RESUMEN
Chinese soft-shelled turtle Pelodiscus sinensis has been an important aquaculture species in Southeast Asian countries. To breed a new variety of soft-shelled turtle with excellent properties and to evaluate the effect of hybridization of two turtle strains with a highly different trait phenotype, inheritance, microsatellite loci, and transcriptome analysis were studied in the hybrid turtles and their parents of P. sinensis Japanese strain and Qingxi black turtle. The genotypic characteristics and economic trait of the hybrid turtles were analyzed and compared to the two parents, showing significant growth vigor. The chromosome number of the hybrid turtle was diploid (2N = 66). The karyotype formulae were 8m+10sm+26t+22mc, with little differences between the two parents. Genotypic segregations of 241 microsatellite loci were screened in 3 populations including 90 species and showed that the specific allele numbers and polymorphic fragments increased in hybrid turtles indicating genetic diversity increased by hybridization. The liver transcriptome analysis of the hybrids and two parents showed similar distribution abundance in the parental and hybrid groups, but the transcripts with high abundance appeared in the hybrid group. There were 274 significant differentially expressed transcripts in the hybrid group compared to the two parental groups, among them 7 differentially expressed genes indicating super-parent expression, and only 2 genes showing low-parent expression. In the differentially expressed genes, expression changes were mainly contributed to regulatory region changes rather than coding region sequences. These results would be important for facilitating successful breeding strategies by hybridization in P. sinensis.
Asunto(s)
Genotipo , Hibridación Genética , Polimorfismo Genético , Tortugas/genética , Animales , Cromosomas/genética , Femenino , Cariotipo , Hígado/metabolismo , Masculino , Repeticiones de Microsatélite , Carácter Cuantitativo Heredable , Transcriptoma , Tortugas/crecimiento & desarrolloRESUMEN
Carotenoid cleavage oxygenases (CCOs) are a family of dioxygenases, which specifically catalyze the cleavage of conjugated double bonds in carotenoids and apocarotenoids in plants. In this study, genome-wide analysis of CCO genes in pepper plants was performed using bioinformatic methods. At least 11 members of the CCO gene family were identified in the pepper genome. Phylogenetic analysis showed that pepper and tomato CCO genes could be divided into two groups (CCDs and NCEDs). The CCD group included five sub-groups (CCD1, CCD4, CCD7, CCD8, and CCD-like). These results indicate that there is a close genetic relationship between the two species. Sequence analysis using the online tool, Multiple Expectation Maximization for Motif Elicitation (MEME), showed that the CCO proteins comprise multiple conserved motifs, with 20 to 41 amino acids. In addition, multiple cis-acting elements in the promoter of CCO genes were identified using the online tool PlantCARE, and were found to be involved in light responsiveness, plant hormone regulation, and biotic and abiotic stresses, suggesting potential roles of these proteins under different conditions. RNA-seq analysis revealed that the CCO genes exhibit distinct patterns of expression in the roots, stems, leaves, and fruit. These findings suggest that the CCO genes have important roles in the vegetative and reproductive development of pepper plants.
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Capsicum/enzimología , Capsicum/genética , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genoma de Planta , Familia de Multigenes , Oxigenasas/genética , Filogenia , Secuencias de Aminoácidos , Secuencia Conservada/genética , Exones/genética , Perfilación de la Expresión Génica , Genes de Plantas , Intrones/genética , Solanum lycopersicum/enzimología , Solanum lycopersicum/genética , Oxigenasas/metabolismo , Regiones Promotoras Genéticas/genética , Alineación de Secuencia , Análisis de Secuencia de ARNRESUMEN
The differentiation deficiencies of osteoclast precursors (pre-OCs) may contribute to osteoporosis. Research on osteoporosis has recently focused on microRNAs (miRNAs) that play crucial roles in pre-OC differentiation. In the current study, we aimed to analyze the expression and function of the glucocorticoid (GC)-associated miRNA-338-3p (miR-338-3p) in osteoclast formation. We found that dexamethasone induced osteoclast differentiation and inhibited miR-338-3p expression. Overexpression of an miR-338-3p mimic in osteoclast precursor cells attenuated GC-induced osteoclast formation and bone resorption, whereas inhibition of miR-338-3p reversed these effects. The expression of the nuclear factor κB ligand RANKL, a potential target gene of miR-338-3p, was inversely correlated with miR-338-3p expression in pre-OCs. Furthermore, we demonstrated that RANKL was directly regulated by miR-338-3p and re-introduction of RANKL reversed the inhibitory effects of miR-338-3p on osteoclast formation and bone resorption. Taken together, these findings demonstrate that miR-338-3p may play a significant role in GC-induced osteoclast differentiation and function by targeting RANKL in osteoclasts.
Asunto(s)
Resorción Ósea/genética , Dexametasona/farmacología , Glucocorticoides/farmacología , MicroARNs/genética , Osteoclastos/efectos de los fármacos , Ligando RANK/genética , Animales , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/metabolismo , Células de la Médula Ósea/patología , Resorción Ósea/metabolismo , Resorción Ósea/patología , Bovinos , Diferenciación Celular/efectos de los fármacos , Técnicas de Cocultivo , Femenino , Fémur/efectos de los fármacos , Fémur/metabolismo , Fémur/patología , Regulación de la Expresión Génica , Ratones , Ratones Endogámicos C57BL , MicroARNs/antagonistas & inhibidores , MicroARNs/metabolismo , Modelos Biológicos , Oligorribonucleótidos Antisentido/genética , Oligorribonucleótidos Antisentido/metabolismo , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Osteoclastos/patología , Cultivo Primario de Células , Ligando RANK/metabolismo , Transducción de SeñalRESUMEN
In this study, the SWKQ series microcomputer automatic incubator was used to study the growth and development of quail in the embryonic stage. Results showed that the embryo shape of became gradually defined as embryo aged. On day 6, the head and body of quail were clearly differentiated, the legs became longer and the wings appeared. At 7 embryo age, the entire embryo of quail was very clear, and the beak has formed. During 3 to 9 days of age, quail embryos length increased quickly, showing a linearly upward trend. At 9 day old, quail embryos length reached 2.2 cm. The regression equation of embryo length to day old was curve regression, giving as following: y=-0.464+0.325x-0.004x2, y: embryo length, x: the age of the embryo.
Asunto(s)
Animales , Coturnix/anatomía & histología , Coturnix/embriología , Desarrollo Embrionario , Análisis de Regresión , Incubadoras/veterinariaRESUMEN
In this study, the SWKQ series microcomputer automatic incubator was used to study the growth and development of quail in the embryonic stage. Results showed that the embryo shape of became gradually defined as embryo aged. On day 6, the head and body of quail were clearly differentiated, the legs became longer and the wings appeared. At 7 embryo age, the entire embryo of quail was very clear, and the beak has formed. During 3 to 9 days of age, quail embryos length increased quickly, showing a linearly upward trend. At 9 day old, quail embryos length reached 2.2 cm. The regression equation of embryo length to day old was curve regression, giving as following: y=-0.464+0.325x-0.004x2, y: embryo length, x: the age of the embryo.(AU)
Asunto(s)
Animales , Coturnix/anatomía & histología , Coturnix/embriología , Desarrollo Embrionario , Incubadoras/veterinaria , Análisis de RegresiónRESUMEN
Polymorphism of three quail communities was analyzed by using 12 microsatellite markers in this paper, aiming to provide scientific references for the evaluation, protection and utilization of quail genetic resources in China. Results demonstrated that the number of observed alleles by 12 microsatellite markers ranges between 4~7. The average polymorphism information contents (PIC) of the Chinese yellow quail, the Chinese black quail and the Korean quail, as detected by 12 microsatellite markers, are 0.6853, 0.6401 and 0.6565,respectively, and average heterozygosity values are 0.7333, 0.6957 and 0.7111, respectively. This indicates that the Chinese yellow quail has the richest genetic polymorphism. According to cluster analysis, the Chinese black quail and the Korean quail have the smallest genetic distance (0.0628), which reflects that they have the closest genetic relationship. The genetic distance between the Chinese yellow quail and the Korean quail is 0.0951. Therefore, the Chinese black quail and the Korean quail are clustered together firstly, and then the Chinese yellow quail.
Asunto(s)
Animales , China , Coturnix/genética , Polimorfismo Genético , Repeticiones de Microsatélite/genética , Variación Genética/fisiología , Alelos , Aves de Corral/genéticaRESUMEN
Polymorphism of three quail communities was analyzed by using 12 microsatellite markers in this paper, aiming to provide scientific references for the evaluation, protection and utilization of quail genetic resources in China. Results demonstrated that the number of observed alleles by 12 microsatellite markers ranges between 4~7. The average polymorphism information contents (PIC) of the Chinese yellow quail, the Chinese black quail and the Korean quail, as detected by 12 microsatellite markers, are 0.6853, 0.6401 and 0.6565,respectively, and average heterozygosity values are 0.7333, 0.6957 and 0.7111, respectively. This indicates that the Chinese yellow quail has the richest genetic polymorphism. According to cluster analysis, the Chinese black quail and the Korean quail have the smallest genetic distance (0.0628), which reflects that they have the closest genetic relationship. The genetic distance between the Chinese yellow quail and the Korean quail is 0.0951. Therefore, the Chinese black quail and the Korean quail are clustered together firstly, and then the Chinese yellow quail.(AU)
Asunto(s)
Animales , Variación Genética/fisiología , Coturnix/genética , Polimorfismo Genético , Repeticiones de Microsatélite/genética , China , Aves de Corral/genética , AlelosRESUMEN
Aiming at accelerating the application of molecular markers in the genetic improvement of quails, six EST-SSR markers were successfully developed using a bioinformatics method. Polymorphisms of three quail populations (Chinese yellow quail, China black quail and Korean quail) were detected. The results showed that there were 2-6 alleles in six EST-SSR markers. Mean polymorphism information contents of Chinese yellow quails, Chinese black quails and Korean quails were determined as 0.5451, 0.4962 and 0.4937, respectively. Average heterozygosity valuesof 0.6134, 0.5759 and 0.5613 were calculated. Among the six EST-SSR markers, three were highly polymorphic, and the other three were moderately polymorphic. The newly-developed six EST-SSR markers may be used to determine the genetic diversity of quails. The six EST-SSR markers identified were related to carbohydrate metabolism and melanin synthesis, but their specific mechanisms need to be further analyzed.
Asunto(s)
Animales , Biología Computacional/instrumentación , Biología Computacional/métodos , Biomarcadores/análisis , Coturnix/genética , Aves de Corral/genética , Carbohidratos/análisis , Melaninas/análisis , Variación GenéticaRESUMEN
Aiming at accelerating the application of molecular markers in the genetic improvement of quails, six EST-SSR markers were successfully developed using a bioinformatics method. Polymorphisms of three quail populations (Chinese yellow quail, China black quail and Korean quail) were detected. The results showed that there were 2-6 alleles in six EST-SSR markers. Mean polymorphism information contents of Chinese yellow quails, Chinese black quails and Korean quails were determined as 0.5451, 0.4962 and 0.4937, respectively. Average heterozygosity valuesof 0.6134, 0.5759 and 0.5613 were calculated. Among the six EST-SSR markers, three were highly polymorphic, and the other three were moderately polymorphic. The newly-developed six EST-SSR markers may be used to determine the genetic diversity of quails. The six EST-SSR markers identified were related to carbohydrate metabolism and melanin synthesis, but their specific mechanisms need to be further analyzed.(AU)
Asunto(s)
Animales , Coturnix/genética , Biomarcadores/análisis , Biología Computacional/instrumentación , Biología Computacional/métodos , Melaninas/análisis , Carbohidratos/análisis , Aves de Corral/genética , Variación GenéticaRESUMEN
The extracellular matrix (ECM) is the major macromolecule in skeletal muscle, which affects meat quality greatly. The remodeling of the ECM is mainly regulated by matrix metalloproteinases (MMPs). The expression patterns of MMP-1, -2, and -8 in longissimus dorsi muscle were explored using quantitative real-time polymerase chain reaction. The results show that the expression of MMP-1, -2, and -8 decreased significantly from 135 days of pregnancy to postnatal 30 months. While the expression of MMP-1, -2, and -8 showed no significant relationships with intramuscular fat contents, MMP-1 and -2 showed significant negative correlations with the shearing force of the longissimus dorsi muscle in cattle. The expression of MMP-1 also showed a significant negative correlation with cooking loss and a positive correlation with water holding capacity. The expression levels of MMP-1 and -2 were usually higher in fat than in skeletal muscle tissue. The expression of MMP-8 was significantly higher in the mammary fat pad and the longissimus dorsi muscle than in all other tissues. This study indicates that the remodeling of the ECM has important effects both on the development of postnatal skeletal muscle and on meat quality.
Asunto(s)
Calidad de los Alimentos , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 8 de la Matriz/metabolismo , Músculos Paraespinales/metabolismo , Carne Roja , Animales , Bovinos , Desarrollo Fetal , Expresión Génica , Metaloproteinasa 1 de la Matriz/genética , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 8 de la Matriz/genética , Músculos Paraespinales/embriología , Músculos Paraespinales/crecimiento & desarrollo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
We investigated azoospermia region microdeletions in male infertility patients with Klinefelter syndrome (KFS), as well as the association between azoospermia symptoms in patients with KFS and Y chromosome microdeletion polymorphisms. A total of 111 cases with male infertility confirmed to have KFS (47, XXY) and 94 fertile men were included in this study. Peripheral blood was drawn and DNA was extracted from these samples. Multiplex polymerase chain reaction was performed to screen the partial deletions of 25 sequence-tagged sites on the Y chromosome. In 111 cases with KFS, 1 case contained the AZFb+d+c deletion. The Gr/Gr deletion was identified in 12 KFS cases and 5 control cases. In addition, the b2/b3 deletion was identified in 13 KFS cases and 6 control cases. There were no significant differences in phenotype and genotype of the 2 partial AZFc deletions between patients and controls (P > 0.05). Our results suggest that patients with KFS may also have Y chromosome microdeletions to varying degrees and that the gr/gr deletion and b2/b3 deletion may not play a role in the susceptible genetic background of azoospermia in patients with KFS in the Sichuan population.
Asunto(s)
Síndrome de Klinefelter/genética , Proteínas de Plasma Seminal/genética , Azoospermia/genética , Deleción Cromosómica , Cromosomas Humanos Y/genética , Eliminación de Gen , Sitios Genéticos/genética , Humanos , Infertilidad Masculina/genética , Masculino , Fenotipo , Lugares Marcados de Secuencia , Aberraciones Cromosómicas Sexuales , Trastornos de los Cromosomas Sexuales del Desarrollo Sexual/genéticaRESUMEN
We examined the association between the methionine synthase reductase (MTRR A66G), methylenetetrahydrofolate reductase (MTHFR C677T and A1298C), and methionine synthase (MS A2756G) genotypes and non-obstructive male infertility in a Chinese population. This case-control study included 162 infertile Chinese patients with azoospermia (N = 100) or oligoasthenozoospermia (N = 62) and 120 fertile men as controls. The polymorphisms MTRR A66G, MTHFR C677T, A1298C, and MS A2756G were identified by direct DNA sequencing and the results were statistically analyzed. We found no association between the incidence of any of these variants in azoospermia patients and control populations. The frequency of the MTRR66 polymorphic genotypes (AG, AG+GG) was significantly higher in the oligoasthenozoospermia group compared to the controls (P = 0.013, 0.012). Our findings revealed an association between the single-nucleotide polymorphism A66G in the MTRR gene and male infertility, particularly in oligoasthenozoospermia males, suggesting that this polymorphism is a genetic risk factor for male infertility in Chinese men.
Asunto(s)
Ferredoxina-NADP Reductasa/genética , Predisposición Genética a la Enfermedad/genética , Infertilidad Masculina/genética , Polimorfismo de Nucleótido Simple , Alelos , Pueblo Asiatico/genética , Azoospermia/etnología , Azoospermia/genética , Secuencia de Bases , Estudios de Casos y Controles , China , Análisis Mutacional de ADN , Frecuencia de los Genes , Predisposición Genética a la Enfermedad/etnología , Genotipo , Humanos , Infertilidad Masculina/etnología , MasculinoRESUMEN
Musculoskeletal embryonic nuclear protein 1 (MUSTN1) gene is involved in myogenic fusion and differentiation in rats. We previously showed the differential expression of MUSTN1 in week (W) 2 and W6 breast muscles of Pekin ducks. In this study, we further investigated its molecular characteristics and expression profiles in different tissues at W7 and in breast and leg muscles at W1, W3, W5, W7, and W9. The relationship between muscle development and muscle fiber areas was also investigated. A 358-bp cDNA sequence was obtained. The coding sequence of duck MUSTN1 cDNA encoded a 78-amino acid sequence, which showed high similarity with those of other species (96% similarity with zebra finch and 94% with chicken). In addition, a 6435-bp genomic DNA sequence of MUSTN1 was obtained. In total, 231 transcription factor-binding sites were found in the promoter region, and many of these transcription factors were involved in the regulation of muscle development. MUSTN1 expression in breast muscle increased from W1 to W5 and then decreased at W9. In leg muscle, the expression increased from W1 to W3 and then decreased. The relative growth rates of breast and leg muscle fibers reached their peaks at W3-W5 and W1-W3, respectively. Since the greatest relative growth rates appeared at the highest expression levels of the MUSTN1 gene, it was thought to play roles in duck muscle development. Our findings would be helpful in understanding the molecular characteristics and functions of the MUSTN1 gene in breast muscle development of ducks.
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Proteínas Aviares/genética , Patos/genética , Regulación del Desarrollo de la Expresión Génica , Desarrollo de Músculos/genética , Músculo Esquelético/crecimiento & desarrollo , Proteínas Nucleares/genética , Secuencia de Aminoácidos , Animales , Proteínas Aviares/metabolismo , Patos/crecimiento & desarrollo , Evolución Molecular , Perfilación de la Expresión Génica , Masculino , Datos de Secuencia Molecular , Músculo Esquelético/metabolismo , Proteínas Nucleares/metabolismo , Especificidad de Órganos , Alineación de SecuenciaRESUMEN
We examined the value of serum procalcitonin (PCT), C-reactive protein (CRP), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) for predicting the survival of patients with early-onset stroke associated pneumonia (EOP). A total of 207 stroke patients were enrolled, and 91 developed EOP. Upon admission, serum PCT, CRP, sTREM-1 levels, clinical pulmonary infection score, and Acute Physiology and Chronic Health Evaluation II score were all significantly higher in patients with EOP than in those without EOP (P < 0.05). Of the 91 patients who developed EOP, 39 (42.9%) died (non-survivors) within 28 days. The Acute Physiology and Chronic Health Evaluation II score on admission was significantly higher in non-survivors than in survivors (P < 0.05). Serum PCT and sTREM-1 levels were slightly elevated on days 1, 3, and 5 in non-survivors and gradually decreased in survivors. Serum PCT, sTREM-1, and CRP levels were all significantly higher in non-survivors than in survivors on days 1, 3, and 5 (P < 0.05). The sensitivity and specificity of PCT for predicting the outcome of EOP were 84.6 and 71.2%, the sensitivity and specificity of sTREM-1 were 71.8 and 92.3%, and the sensitivity and specificity of sTREM-1 combined with PCT were 74.4 and 96.2%. Serum PCT combined with sTREM-1 accurately predicted the outcome of EOP patients, and dynamic monitoring of serum PCT and sTREM-1 levels is necessary.
Asunto(s)
Biomarcadores/sangre , Proteína C-Reactiva/metabolismo , Calcitonina/sangre , Células Mieloides/metabolismo , Neumonía/complicaciones , Precursores de Proteínas/sangre , Receptores Inmunológicos/metabolismo , Accidente Cerebrovascular/complicaciones , Adulto , Edad de Inicio , Anciano , Anciano de 80 o más Años , Péptido Relacionado con Gen de Calcitonina , Femenino , Humanos , Masculino , Persona de Mediana Edad , Neumonía/sangre , Accidente Cerebrovascular/sangre , Tasa de Supervivencia , Adulto JovenRESUMEN
The enhanced green fluorescent protein (EGFP) pEGFP-N1-P53 eukaryotic expression vector, which contains the human tumor suppressor p53, was constructed and transfected into chicken fibroblast cells and stage-X blastoderm to analyze the transfection efficiency. The complementary DNA of the human p53 gene was cloned by reverse transcription-polymerase chain reaction from human peripheral blood and inserted into the pEGFP-N1 vector by HindIII and BamHI double digestion. The pEGFP-N1-P53 vector was transfected into chicken embryo fibroblasts by Lipofectamine 2000 liposomes, and the transfection efficiency was analyzed by fluorescence microscope after 36 h of transfection. The stage-X blastoderm was also transfected by blastoderm injection using Lipofectamine 2000 liposomes at room temperature after 12-24 h; then hatching occurred until seventh day, and the transfection efficiency was analyzed by fluorescence microscope in the dead embryo. A total of 90 hatching eggs were transfected by the pEGFP-N1-P53 vector, and 20 chicken embryos expressed the reporter gene, which indicated that recombinant pEGFP-N1-P53 could be transfected and expressed in stage-X blastoderm by liposomes. Chicken embryo fibroblasts were transfected and expressed the reporter gene. The pEGFP-N1-P53 vector was constructed successfully and could be transfected and expressed in chicken embryo fibroblasts and stage-X blastoderms efficiently.
Asunto(s)
Vectores Genéticos , Proteínas Fluorescentes Verdes/genética , Proteínas Recombinantes de Fusión/genética , Proteína p53 Supresora de Tumor/genética , Animales , Blastodermo/crecimiento & desarrollo , Blastodermo/metabolismo , Embrión de Pollo , Fibroblastos/metabolismo , Regulación del Desarrollo de la Expresión Génica , Humanos , Proteínas Recombinantes de Fusión/biosíntesis , Proteína p53 Supresora de Tumor/biosíntesisRESUMEN
OBJECTIVES: The present study is to evaluate the expression level of enhancer of zeste homolog 2 (EZH2) and vascular endothelial growth factor (VEGF), and analyze their correlations with clinicopathological characteristics and survival in patients with clear cell renal cell carcinoma (CCRCC). The effect of EZH2 on apoptosis and cell proliferation in 786-O renal cancer cell line is investigated. METHODS: The expression level of EZH2 and VEGF was detected in 185 primary CCRCC patients' tissues using tissue microarray and immunohistochemistry. Small interfering RNA or enhanced green fluorescent protein transfection was employed to investigate the effect of EZH2 inhibition or overexpression on VEGF expression, apoptosis and cell proliferation in 786-O cells using flow cytometry, immunofluorescence microscopy, quantitative real-time reverse-transcription polymerase chain reaction and Western blot analysis. RESULTS: High expression level of EZH2 and VEGF was observed in advanced CCRCC and correlated with the TNM stage (p = 0.013, p = 0.001) and distant metastasis (p = 0.011, p = 0.038), respectively. EZH2 was positively correlated with VEGF in CCRCC tissues (correlation coefficient = 0.850, p < 0.001). Kaplan-Meier survival analysis revealed that patients with positive EZH2 expression had a shorter overall survival time compared to patients with negative EZH2 expression (34.3 vs. 67.2, p < 0.001). In 786-O cells, EZH2 silencing inhibited VEGF expression and cell proliferation while increasing apoptosis (p < 0.001). EZH2 overexpression promoted VEGF expression and cell proliferation while inhibiting apoptosis (p < 0.001). CONCLUSIONS: EZH2 correlates positively with VEGF and associates with adverse clinicopathologic characteristics and shorter survival time in CCRCC patients. EZH2 accelerates antiapoptosis and cell cycle in 786-O cells.
Asunto(s)
Carcinoma de Células Renales/metabolismo , Regulación Neoplásica de la Expresión Génica , Neoplasias Renales/metabolismo , Complejo Represivo Polycomb 2/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Anciano , Apoptosis , Ciclo Celular , Línea Celular Tumoral , Proliferación Celular , Progresión de la Enfermedad , Proteína Potenciadora del Homólogo Zeste 2 , Femenino , Citometría de Flujo , Silenciador del Gen , Proteínas Fluorescentes Verdes/química , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , ARN Interferente Pequeño/metabolismo , Análisis de Matrices TisularesRESUMEN
Development and selection of an ideal scaffold is of importance for tissue engineering. Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) is a biocompatible bioresorbable copolymer that belongs to the polyhydroxyalkanoate family. Because of its good biocompatibility, PHBHHx has been widely used as a cell scaffold for tissue engineering. This review focuses on the utilization of PHBHHx-based scaffolds in tissue engineering. Advances in the preparation, modification, and application of PHBHHx scaffolds are discussed.
Asunto(s)
Ácido 3-Hidroxibutírico/química , Materiales Biocompatibles/química , Caproatos/química , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Ácido 3-Hidroxibutírico/uso terapéutico , Materiales Biocompatibles/uso terapéutico , Huesos/fisiología , Caproatos/uso terapéutico , Cartílago/fisiología , Liofilización , Humanos , Músculo Liso/fisiología , Regeneración , Propiedades de SuperficieRESUMEN
Development and selection of an ideal scaffold is of importance for tissue engineering. Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) is a biocompatible bioresorbable copolymer that belongs to the polyhydroxyalkanoate family. Because of its good biocompatibility, PHBHHx has been widely used as a cell scaffold for tissue engineering. This review focuses on the utilization of PHBHHx-based scaffolds in tissue engineering. Advances in the preparation, modification, and application of PHBHHx scaffolds are discussed.
Asunto(s)
Humanos , /química , Materiales Biocompatibles/química , Caproatos/química , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , /uso terapéutico , Materiales Biocompatibles/uso terapéutico , Huesos/fisiología , Caproatos/uso terapéutico , Cartílago/fisiología , Liofilización , Músculo Liso/fisiología , Regeneración , Propiedades de SuperficieRESUMEN
SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.
Asunto(s)
Humanos , Diferenciación Celular/genética , Condrogénesis/genética , Sangre Fetal/citología , Células Madre Mesenquimatosas/citología , Factor de Transcripción SOX9/genética , Agrecanos/biosíntesis , Western Blotting , Cartílago/metabolismo , Proliferación Celular/genética , Condrocitos/metabolismo , Colágeno Tipo II/biosíntesis , Citometría de Flujo , Proteínas Fluorescentes Verdes , Regulación de la Expresión Génica/fisiología , Células Endoteliales de la Vena Umbilical Humana/citología , Inmunohistoquímica , Inmunofenotipificación , Cultivo Primario de Células , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ingeniería de Tejidos , TransfecciónRESUMEN
SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.