Objective To evaluate the short-term effectiveness of talonavicular arthrodesis for Müller-Weiss disease. Methods Between May 2013 and February 2015, 13 patients with Müller-Weiss disease were treated with talonavicular arthrodesis. There were 11 females and 2 males with an average age of 59 years (range, 42-67 years). The disease duration was 8-20 years (mean, 13 years). According to Maceira stage, there were 7 cases of stage Ⅲ, 6 cases of stage Ⅳ. The foot longitudinal arch height measured on weight-bearing X-ray films was (43.1±1.8) mm; the Meary angle and talocalcaneal angle measured on lateral X-ray films were (–2.8±2.3)° and (5.8±2.4)°, respectively; the calcaneal valgus angle measured on Saltzman position X-ray films was (–2.0±0.7)°. The American Orthopaedic Foot and Ankle Society (AOFAS) score was 43.5±12.4, and visual analogue scale (VAS) score was 7.3±1.5. Results All the patients were followed up 14-39 months (mean, 20 months). The symptoms of foot pain and intermittent claudication disappeared in all patients. All cases achieved bony union, the fusion time was 12-16 weeks (mean, 13 weeks). There was no complications such as wound infection, skin necrosis, or internal fixator broken. At last follow-up, the foot longitudinal arch height, Meary angle, talocalcaneal angle, and calcaneal valgus angle were (52.5±2.2) mm, (1.3±2.2)°, (16.5±3.7)°, and (0.4±0.7)°, respectively; the AOFAS score and VAS score were 83.8±9.1 and 1.0±0.4, respectively; all were significantly improved when compared with preoperative ones (P<0.05). Conclusion If the subtalar and calcaneocuboid joints are relatively healthy, talonavicular arthrodesis may be a reliable and effective surgical option for Müller-Weiss disease that is resistant to conservative treatment.
Müller cells are glial cells of the retina, whose major processes cross the internal and external limiting membranes of the retina, maintaining the function and metabolism of retinal photoreceptors and neurons. Their structure and function are closely related to the development of macular hole (MH). Müller cells are involved in the formation and recovery of MH from the aspect of traction and protein, and their morphology and biological function also influence the regression of MH. The current treatment modality for MH is vitrectomy combined with internal limiting membrane (ILM) peeling, in which Müller cells play a dual role after ILM peeling in different stages of MH. And its potential to re-acquire a progenitor-like state following retinal injury with the ability to proliferate and generate new neurons making it a current research hot topic, which can be a reference and inspiration for clinical treatment.
Ischemic retinopathy, resulting in multiple lesions like microvasculature damage, inflammation and neovascularization, is a major contributor of vision damage. In these pathological changes, retinal glia cannot be ignored in the development of retinopathy. They constitute a highly versatile population that interacts with various cells to maintain homeostasis and limit disease. Therefore, glial activation and gliosis are strikingly ubiquitous responses to almost every form of retinal disease. Both of microglial cells and Müller cells are major intrinsic retinal glial cells and they are in close relationship, which means they can influence each other, make joint action or even become interdependent. They exhibit morphological and functional changes to have an impact on degree of retinal injury through different responses, which mediated by glial cells are important not only for course of disease progression, but also for the maintenance of neuronal and photoreceptor survival. Thus, defining the mechanisms that underlie communications between microglial cells and Müller cells could enable the development of more selective therapeutic targets, with great potential clinical applications.
Objective To study the effects of connective tissue growth factor (CTGF) on retinal Müller cells based on transcriptome analysis of RNA-seq technology.MethodsRetinal Müller cells were divided into the control group and the CTGF treatment group which was continuously cultured with 10 ng/ml of CTGF for 24 h. The influence of CTGF on the migration of Müller cells were tested by scratching experiments. The RNA transcriptome analysis was applied to complete transcriptome sequencing, differentially expressed genes and functional enrichment analysis of the two groups of cells. HiSeq sequencing technology was used to sequence the whole transcriptome of the two groups of cells to obtain biological big data, and analyze the differentially expressed miRNAs on this basis. The functions and signal pathways of differential miRNAs were analyzed through gene annotation (GO) functional significance enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway significant enrichment analysis. Based on transcriptome data, genes with differential expression multiples in the top ten between the two groups were screened out, and the expression of bone morphogenetic protein 4 (BMP4) gene was verified by real time fluorescence quantification PCR (qRT-PCR), immunofluorescence and Western blot.ResultsAfter CTGF stimulation of Müller cells, cell viability and mobility which compared with the control group were significantly increased, with statistically significant differences (t=3.453, P＜0.05). The differential gene expression profile of CTGF induced Müller cells was obtained by RNA transcriptome analysis. Comparing the sequencing results of the two groups, it was found that 325 differentially expressed genes included 152 up-regulated genes and 173 down-regulated genes. The results of GO functional significance enrichment analysis showed that the functions of differential miRNA were mainly divided into three categories: biological processes, cellular components, and molecular functions. These differentially expressed genes were involved in signaling between nervous systems, adhesion between cells, and the interaction between cytokines and their receptors. These differentially expressed genes were involved in different metabolic pathways and biological processes such as tissue inflammation and fibrosis. BMP4 gene was seected for verification through immunofluorescence, qRT-PCR and western blot. The results showed that the expression of BMP4 was significantly higher than that in the control group, and the difference was statistically significant (t=39.490, 10.110, 5.470; P=0.004, 0.001, 0.006).ConclusionCTGF promotes cell proliferation and migration by up-regulating the expression of BMP4 in Müller cells, leading to tissue fibrosis and inducing inflammation.
ObjectiveTo investigate the short-term effectiveness of talonavicular joint arthrodesis and calcaneus osteotomy in the treatment of Müller-Weiss disease. MethodsBetween June 2015 and February 2017, 14 patients diagnosed Müller-Weiss disease, who were ineffective on conservative treatment, were treated with talonavicular joint arthrodesis and calcaneus osteotomy. There are 3 males and 11 females, with an average age of 46.2 years (range, 35-56 years). According to the Maceira grading criteria, 5 patients were rated as stage Ⅲ and 9 patients as stage Ⅳ. The disease duration ranged from 4 to 12 years (mean, 7 years). Preoperative X-ray films showed that all patients were not accompanied with adjacent joint arthritis. The hindfoot axis on Saltzman view was (9.8±2.8)°, calcaneal pitch angle (CPA) on lateral position was (14.7±5.1)°, Meary angle on lateral position was (4.8±2.8)°, and talar 1 meta-tarsal angle (T1MA) on anteroposterior position was (25.0±7.3)°. Preoperative visual analogue scale (VAS) score was 5.9±1.5, American Orthopedic Foot Ankle Society (AOFAS) ankle-hindfoot score was 58.8±17.6. ResultsAll patients were followed up 14-27 months (mean, 22.3 months). Medial numbness and incision infection occurred in 2, 2 cases, respectively. The other patients had no obvious discomfort. At last follow-up, VAS score was 1.6±1.3 and AOFAS score was 90.6±2.7, showing significant differences when compared with preoperative ones (t=8.18, P=0.00; t=–6.95, P=0.00). X-ray films showed that the talonavicular joint and calcaneus osteotomy achieved bony healing. The hindfoot axis on Saltzman view was (–2.5±2.7)°, CPA on lateral position was (25.0±5.2) °, Meary angle on lateral position was (2.6±2.1)°, T1MA on anteroposterior position was (8.1±3.8)°. There was no significant difference in Meary Angle between pre- and post-operation (t=1.53, P=0.15). And there were significant differences in the hindfoot axis, CPA, and T1MA between pre- and post-operation (t=11.93, P=0.00; t=–8.89, P=0.00; t=8.05, P=0.00). ConclusionFor Müller-Weiss disease patients without adjacent joint arthritis, who are ineffective on conservative treatment, the satisfied short-term effectiveness can be obtained when treated by talonavicular joint arthrodesis and calcaneus osteotomy.
Objective To develop a method for the primary culture of retinal Muuml;ller cells of adult rabbit in vitro. Methods Retina was isolated from adult rabbit, cut into 1 mm times; 1 mm pieces, and placed into Dulbecco modified Eagle medium/F12 containing 20% fetal bovine serum to culture. Cultured cells were identified by inverted phase contrast microscope, transmissim electron microscope and immunohistochemistry staining method. Results Visible cell processes grew out from the retinal tissues after three days culture, and more cells grew radically around the retina after seven days culture. The cultured cells were often inflated at one side and had one long process at another side, and the nuclei were elliptical and there were two or more than two nucleoli under inverted phase contrast microscope. The cytoplasm was rich and contained abundant microfilaments in eight to ten nanometers under transmission electron microscope. Immunohistochemistry assay showed that 95% of the cells were positive for glial fibrillary acidic protein and cellular retinaldehydebinding protein. Conclusion Rabbit retinal Muuml;ller cells can be cultured by the explant culture method.
ObjectiveTo observe the effect of polypyramidine tract binding protein-associated splicing factor (PSF) towards advanced glycation end products (AGEs) induced the apoptosis of Müller cells in vitro.MethodsExperimental study. Müller cells were cultured and divided into groups according to the project design, plasmid enhanced green fluorescent protein-PSF were transfected into the cells to achieve the overexpression of PSF Müller cells in vitro, then cells were exposed to AGEs and the Morphological changes were observed by HE staining and Hoechst 33258 staining while the survival rate of cells were detected by MTT assay. The effects of PSF on AGEs-induced Müller apoptosis was measured by Cell Death Detection ELISA kit. Meanwhile, 2′,7′-dichlorofluorescin diacetate staining was performed to monitor the protective effects of PSF on AGEs-induced Müller cells ROS.ResultsThe morphology of cells in normal group was full and the cytoplasm staining was uniform. In N+AGEs group and Vec+AGEs group, cell volume decreased, cytoplasm was dense and concentrated, and eosinophilic staining was enhanced. The cell morphology of PSF+AGEs group was still full, with uniform cytoplasm staining and uniform nucleus staining. The viability of N+AGEs group, Vec+AGEs group and PSF+AGEs group were 0.42±0.11, 0.35±0.12 and 0.68±0.12. The apoptosis values were 1.08±0.16, 0.96±0.20 and 0.44±0.08. The intracellular ROS levels were 28 833.67±3 550.06, 28 356.67±4 854.81, 186 163.00±382.54. Compared with N+AGEs group and Vec+AGEs group, the cell viability of PSF+AGEs group was significantly improved (F=20.65, P=0.000), cell apoptosis value (F=43.43, P=0.000) and intracellular ROS level (F=18.86, P=0.000).ConclusionPSF overexpression play a protective role in AGEs-induced apoptosis by inhibiting the production of ROS in Müller cells.
ObjectiveTo observe the protective effect of dl-3-n-Butylphthalide (NBP) on apoptosis of retinal Müller cells induced by hydrogen peroxide (H2O2).MethodsHuman retinal Müller cells cultured in vitro were divided into normal control group, model group (H2O2 group) and experimental group (H2O2+NBP group). The cells in the H2O2 group and H2O2+NBP group were cultured with 200 μmol/L H2O2 for 2 h. Then the culture solution of the H2O2 group replace with complete medium and the H2O2+NBP group replace with complete medium containing 1 μmol/L NBP. The normal control group was a conventional cultured cells. Müller cells were identified by immunofluorescence staining. Hematoxylin-eosin (HE) staining was used to observe the apoptosis morphological changes. MTT assay was used to detect the activity of of retinal Müller cells after after 24 h and 48 h of NBP intervention. Hoechst33258 staining was used to observe the apoptosis. LIVE/DEAD ® cell activity/cytotoxicity kit was used to detect cell viability. Dichlorofluorescein diacetate (DCFH-DA) + endoplasmic reticulum (ER) red fluorescent probe (ER-Tracker Red) double staining was used to observe the expression level of reactive oxygen species (ROS) in ER of cells. One-way ANOVA combined with Dunnett statistical method were used for data analysis.ResultsHE staining showed that the number of cells in H2O2+NBP group was higher than that in H2O2 group. MTT assay showed that after 24 h and 48 h of NBP intervention, the differences in cell viability between the normal control group and the H2O2 group, the H2O2 group and the H2O2+NBP group were statistically significant (t=28.96, 3.658, 47.58, 20.33; P＜0.001, 0.022). The results of Hoechst33258 showed that the nuclear nucleus of a few cells in the H2O2+NBP group was crescent-shaped and the nuclear fragmentation was reduced, and the blue fluorescence of the remaining cells was uniform. The LIVE/DEAD ® cell activity/cytotoxicity kit showed that the number of dead cells with red fluorescence in the H2O2 group increased significantly, and the number of viable cells with green fluorescence decreased significantly. In the H2O2+NBP group, the number of viable cells with green fluorescence increased, and the number of dead cells with red fluorescence decreased. The double staining results of DCFH-DA+ER-Tracker Red showed that the green fluorescence intensity of H2O2 group was significantly enhanced; the green fluorescence intensity of H2O2+NBP group was lower than that of H2O2 group.ConclusionNBP alleviates H2O2-induced apoptosis of human retinal Müller cells by inhibiting ROS production.
Objective To investigate the effect of astragaloside A (AS-A) on the photoreceptor degeneration induced by sodium iodate (NaIO3) and its related mechanism. MethodsSixty healthy male C57BL/6J mice, aged 6-8 weeks, were randomly divided into normal control (NC) group, NaIO3 group, and AS-A group, with twenty mice in each group. 30 min before modeling, AS-A group mice were intraperitoneally injected with 100 μl AS-A at a dose of 100 mg/kg body weight. 30 min later, mice in NaIO3 group and AS-A group were intraperitoneally injected with 100 μl NaIO3 at a dose of 30 mg/kg body weight. Subsequently, AS-A group mice were administered AS-A twice daily at 12 h intervals until the end of the experiment. On day 1 post-modeling, zonula occludens-1 (ZO-1) immunohistochemistry was performed to observe the structure of retinal pigment epithelium (RPE) cells; real-time quantitative polymerase chain reaction (qPCR) was conducted to detect the mRNA expression of various retinal chemokine ligand-2 (Ccl2), interleukin-1 beta (Il-1β), mixed lineage kinase domain-like protein (Mlkl), receptor-interacting protein kinase 3 (Ripk3), and tumor necrosis factor (Tnf). On day 3 post-modeling, immunohistochemistry was performed to observe the expression of ionized calcium binding adaptor molecule 1 (Iba1) and glial fibrillary acid protein (GFAP) in the retina; TdT-mediated dUTP nick-end labeling (TUNEL) assay was used to detect photoreceptor cell death in each group. On day 4 post-modeling, fundus morphology of mice in each group was observed by fundus color photography and optical coherence tomography (OCT). Hematoxylin-eosin staining (HE) was used to observe the morphological structure of the retina in each group. Inter-group comparisons between two groups were conducted using independent samples t-test, while comparisons among three groups were performed using one-way ANOVA. ResultsFundus color photography and OCT examination showed that a large number of scattered yellow-white subretinal nodular structures in the fundus of NaIO3 group mice, and a large number of strong reflection areas in the RPE layer. The number of strong reflection areas in the RPE layer was reduced in the AS-A group. Immunohistochemical analysis of ZO-1 showed that ZO-1 was largely lost on the RPE cell membrane in that NaIO3 group; whereas in the AS-A group, ZO-1 was evenly distributed on the RPE cell membrane. HE staining results showed circular black deposits were visible in the RPE layer of the NaIO3 group, and the inner and outer segments of photoreceptors were severely damaged, with a significant decrease in the number of outer nuclear layer (ONL) cell nuclei; whereas in the AS-A group, the RPE layer pigments were orderly, the inner and outer segments of photoreceptors were intact, and the number of ONL cell nuclei significantly increased. The results of TUNEL staining show that numerous TUNEL-positive cell nuclei were observed in the ONL of the retina in the NaIO3 group, while the number of TUNEL-positive cell nuclei in the ONL of the retina was significantly reduced in the AS-A group, with statistically significant differences (t=2.66, P＜0.05). The analysis of qPCR data showed that compared with the AS-A group, the relative expression levels of Mlkl, Ripk3, Ccl2, Il-1β and Tnf mRNA in the retina were significantly increased in the NaIO3 group, with statistically significant differences (F=39.18, 10.66, 53.51, 41.40, 24.13; P＜0.001). Immunohistochemical staining results showed that compared with NC group and AS-A group, the positive expression of GFAP in retina of NaIO3 group was significantly increased, and the difference was statistically significant (F=9.62, P＜0.05). ConclusionAS-A antagonizes NaIO3-induced photoreceptor degeneration in part by inhibiting photoreceptor cell death and neuroinflammation. Meanwhile, AS-A treatment protects against NaIO3-triggered perturbation of retinal homeostasis.
Objective To compare the effectiveness of talonavicular-cuneiform joint fusion with iliac bone grafting and without bone grafting in the treatment of Müller-Weiss diseases (MWD). Methods The clinical data of 44 patients (44 feet) with MWD who received talonavicular-cuneiform joint fusion between January 2017 and November 2022 and met the selection criteria was retrospectively analyzed. Among them, 25 patients were treated with structural iliac bone grafting (bone grafting group) and 19 patients without bone grafting (non-bone grafting group). There was no significant difference (P>0.05) in age, gender composition, body mass index, disease duration, affected side, Maceira stage, and preoperative American Orthopaedic Foot and Ankle Society (AOFAS) score, visual analogue scale (VAS) score, anteroposterior/lateral Meary angle, and Pitch angle between the two groups. Operation time, operation cost, and postoperative complications were recorded in the two groups. AOFAS and VAS scores were used to evaluate the function and pain degree of the affected foot. Meary angle and Pitch angle were measured on the X-ray film, and the joint fusion was observed after operation. The difference (change value) of the above indexes before and after operation was calculated for comparison between groups to evaluate the difference in effectiveness. Results The operation was successfully completed in both groups, and the incisions in the two groups healed by first intention. The operation time and cost in the bone grafting group were significantly more than those in the non-bone grafting group (P<0.05). All patients were followed up. The median follow-up time was 41.0 months (range, 16-77 months) in the non-bone grafting group and 40.0 months (range, 16-80 months) in the bone grafting group. There was skin numbness of the medial dorsalis of the foot in 1 case, internal fixation stimulation in 2 cases, and pain at the iliac bone harvesting area in 1 case of the bone grafting group. There was skin numbness of the medial dorsalis of the foot in 1 case and muscle atrophy of the lower limb in 1 case of the non-bone grafting group. There was no significant difference in the incidence of complications between the two groups (P>0.05). At last follow-up, the AOFAS scores of the two groups significantly improved when compared with those before operation, while the VAS scores significantly decreased, the anteroposterior/lateral Meary angle and Pitch angle significantly improved, and the differences were significant (P<0.05). There was no significant difference in the change values of outcome indicators between the two groups (P>0.05). There was no delayed bone union or bone nonunion in both groups, and joint fusion was achieved at last follow-up. Conclusion In the treatment of MWD, there is no significant difference in effectiveness and imaging improvement of talonavicular-cuneiform joint fusion combined with or without bone grafting. However, non-bone grafting can shorten the operation time, reduce the cost, and may avoid the complications of bone donor site.