BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an ...BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an essential physiological process that occurs during embryonic development and tissue regeneration.Angiogenesis is the growth of new capillaries from pre-existing blood vessels,which is observed both prenatally and postnatally.The placenta is an organ composed of a variety of fetal-derived cells,including ECFCs,and therefore has significant potential as a source of fetal ECFCs for tissue engineering.AIM To investigate the possibility of isolating clonal ECFCs from human early gestation chorionic villi(CV-ECFCs)of the placenta,and assess their potential for tissue engineering.METHODS The early gestation chorionic villus tissue was dissociated by enzyme digestion.Cells expressing CD31 were selected using magnetic-activated cell sorting,and plated in endothelial-specific growth medium.After 2-3 wks in culture,colonies displaying cobblestone-like morphology were manually picked using cloning cylinders.We characterized CV-ECFCs by flow cytometry,immunophenotyping,tube formation assay,and Dil-Ac-LDL uptake assay.Viral transduction of CVECFCs was performed using a Luciferase/tdTomato-containing lentiviral vector,and transduction efficiency was tested by fluorescent microscopy and flow cytometry.Compatibility of CV-ECFCs with a delivery vehicle was determined using an FDA approved,small intestinal submucosa extracellular matrix scaffold.RESULTS After four passages in 6-8 wks of culture,we obtained a total number of 1.8×107 CV-ECFCs using 100 mg of early gestational chorionic villus tissue.Immunophenotypic analyses by flow cytometry demonstrated that CV-ECFCs highly expressed the endothelial markers CD31,CD144,CD146,CD105,CD309,only partially expressed CD34,and did not express CD45 and CD90.CV-ECFCs were capable of acetylated low-density lipoprotein uptake and tube formation,similar to cord blood-derived ECFCs(CB-ECFCs).CV-ECFCs can be transduced with a Luciferase/tdTomato-containing lentiviral vector at a transduction efficiency of 85.1%.Seeding CV-ECFCs on a small intestinal submucosa extracellular matrix scaffold confirmed that CV-ECFCs were compatible with the biomaterial scaffold.CONCLUSION In summary,we established a magnetic sorting-assisted clonal isolation approach to derive CV-ECFCs.A substantial number of CV-ECFCs can be obtained within a short time frame,representing a promising novel source of ECFCs for fetal treatments.展开更多
The organization of the compartment of mesenchymal stem cells is still obscure. Two types of human stromal precursor cells are known. Both of them are analyzed in in vitro system: mesenchymal multipotent stromal cells...The organization of the compartment of mesenchymal stem cells is still obscure. Two types of human stromal precursor cells are known. Both of them are analyzed in in vitro system: mesenchymal multipotent stromal cells (MMSC) and fibroblast colony forming units (CFU-F). The aim of this study was to compare the main characteristics of MMSC and CFU-F derived from the bone marrow of 24 healthy donors. Growth and differentiation parameters, as well as relative expression levels of different genes were analyzed in MMSC and CFU-F. MMSC were cultivated for 5 passages. CFU-F concentration was determined for each bone marrow sample. The data obtained demonstrated the heterogeneity and hierarchical organization of both studied populations of stromal precursor cells-MMSC and CFU-F. These two types of stromal precursor cells turned to be different in most parameters studied. Altogether MMSC seemed to be more immature cells than CFU-F and took up the higher position in hierarchical tree of mesenchymal stem cells. The rate of differentiation and proliferative potential decreased with the donor’s age in both populations MMSC and CFU-F.展开更多
The mold pressing process was applied to investigate the formability of closed-cell aluminum foam in solid–liquid–gas coexisting state.Results show that the shape formation of closed-cell aluminum foam in the solid...The mold pressing process was applied to investigate the formability of closed-cell aluminum foam in solid–liquid–gas coexisting state.Results show that the shape formation of closed-cell aluminum foam in the solid–liquid–gas coexisting state was realized through cell wall deformation and cell movement caused by primary α-Al grains that slid,rotated,deformed,and ripened within cell walls.During formation,characteristic parameters of closed-cell aluminum foam were almost unchanged.Under proper forming conditions,shaped products of closed-cell aluminum foam could be fabricated through mold pressing.展开更多
Hot plate forming using a cell-typed die is a process for forming a large thick plate with a spherical shape for the manufacture of a large spherical LNG tank.Cell-typed upper and lower dies made of a framework of ste...Hot plate forming using a cell-typed die is a process for forming a large thick plate with a spherical shape for the manufacture of a large spherical LNG tank.Cell-typed upper and lower dies made of a framework of steel plates fitted to make a grid pattern are used in this process,and an air-cooling device is separately installed inside the lower die.A finite element analysis (FEA) technique was developed,which included hot forming,air flow,cooling and thermal deformation analysis for the hot plate forming process using the cell-typed die.Further,the convective and interface heat transfer coefficients were used to reproduce analytically the effects of the cooling device in the hot plate forming analysis.A small-scale model test of the process was conducted to verify the FEA technique.The analysis results show that the curvature of the final plate agrees well with that of the designed experiment within a maximum relative error of 0.03% at the corner of the plate.展开更多
Cell-based technologies are used as a therapeutic strategy in spinal cord injury(SCI). Mesenchymal stem cells(MSCs), which secrete various neurotrophic factors and cytokines, have immunomodulatory, anti-apoptotic and ...Cell-based technologies are used as a therapeutic strategy in spinal cord injury(SCI). Mesenchymal stem cells(MSCs), which secrete various neurotrophic factors and cytokines, have immunomodulatory, anti-apoptotic and anti-inflammatory effects, modulate reactivity/phenotype of astrocytes and the microglia, thereby promoting neuroregeneration seem to be the most promising. The therapeutic effect of MSCs is due to a paracrine mechanism of their action, therefore the survival of MSCs and their secretory phenotype is of particular importance. Nevertheless, these data are not always reported in efficacy studies of MSC therapy in SCI. Here, we provide a review with summaries of preclinical trials data evaluating the efficacy of MSCs in animal models of SCI. Based on the data collected, we have tried(1) to establish the behavior of MSCs after transplantation in SCI with an evaluation of cell survival, migration potential, distribution in the area of injured and intact tissue and possible differentiation;(2) to determine the effects MSCs on neuronal microenvironment and correlate them with the efficacy of functional recovery in SCI;(3) to ascertain the conditions under which MSCs demonstrate their best survival and greatest efficacy.展开更多
Biofilm-associated microorganisms play crucial roles in terrestrial and aquatic nutrient cycling and in the biodegradation of environmental pollutants.Biofilm formation was determined for a total of 18 bacterial isola...Biofilm-associated microorganisms play crucial roles in terrestrial and aquatic nutrient cycling and in the biodegradation of environmental pollutants.Biofilm formation was determined for a total of 18 bacterial isolates obtained from the biofilms of wastewater treatment systems and of little carpolite in soil.Among these isolates,seven showed strong biofilm-forming capacity.The phylogenetic affiliation of the isolates showing high biofilm formation capacity was determined through 16S rDNA sequencing and the isolates were grouped into 7 bacterial species including Pseudomonas sp.,Pseudomonas putida,Aeromonas caviae,Bacillus cereus,Pseudomonas plecoglossicida,Aeromonas hydrophila,and Comamonas testosteroni.The biofilm-forming capacity was closely related with flagella,exopolysaccharide,and extracel-lular protein.According to the coefficient of determination,the relative importance of the five biological characteristics to biofilm formation was,in order from greatest to least,exopolysaccharide > flagella > N-acyl-homoserine lactones(AHLs) signaling molecules > extracellular protein > swarming motility.展开更多
The mechanism of attachment and leaching of thiobacillus ferrooxidans（T.f.） on chalcopyrite were studied. The shaking flasks with bacteria were observed by SEM. The process of T.f attached to the surface of the mine...The mechanism of attachment and leaching of thiobacillus ferrooxidans（T.f.） on chalcopyrite were studied. The shaking flasks with bacteria were observed by SEM. The process of T.f attached to the surface of the mineral sample and the biofilm forming were described. The promoting role of the biofilm for bioleaching was discussed. The existence of Fe2+ in the exopolysaccharide layer of T.f was demonstrated by EM（electronic microscope）cell-chemistry analysis. These results show that under the proper growth condition of bacteria, bioleaching of chalcopyrite results in the formation of complete biofilm after 23 weeks. There are iron ions in the outer layer polymer of T.f., which provides the micro-environment for themselves, and can guaruntee the energy needed for the bacteria growth in the biofilm. At the same time, Fe3+ions produced oxidize sulfide which brings about the increase of both growth rate of the bacterial and leaching rate of sulfide minerals.展开更多
Non-adherent bone marrow cell-derived mesenchymal stem cells from C57BL/6J mice were sepa- rated and cultured using the ＂pour-off＂ method. Non-adherent bone marrow cell-derived mesen- chymal stem ceils developed col...Non-adherent bone marrow cell-derived mesenchymal stem cells from C57BL/6J mice were sepa- rated and cultured using the ＂pour-off＂ method. Non-adherent bone marrow cell-derived mesen- chymal stem ceils developed colony-forming unit-fibroblasts, and could be expanded by supple- mentation with epidermal growth factor. Immunocytochemistry showed that the non-adherent bone marrow cell-derived mesenchymal stem cells exposed to basic fibroblast growth factor/epidermal growth factor/nerve growth factor expressed the neuron specific markers, neurofilament-200 and NeuN, in vitro. Non-adherent bone marrow cell-derived mesenchymal stem cells from 13-galactosidase transgenic mice were also transplanted into focal ischemic brain （right corpus striatum） of C57BL/6J mice. At 8 weeks, cells positive for LacZ and 13-galactosidase staining were observed in the ischemic tissues, and cells co-labeled with both 13-galactosidase and NeuN were seen by double immunohistochemical staining. These findings suggest that the non-adherent bone marrow cell-derived mesenchymal stem cells could differentiate into neuronal-like cells in vitro and in vivo.展开更多
AIM: To determine the proliferative potential and the maintenance of stem cell activity in stored human limbal tissues, and correlate this with the preservation time, cell viability and the expression of stem cell mar...AIM: To determine the proliferative potential and the maintenance of stem cell activity in stored human limbal tissues, and correlate this with the preservation time, cell viability and the expression of stem cell markers. METHODS: Thirty limbal rims were split into 4 parts and stored in corneal preservation medium at 4 degrees C for 0, 1, 4, or 7 days. The limbal stem cell and mitotic markers P63, CK19, proliferating cell nuclear antigen (PCNA), and Ki67 were determined by immunohistochemical staining. The proliferative potential of limbal epithelial cells was assessed by cell viability, the ability of generating stratified epithelium, and colony forming assay. RESULTS: The stored tissues maintained limbal stratified structure to 7 days and exhibited comparable expression level of stem cell and mitotic markers. The proportion of viable cells decreased with the prolonged preservation time, while colony forming efficiency decreased from the 1st day and disappeared at the 4th day. When inoculated on amniotic membrane, the cells preserved for 1 day formed a stratified epithelium, while the cells from 4 days' preservation formed a discontinuous layer. CONCLUSION: The colony forming efficiency of limbal epithelial stem/progenitor cells decreased rapidly with the increasing preservation time, while the expression level of markers and capacity of forming epithelial monolayer on amniotic membrane decreased gradually. The limbal epithelial stem cells lost their function earlier than the lost expression level of stem cell markers. This may help us to better choose the appropriate preservation grafts for future limbal stem cell transplantation.展开更多
AIM:To investigate the effect of intravitreal injection of DL-alpha-aminoadipic acid (DL-α-AAA) on ocular refractive state and retinal dopamine, transforming growth factor-β2 (TGFβ2 ), vasoactive intestinal polypep...AIM:To investigate the effect of intravitreal injection of DL-alpha-aminoadipic acid (DL-α-AAA) on ocular refractive state and retinal dopamine, transforming growth factor-β2 (TGFβ2 ), vasoactive intestinal polypeptide (VIP) in guinea pig form-deprived myopia. METHODS:Four-week-old pigmented guinea pigs were randomly assigned to 4 groups:normal control, deprivation, deprivation plus DL-α-AAA, deprivation plus saline. Form deprivation was induced with the self-made translucent eye shields, and lasted for 14 days. 8μg DL-α-AAA was injected into the vitreous chamber of deprived eyes. The corneal radius of curvature, refraction and axial length were measured. Retinal dopamine content was evaluated by the high-performance liquid chromatography with electrochemical detection, and TGFβ2 and VIP protein were detected by Western blotting. RESULTS:Fourteen days of eye occlusion caused the axial length to elongate and become myopic in the form-deprived eyes, with the decrease of retinal dopamine and the increase of TGFβ 2 and vasoactive intestinal polypeptide (VIP) protein. Intravitreal injection of DL-α-AAA could inhibit the myopic shift from (-3.65±1.06)D to (-1.48 ±0.63)D, P 【0.01 due to goggles occluding and cause the decrease of retinal TGFβ2 protein in the deprived eyes. However, intravitreal injection of DL-α-AAA had no significant effect on retinal dopamine and VIP protein in deprived eyes. Retinal TGFβ2 protein correlated highly with the ocular refraction (y =-3.34 + 0.31/x , F =74.75, P 【0.001) and axial length (y =8.39-0.02/ x , F =48.32, P 【0.001) in different treatment groups. ·CONCLUSION:Intravitreal injection of DL-α-AAA is effectively able to suppress the development of form deprivation myopia, which may be associated with retinalTGFβ2 protein in guinea pigs.展开更多
基金the Shriners Hospital for Children Postdoctoral Research Fellowship award,No.84704-NCA-19UC Davis School of Medicine Dean’s Fellowship award and funding from the NIH,No.5R01NS100761-02 and No.R03HD091601-01+2 种基金the California Institute of Regenerative Medicine,No.PC1-08103 and No.CLIN1-11404Shriners Hospitals for Children,No.85120-NCA-16,No.85119-NCA-18,No.85108-NCA-19 and No.87200-NCA-19March of Dimes Foundation,No.5FY1682
文摘BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an essential physiological process that occurs during embryonic development and tissue regeneration.Angiogenesis is the growth of new capillaries from pre-existing blood vessels,which is observed both prenatally and postnatally.The placenta is an organ composed of a variety of fetal-derived cells,including ECFCs,and therefore has significant potential as a source of fetal ECFCs for tissue engineering.AIM To investigate the possibility of isolating clonal ECFCs from human early gestation chorionic villi(CV-ECFCs)of the placenta,and assess their potential for tissue engineering.METHODS The early gestation chorionic villus tissue was dissociated by enzyme digestion.Cells expressing CD31 were selected using magnetic-activated cell sorting,and plated in endothelial-specific growth medium.After 2-3 wks in culture,colonies displaying cobblestone-like morphology were manually picked using cloning cylinders.We characterized CV-ECFCs by flow cytometry,immunophenotyping,tube formation assay,and Dil-Ac-LDL uptake assay.Viral transduction of CVECFCs was performed using a Luciferase/tdTomato-containing lentiviral vector,and transduction efficiency was tested by fluorescent microscopy and flow cytometry.Compatibility of CV-ECFCs with a delivery vehicle was determined using an FDA approved,small intestinal submucosa extracellular matrix scaffold.RESULTS After four passages in 6-8 wks of culture,we obtained a total number of 1.8×107 CV-ECFCs using 100 mg of early gestational chorionic villus tissue.Immunophenotypic analyses by flow cytometry demonstrated that CV-ECFCs highly expressed the endothelial markers CD31,CD144,CD146,CD105,CD309,only partially expressed CD34,and did not express CD45 and CD90.CV-ECFCs were capable of acetylated low-density lipoprotein uptake and tube formation,similar to cord blood-derived ECFCs(CB-ECFCs).CV-ECFCs can be transduced with a Luciferase/tdTomato-containing lentiviral vector at a transduction efficiency of 85.1%.Seeding CV-ECFCs on a small intestinal submucosa extracellular matrix scaffold confirmed that CV-ECFCs were compatible with the biomaterial scaffold.CONCLUSION In summary,we established a magnetic sorting-assisted clonal isolation approach to derive CV-ECFCs.A substantial number of CV-ECFCs can be obtained within a short time frame,representing a promising novel source of ECFCs for fetal treatments.
文摘The organization of the compartment of mesenchymal stem cells is still obscure. Two types of human stromal precursor cells are known. Both of them are analyzed in in vitro system: mesenchymal multipotent stromal cells (MMSC) and fibroblast colony forming units (CFU-F). The aim of this study was to compare the main characteristics of MMSC and CFU-F derived from the bone marrow of 24 healthy donors. Growth and differentiation parameters, as well as relative expression levels of different genes were analyzed in MMSC and CFU-F. MMSC were cultivated for 5 passages. CFU-F concentration was determined for each bone marrow sample. The data obtained demonstrated the heterogeneity and hierarchical organization of both studied populations of stromal precursor cells-MMSC and CFU-F. These two types of stromal precursor cells turned to be different in most parameters studied. Altogether MMSC seemed to be more immature cells than CFU-F and took up the higher position in hierarchical tree of mesenchymal stem cells. The rate of differentiation and proliferative potential decreased with the donor’s age in both populations MMSC and CFU-F.
基金financially supported by the National Natural Science Foundations of China (No.51371104)
文摘The mold pressing process was applied to investigate the formability of closed-cell aluminum foam in solid–liquid–gas coexisting state.Results show that the shape formation of closed-cell aluminum foam in the solid–liquid–gas coexisting state was realized through cell wall deformation and cell movement caused by primary α-Al grains that slid,rotated,deformed,and ripened within cell walls.During formation,characteristic parameters of closed-cell aluminum foam were almost unchanged.Under proper forming conditions,shaped products of closed-cell aluminum foam could be fabricated through mold pressing.
基金Project(2010-0008-277)supported by the NCRC(National Core Research Center)Program through the National Research Foundation of Korea,funded by the Ministry of Education,Science,and TechnologyProject supported by R&D for Technology Development Program of Ministry of Knowledge Economy,Korea
文摘Hot plate forming using a cell-typed die is a process for forming a large thick plate with a spherical shape for the manufacture of a large spherical LNG tank.Cell-typed upper and lower dies made of a framework of steel plates fitted to make a grid pattern are used in this process,and an air-cooling device is separately installed inside the lower die.A finite element analysis (FEA) technique was developed,which included hot forming,air flow,cooling and thermal deformation analysis for the hot plate forming process using the cell-typed die.Further,the convective and interface heat transfer coefficients were used to reproduce analytically the effects of the cooling device in the hot plate forming analysis.A small-scale model test of the process was conducted to verify the FEA technique.The analysis results show that the curvature of the final plate agrees well with that of the designed experiment within a maximum relative error of 0.03% at the corner of the plate.
基金supported by a grant from the Russian Foundation for Basic Research,No.16-34-60101(to YOM)a grant from the Ministry of Education and Science of the Russian Federation,No.20.5175.2017/6.7(to AAR)performed in accordance with the Program of Competitive Growth of the Kazan Federal University
文摘Cell-based technologies are used as a therapeutic strategy in spinal cord injury(SCI). Mesenchymal stem cells(MSCs), which secrete various neurotrophic factors and cytokines, have immunomodulatory, anti-apoptotic and anti-inflammatory effects, modulate reactivity/phenotype of astrocytes and the microglia, thereby promoting neuroregeneration seem to be the most promising. The therapeutic effect of MSCs is due to a paracrine mechanism of their action, therefore the survival of MSCs and their secretory phenotype is of particular importance. Nevertheless, these data are not always reported in efficacy studies of MSC therapy in SCI. Here, we provide a review with summaries of preclinical trials data evaluating the efficacy of MSCs in animal models of SCI. Based on the data collected, we have tried(1) to establish the behavior of MSCs after transplantation in SCI with an evaluation of cell survival, migration potential, distribution in the area of injured and intact tissue and possible differentiation;(2) to determine the effects MSCs on neuronal microenvironment and correlate them with the efficacy of functional recovery in SCI;(3) to ascertain the conditions under which MSCs demonstrate their best survival and greatest efficacy.
基金supported by the National Natural Science Foundation of China (No.30600016)the Environment Protection Department of Jiangsu Province,China (No.2004007)
文摘Biofilm-associated microorganisms play crucial roles in terrestrial and aquatic nutrient cycling and in the biodegradation of environmental pollutants.Biofilm formation was determined for a total of 18 bacterial isolates obtained from the biofilms of wastewater treatment systems and of little carpolite in soil.Among these isolates,seven showed strong biofilm-forming capacity.The phylogenetic affiliation of the isolates showing high biofilm formation capacity was determined through 16S rDNA sequencing and the isolates were grouped into 7 bacterial species including Pseudomonas sp.,Pseudomonas putida,Aeromonas caviae,Bacillus cereus,Pseudomonas plecoglossicida,Aeromonas hydrophila,and Comamonas testosteroni.The biofilm-forming capacity was closely related with flagella,exopolysaccharide,and extracel-lular protein.According to the coefficient of determination,the relative importance of the five biological characteristics to biofilm formation was,in order from greatest to least,exopolysaccharide > flagella > N-acyl-homoserine lactones(AHLs) signaling molecules > extracellular protein > swarming motility.
文摘The mechanism of attachment and leaching of thiobacillus ferrooxidans（T.f.） on chalcopyrite were studied. The shaking flasks with bacteria were observed by SEM. The process of T.f attached to the surface of the mineral sample and the biofilm forming were described. The promoting role of the biofilm for bioleaching was discussed. The existence of Fe2+ in the exopolysaccharide layer of T.f was demonstrated by EM（electronic microscope）cell-chemistry analysis. These results show that under the proper growth condition of bacteria, bioleaching of chalcopyrite results in the formation of complete biofilm after 23 weeks. There are iron ions in the outer layer polymer of T.f., which provides the micro-environment for themselves, and can guaruntee the energy needed for the bacteria growth in the biofilm. At the same time, Fe3+ions produced oxidize sulfide which brings about the increase of both growth rate of the bacterial and leaching rate of sulfide minerals.
基金supported by the National Natural Science Foundation of China,No.30471836
文摘Non-adherent bone marrow cell-derived mesenchymal stem cells from C57BL/6J mice were sepa- rated and cultured using the ＂pour-off＂ method. Non-adherent bone marrow cell-derived mesen- chymal stem ceils developed colony-forming unit-fibroblasts, and could be expanded by supple- mentation with epidermal growth factor. Immunocytochemistry showed that the non-adherent bone marrow cell-derived mesenchymal stem cells exposed to basic fibroblast growth factor/epidermal growth factor/nerve growth factor expressed the neuron specific markers, neurofilament-200 and NeuN, in vitro. Non-adherent bone marrow cell-derived mesenchymal stem cells from 13-galactosidase transgenic mice were also transplanted into focal ischemic brain （right corpus striatum） of C57BL/6J mice. At 8 weeks, cells positive for LacZ and 13-galactosidase staining were observed in the ischemic tissues, and cells co-labeled with both 13-galactosidase and NeuN were seen by double immunohistochemical staining. These findings suggest that the non-adherent bone marrow cell-derived mesenchymal stem cells could differentiate into neuronal-like cells in vitro and in vivo.
基金National Natural Science Foundation of China (No.81170816)Specialized Research Fund for the Doctoral Program of Higher Education (No.20113706110004)Qingjun Zhou is partially supported by the TaishanScholar Program of Jinan City, China (No.20081148)
文摘AIM: To determine the proliferative potential and the maintenance of stem cell activity in stored human limbal tissues, and correlate this with the preservation time, cell viability and the expression of stem cell markers. METHODS: Thirty limbal rims were split into 4 parts and stored in corneal preservation medium at 4 degrees C for 0, 1, 4, or 7 days. The limbal stem cell and mitotic markers P63, CK19, proliferating cell nuclear antigen (PCNA), and Ki67 were determined by immunohistochemical staining. The proliferative potential of limbal epithelial cells was assessed by cell viability, the ability of generating stratified epithelium, and colony forming assay. RESULTS: The stored tissues maintained limbal stratified structure to 7 days and exhibited comparable expression level of stem cell and mitotic markers. The proportion of viable cells decreased with the prolonged preservation time, while colony forming efficiency decreased from the 1st day and disappeared at the 4th day. When inoculated on amniotic membrane, the cells preserved for 1 day formed a stratified epithelium, while the cells from 4 days' preservation formed a discontinuous layer. CONCLUSION: The colony forming efficiency of limbal epithelial stem/progenitor cells decreased rapidly with the increasing preservation time, while the expression level of markers and capacity of forming epithelial monolayer on amniotic membrane decreased gradually. The limbal epithelial stem cells lost their function earlier than the lost expression level of stem cell markers. This may help us to better choose the appropriate preservation grafts for future limbal stem cell transplantation.
基金National Natural Science Foundation of China (No. 30600694)
文摘AIM:To investigate the effect of intravitreal injection of DL-alpha-aminoadipic acid (DL-α-AAA) on ocular refractive state and retinal dopamine, transforming growth factor-β2 (TGFβ2 ), vasoactive intestinal polypeptide (VIP) in guinea pig form-deprived myopia. METHODS:Four-week-old pigmented guinea pigs were randomly assigned to 4 groups:normal control, deprivation, deprivation plus DL-α-AAA, deprivation plus saline. Form deprivation was induced with the self-made translucent eye shields, and lasted for 14 days. 8μg DL-α-AAA was injected into the vitreous chamber of deprived eyes. The corneal radius of curvature, refraction and axial length were measured. Retinal dopamine content was evaluated by the high-performance liquid chromatography with electrochemical detection, and TGFβ2 and VIP protein were detected by Western blotting. RESULTS:Fourteen days of eye occlusion caused the axial length to elongate and become myopic in the form-deprived eyes, with the decrease of retinal dopamine and the increase of TGFβ 2 and vasoactive intestinal polypeptide (VIP) protein. Intravitreal injection of DL-α-AAA could inhibit the myopic shift from (-3.65±1.06)D to (-1.48 ±0.63)D, P 【0.01 due to goggles occluding and cause the decrease of retinal TGFβ2 protein in the deprived eyes. However, intravitreal injection of DL-α-AAA had no significant effect on retinal dopamine and VIP protein in deprived eyes. Retinal TGFβ2 protein correlated highly with the ocular refraction (y =-3.34 + 0.31/x , F =74.75, P 【0.001) and axial length (y =8.39-0.02/ x , F =48.32, P 【0.001) in different treatment groups. ·CONCLUSION:Intravitreal injection of DL-α-AAA is effectively able to suppress the development of form deprivation myopia, which may be associated with retinalTGFβ2 protein in guinea pigs.
