Osteoporosis is a major cause of bone fracture and can be characterised by both mass loss and microstructure deterioration of the bone.The modern way of osteoporosis assessment is through the measurement of bone miner...Osteoporosis is a major cause of bone fracture and can be characterised by both mass loss and microstructure deterioration of the bone.The modern way of osteoporosis assessment is through the measurement of bone mineral density,which is not able to unveil the pathological condition from the mesoscale aspect.To obtain mesoscale information from computed tomography(CT),the super-resolution(SR)approach for volumetric imaging data is required.A deep learning model AESR3D is proposed to recover high-resolution(HR)Micro-CT from low-resolution Micro-CT and implement an unsupervised segmentation for better trabecular observation and measurement.A new regularisation overcomplete autoencoder framework for the SR task is proposed and theoretically analysed.The best performance is achieved on structural similarity measure of trabecular CT SR task compared with the state-of-the-art models in both natural and medical image SR tasks.The HR and SR images show a high correlation(r=0.996,intraclass correlation coefficients=0.917)on trabecular bone morphological indicators.The results also prove the effectiveness of our regularisation framework when training a large capacity model.展开更多
BACKGROUND Clinical studies using Trabecular Titanium™acetabular cups have shown promising short and medium-term results.This material,due to its macro and micro surface roughness,provides a substrate for osseointegra...BACKGROUND Clinical studies using Trabecular Titanium™acetabular cups have shown promising short and medium-term results.This material,due to its macro and micro surface roughness,provides a substrate for osseointegration and enhances implant stability.However,there is a lack of evidence in the literature on the use of this material in patients with femoral neck fracture.AIM To evaluate the short-term clinical-functional and radiographic outcomes in patients with femoral neck fractures undergoing total hip arthroplasty(THA)with Trabecular Titanium™acetabular cup implants.METHODS The study included 104 patients with medial femoral neck fractures who underwent THA between January 2020 and December 2020 with the Delta TT acetabular cup(Lima Corporate,Villanova di San Daniele del Friuli,Italy).The mean age of the patients was 69.57±10.16 years(range:36-85 years).The followup period ranged from a minimum of 3 to a maximum of 4 years.Three questionnaires(Harris Hip Score,Oxford Hip Score,and EQ5D)were administered along with radiographic evaluations.Statistical methods included the Student's t-test and one-way analysis of variance for comparisons(with significance set at 0.05),and the Kaplan-Meier curve for prosthetic implant survival.RESULTS The mean follow-up was 41.5 months.The Harris Hip Score(HHS)showed a mean increase of 2.74 points(mean HHS 88.52 at 6 months postoperatively and mean HHS 91.26 at the last follow-up)with statistical significance.Similarly,the Oxford Hip Score demonstrated a statistically significant difference between follow-up groups.However,the EQ5D did not show statistically significant differences among the three groups(preoperative,6-month follow-up,and last follow-up).Revision surgery was required in 6 patients.According to Moore's criteria,96%of the acetabular components were radiographically stable and well-integrated at the last follow-up.The Kaplan-Meier curve showed a 96%survival rate.CONCLUSION The clinical and radiographic results obtained in the short to medium term confirm the excellent performance of the Delta TT acetabular cup in terms of osseointegration,providing an optimal solution both for young patients with high functional recovery demands and for fragile patients requiring optimal stability of the acetabular component to reduce the risk of implant failure.展开更多
Background: When applied to trabecular bone X-ray images, the anisotropic properties of trabeculae located at ultra-distal radius were investigated by using the trabecular bone scores (TBS) calculated along directions...Background: When applied to trabecular bone X-ray images, the anisotropic properties of trabeculae located at ultra-distal radius were investigated by using the trabecular bone scores (TBS) calculated along directions parallel and perpendicular to the forearm. Methodology: Data from more than two hundred subjects were studied retrospectively. A DXA (GE Lunar Prodigy) scan of the forearm was performed on each subject to measure the bone mineral density (BMD) value at the location of ultra-distal radius, and an X-ray digital image of the same forearm was taken on the same day. The values of trabecular bone score along the direction perpendicular to the forearm, TBS<sub>x</sub>, and along the direction parallel to the forearm, TBS<sub>y</sub>, were calculated respectively. The statistics of TBS<sub>x</sub> and TBS<sub>y</sub> were calculated, and the anisotropy of the trabecular bone, which was defined as the ratio of TBS<sub>y</sub> to TBS<sub>x</sub> and changed with subjects’ BMD and age, was reported and analyzed. Results: The results show that the correlation coefficient between TBS<sub>x</sub> and TBS<sub>y</sub> was 0.72 (p BMD and age was reported. The results showed that decreased trabecular bone anisotropy was associated with deceased BMD and increased age in the subject group. Conclusions: This study shows that decreased trabecular bone anisotropy was associated with decreased BMD and increased age.展开更多
AIM：To describe a novel technique of creating a landing strip within the trabecular meshwork to guide trabecular micro-bypass stent（iStent） implantation in patients who underwent phacoemulsification.METHODS：Thirty...AIM：To describe a novel technique of creating a landing strip within the trabecular meshwork to guide trabecular micro-bypass stent（iStent） implantation in patients who underwent phacoemulsification.METHODS：Thirty-four eyes from 30 patients who underwent i Stent implantation after phacoemulsification from May 2014 to February 2015 were included in our retrospective study. All iStents were implanted via the ＂landing strip＂ technique. A 25-gauge microvitreoretinal blade was used to bisect the trabecular meshwork to less than 1 clock-hour, effectively creating a landing strip. The iStent applicator was pressed along the landing strip and then the stent was released into the trabecular meshwork. RESULTS：Of the 34 eyes with iStent implantation, 27（79.4%） eyes had primary open-angle glaucoma, 6（17.6%） eyes had pseudoexfoliation glaucoma, and 1（2.9%） eye had ocular hypertension. At 6-month follow-up（n=17）, the mean number of hypotensive medications decreased from 2.2±1.2 at baseline to 0.8±1.3（P=0.05） and mean intraocular pressure decreased from 19.7±4.1 mm Hg at baseline to 16.7±2.1 mm Hg（P=0.58）. Two eyes（5.9%） required subsequent trabeculectomy. CONCLUSION：The ＂landing strip＂ technique appears to be an effective way to assist with iS tent implantation.展开更多
The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were stu...The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.展开更多
As the aqueous humor leaves the eye, it first passes through the trabecular meshwork （TM）. Increased flow resistance in this region causes elevation of intraocular pressure （IOP）, which leads to the occurrence of ...As the aqueous humor leaves the eye, it first passes through the trabecular meshwork （TM）. Increased flow resistance in this region causes elevation of intraocular pressure （IOP）, which leads to the occurrence of glaucoma. To quantitatively evaluate the effect of high IOP on the configuration and hydraulic permeability of the TM, second harmonic generation （SHG） microscopy was used to image the microstructures of the TM and adjacent tissues in control （normal） and high IOP conditions. Enucleated rabbit eyes were perfused at a pressure of 60 mmHg to achieve the high lOP. Through the anterior chamber of the eye, in situ images were obtained from different depths beneath the surface of the TM. Porosity and specific surface area of the TM in control and high IOP conditions were then calculated to estimate the effect of the high pressure on the permeability of tissue in different depths. We further photographed the histological sections of the TM and compared the in situ images. The following results were obtained in the control condition, where the region of depth was less than 55 μm with crossed branching beams and large pores in the superficial TM. The deeper meshwork is a silk-like tissue with abundant fluorescence separating the small size of pores. The total thickness of pathway tissues composed of TM and juxtacanalicular （JCT） is more than 100 p.m. After putting a high pressure on the inner wall of the eye, the TM region progressively collapses and decreases to be less than 40 μm. Fibers of the TM became dense, and the porosity at 34 μm in the high IOP condition is comparable to that at 105 μm in the control condition. As a consequent result, the permeability of the superficial TM decreases rapidly from 120 μm2 to 49.6 μm2 and that of deeper TM decreases from 1.66 μm2 to 0.57 μm2. Heterogeneity reflected by descent in permeability reduces from 12.4 μm of the control condition to 3.74 μm of the high IOP condition. The persistently high IOP makes the TM region collapse from its normal state, in which the collagen fibers of the TM are arranged in regular to maintain the physiological permeability of the outflow pathway. In the scope of pathologically high IOP, the microstructure of the TM is sensitive to pressure and hydraulic permeability can be significantly affected by IOP.展开更多
This study compared the efficacy of non-penetrating trabecular surgery and trabeculectomy for the treatment of open angle glaucoma. We searched the Cochrane Library, PUBMED （1966 to 2009）, Embase （1980 to 2009） an...This study compared the efficacy of non-penetrating trabecular surgery and trabeculectomy for the treatment of open angle glaucoma. We searched the Cochrane Library, PUBMED （1966 to 2009）, Embase （1980 to 2009） and CMB-disk （1979 to 2009） for the randomized clinical trials （RCT） concerning the two treatment strategies. The reports, including the papers listed in bibliographies, were evaluated against a set of quality criteria and the RCTs that satisfied the criteria were selected and subjected to Meta analysis by employing the Cochrane Collaboration＇s RevMan 4.5 software package. A total of nine RCTs were included in the study. The analyses of the reports showed that, 12 months after surgery, there was significant difference in the reduction of interocular pressure （IOP） between non-penetrating trabecular surgery and trabeculectomy （Z=6.05 P0.00001）. There also existed statistically significant difference in the reduction of IOP at the censored time between the two procedures （Z=4.92, P0.00001）. Difference in the success rate was also found between the two surgeries （Z=3.82, P=0.0001）. It is concluded that, compared with the non-penetrating trabeculectomy, the traditional trabeculectomy could reduce IOP more and had higher success rate while the non-penetrating trabecular surgery is associated with lower postoperative complications.展开更多
Osteoporosis represents one major health condition for our growing elderly population. It accounts for severe morbidity and increased mortality in postmenopausal women and it is becoming an emerging health concern eve...Osteoporosis represents one major health condition for our growing elderly population. It accounts for severe morbidity and increased mortality in postmenopausal women and it is becoming an emerging health concern even in aging men. Screening of the population at risk for bone degeneration and treatment assessment of osteoporotic patients to prevent bone fragility fractures represent useful tools to improve quality of life in the elderly and to lighten the related socio-economic impact. Bone mineral density(BMD) estimate by means of dual-energy X-ray absorptiometry is normally used in clinical practice for osteoporosis diagnosis. Nevertheless, BMD alone does not represent a good predictor of fracture risk. From a clinical point of view, bone microarchitecture seems to be an intriguing aspect to characterize bone alteration patterns in aging and pathology. The widening into clinical practice of medical imaging techniques and the impressive advances in information technologies together with enhanced capacity of power calculation have promoted proliferation of new methods to assess changes of trabecular bone architecture(TBA) during aging and osteoporosis. Magnetic resonance imaging(MRI) has recently arisen as a useful tool to measure bone structure in vivo. In particular, high-resolution MRI techniques have introduced new perspectives for TBA characterization by non-invasive non-ionizing methods. However, texture analysis methods have not found favor with clinicians as they produce quite a few parameters whose interpretation is difficult. The introduction in biomedical field of paradigms, such as theory of complexity, chaos, and fractals, suggests new approaches and provides innovative tools to develop computerized methods that, by producing a limited number of parameters sensitive to pathology onset and progression, would speed up their application into clinical practice. Complexity of living beings and fractality of several physio-anatomic structures suggest fractal analysis as a promising approach to quantify morphofunctional changes in both aging and pathology. In this particular context, fractal lacunarity seems to be the proper tool to characterize TBA texture as it is able to describe both discontinuity of bone network and sizes of bone marrow spaces, whose changes are an index of bone fracture risk. In this paper, an original method of MRI texture analysis, based on TBA fractal lacunarity is described and discussed in the light of new perspectives for early diagnosis of osteoporotic fractures.展开更多
AIM:To compare the trabecular meshwork(TM)and iris apoptosis of treated and untreated primary open angle glaucoma(POAG)patients.METHODS:Eight treatment-naive,newly diagnosed(group 1)and 11 medlcaiy treated(group 2)pat...AIM:To compare the trabecular meshwork(TM)and iris apoptosis of treated and untreated primary open angle glaucoma(POAG)patients.METHODS:Eight treatment-naive,newly diagnosed(group 1)and 11 medlcaiy treated(group 2)patients with POAG were included in the study.Each patient underwent a limbus-based trabeculectomy.The TM and peripheral iris specimens were dissected out and were snap-frozen in liquid nitrogen and stored at-80t until they were assayed.Apoptosis in each group was assesed by TUNEL method.RESULTS:The mean patient age was 60.6±5.8 years(53-68 years)vs 58.9±8.9 years(47-70 years)in group 1and group 2(P=0.859).The mean treatment time in group 2 was 22.2±7.3 months(12-34 months).Apoptotic indexes in TM and iris were significantly higher in POAG patients using medication(group 2)compared to treatment-naive POAG patients(group 1)(P=0.004,0.015;respectively).CONCLUSION:Long term administration of topical antiglaucoma medications causes additional toxic effects on TM.展开更多
The effects of suppression of CD44 by CD44-specific antisense oligonucleotide on attachment of human trabecular meshwork cells to hyaluronic acid (HA) were observed and the possible relationship between CD44 and prim...The effects of suppression of CD44 by CD44-specific antisense oligonucleotide on attachment of human trabecular meshwork cells to hyaluronic acid (HA) were observed and the possible relationship between CD44 and primary open-angle glaucoma (POAG) investigated. CD44-specific antisense oligonucleotide was delivered with cationic lipid to cultured human trabecular meshwork cells. The expression of CD44 suppressed by CD44-specific antisense oligonucleotide was detected by RT-PCR and Western blotting. The effect of CD44 suppression by specific antisense oligonucleotide on attachment of trabecular meshwork cells to HA was measured by MTT assay. Results showed that expression of CD44 was suppressed by CD44-specific antisense oligonucleotide. Antisense oligonucleotide also suppressed the adhesion of human trabecular meshwork cells to HA in a concentration dependent manner. It was concluded that attachment of human trabecular meshwork cells to HA was decreased when CD44 was suppressed by specific antisense oligonucleotide. CD44 might play a role in pathogenesis of POAG by affecting the adhesion of trabecular meshwork cells to HA.展开更多
To confirm the existence of heme oxygenase (HO)-carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HTMCs) in vitro, and to evaluate the inductive role...To confirm the existence of heme oxygenase (HO)-carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HTMCs) in vitro, and to evaluate the inductive role of hemin on this pathway, HTMCs of the third to fourth generation were cultured in vitro. Reverse transcripase-polymerase chain reaction (RT-PCR) was employed for detection of HO-1 and HO-2 mRNA. Immunohistochemical staining was used to detect HO-1 and HO-2 proteins. Hemin was added into the culture solution. The HO-1 mRNA levels were quantified by RT-PCR. The relative amount of carbon monoxide released into the media was measured with the quantifying carbon monoxide hemoglobin (HbCO) by spectrophotometry. Radioimmunoassay was used to determine changes of cGMP in HTMCs. The results showed that cultured cells had the specific characteristics of HTMCs. Both HO-1 and HO-2 genes were expressed in HTMCs, as well as HO-1 and HO-2 proteins in HTMCs. Hemin induced HO-1 mRNA, HbCO and cGMP in a dose-dependent manner. In conclusion, HO-CO-cGMP pathway exists in the cultured HTMCs and can be induced by hemin. Pharmacological stimulation of HO-CO-cGMP pathway may constitute a novel therapeutic approach to rescuing glaucoma.展开更多
The objective of this study was to study the age-related adaptation of lumbar vertebral trabecular bone at the apparent level, as well as the tissue level in three orthogonal directions. Ninety trabecular specimens we...The objective of this study was to study the age-related adaptation of lumbar vertebral trabecular bone at the apparent level, as well as the tissue level in three orthogonal directions. Ninety trabecular specimens were obtained from six normal L4 vertebral bodies of six male cadavers in two age groups, three aged 62 years and three aged 69 years, and were scanned using a high-resolution micro-computed tomography （micro-CT） system, then converted to micro- finite element models to do micro-finite element analyses. The relationship between apparent stiffness and bone volume fraction, and the tissue level yon Mises stress distribution for each trabecular specimen when compressed separately in the longitudinal direction, medial-lateral and anterior-posterior directions （transverse directions） were derived and compared between two age groups. The results showed that at the apparent level, trabecular bones from 69-year group had stiffer bone structure relative to their volume fractions in all three directions, and in both age groups, changes in bone volume fraction could explain more variations in apparent stiffness in the longitudinal direction than the transverse directions; at the tissue level, aging had little effect on the tissue von Mises stress distributions for the compressions in all the three directions. The novelty of the present study was that it provided quantitative assessments on the age and direction- related adaptation of Chinese male lumbar vertebral trabecular bone from two different levels： stiffness at the apparent level and stress distribution at the tissue level. It may help to understand the failure mechanisms and fracture risks of vertebral body associated with aging and direction for the prevention of fracture risks in elder individuals.展开更多
AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells. METHODS:...AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells. METHODS: HTM cells were cultured to confluent and incubated with 0.4 mu mol/L Dex and/or 0.05 mu mol/L LAT-A. FN expression in HTM cells was evaluated by Western blot and immunofluorescence microscopy. RESULTS: Dex up-regulated FN production in HTM cells, failed to do so when co-incubated with LAT-A. LAT-A decreased production of FN in cultured HTM cells. CONCLUSION: This study indicated that LAT-A may modulate the expression of fibronectin in trabecular meshwork to achieve treatment for steroids and other types of glaucoma. It has an important prospect as an intraocular pressure-lowering drug.展开更多
AIM: To investigate changes in extracellular matrix(ECM) gene expression in human trabecular meshwork(HTM) cells in response to mechanical fluid flow stimulation.METHODS: HTM cells were grown on a glass plate coated w...AIM: To investigate changes in extracellular matrix(ECM) gene expression in human trabecular meshwork(HTM) cells in response to mechanical fluid flow stimulation.METHODS: HTM cells were grown on a glass plate coated with 0.02% type Ⅰ collagen(COL) and exposed to shear stress(0, 0.2, 1.0 dyne/cm;) for 12 h.Changes in genes related to the ECM were evaluated by real-time reverse transcriptase-polymerase chain reaction.Phosphorylation of Smad2 protein was investigated by Western blotting.RESULTS: After mechanical stimulation, COL type 4 alpha 2, COL type 6 alpha 1, and fibronectin-1 mRNA were significantly higher than the static control(P<0.05, <0.05, and <0.01, respectively).The metalloproteinase-2 and plasminogen activator inhibitor-1 mRNA were significantly higher than the static control(P<0.05 and <0.01, respectively), while the differences in the tissue inhibitors of metalloproteinases-2 mRNA were not significant.The phosphorylation of Smad2 levels was significantly higher compared to the static control cells.CONCLUSION: Changes in the expressions of genes associated ECM metabolism result in HTM cells after mechanical stimulation.The mechanical stimulation of the aqueous humor to the trabecular meshwork may promote ECM turnover and contribute to intraocular pressure homeostasis.展开更多
AIM:To observe the changes in Schlemm’s canal(SC),trabecular meshwork(TM),and anterior chamber relevant parameters after small incision lenticule extraction(SMILE) of myopia patients.METHODS:A total of 58 eyes from 3...AIM:To observe the changes in Schlemm’s canal(SC),trabecular meshwork(TM),and anterior chamber relevant parameters after small incision lenticule extraction(SMILE) of myopia patients.METHODS:A total of 58 eyes from 30 patients who underwent SMILE were divided into a low and moderate myopia group(group A,32 eyes) and a high myopia group(group B,26 eyes).The diameter and area of the SC,the width and thickness of TM obtained by CIRRUS HD-OCT5000,and the related anterior chamber parameters obtained by Pentacam anterior segment analysis system,accommodation amplitude(AMP) were observed pre-and postoperatively.The preoperative intraocular pressure(IOP) and postoperative correction of intraocular pressure(IOPcc) were measured.RESULTS:The diameter and area of the SC in the two groups were significantly increased postoperatively(all P<0.01).The TM width of the patients in the two groups were increased at 1mo after surgery(both P<0.01),but the TM thickness did not change(P>0.05).The corneal curvature,central anterior chamber depth,and anterior chamber volume decreased after SMILE surgery(all P<0.01).There was a weak negative correlation between the SC area change and AMP change in group A(r=-0.362,P<0.01).The postoperative IOP decreased after correction by Shah formula(P<0.05).CONCLUSION:SC and TM in myopia patients change in the early postoperative stage of SMILE and the IOP is decline.展开更多
AIM:To investigate potential gene changes in trabecular meshwork(TM)induced by dexamethasone(DEX)in steroidinduced glaucoma(SIG).METHODS:The expression data of 24 cases from a public functional genomics data were sort...AIM:To investigate potential gene changes in trabecular meshwork(TM)induced by dexamethasone(DEX)in steroidinduced glaucoma(SIG).METHODS:The expression data of 24 cases from a public functional genomics data were sorted to identify the mechanisms of action of DEX on the TM.The relationships of the differentially expressed genes(DEGs)were enriched using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.In addition,the hub genes were screened by the Search Tool for the Retrieval of Interacting Genes Database(STRING)and Cytoscape tools.Finally,human TM cells(HTMCs)were treated with DEX to preliminarily explore the function of hub genes.RESULTS:Totally 47 DEGs,including 21 downregulated and 26 upregulated genes were identified.The primary enriched results of the DEGs consisted of inflammatory response,extracellular matrix(ECM),negative regulation of cell proliferation,TNF signalling pathway and the regulation of tr yptophan channels by inflammator y mediators.Subsequently,pro-melanin-enriched hormone(PMCH)and Bradykinin B1 receptor(BDKRB1)were screened as hub genes.It is verified in GSE37474 data set.Western blot and quantitative real-time polymerase chain reaction(q PCR)results showed that protein and RNA expression levels of BDKRB1 were significantly decreased after DEX treatment,while PMCH was not significantly changed.CONCLUSION:BDKRB1 may be a key gene involved in SIG onset,providing a suitable therapeutic target for improving the prognosis of SIG patients.展开更多
Whether transforming growth factor β 2 (TGF β 2) induces apoptosis of human trabecular meshwork cells was investigated in vitro . Cultured 3 5 passage human trabecular meshwork cells were treated with 0 (con...Whether transforming growth factor β 2 (TGF β 2) induces apoptosis of human trabecular meshwork cells was investigated in vitro . Cultured 3 5 passage human trabecular meshwork cells were treated with 0 (control), 0.32, 1, 3.2 ng/ml TGF β 2 for 48 h and divided into control group and experimental group. The apoptosis of human trabecular meshwork cells was examined by transmisson electron microscopy, TUNEL technique and flow cytometry. The results showed characteristic morphologic changes of apoptotic cells were observed under transmission electron microscopy. DNA fragmentation of human trabecular meshwork cells was found by TUNEL technique. Quantitative analysis of flow cytometry showed that percentages of apoptotic human trabecular meshwork cells were (2.79±0.44) %, (4.43 ±1.17) % and (9.60±2.05) % respectively with different concentrations [1 ng/ml ( P< 0.05), 3.2 ng/ml ( P< 0.01)] of TGF β 2 with the difference being significant between experimental group and control group[(1.41±0.34) %]. It was concluded that TGF β 2 can induce apoptosis of human trabecular meshwork cells in vitro and may be involved in the decrease of trabecular meshwork cells in the patients with primary open angle glaucoma and aging of normal people.展开更多
AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-compleme...AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2(scAAV2)vector technologies.METHODS:An scAAV2 vector with C3 transferase(C3)as the reporter gene(scAAV2-C3)was selected.The scAAV2-C3 vectors were driven by Ch3L1(scAAV2-Ch3L1-C3),MGP(scAAV2-MGP-C3),enhanced MGP(scAAV2-eMGP-C3)and cytomegalovirus(scAAV2-CMV-C3),respectively.The cultured primary human TM cells were treated with each vector at different multiplicities of infections.Changes in cell morphology were observed by phase contrast microscopy.Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining,real-time quantitative polymerase chain reaction and Western blot.Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively.In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope.Intraocular pressure(IOP)was monitored using a rebound tonometer.Ocular responses were evaluated by slit-lamp microscopy.Ocular histopathology analysis was examined by hematoxylin and eosin staining.RESULTS:In TM cell culture studies,the vectormediated C3 expression induced morphologic changes,disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rhoassociated protein kinase signaling pathway.At the same dose,these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3,but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3.At lowinjected dose,the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGPC3-injected and scAAV2-eMGP-C3-injected eyes.At highinjected dose,significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes.Similar to scAAV2-CMV-C3,scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium.In anterior segment tissues of scAAV2-eMGP-C3-injected eyes,no obvious morphological changes were found except for the TM.Inflammation was absent.CONCLUSION:In scAAV2-transduced TM cells,the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus,but obviously higher than that of MGP.In the anterior chamber of rat eye,the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter,but not by Ch3L1 promoter.These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector.展开更多
The mechanical properties of the pelvic trabecular bone have been studied at the continuum level. However, nothing is known about the tissue-level damage in the trabecular bone of the healthy human acetabulum at appar...The mechanical properties of the pelvic trabecular bone have been studied at the continuum level. However, nothing is known about the tissue-level damage in the trabecular bone of the healthy human acetabulum at apparent small strains characteristic of habitual. By a DAWING 4000 A supercomputer, nonlinear micro-finite element (μFE) analysis was performed to quantify tissue-level damage accumulation in trabecular bone at small strains. The data indicate that damage in trabecular bone commence at 0.2% apparent strain. The findings imply that tissue yielding can initiate at very low strains in the trabecular bone of the healthy acetabulum and that this local failure has negative consequences on the apparent mechanical properties of trabecular bone.展开更多
AIM: To investigate whether the enhanced green fluorescent protein(EGFP) reporter gene could be transferred into human trabecular meshwork(HTM) cells by a HIV-based lentivirus both in vitro and ex vivo.METHODS: The HI...AIM: To investigate whether the enhanced green fluorescent protein(EGFP) reporter gene could be transferred into human trabecular meshwork(HTM) cells by a HIV-based lentivirus both in vitro and ex vivo.METHODS: The HIV-based lentivirus that contains an EF1-α promoter driving EGFP expression cassette was constructed following the standard molecular cloning methods. The cultured HTM cells were transduced at a range of multiplicity of infection(MOI) with HIV-based lentivirus. EGFP positive cell populations were detected by flow cytometry. Human anterior eye segments were cultured with perfusion culture system and transfected by HIV-based lentivirus with a 1 ×108transducing unit(TU) virus in perfusion liquid. The intraocular pressure was recorded every 8h for 21 d. The expression of EGFP in the anterior segment of the human eye was detected by fluorescence microscopy. Furthermore, the distribution of EGFP expression was confirmed by anti-EGFP immunohistochemical staining.RESULTS: The HIV-based lentivirus which contains an EF1-α promoter driving EGFP expression cassette was constructed successfully. After HTM cells were transduced with HIV-based lentivirus containing EGFP in vitro, the ratio of EGFP positive cells to the total cell number reached 92.3%, with the MOI of 15. After the lentivirus containing EGFP were used to transduce human anterior eye segments, the EGFP could be directly detected by fluorescence microscopy in vivo.Immunohistochemistry staining revealed that 88.19%EGFP-positive trabecular meshwork(TM) cells were observed in the human anterior segment. Nevertheless,the intraocular pressure in the lentivirus-transduced group kept constant when compared with control group(P 】0.05).CONCLUSION: EGFP gene could be efficiently transferred into HTM cells both in vitro and ex vivo by using HIV-based lentivirus.展开更多
基金Beijing Natural Science Foundation-Haidian original Innovation Joint Foundation,Grant/Award Number:L192016Joint Funds of the National Natural Science Foundation of China,Grant/Award Number:U21A20489+3 种基金National Natural Science Foundation of China,Grant/Award Number:62003330Shenzhen Fundamental Research Funds,Grant/Award Numbers:JCYJ20220818101608019,JCYJ20190807170407391,JCYJ20180507182415428Natural Science Foundation of Guangdong Province,Grant/Award Number:2019A1515011699Guangdong-Hong Kong-Macao Joint Laboratory of Human-Machine Intelligence-Synergy Systems,Shenzhen Institute of Advanced Technology。
文摘Osteoporosis is a major cause of bone fracture and can be characterised by both mass loss and microstructure deterioration of the bone.The modern way of osteoporosis assessment is through the measurement of bone mineral density,which is not able to unveil the pathological condition from the mesoscale aspect.To obtain mesoscale information from computed tomography(CT),the super-resolution(SR)approach for volumetric imaging data is required.A deep learning model AESR3D is proposed to recover high-resolution(HR)Micro-CT from low-resolution Micro-CT and implement an unsupervised segmentation for better trabecular observation and measurement.A new regularisation overcomplete autoencoder framework for the SR task is proposed and theoretically analysed.The best performance is achieved on structural similarity measure of trabecular CT SR task compared with the state-of-the-art models in both natural and medical image SR tasks.The HR and SR images show a high correlation(r=0.996,intraclass correlation coefficients=0.917)on trabecular bone morphological indicators.The results also prove the effectiveness of our regularisation framework when training a large capacity model.
文摘BACKGROUND Clinical studies using Trabecular Titanium™acetabular cups have shown promising short and medium-term results.This material,due to its macro and micro surface roughness,provides a substrate for osseointegration and enhances implant stability.However,there is a lack of evidence in the literature on the use of this material in patients with femoral neck fracture.AIM To evaluate the short-term clinical-functional and radiographic outcomes in patients with femoral neck fractures undergoing total hip arthroplasty(THA)with Trabecular Titanium™acetabular cup implants.METHODS The study included 104 patients with medial femoral neck fractures who underwent THA between January 2020 and December 2020 with the Delta TT acetabular cup(Lima Corporate,Villanova di San Daniele del Friuli,Italy).The mean age of the patients was 69.57±10.16 years(range:36-85 years).The followup period ranged from a minimum of 3 to a maximum of 4 years.Three questionnaires(Harris Hip Score,Oxford Hip Score,and EQ5D)were administered along with radiographic evaluations.Statistical methods included the Student's t-test and one-way analysis of variance for comparisons(with significance set at 0.05),and the Kaplan-Meier curve for prosthetic implant survival.RESULTS The mean follow-up was 41.5 months.The Harris Hip Score(HHS)showed a mean increase of 2.74 points(mean HHS 88.52 at 6 months postoperatively and mean HHS 91.26 at the last follow-up)with statistical significance.Similarly,the Oxford Hip Score demonstrated a statistically significant difference between follow-up groups.However,the EQ5D did not show statistically significant differences among the three groups(preoperative,6-month follow-up,and last follow-up).Revision surgery was required in 6 patients.According to Moore's criteria,96%of the acetabular components were radiographically stable and well-integrated at the last follow-up.The Kaplan-Meier curve showed a 96%survival rate.CONCLUSION The clinical and radiographic results obtained in the short to medium term confirm the excellent performance of the Delta TT acetabular cup in terms of osseointegration,providing an optimal solution both for young patients with high functional recovery demands and for fragile patients requiring optimal stability of the acetabular component to reduce the risk of implant failure.
文摘Background: When applied to trabecular bone X-ray images, the anisotropic properties of trabeculae located at ultra-distal radius were investigated by using the trabecular bone scores (TBS) calculated along directions parallel and perpendicular to the forearm. Methodology: Data from more than two hundred subjects were studied retrospectively. A DXA (GE Lunar Prodigy) scan of the forearm was performed on each subject to measure the bone mineral density (BMD) value at the location of ultra-distal radius, and an X-ray digital image of the same forearm was taken on the same day. The values of trabecular bone score along the direction perpendicular to the forearm, TBS<sub>x</sub>, and along the direction parallel to the forearm, TBS<sub>y</sub>, were calculated respectively. The statistics of TBS<sub>x</sub> and TBS<sub>y</sub> were calculated, and the anisotropy of the trabecular bone, which was defined as the ratio of TBS<sub>y</sub> to TBS<sub>x</sub> and changed with subjects’ BMD and age, was reported and analyzed. Results: The results show that the correlation coefficient between TBS<sub>x</sub> and TBS<sub>y</sub> was 0.72 (p BMD and age was reported. The results showed that decreased trabecular bone anisotropy was associated with deceased BMD and increased age in the subject group. Conclusions: This study shows that decreased trabecular bone anisotropy was associated with decreased BMD and increased age.
文摘AIM：To describe a novel technique of creating a landing strip within the trabecular meshwork to guide trabecular micro-bypass stent（iStent） implantation in patients who underwent phacoemulsification.METHODS：Thirty-four eyes from 30 patients who underwent i Stent implantation after phacoemulsification from May 2014 to February 2015 were included in our retrospective study. All iStents were implanted via the ＂landing strip＂ technique. A 25-gauge microvitreoretinal blade was used to bisect the trabecular meshwork to less than 1 clock-hour, effectively creating a landing strip. The iStent applicator was pressed along the landing strip and then the stent was released into the trabecular meshwork. RESULTS：Of the 34 eyes with iStent implantation, 27（79.4%） eyes had primary open-angle glaucoma, 6（17.6%） eyes had pseudoexfoliation glaucoma, and 1（2.9%） eye had ocular hypertension. At 6-month follow-up（n=17）, the mean number of hypotensive medications decreased from 2.2±1.2 at baseline to 0.8±1.3（P=0.05） and mean intraocular pressure decreased from 19.7±4.1 mm Hg at baseline to 16.7±2.1 mm Hg（P=0.58）. Two eyes（5.9%） required subsequent trabeculectomy. CONCLUSION：The ＂landing strip＂ technique appears to be an effective way to assist with iS tent implantation.
文摘The changes in the expression of aquaporin-1 （AQP1） mRNA and protein in cultured human trabecular meshwork （HTM） cells treated with dexamethasone and transfected with antisense oligonucleotides （AS-ODN） were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0.04 μg/mL and 0.4 μg/mL groups, the levels of AQP1 were higher than in control group （0 μg/mL）. In the 4 μg/ mL and 40 μg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down-regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 μg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down-regulate the AQP1 expression in HTM cells to some extent.
基金supported by the National Natural Science Foundation of China(Grant Nos.31070840,10802053,and 81201123)the Natural Science Foundation of Beijing,China(Grant No.3122010)+2 种基金the Natural Science Foundation of Guangdong Province,China(Grant No.S2012040006576)the Shenzhen Science and Technology Innovation Committee,China(Grant No.KQCX20120816155352228)the Funding Project for Academic Human Resources Development in Institutions of Higher Learning under the Jurisdiction of Beijing Municipality,China(Grant No.PHR201110506)
文摘As the aqueous humor leaves the eye, it first passes through the trabecular meshwork （TM）. Increased flow resistance in this region causes elevation of intraocular pressure （IOP）, which leads to the occurrence of glaucoma. To quantitatively evaluate the effect of high IOP on the configuration and hydraulic permeability of the TM, second harmonic generation （SHG） microscopy was used to image the microstructures of the TM and adjacent tissues in control （normal） and high IOP conditions. Enucleated rabbit eyes were perfused at a pressure of 60 mmHg to achieve the high lOP. Through the anterior chamber of the eye, in situ images were obtained from different depths beneath the surface of the TM. Porosity and specific surface area of the TM in control and high IOP conditions were then calculated to estimate the effect of the high pressure on the permeability of tissue in different depths. We further photographed the histological sections of the TM and compared the in situ images. The following results were obtained in the control condition, where the region of depth was less than 55 μm with crossed branching beams and large pores in the superficial TM. The deeper meshwork is a silk-like tissue with abundant fluorescence separating the small size of pores. The total thickness of pathway tissues composed of TM and juxtacanalicular （JCT） is more than 100 p.m. After putting a high pressure on the inner wall of the eye, the TM region progressively collapses and decreases to be less than 40 μm. Fibers of the TM became dense, and the porosity at 34 μm in the high IOP condition is comparable to that at 105 μm in the control condition. As a consequent result, the permeability of the superficial TM decreases rapidly from 120 μm2 to 49.6 μm2 and that of deeper TM decreases from 1.66 μm2 to 0.57 μm2. Heterogeneity reflected by descent in permeability reduces from 12.4 μm of the control condition to 3.74 μm of the high IOP condition. The persistently high IOP makes the TM region collapse from its normal state, in which the collagen fibers of the TM are arranged in regular to maintain the physiological permeability of the outflow pathway. In the scope of pathologically high IOP, the microstructure of the TM is sensitive to pressure and hydraulic permeability can be significantly affected by IOP.
文摘This study compared the efficacy of non-penetrating trabecular surgery and trabeculectomy for the treatment of open angle glaucoma. We searched the Cochrane Library, PUBMED （1966 to 2009）, Embase （1980 to 2009） and CMB-disk （1979 to 2009） for the randomized clinical trials （RCT） concerning the two treatment strategies. The reports, including the papers listed in bibliographies, were evaluated against a set of quality criteria and the RCTs that satisfied the criteria were selected and subjected to Meta analysis by employing the Cochrane Collaboration＇s RevMan 4.5 software package. A total of nine RCTs were included in the study. The analyses of the reports showed that, 12 months after surgery, there was significant difference in the reduction of interocular pressure （IOP） between non-penetrating trabecular surgery and trabeculectomy （Z=6.05 P0.00001）. There also existed statistically significant difference in the reduction of IOP at the censored time between the two procedures （Z=4.92, P0.00001）. Difference in the success rate was also found between the two surgeries （Z=3.82, P=0.0001）. It is concluded that, compared with the non-penetrating trabeculectomy, the traditional trabeculectomy could reduce IOP more and had higher success rate while the non-penetrating trabecular surgery is associated with lower postoperative complications.
文摘Osteoporosis represents one major health condition for our growing elderly population. It accounts for severe morbidity and increased mortality in postmenopausal women and it is becoming an emerging health concern even in aging men. Screening of the population at risk for bone degeneration and treatment assessment of osteoporotic patients to prevent bone fragility fractures represent useful tools to improve quality of life in the elderly and to lighten the related socio-economic impact. Bone mineral density(BMD) estimate by means of dual-energy X-ray absorptiometry is normally used in clinical practice for osteoporosis diagnosis. Nevertheless, BMD alone does not represent a good predictor of fracture risk. From a clinical point of view, bone microarchitecture seems to be an intriguing aspect to characterize bone alteration patterns in aging and pathology. The widening into clinical practice of medical imaging techniques and the impressive advances in information technologies together with enhanced capacity of power calculation have promoted proliferation of new methods to assess changes of trabecular bone architecture(TBA) during aging and osteoporosis. Magnetic resonance imaging(MRI) has recently arisen as a useful tool to measure bone structure in vivo. In particular, high-resolution MRI techniques have introduced new perspectives for TBA characterization by non-invasive non-ionizing methods. However, texture analysis methods have not found favor with clinicians as they produce quite a few parameters whose interpretation is difficult. The introduction in biomedical field of paradigms, such as theory of complexity, chaos, and fractals, suggests new approaches and provides innovative tools to develop computerized methods that, by producing a limited number of parameters sensitive to pathology onset and progression, would speed up their application into clinical practice. Complexity of living beings and fractality of several physio-anatomic structures suggest fractal analysis as a promising approach to quantify morphofunctional changes in both aging and pathology. In this particular context, fractal lacunarity seems to be the proper tool to characterize TBA texture as it is able to describe both discontinuity of bone network and sizes of bone marrow spaces, whose changes are an index of bone fracture risk. In this paper, an original method of MRI texture analysis, based on TBA fractal lacunarity is described and discussed in the light of new perspectives for early diagnosis of osteoporotic fractures.
文摘AIM:To compare the trabecular meshwork(TM)and iris apoptosis of treated and untreated primary open angle glaucoma(POAG)patients.METHODS:Eight treatment-naive,newly diagnosed(group 1)and 11 medlcaiy treated(group 2)patients with POAG were included in the study.Each patient underwent a limbus-based trabeculectomy.The TM and peripheral iris specimens were dissected out and were snap-frozen in liquid nitrogen and stored at-80t until they were assayed.Apoptosis in each group was assesed by TUNEL method.RESULTS:The mean patient age was 60.6±5.8 years(53-68 years)vs 58.9±8.9 years(47-70 years)in group 1and group 2(P=0.859).The mean treatment time in group 2 was 22.2±7.3 months(12-34 months).Apoptotic indexes in TM and iris were significantly higher in POAG patients using medication(group 2)compared to treatment-naive POAG patients(group 1)(P=0.004,0.015;respectively).CONCLUSION:Long term administration of topical antiglaucoma medications causes additional toxic effects on TM.
文摘The effects of suppression of CD44 by CD44-specific antisense oligonucleotide on attachment of human trabecular meshwork cells to hyaluronic acid (HA) were observed and the possible relationship between CD44 and primary open-angle glaucoma (POAG) investigated. CD44-specific antisense oligonucleotide was delivered with cationic lipid to cultured human trabecular meshwork cells. The expression of CD44 suppressed by CD44-specific antisense oligonucleotide was detected by RT-PCR and Western blotting. The effect of CD44 suppression by specific antisense oligonucleotide on attachment of trabecular meshwork cells to HA was measured by MTT assay. Results showed that expression of CD44 was suppressed by CD44-specific antisense oligonucleotide. Antisense oligonucleotide also suppressed the adhesion of human trabecular meshwork cells to HA in a concentration dependent manner. It was concluded that attachment of human trabecular meshwork cells to HA was decreased when CD44 was suppressed by specific antisense oligonucleotide. CD44 might play a role in pathogenesis of POAG by affecting the adhesion of trabecular meshwork cells to HA.
文摘To confirm the existence of heme oxygenase (HO)-carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HTMCs) in vitro, and to evaluate the inductive role of hemin on this pathway, HTMCs of the third to fourth generation were cultured in vitro. Reverse transcripase-polymerase chain reaction (RT-PCR) was employed for detection of HO-1 and HO-2 mRNA. Immunohistochemical staining was used to detect HO-1 and HO-2 proteins. Hemin was added into the culture solution. The HO-1 mRNA levels were quantified by RT-PCR. The relative amount of carbon monoxide released into the media was measured with the quantifying carbon monoxide hemoglobin (HbCO) by spectrophotometry. Radioimmunoassay was used to determine changes of cGMP in HTMCs. The results showed that cultured cells had the specific characteristics of HTMCs. Both HO-1 and HO-2 genes were expressed in HTMCs, as well as HO-1 and HO-2 proteins in HTMCs. Hemin induced HO-1 mRNA, HbCO and cGMP in a dose-dependent manner. In conclusion, HO-CO-cGMP pathway exists in the cultured HTMCs and can be induced by hemin. Pharmacological stimulation of HO-CO-cGMP pathway may constitute a novel therapeutic approach to rescuing glaucoma.
基金The project supported by the Hong Kong Polytechnic University Research Grants(G-U273) the National Natural Science Foundation of China(10502021 and 10529202)
文摘The objective of this study was to study the age-related adaptation of lumbar vertebral trabecular bone at the apparent level, as well as the tissue level in three orthogonal directions. Ninety trabecular specimens were obtained from six normal L4 vertebral bodies of six male cadavers in two age groups, three aged 62 years and three aged 69 years, and were scanned using a high-resolution micro-computed tomography （micro-CT） system, then converted to micro- finite element models to do micro-finite element analyses. The relationship between apparent stiffness and bone volume fraction, and the tissue level yon Mises stress distribution for each trabecular specimen when compressed separately in the longitudinal direction, medial-lateral and anterior-posterior directions （transverse directions） were derived and compared between two age groups. The results showed that at the apparent level, trabecular bones from 69-year group had stiffer bone structure relative to their volume fractions in all three directions, and in both age groups, changes in bone volume fraction could explain more variations in apparent stiffness in the longitudinal direction than the transverse directions; at the tissue level, aging had little effect on the tissue von Mises stress distributions for the compressions in all the three directions. The novelty of the present study was that it provided quantitative assessments on the age and direction- related adaptation of Chinese male lumbar vertebral trabecular bone from two different levels： stiffness at the apparent level and stress distribution at the tissue level. It may help to understand the failure mechanisms and fracture risks of vertebral body associated with aging and direction for the prevention of fracture risks in elder individuals.
基金National Natural Science Foundation of China (No.30772379)Education Department Project of Hunan Province, China(No.04A049)
文摘AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells. METHODS: HTM cells were cultured to confluent and incubated with 0.4 mu mol/L Dex and/or 0.05 mu mol/L LAT-A. FN expression in HTM cells was evaluated by Western blot and immunofluorescence microscopy. RESULTS: Dex up-regulated FN production in HTM cells, failed to do so when co-incubated with LAT-A. LAT-A decreased production of FN in cultured HTM cells. CONCLUSION: This study indicated that LAT-A may modulate the expression of fibronectin in trabecular meshwork to achieve treatment for steroids and other types of glaucoma. It has an important prospect as an intraocular pressure-lowering drug.
基金Supported by JSPS KAKENHI (No.15K10857No.17H00695No.19K18865)。
文摘AIM: To investigate changes in extracellular matrix(ECM) gene expression in human trabecular meshwork(HTM) cells in response to mechanical fluid flow stimulation.METHODS: HTM cells were grown on a glass plate coated with 0.02% type Ⅰ collagen(COL) and exposed to shear stress(0, 0.2, 1.0 dyne/cm;) for 12 h.Changes in genes related to the ECM were evaluated by real-time reverse transcriptase-polymerase chain reaction.Phosphorylation of Smad2 protein was investigated by Western blotting.RESULTS: After mechanical stimulation, COL type 4 alpha 2, COL type 6 alpha 1, and fibronectin-1 mRNA were significantly higher than the static control(P<0.05, <0.05, and <0.01, respectively).The metalloproteinase-2 and plasminogen activator inhibitor-1 mRNA were significantly higher than the static control(P<0.05 and <0.01, respectively), while the differences in the tissue inhibitors of metalloproteinases-2 mRNA were not significant.The phosphorylation of Smad2 levels was significantly higher compared to the static control cells.CONCLUSION: Changes in the expressions of genes associated ECM metabolism result in HTM cells after mechanical stimulation.The mechanical stimulation of the aqueous humor to the trabecular meshwork may promote ECM turnover and contribute to intraocular pressure homeostasis.
基金Supported by the National Natural Science Foundation of China (No.81974133)。
文摘AIM:To observe the changes in Schlemm’s canal(SC),trabecular meshwork(TM),and anterior chamber relevant parameters after small incision lenticule extraction(SMILE) of myopia patients.METHODS:A total of 58 eyes from 30 patients who underwent SMILE were divided into a low and moderate myopia group(group A,32 eyes) and a high myopia group(group B,26 eyes).The diameter and area of the SC,the width and thickness of TM obtained by CIRRUS HD-OCT5000,and the related anterior chamber parameters obtained by Pentacam anterior segment analysis system,accommodation amplitude(AMP) were observed pre-and postoperatively.The preoperative intraocular pressure(IOP) and postoperative correction of intraocular pressure(IOPcc) were measured.RESULTS:The diameter and area of the SC in the two groups were significantly increased postoperatively(all P<0.01).The TM width of the patients in the two groups were increased at 1mo after surgery(both P<0.01),but the TM thickness did not change(P>0.05).The corneal curvature,central anterior chamber depth,and anterior chamber volume decreased after SMILE surgery(all P<0.01).There was a weak negative correlation between the SC area change and AMP change in group A(r=-0.362,P<0.01).The postoperative IOP decreased after correction by Shah formula(P<0.05).CONCLUSION:SC and TM in myopia patients change in the early postoperative stage of SMILE and the IOP is decline.
文摘AIM:To investigate potential gene changes in trabecular meshwork(TM)induced by dexamethasone(DEX)in steroidinduced glaucoma(SIG).METHODS:The expression data of 24 cases from a public functional genomics data were sorted to identify the mechanisms of action of DEX on the TM.The relationships of the differentially expressed genes(DEGs)were enriched using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis.In addition,the hub genes were screened by the Search Tool for the Retrieval of Interacting Genes Database(STRING)and Cytoscape tools.Finally,human TM cells(HTMCs)were treated with DEX to preliminarily explore the function of hub genes.RESULTS:Totally 47 DEGs,including 21 downregulated and 26 upregulated genes were identified.The primary enriched results of the DEGs consisted of inflammatory response,extracellular matrix(ECM),negative regulation of cell proliferation,TNF signalling pathway and the regulation of tr yptophan channels by inflammator y mediators.Subsequently,pro-melanin-enriched hormone(PMCH)and Bradykinin B1 receptor(BDKRB1)were screened as hub genes.It is verified in GSE37474 data set.Western blot and quantitative real-time polymerase chain reaction(q PCR)results showed that protein and RNA expression levels of BDKRB1 were significantly decreased after DEX treatment,while PMCH was not significantly changed.CONCLUSION:BDKRB1 may be a key gene involved in SIG onset,providing a suitable therapeutic target for improving the prognosis of SIG patients.
文摘Whether transforming growth factor β 2 (TGF β 2) induces apoptosis of human trabecular meshwork cells was investigated in vitro . Cultured 3 5 passage human trabecular meshwork cells were treated with 0 (control), 0.32, 1, 3.2 ng/ml TGF β 2 for 48 h and divided into control group and experimental group. The apoptosis of human trabecular meshwork cells was examined by transmisson electron microscopy, TUNEL technique and flow cytometry. The results showed characteristic morphologic changes of apoptotic cells were observed under transmission electron microscopy. DNA fragmentation of human trabecular meshwork cells was found by TUNEL technique. Quantitative analysis of flow cytometry showed that percentages of apoptotic human trabecular meshwork cells were (2.79±0.44) %, (4.43 ±1.17) % and (9.60±2.05) % respectively with different concentrations [1 ng/ml ( P< 0.05), 3.2 ng/ml ( P< 0.01)] of TGF β 2 with the difference being significant between experimental group and control group[(1.41±0.34) %]. It was concluded that TGF β 2 can induce apoptosis of human trabecular meshwork cells in vitro and may be involved in the decrease of trabecular meshwork cells in the patients with primary open angle glaucoma and aging of normal people.
基金Supported by the National Natural Science Foundation of China(No.81900829,No.82070963)the Xiamen Medical and Health Guiding Project Fund Project(No.3502Z20214ZD1214)+1 种基金the Guangdong Basic and Applied Basic Research Foundation(No.2019A1515011234)the Science and Technology Innovation Committee of Shenzhen(No.JCYJ20210324125614039)。
文摘AIM:To evaluate the potential of two trabecular meshwork(TM)-specific promoters,Chitinase 3-like 1(Ch3L1)and matrix gla protein(MGP),for improving specificity and safety in glaucoma gene therapy based on self-complementary AAV2(scAAV2)vector technologies.METHODS:An scAAV2 vector with C3 transferase(C3)as the reporter gene(scAAV2-C3)was selected.The scAAV2-C3 vectors were driven by Ch3L1(scAAV2-Ch3L1-C3),MGP(scAAV2-MGP-C3),enhanced MGP(scAAV2-eMGP-C3)and cytomegalovirus(scAAV2-CMV-C3),respectively.The cultured primary human TM cells were treated with each vector at different multiplicities of infections.Changes in cell morphology were observed by phase contrast microscopy.Actin stress fibers and Rho GTPases/Rho-associated protein kinase pathway-related molecules were assessed by immunofluorescence staining,real-time quantitative polymerase chain reaction and Western blot.Each vector was injected intracamerally into the one eye of each rat at low and high doses respectively.In vivo green fluorescence was visualized by a Micron III Retinal Imaging Microscope.Intraocular pressure(IOP)was monitored using a rebound tonometer.Ocular responses were evaluated by slit-lamp microscopy.Ocular histopathology analysis was examined by hematoxylin and eosin staining.RESULTS:In TM cell culture studies,the vectormediated C3 expression induced morphologic changes,disruption of actin cytoskeleton and reduction of fibronectin expression in TM cells by inhibiting the Rho GTPases/Rhoassociated protein kinase signaling pathway.At the same dose,these changes were significant in TM cells treated with scAAV2-CMV-C3 or scAAV2-Ch3L1-C3,but not in cells treated with scAAV2-eMGP-C3 or scAAV2-MGP-C3.At lowinjected dose,the IOP was significantly decreased in the scAAV2-Ch3L1-C3-injected eyes but not in scAAV2-MGPC3-injected and scAAV2-eMGP-C3-injected eyes.At highinjected dose,significant IOP reduction was observed in the scAAV2-eMGP-C3-injected eyes but not in scAAV2-MGP-C3-injected eyes.Similar to scAAV2-CMV-C3,scAAV2-Ch3L1-C3 vector showed efficient transduction both in the TM and corneal endothelium.In anterior segment tissues of scAAV2-eMGP-C3-injected eyes,no obvious morphological changes were found except for the TM.Inflammation was absent.CONCLUSION:In scAAV2-transduced TM cells,the promoter-driven efficiency of Ch3L1 is close to that of cytomegalovirus,but obviously higher than that of MGP.In the anterior chamber of rat eye,the transgene expression pattern of scAAV2 vector is presumably affected by MGP promoter,but not by Ch3L1 promoter.These findings would provide a useful reference for improvement of specificity and safety in glaucoma gene therapy using scAAV2 vector.
基金the National High Technology Research and Development Program (863) of China(No. 2006AA02A137the Postgraduate Creativity Foundation of Shanghai Jiaotong University (No. BXJ0730)
文摘The mechanical properties of the pelvic trabecular bone have been studied at the continuum level. However, nothing is known about the tissue-level damage in the trabecular bone of the healthy human acetabulum at apparent small strains characteristic of habitual. By a DAWING 4000 A supercomputer, nonlinear micro-finite element (μFE) analysis was performed to quantify tissue-level damage accumulation in trabecular bone at small strains. The data indicate that damage in trabecular bone commence at 0.2% apparent strain. The findings imply that tissue yielding can initiate at very low strains in the trabecular bone of the healthy acetabulum and that this local failure has negative consequences on the apparent mechanical properties of trabecular bone.
基金Supported by Natural Science Foundation of China(No.30901395)the Doctoral Fund of Ministry of Education of China(No.20090142120012,20110142120021)
文摘AIM: To investigate whether the enhanced green fluorescent protein(EGFP) reporter gene could be transferred into human trabecular meshwork(HTM) cells by a HIV-based lentivirus both in vitro and ex vivo.METHODS: The HIV-based lentivirus that contains an EF1-α promoter driving EGFP expression cassette was constructed following the standard molecular cloning methods. The cultured HTM cells were transduced at a range of multiplicity of infection(MOI) with HIV-based lentivirus. EGFP positive cell populations were detected by flow cytometry. Human anterior eye segments were cultured with perfusion culture system and transfected by HIV-based lentivirus with a 1 ×108transducing unit(TU) virus in perfusion liquid. The intraocular pressure was recorded every 8h for 21 d. The expression of EGFP in the anterior segment of the human eye was detected by fluorescence microscopy. Furthermore, the distribution of EGFP expression was confirmed by anti-EGFP immunohistochemical staining.RESULTS: The HIV-based lentivirus which contains an EF1-α promoter driving EGFP expression cassette was constructed successfully. After HTM cells were transduced with HIV-based lentivirus containing EGFP in vitro, the ratio of EGFP positive cells to the total cell number reached 92.3%, with the MOI of 15. After the lentivirus containing EGFP were used to transduce human anterior eye segments, the EGFP could be directly detected by fluorescence microscopy in vivo.Immunohistochemistry staining revealed that 88.19%EGFP-positive trabecular meshwork(TM) cells were observed in the human anterior segment. Nevertheless,the intraocular pressure in the lentivirus-transduced group kept constant when compared with control group(P 】0.05).CONCLUSION: EGFP gene could be efficiently transferred into HTM cells both in vitro and ex vivo by using HIV-based lentivirus.
