ZOU ; Wenjun WEI ; Zhiqiang QIN ; Yang HONG ; Shiqing ZHANG ; Jing XU
Chinese Journal of Schistosomiasis Control 2025;37(5):475-481
Objective To investigate the prevalence of Schistosoma japonicum infections in wild rodents in schistosomiasis-endemic areas of China, so as to provide insights into formulation of technical guidelines for monitoring of and the precise control strategy for S. japonicum infections in wild rodents during the elimination phase. Methods Two administrative villages where schistosomiasis was historically highly prevalent were selected each from Dongzhi County, Anhui Province, and Duchang County, Jiangxi Province as study villages. Wild rodents were captured from study villages with baited traps or cages at night in June and September, 2021. The number of rodents captured was recorded, and the rodent species was characterized based on morphologi-cal characteristics. Liver tissues were sampled from captured rodents for macroscopical observation of the presence of egg granu- lomas, and S. japonicum infection was detected simultaneously using liver tissue homogenate microscopy, examinations of mesenteric tissues for parasites, and modified Kato-Katz thick smear technique (Kato-Katz technique). A positive S. japonicum infection was defined as detection of S. japonicum eggs or adult worms by any of these methods. The rate of wild rodent capture and prevalence of S. japonicum infections in wild rodents were compared in different study villages and at different time periods, and the detection of S. japonicum infections in wild rodents was compared by different assays. Results The overall rate of wild ro- dent capture was 8.28% (237/2 861) in Dongzhi County, and the wild rodent capture rates were 9.24% (133/1 439) and 7.31% (104/1 422) in two study villages (χ2 = 3.503, P = 0.061), and were 8.59% (121/1 409) and 7.99% (116/1 452) in June and September, 2021, respectively (χ2 = 0.337, P = 0.561). The overall rate of wild rodent capture was 3.72% (77/2 072) in Duchang County, and the wild rodent capture rates were 6.91% (67/970) and 0.91% (10/1 102) in two study villages (χ2 = 51.901, P < 0.001), and were 4.13% (39/945) and 3.37% (38/1 127) in June and September, 2021, respectively (χ2 = 0.815, P = 0.365). Rattus norvegicus was the predominant rodent species captured in both counties, accounting for 70.04% (166/237) of all captured wild rodents in Dongzhi County and 88.31% (68/77) in Duchang County. No S. japonicum infection was detected in wild rodents captured in Duchang County. Nevertheless, the overall prevalence of S. japonicum infections was 51.05% (121/237) in wild rodents captured in Dongzhi County, with prevalence rates of 50.38% (67/133) and 51.92% (54/104) in two study villages (χ2 = 0.098, P = 0.755), and 54.31% (63/116) and 47.93% (58/121) in September and June, 2021, respectively (χ2 = 0.964, P = 0.326). Of 237 wild rodents captured in Dongzhi County, there were 140 (59.07%) rodents with visible hepatic egg granulomas, 117 (49.47%) tested positive for S. japonicum eggs by liver tissue homogenate microscopy, 34 (14.35%) tested positive for S. japonicum eggs with Kato-Katz technique; however, no adult S. japonicum worms were detected in mesenteric tissues. In addition, hepatic egg granulomas were found in all wild rodents tested positive for S. japonicum eggs with liver tissue homogenate microscopy. Conclusions The rate of wild rodent capture and prevalence of S. japonicum infection in wild rodents vary greatly in schistosomiasis-endemic areas of China, and the prevalence of S. japonicum infection is slightly higher in wild rodents captured in autumn than in summer. Liver tissue is recommended as the preferred sample for surveillance of S. japonicum infection in wild rodents, and a combination of macroscopical observation of hepatic egg granulomas and liver tissue homogenate microscopy may be a standard method for surveillance of S. japonicum infection in wild rodents.
8.Comparison of differences in dosimetry and treatment efficiency of modified radiotherapy plans after left-sided breast-conserving surgery
Jian-hai LIN ; Jing FENG ; Zhong-hua CHEN ; Zhi-chao FU ; Jie CHEN ; Nan-bao ZHONG
Chinese Medical Equipment Journal 2025;46(4):45-51
Objective To compare the differences in dosimetry and treatment efficiency of three radiotherapy plans after left-sided breast-conserving surgery,including modified intensity-modulated radiation therapy(IMRT),cross-field volume-modulated arc therapy(VMAT)or improved VMAT,so as to provide references for clinical practice.Methods Three radiotherapy plans of modified IMRT,cross-field VMAT and improved VMAT were designed for 12 patients after left-sided-breast-conserving surgery.The modified IMRT with five irradiation fields and the improved VMAT with two arcs were modified by not setting cross-fields while determining the start and end angles with the rays passing through the least lung area.The cross-field VMAT had its start and end angles set based on the cross-fields.The doses to the target areas,peripheral organs at risk,heart and its substructures were evaluated,and dose verification was carried out.The three plans were compared in terms of treatment efficiency and gamma pass rate.SPSS 22.0 was used for statistical analysis.Results All the three plans behaved well in dose distribution.In terms of planning gross tumor volume dosimetry dosimetry,the improved VMAT and modified IMRT gained advantages than others in CI and D50,respectively,with the differences being significant(all P<0.05).In terms of planning target volume dosimetry,the modified IMRT had the V107 and D50 lower than those of the others,with the differences being significant(P<0.05).In terms of the protection of peripheral organs at risk,V5 of the left lung,Dmean of the right lung and Dmean of the healthy breast were lower in the modified IMRT plan than in the other 2 plans,with statistically significant differences(P<0.05);V20,V30,V35 and V40 of the left lung were lower in the modified VMAT plan than in the other 2 plans,with statistically significant differences(P<0.05).In terms of protection of heart and its substructures,the left ventricle V20,V30 and Dmean of the improved VMAT plan behaved better than those of other 2 plans,and the difference was statistically significant(all P<0.05).In terms of treatment efficiency,the cross-field VMAT plan had the lowest MU while highest treatment efficiency;the improved VMAT plan had the MU higher while the treatment efficiency lower than the cross-field VMAT plan;the modified IMRT plan had the highest MU while the lowest gamma pass rate,and the differences in the MUs and gamma pass rates among the three plans were statistically significant(P<0.05).Conclusion Under the same standard conditions,the cross-field VMAT and improved VMAT plans show technical advantages.Though the improved VMAT plan has the treatment efficiency lower than the cross-field VMAT,it decreases the possibility of radiocardiac injury in terms of dosimetry and thus can be used for radiotherapy after left-sided breast-conserving surgery.[Chinese Medical Equipment Journal,2025,46(4):45-51]
9.Endovascular intervention technology for constructing canine models of hepatic vein obstruction type Budd-Chiari syndrome
Xuedong SUN ; Feng DUAN ; Chao MA ; Zhenfei SONG ; Maoqiang WANG ; Long JIN
Chinese Journal of Interventional Imaging and Therapy 2025;22(3):205-209
Objective To explore the feasibility of endovascular intervention technology for constructing canine models of hepatic vein(HV)obstruction type Budd-Chiari syndrome(BCS).Methods HV obstruction type BCS models were established through double approaches including external jugular vein and percutaneous transhepatic vein puncture,blocking the main HV with double balloon and injecting anhydrous ethanol using 10 Beagle dogs.Laboratory indexes before and after modeling were compared.Liver ultrasound was performed 2,4 and 8 weeks after modeling,respectively,while intravascular ultrasound(IVUS)and digital subtracting angiography(DSA)were performed after the last time ultrasound to observe the obstruction degree,lumen and blood flow of left hepatic vein(LHV).The stenosis rate of LHV was evaluated based on DSA findings,and successful establishment of HV obstruction type BCS canine model was regarded as stenosis rate>30%and incomplete occlusion of LHV,then the success rate of modeling was recorded.Results No significant difference of laboratory indexes was found before and after modeling(all P>0.05).Ultrasound,IVUS and DSA showed that after modeling,different degrees of LHV stenosis complicated with intrahepatic collateral circulation were found in 9 dogs,while complete occlusion of LHV was noticed in 1 dog.The success rate of modeling was 90.00%(9/10).Pathological results showed the target HV lumen narrow with thickened endovascular wall,and the latter mainly composed of fibroblast and collagen fiber proliferation.Conclusion Endovascular intervention technology was feasible for constructing canine models of HV obstruction type BCS.
10.Iron overload induces ferroptosis in osteoblast precursor cells and inhibits osteogenic differentiation
Yu PAN ; Renfeng ZHAO ; Xingping LI ; Chengdong ZHANG ; Feng SHI ; Chao PU ; Xuwei LUO ; Dongqin XIAO
Chinese Journal of Tissue Engineering Research 2025;29(30):6381-6390
BACKGROUND:Iron overload is an independent factor inducing osteoporosis,but the action mechanism is currently unclear.Therefore,exploring the effects of iron overload on osteoblast-related cells will help to deeply understand the pathogenesis of osteoporosis and provide potential strategies for osteoporosis treatment.OBJECTIVE:To explore the effects of iron overload environment on osteoblast precursor cell activity,ferroptosis,and osteogenic differentiation.METHODS:Osteoblast precursor cells(MC3T3-E1 cells)were divided into blank group,iron overload group,fer-1 group,and deferoxamine group.The iron overload group was treated with 300 μmol/L ammonium ferric citrate in the culture medium for 48 hours to simulate the iron overload microenvironment.The cells in fer-1 group and deferoxamine group were pretreated with 5 μmol/L antioxidant fer-1 and 5 μmol/L deferoxamine for 8 hours,respectively,and then added with 300 μmol/L ammonium ferric citrate for 48 hours.CCK-8 assay was used to determine the cell viability.Intracellular reactive oxygen species levels were detected employing a reactive oxygen species fluorescent probe.Changes in mitochondrial membrane potential were monitored with a mitochondrial membrane potential fluorescent probe.Mitochondrial morphology was observed employing transmission electron microscopy.Cellular glutathione levels were measured with a reduced glutathione colorimetric assay kit.Lipid peroxidation levels were assessed with a malondialdehyde colorimetric assay kit.Cellular ferrous ion levels were determined with a ferrous ion colorimetric assay kit.The osteogenic and mineralization capabilities of the cells were verified by alkaline phosphatase staining and alizarin red staining.Collagen secretion ability was detected using Sirius Red staining.The expression of osteogenic/ferroptosis-related genes and proteins was examined through reverse transcription quantitative polymerase chain reaction and western blot analysis.RESULTS AND CONCLUSION:(1)In an iron-overload environment,the mitochondrial membrane potential of cells decreased and their structure was compromised,with an elevation in intracellular lipid peroxidation levels and a downregulation of genes and proteins associated with ferroptosis resistance.However,pretreatment with fer-1 and deferoxamine led to an increase in mitochondrial membrane potential and partial restoration of morphology,a reduction in intracellular lipid peroxidation levels,and an upregulation of genes and proteins related to ferroptosis resistance.(2)In an iron-overload environment,the levels of cellular alkaline phosphatase,the formation of mineralized nodules,and the synthesis of collagen fibers were all found to be decreased.Pretreatment with fer-1 and deferoxamine was observed to upregulate the expression of osteogenic differentiation in cells.(3)In summary,iron overload could increase intracellular oxidative stress levels,mediate ferroptosis in MC3T3-E1 cells and inhibit osteogenic differentiation,thereby inducing osteoporosis.Therefore,maintaining iron homeostasis and inhibiting osteogenesis-related ferroptosis may be potential strategies to prevent or treat osteoporosis.

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