1.Finite element analysis of optimization of femoral prosthesis implantation position in unicompartmental knee arthroplasty in osteoporotic patients
Mengfei LIU ; Gang CHEN ; Yihan SHI ; Lin ZENG ; Kan JIANG ; Yilihamujiang·Wusiman
Chinese Journal of Tissue Engineering Research 2025;29(3):464-470
BACKGROUND:The reasonable implantation range of femoral prosthesis in unicompartmental knee arthroplasty in patients with osteoporosis has not been investigated,and previous studies have often been based on unicompartmental knee arthroplasty models in normal bone,with fewer mechanical studies in models with non-normal bone.Complications after unicompartmental knee arthroplasty have been shown to be highly associated with osteoporosis. OBJECTIVE:To analyze the biomechanical effects of the coronal inclination of the Sled fixed platform femoral prosthesis on unicompartmental knee arthroplasty in patients with osteoporosis and to find the correlation between osteoporosis and mid-and long-term complications after unicompartmental knee arthroplasty. METHODS:Based on the digital imaging technology to obtain the data of the knee joint and prosthesis,a normal bone knee model is then created by using specialized software such as Mimics and Geomagic studio.Based on a validated normal bone knee model,an osteoporotic knee model was created by changing the material parameters.Totally 14 unicompartmental knee arthroplasty finite element models were created using Sled fixed platform femoral prosthesis:standard position(0°),varus and valgus angles:3°,6°,9° in the normal bone and osteoporosis groups.Stress changes on the surface of polyethylene liner,cancellous bone under tibial prosthesis,and cortical bone were calculated and analyzed in all unicompartmental knee arthroplasty models. RESULTS AND CONCLUSION:(1)In the osteoporotic models,the high stress values of the polyethylene liner surface and the cancellous bone under the tibial prosthesis increased with the increase of the tilt angle of the femoral prosthesis,and the high stress values of the cortical bone surface under the tibial prosthesis increased with the increase of the prosthesis valgus angles and decreased with the increase of the varus angles.(2)For the polyethylene liner surface as well as the subcortical bone surface of the tibial prosthesis,the high stress values of the models for each inclination angle in the osteoporosis group were greater than those of the corresponding models in the normal bone group.For the surface of the cancellous bone under the tibial prosthesis,the high stress values of the tilt angle models of the osteoporosis groups were smaller than those of the normal bone groups.(3)Osteoporosis may cause biomechanical abnormalities in the internal structures of the knee after unicondylar replacement,increasing the potential risk of postoperative aseptic loosening of the prosthesis and periprosthetic fractures.Varus and valgus of the femoral prosthesis in the coronal plane should be avoided as much as possible when performing medial unicompartmental knee arthroplasty with a Sled fixation platform in osteoporotic knees.
2.Effects of polylactic acid-glycolic acid copolymer/lysine-grafted graphene oxide nanoparticle composite scaffolds on osteogenic differentiation of MC3T3 cells
Shuangqi YU ; Fan DING ; Song WAN ; Wei CHEN ; Xuejun ZHANG ; Dong CHEN ; Qiang LI ; Zuoli LIN
Chinese Journal of Tissue Engineering Research 2025;29(4):707-712
BACKGROUND:How to effectively promote bone regeneration and bone reconstruction after bone injury has always been a key issue in clinical bone repair research.The use of biological and degradable materials loaded with bioactive factors to treat bone defects has excellent application prospects in bone repair. OBJECTIVE:To investigate the effect of polylactic acid-glycolic acid copolymer(PLGA)composite scaffold modified by lysine-grafted graphene oxide nanoparticles(LGA-g-GO)on osteogenic differentiation and new bone formation. METHODS:PLGA was dissolved in dichloromethane and PLGA scaffold was prepared by solvent evaporation method.PLGA/GO composite scaffolds were prepared by dispersing graphene oxide uniformly in PLGA solution.LGA-g-GO nanoparticles were prepared by chemical grafting method,and the PLGA/LGA-g-GO composite scaffolds were constructed by blending LGA-g-GO nanoparticles at different mass ratios(1%,2%,and 3%)with PLGA.The micromorphology,hydrophilicity,and protein adsorption capacity of scaffolds of five groups were characterized.MC3T3 cells were inoculated on the surface of scaffolds of five groups to detect cell proliferation and osteogenic differentiation. RESULTS AND CONCLUSION:(1)The surface of PLGA scaffolds was smooth and flat under scanning electron microscope,while the surface of the other four scaffolds was rough.The surface roughness of the composite scaffolds increased with the increase of the addition of LGA-g-GO nanoparticles.The water contact angle of PLGA/LGA-g-GO(3%)composite scaffolds was lower than that of the other four groups(P<0.05).The protein adsorption capacity of PLGA/LGA-g-GO(1%,2%,and 3%)composite scaffolds was stronger than PLGA and PLGA/GO scaffolds(P<0.05).(2)CCK-8 assay showed that PLGA/LGA-g-GO(2%,3%)composite scaffold could promote the proliferation of MC3T3 cells.Alkaline phosphatase staining and alizarin red staining showed that the cell alkaline phosphatase activity in PLGA/LGA-g-GO(2%,3%)group was higher than that in the other three groups(P<0.05).The calcium deposition in the PLGA/GO and PLGA/LGA-g-GO(1%,2%,and 3%)groups was higher than that in the PLGA group(P<0.05).(3)In summary,PLGA/LGA-g-GO composite scaffold can promote the proliferation and osteogenic differentiation of osteoblasts,and is conducive to bone regeneration and bone reconstruction after bone injury.
3.Stress analysis of computer aided design/computer aided manufacture post-core materials with different elastic moduli
Liangwei XU ; Xitian TIAN ; Lin CHEN ; Hongyan GAO ; Xian ZHU ; Guican YANG ; Yinghao CHEN
Chinese Journal of Tissue Engineering Research 2025;29(10):2061-2066
BACKGROUND:Post and core restoration is a common choice for tooth defects,but the repair effects of various post and core materials are different. OBJECTIVE:To evaluate the stress distribution at the post and core,tooth root,and bonding agent site of post and core models made of different elastic modulus post and core materials using finite element method. METHODS:A three-dimensional root canal treated maxillary central incisor model was built using three-dimensional modeling software,which was restored with a full ceramic crown.The post and core materials in the restoration used nanoceramic resin(elastic modulus=12.8 GPa),composite resin(elastic modulus=16 GPa),hybrid ceramic(elastic modulus=34.7 GPa),glass ceramic(elastic modulus=95 GPa),titanium alloy(elastic modulus=112 GPa),and zirconia(elastic modulus=209.3 GPa).The model was fixed in cortical bone.A 100 N concentrated force of 45° from the long axis of the tooth was applied to 1/3 of the crown and tongue side of the central incisor.The stress distribution of the post and core,dentin,and tooth-root bonding agent in the model was repaired by the maximum principal stress criterion. RESULTS AND CONCLUSION:(1)When the post and core materials with higher elastic modulus was used,the post-core stress in the repair model was more concentrated.When the elastic modulus of the post and core materials(nanoceramic resin and composite resin)was close to dentin,the stress distribution of the post and core was more uniform.The stress distribution of dentin in all restoration models was similar regardless of post and core materials.When the post and core with higher elastic modulus was used,more stress concentration was shown at the post and root bonding agent in the repair model.(2)The maximum stress values at the post and core,tooth root,and the bonding agent site of post and tooth root in the nanoceramic resin model were 31.00,33.21,and 0.51 MPa,respectively.The maximum stress values at the post and core,tooth root,and the bonding agent between the post and tooth root in the composite resin model were 36.84,33.14,and 0.59 MPa,respectively.In the mixed ceramic model,the maximum stress values at the post and core,tooth root,and the bonding agent between the post and tooth root were 64.05,32.83,and 1.00 MPa,respectively.In the glass ceramic model,the maximum stress values at the post and core,tooth root,and the bonding agent between the post and tooth root were 112.30,32.69,and 1.73 MPa,respectively.In the titanium alloy model,the maximum stress values of the post and core,tooth root,and the bonding agent between the post and tooth root were 120.00,32.17,and 1.86 MPa,respectively.In the zirconia model,the maximum stress values of the post and core,tooth root,and the bonding agent between the post and tooth root were 148.80,31.85,and 2.28 MPa,respectively.(3)The higher the elastic modulus of the post and core material,the higher the maximum stress at the post and core during restoration.The elastic modulus of the post and core material had no significant effect on the maximum stress of the dental bonding agent and dentin.
4.The multi-center mid-term clinical outcomes of combined complete preservation of chordal structure mitral valve replacement with total anatomic complete arterial myocardial revascularization for coronary patients with moderate-to-severe or severe ischemic mitral regurgitation
Ke GUO ; Xujun CHEN ; Baoshi ZHENG ; Chao SHI ; Keli HUANG ; Yong CAO ; Chengquan LIAO ; Jingwei CHEN ; Yu LIN ; Chengxin LIU ; Quansheng CAO ; Lin SHEN ; Zhendong WANG
Chinese Journal of Surgery 2025;63(1):58-67
Objective:To evaluate the clinical outcomes of combined complete preservation of chordal structure mitral valve replacement (C-MVR) with total anatomical arterial myocardial revascularization (TACR) in coronary patients with moderate-to-severe or severe ischemic mitral regurgitation (IMR).Methods:This is a retrospective multi-center case series study. Data were retrospectively collected from 127 patients with coronary artery disease with moderate to severe or severe IMR who received TACR with C-MVR from July 2015 to April 2024 in 13 hospitals in China. There were 90 males and 37 females, aged (56.5±10.7) years (range: 33 to 74 years). Perioperative data and follow-up data including left ventricular ejection fraction, left ventricular end-diastolic diameter, and patency rate of arterial grafts of patients were collected. Comparisons were made using paired sample t-test or χ2 test. Results:In this cohort of 127 patients, 67 underwent concurrent tricuspid valve repair. During surgery, 113 grafts of the left internal mammary artery (LIMA), 127 grafts of the left radial artery, 80 grafts of the right radial artery, and 110 grafts of the right internal mammary artery (RIMA) were harvested. The number of the distal anastomosis was 4.2±0.4 (range: 3 to 5). The aortic cross-clamp time and cardiopulmonary bypass time were (97.5±23.4) minutes (range: 90 to 161 minutes) and (145.4±19.2) minutes (range: 101 to 210 minutes), respectively. There was one operative death. Intraoperative placement of an intra-aortic balloon pump was performed in 21 patients to improve the left ventricular ejection. No sternal ischemic occurred. All patients completed follow-up, with a mean follow-up period of (64.3±7.5) months (range: 4 to 110 months). No major cerebrovascular events occurred during the follow-up period, and all patients survived. Left ventricular ejection fraction improved postoperatively (55.0%±5.3% vs. 41.0%±15.3%, t=17.23, P<0.01). The proportion of patients with New York Heart Association functional class ≤2 increased postoperatively (23.6% (30/127) vs. 87.3% (110/126), χ2=103.77, P<0.01). The proportion of patients with Canadian Cardiovascular Society Angina Classification ≤3 decreased postoperatively (4.8% (6/126) vs. 78.7% (100/127), χ2=142.19, P<0.01). The left ventricular end-diastolic diameter decreased postoperatively ((5.70±4.50) cm vs. (6.10±0.23) cm, t=12.15, P<0.01). Coronary multi-detector computed tomography angiography (MDCTA) follow-up was conducted for (60.5±11.7) months (range: 6 to 109 months) postoperatively. MDCTA confirmed the patency rates of the grafts: 96.4% (108/112) for the LIMA grafts, 88.9% (112/126) for the left radial artery grafts, 93.7% (74/79) for the right radial artery grafts, and 90.9% (100/110) for the free RIMA grafts. No significant differences in graft patency rates were observed between the arterial grafts ( χ2=5.24, P=0.155). Conclusion:The results of this multi-centre study demonstrate satisfactory mid-term results of C-MVR with TACR for the treatment of coronary artery disease with moderate to severe or severe IMR.
5.Effect of Serum Containing Zhenwutang on Apoptosis of Myocardial Mast Cells and Mitochondrial Autophagy
Wei TANG ; Meiqun ZHENG ; Xiaolin WANG ; Zhiyong CHEN ; Chi CHE ; Zongqiong LU ; Jiashuai GUO ; Xiaomei ZOU ; Lili XU ; Lin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):11-21
ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ (AngⅡ) and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment, AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h, and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 (CCK-8) to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of B-type natriuretic peptide (BNP) in cell culture supernatant, and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently, the experiment was divided into a blank group (20% blank serum), a model group (20% blank serum + 5×10-5 mol·L-1 AngⅡ), low-, medium-, and high-dose (5%, 10% and 20%) serum containing Zhenwutang groups, an autophagy inhibitor group (1×10-4 mol·L-1 3-MA), and autophagy inducer group (1×10-7 mol·L-1 rapamycin). The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe (Lyso Tracker) and mitochondrial probe (Mito Tracker) co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax), and cytochrome C (Cyt C) in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase (Parkin), phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1), and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3, cleaved Caspase-9, Bax, Bcl-2, and Cyt C in apoptosis-related pathways, phosphorylated ubiquitin ligase (p-Parkin), phosphorylated PTEN-induced kinase 1 (p-PINK1), p62, and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways, and the change of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10-5 mol·L-1, the cell activity was the lowest, and there was no cytotoxicity. At this concentration, the surface area of cardiomyocytes was significantly increased (P<0.01), and the content of BNP in the supernatant of culture medium was significantly increased (P<0.05). Therefore, AngⅡ with a concentration of 5×10-5 mol·L-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group, the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate (P<0.01) and significantly decreased mitochondrial membrane potential (P<0.01). The results of immunofluorescence co-localization showed that compared with the blank group, the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group, the relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 in the model group was significantly up-regulated (P<0.01), while the relative mRNA expression of Bcl-2, Parkin, and PINK1 was significantly down-regulated (P<0.01). In addition, the relative protein expression of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 was significantly up-regulated (P<0.01). The LC3Ⅱ/Ⅰ was significantly decreased, and the relative protein expression of Bcl-2, p-Parkin, p-PINK1, and Beclin1 was significantly down-regulated (P<0.01). Compared with the model group, the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate (P<0.01), and the decrease ratio of mitochondrial membrane potential is significantly lowered (P<0.01) in a dose-dependent manner. Additionally, both red and green fluorescence spots became more in these groups. In the 3-MA group, the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 was significantly down-regulated (P<0.05, P<0.01), while that of Bcl-2, Parkin, and PINK1 was significantly up-regulated (P<0.01). In the serum containing Zhenwutang groups, the relative protein expression levels of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 were significantly down-regulated (P<0.05,P<0.01). The LC3Ⅱ/Ⅰ was significantly increased, and the relative protein expression levels of Bcl-2, p-Parkin, p-PINK1, and Beclin1 were significantly up-regulated (P<0.01). ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.
6.Herbal Textual Research and Modern Research Progress of Ostreae Concha
Hongyi ZHANG ; Bin WANG ; Jiawen LIU ; Yuan HU ; Lin CHEN ; Youping LIU ; Hongping CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):223-234
By consulting relevant literature of ancient herbal books and processing specifications, this paper made a systematic research and analysis of Ostreae Concha, including the name, producing area, harvesting, quality, historical evolution of processing, relevant processing specifications, modern processing technology, and changes in chemical composition and pharmacological effects before and after processing, in order to provide documentary evidence for the research on processing technology and the establishment of quality standards. According to the textual research, it is known that Ostreae Concha has a long history of being used in medicine, and there have been many aliases and local names in each historical period. Shennong's Classic of the Materia Medica(Shennong Bencaojing) began to use Muli as the correct name, which has continued to use to today, and there were also aliases such as Muge, Zuogu Muli and Haoke. Ostreae Concha has a wide range of localities and irregular harvesting periods. The ancients believed that its left shell was of superior quality, but this has not been seen in modern. And there were many kinds of processing methods of Ostreae Concha, such as grinding, roasting, calcining, frying, simmering, quenching and so on, and the calcining was still in use. The different editions of Chinese Pharmacopoeia from 1963 to 2020 contain only calcined Ostreae Concha, and the local processing specifications mainly include three kinds of processed products(calcined products, salt-soaked products and vinegar-soaked products). Modern processing research mainly focuses on process optimization, changes in chemical composition and pharmacological effects, and the research methods are relatively single. Overall, there are currently issues such as inconsistent processing standards, unclear process parameters and imperfect quality standards, which are not conducive to the quality control and standardized clinical use of Ostreae Concha. Therefore, it is necessary to further investigate the pharmacological substance basis of Ostreae Concha and its processed products in order to elucidate the processing mechanism, standardize the processing technology and improve the quality standard.
7.Simultaneous TAVI and McKeown for esophageal cancer with severe aortic regurgitation: A case report
Liang CHENG ; Lulu LIU ; Xin XIAO ; Lin LIN ; Mei YANG ; Jingxiu FAN ; Hai YU ; Longqi CHEN ; Yingqiang GUO ; Yong YUAN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(02):277-280
A 71-year-old male presented with esophageal cancer and severe aortic valve regurgitation. Treatment strategies for such patients are controversial. Considering the risks of cardiopulmonary bypass and potential esophageal cancer metastasis, we successfully performed transcatheter aortic valve implantation and minimally invasive three-incision thoracolaparoscopy combined with radical resection of esophageal cancer (McKeown) simultaneously in the elderly patient who did not require neoadjuvant treatment. This dual minimally invasive procedure took 6 hours and the patient recovered smoothly without any surgical complications.
8.Changes in hemoglobin and related influencing factors in patients with liver failure undergoing artificial liver support therapy
Ying LIN ; Li CHEN ; Fei PENG ; Jianhui LIN ; Chuanshang ZHUO
Journal of Clinical Hepatology 2025;41(1):104-109
ObjectiveTo investigate the changing trend of hemoglobin (Hb) and related influencing factors in patients with liver failure after artificial liver support system (ALSS) therapy. MethodsA total of 106 patients with liver failure who were hospitalized and received ALSS therapy in our hospital from January to December 2018 were enrolled and analyzed in terms of clinical data and red blood cell parameters such as Hb, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and red blood cell distribution width-coefficient of variation (RDW-CV). A one-way repeated-measures analysis of variance was used for comparison of continuous data with repeated measurement between groups, and the paired t-test was used for comparison between two groups. The Kruskal-Wallis H test was used for comparison of continuous data with skewed distribution between multiple groups, the Mann-Whitney U test was used for further comparison between two groups. Univariate and multivariate linear regression analyses were used to identify the influencing factors for the reduction in Hb after ALSS therapy. ResultsThe 106 patients with liver failure received 606 sessions of ALSS therapy, and Hb was measured for 402 sessions before and after treatment. There was a significant reduction in Hb after ALSS therapy in the patients with liver failure (97.49±20.51 g/L vs 109.38±20.22 g/L, t=32.764, P<0.001). Longitudinal observation was further performed for 14 patients with liver failure, and the results showed that the level of Hb was 108.50±21.61 g/L before the last session of ALSS therapy, with certain recovery compared with the level of Hb (103.14±19.15 g/L) on the second day after ALSS, and there was an increase in Hb on day 3 (102.57±21.73 g/L) and day 7 (105.57±22.04 g/L) after surgery. The level of Hb in patients with liver failure on the second day after ALSS decreased with the increase in the number of ALSS sessions (F=8.996, P<0.001), while MCV and MCH gradually increased with the increase in the number of ALSS sessions (F=9.154 and 13.460, P=0.004 and P<0.001), and RDW-CV first gradually increased and then gradually decreased (F=4.520, P=0.032); MCHC showed fluctuations with no clear trend (F=0.811, P=0.494). The multivariate linear regression analysis showed that the duration of ALSS therapy, the mode of ALSS therapy, and initial treatment were independent risk factors for the reduction in Hb after ALSS therapy. ConclusionALSS therapy can influence the level of peripheral blood Hb in patients with liver failure, and patient blood management should be strengthened for patients with liver failure who are receiving ALSS therapy.
9.Role of Macrophage Activation and Polarization in Diabetes Mellitus and Its Related Complications and Traditional Chinese Medicine Intervention
Zhichao CHEN ; Qiaoni LIN ; Liya SUN ; Jinxi WANG ; Zishan FU ; Yufeng YANG ; Yan SHI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):311-320
The occurrence of diabetes mellitus (DM) is closely related to insulin resistance and islet β cell dysfunction. Modern studies have found that macrophages are widely present in the liver,fat,skeletal muscle,islets, and other tissues and organs. Macrophage M1/M2 polarization plays an important role in the occurrence and development of diabetes mellitus and its related complications by intervening in inflammatory response,improving insulin resistance,and promoting tissue repair. Most of the traditional Chinese medicines that regulate the activation and polarization of macrophages are Qi-replenishing and Yin-nourishing,heat-clearing, and detoxicating medicinal,which are consistent with the etiology and pathogenesis of diabetes and its related complications. Therefore,by summarizing the mechanisms between macrophage activation,polarization, and insulin resistance in various tissues,this paper reviewed traditional Chinese medicine and its effective components and compounds in improving diabetes mellitus and its related complications through multi-channel regulation of macrophage polarization and regulation of M1/M2 ratio,providing references for the future treatment of DM and its related complications with traditional Chinese medicine.
10.The mechanism of Prim-O-glucosylcimifugin in improving cholesterol metabolism in osteoarthritis chondrocytes via lncRNA NEAT1/miR-128-3p
Yanming LIN ; Haishui TU ; Shujie LAN ; Chao LI ; Shiyu LU ; Yue CHEN ; Changlong FU
Journal of Beijing University of Traditional Chinese Medicine 2025;48(1):55-67
Objective:
To investigate the mechanism of action of Prim-O-glucosylcimifugin (POG) to improve cholesterol metabolism in osteoarthritic (OA) chondrocytes based on the long noncoding RNA nuclear-enriched transcript 1 (lncRNA NEAT1)/microRNA-128-3p (miR-128-3p) pathway.
Methods:
For in vivo experiments, 60 mice were divided into the normal, sham operation, model, and POG groups using the random number table method, with 15 mice per group. The osteoarthritis mouse model was constructed using the modified Hulth method in the model and POG groups. Mice in the POG group were administered 30 mg/(kg·d)POG by gavage. The other groups were administered an equal amount of normal saline for 8 weeks. The cartilage tissue structure of mice in each group was observed using hematoxylin and eosin staining. Real-time PCR was used to detect changes in the lncRNA NEAT1 and miR-128-3p mRNA expression levels in the cartilage tissues of mice. Western blotting was used to detect the protein expressions of ATP-binding cassette transporter A1 (ABCA1), liver X receptor β (LXRβ), matrix metalloprotein-3 (MMP-3), and B-lymphoblastoma-2-associated X protein (Bax) in articular cartilage of mice. An enzyme-linked immunosorbent assay was used to measure the tumor necrosis factor-α (TNF-α) content in the synovial fluid of mice. A biochemical microplate assay was used to measure the total cholesterol level in the synovial fluid of mice. The in vitro experiments were divided into the negative control, interleukin-1β(IL-1β), IL-1β+ POG, IL-1β+ oe-lncRNA NEAT1, IL-1β+ oe-lncRNA NEAT1 + POG, IL-1β + miR-128-3p inhibition, and IL-1β+ miR-128-3p inhibition+ POG groups. An OA model was established by inducing chondrocytes with IL-1β for 24 h, and 90 mg/L of POG and miR-128-3p inhibitor(50 nmol/L) were administered for 48 h as an intervention. lncRNA NEAT1 expression in chondrocytes was detected using fluorescence in situ hybridization. A dual luciferase assay was used to detect the targeting relationship between lncRNA NEAT1 and miR-128-3p. Lentiviral plasmids overexpressing lncRNA NEAT1 were used to transfect mouse chondrocytes. Real-time PCR was used to detect the effect of lncRNA NEAT1 overexpression on the mRNA level of miR-128-3p in chondrocytes. Western blotting was used to detect ABCA1, LXRβ, MMP-3, and Bax protein expression in chondrocytes after lncRNA NEAT1 overexpression and miR-128-3p inhibition.
Results:
POG significantly reduced OA cartilage tissue damage. Compared with the model group, the lncRNA NEAT1 mRNA level decreased, whereas the miR-128-3p mRNA level increased in the cartilage tissue of the POG group (P<0.05). Compared with the model group, ABCA1 and LXRβ protein expression increased in the POG group, whereas MMP-3 and Bax protein expression decreased (P<0.05). The TNF-α levels decreased in the POG group compared to the model group (P<0.05). Compared with the model group, the total cholesterol level in the synovial fluid of the joint of mice in the POG group decreased (P<0.05). The mean fluorescence intensity of lncRNA NEAT1 in the IL-1β+ POG group decreased compared with the IL-1β group (P<0.05). The relative luciferase activity in the miR-128-3p mimics group bound to the lncRNA NEAT1-WT plasmid decreased compared with the miR-128-3p negative control group (P<0.05). The lncRNA NEAT1 mRNA levels decreased, whereas the miR-128-3p mRNA levels increased in the IL-1β+ oe-lncRNA NEAT1 + POG group compared with the IL-1β+ oe-lncRNA NEAT1 group (P<0.05). Compared with the IL-1β+ POG group, ABCA1 and LXRβ protein expression decreased, whereas MMP-3 and Bax protein expression increased (P<0.05).
Conclusion
POG mediates lncRNA NEAT1/miR-128-3p to improve cholesterol metabolism in OA chondrocytes.


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