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1.
J Cell Physiol ; 234(7): 11401-11410, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30623407

RESUMO

Multiple sclerosis (MS) patients should take medication such as fingolimod (FTY-720) for a long time, hence pharmaceutical effects on other neural cells such as dopaminergic cells are important. Dopaminergic cell line, BE(2)-M17, was treated by FTY-720 and then cell viability and genes involve in neurosurvival were investigated. It was disclosed that FTY-720 significantly stimulates Bcl2 overexpression. Whereas, it decreased intracellular reactive oxygen species production and cell membrane damage of dopaminergic cells. The increase in Bcl2/Bax ratio increased the cell metabolic activity and decreased propidium iodide-positive cells. Besides, FTY-720 induced the overexpression of CACNA1C, nNOS gene, and nitric oxide production. However, FTY-720 induced GABARA1 overexpression and eventually it could overcame to the cytotoxic effect of intracellular calcium. This cascade led to tyrosine hydroxylase and BDNF genes overexpression whereas FTY-720 did not change GDNF concentration in BE(2)-M17 cells. Concluding, it might be said that taking FTY-720 in MS patients did not induce adverse effect on dopaminergic cells.


Assuntos
Neurônios Dopaminérgicos/metabolismo , Cloridrato de Fingolimode/farmacologia , Esfingosina/metabolismo , Canais de Cálcio Tipo L/genética , Canais de Cálcio Tipo L/metabolismo , Linhagem Celular , Sobrevivência Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Humanos , Imunossupressores/farmacologia , L-Lactato Desidrogenase/metabolismo , Óxido Nítrico/metabolismo , Propídio , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio , Receptores de GABA-A/genética , Receptores de GABA-A/metabolismo , Tirosina 3-Mono-Oxigenase/genética , Tirosina 3-Mono-Oxigenase/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
2.
Cell Biol Int ; 37(11): 1181-9, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23765607

RESUMO

Bone matrix consists of two major phases at the nanoscale: organic and hydroxyapatite. Nanotechnology as a diverse and interdisciplinary area of research has the capacity to revolutionise many areas of applications such as bone tissue engineering. Nanohydroxyapatite/gelatin composite has higher osteoblast attachment and proliferation than micro-sized ones, and shorter culturing period and lower cell seeding density compared to pure gelatin. A nanostructured scaffold was fabricated by three methods for bone repair using nanohydroxyapatite and gelatin as the main components. Its biocompatibility, alizarin red test on the 14th and 21st days, gene expression on the 21st day in in vitro using and histomorphometry after 4 and 8 weeks post-implantation in the rat were investigated. Cultured unrestricted somatic stem cells used for in vitro study showed an excellent level of cell attachment to the scaffold. Cells induced more osteoblast differentiation on the scaffold than in 2D cell culture. Osteoblast differentiation and bone regeneration results of in vitro and in vivo investigation on scaffold were extremely significant, better than control and treatment groups. These effects could be attributed to the shape and size of nanoHA particles and good architecture of the scaffold. The results confirm the feasibility of bone regeneration using synthesised scaffold as a temporary bone substitute.


Assuntos
Durapatita/farmacologia , Gelatina/farmacologia , Nanocompostos/química , Osteogênese/efeitos dos fármacos , Células-Tronco/citologia , Alicerces Teciduais/química , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Materiais Biocompatíveis/farmacologia , Regeneração Óssea/efeitos dos fármacos , Regeneração Óssea/genética , Cálcio/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Nanocompostos/ultraestrutura , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Osteogênese/genética , Implantação de Prótese , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo
3.
Daru ; 27(1): 159-168, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30875026

RESUMO

BACKGROUND: Since women pay more attention to their skin's health, pharmaceutical companies invest heavily on skin care product development. Further, the success of drug nano-carriers in passing through the skin justifies the need to conduct studies at the nano-scale. ß1-integrin down regulation has been proposed as a sign of skin aging. METHODS: Six drug nano-carriers (50 and 75 nm) were prepared at three ethanol concentrations (0, 3,and 5%) and different temperatures. Then, the impact of Nanocarriers on fibroblasts were investigated. RESULTS: DLS showed that increasing ethanol concentration decreased the surface tension that caused a decrease in the particle size in non-temperature formulations while increasing the temperature to 60 °C to lower Gibbs free energy increased the particle size. Ethanol addition decreased ß1-integrin over-expression, whereas larger nano-carriers induced an over-expression of ß1-integrin, Bcl2/Bax ratio, and an increase in live cell number. ß1-integrin over-expression did not correlate with the rate of fibroblast proliferation and NFκB expression. An increase in fibroblast mortality in relation to smaller nano-carriers was not only due to the increase in Bax ratio, but was related to NFκB over-expression. CONCLUSION: The development of a regenerative pharmaceutical approach in skin repair was based on the effect of particle size and ethanol concentration of the drug nano-carriers on the expression of ß1-integrin in fibroblasts. A curcumin nanoformulation sized 77 nm and containing of 3% ethanol was more effective in increasing ß1-integrin gene over-expression, anti-apoptosis of fibroblast cells (Bcl2/Bax ratio), and in decreasing Bax and NFκB gene expression than that with a particle size of 50 nm. Such a formulation may be considered a valuable candidate in anti-aging and wound-healing formulations. Graphical abstract The effect of particle size on Bcl2/Bax ratio and NFκ-B gene expression through the cell surface receptor of ß1- integrin. Bigger nanocarriers induce over-expression of integrin ß1 gene and also lead to an increase in Bcl2/Bax ratio along with a decrease in NFκ-B, unlike the smaller nanocarriers.


Assuntos
Curcumina/farmacologia , Etanol/química , Fibroblastos/citologia , Integrina beta1/genética , Envelhecimento da Pele/efeitos dos fármacos , Regulação para Cima , Animais , Linhagem Celular , Proliferação de Células , Sobrevivência Celular/efeitos dos fármacos , Curcumina/química , Feminino , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Camundongos , NF-kappa B/genética , Nanopartículas , Tamanho da Partícula , Proteína X Associada a bcl-2/genética
4.
Mater Sci Eng C Mater Biol Appl ; 101: 148-158, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31029307

RESUMO

Recent studies suggest that nanotopography can trigger colocalization of integrins and bone morphogenetic protein 2 (BMP2) receptors (e.g., BMPR1A), thereby leading to osteogenesis. In this study, the bone marrow homing peptide 1 (BMHP1) motif was bound to a self-assembling peptide core to form a hydrogel-based nanofiber (R-BMHP1). The docking and molecular dynamic study revealed that the R-BMHP1 sequence induced a stronger electrostatic interaction than BMP2 through arginines in the RADA core sequence and through lysine24 in the BMHP1 motif with BMPR1A. Notably, decrease of polar solvation binding energy will enhance the total binding energy and increases bone regeneration even more than BMP2 The enhanced osteogenesis and bone repair potential of R-BMHP1 nanofiber might be related to its chemical interaction with BMPR1A, which triggered downstream signal transduction through osteogenic genes overexpression in osteo-differentiated mesenchymal stem cells (MSCs), as well as implanted critical-sized bone defects in rats. Following that, calcium deposition occurred by osteoblast-like cells, ALP activity increased in osteodifferentiation MSCs and rat serum, and calcium density improved in bone defects (X-ray). The nanofiber was biocompatible and enhanced the cell viability of MSCs, without multinuclear cell infiltration into the defect site. Taking everything into account, not only does nanotopography induce osteogenesis through colocalization of BMPRs and integrins, but also R-BMHP1 nanofibers (considering their chemical structure) induce cell proliferation, osteogenesis, and bone repair through strong electrostatic interaction with BMPR1A and downstream signaling. The entire outcome of this study manifests the plausibility of R-BMHP1 for spine and spinal cord injury repair.


Assuntos
Proteína Morfogenética Óssea 2/química , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/química , Nanofibras/química , Peptídeos/química , Eletricidade Estática , Fosfatase Alcalina/metabolismo , Motivos de Aminoácidos , Animais , Biomarcadores/metabolismo , Osso e Ossos/patologia , Membrana Celular/metabolismo , Modelos Animais de Doenças , Regulação da Expressão Gênica , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Óxido Nítrico/biossíntese , Osteogênese , Ratos , Alicerces Teciduais/química
5.
Mol Neurobiol ; 54(6): 4609-4616, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-27389777

RESUMO

Tyrosine hydroxylase (TH), a rate-limiting step in catecholamine synthesis in which its activity influences Alzheimer disease, Parkinson disease, and IQ of schizophrenia patients, has been studied for a long time. In the meantime, the present investigation assessed the effect of noggin and type of self-assembling nanofibers in TH gene over-expression by neuron-like cells derived from human endometrial-derived stromal cells (hEnSCs). Neuroblastoma cells and hEnSCs encapsulated into nanofibers including Matrigel, (RADA)4, laminin, and BMHP-1 motif bounded to (RADA)4 and their cell viability were studied for 48 h and 18 days in basal and neurogenic media, respectively, in noggin-rich media. Then, expression of neural genes and proteins has been investigated by immunocytochemistry (ICC) and real-time PCR methods, respectively. The results indicated that neuroblastoma cell and hEnSC viability is in good agreement with the level of Bcl2 and ß-tubulin III gene expression; however, -BMHP-1 and -laminin nanofibers exhibited significantly higher cell viability eventually through Wnt/ß-catenin signaling pathway as compared to others, respectively. The gene expression analysis of nanofibers showed that none of them induced gamma-aminobutyric acid (GABA) gene expression while glial fibrillary acidic protein (GFAP) gene just over-expressed in cells encapsulated into Matrigel with a low level of Bcl2 gene expression. However, the TH gene just had been over-expressed in cells encapsulated into -laminin nanofiber and 2D cell culture. In the absence of noggin with -laminin nanofibers, TH gene expression was suppressed. It might be concluded that although noggin through anti-BMP pathways resulted in GFAP decrement and TH gene increment, the type of scaffold that defined the final fate of cells and -laminin accompaniment might be useful for the recovery of Alzheimer and Parkinson disease patients.


Assuntos
Proteínas de Transporte/farmacologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Laminina/química , Nanofibras/química , Peptídeos/farmacologia , Tirosina 3-Mono-Oxigenase/genética , Linhagem Celular Tumoral , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Endométrio/citologia , Feminino , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Células Estromais/citologia , Células Estromais/efeitos dos fármacos , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo , Ácido gama-Aminobutírico/metabolismo
6.
Artif Cells Nanomed Biotechnol ; 45(4): 817-823, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27160016

RESUMO

Recently, it has been disclosed that silver nanoparticles (AgNPs) have the potential to inhibit infection and cancerous cells and eventually penetrate through injected site into the capillary due to their small size. This study focuses on the effect of size and zeta potential of bare and citrate-coated AgNPs on human umbilical vein endothelial cells (HUVECs) as main capillary cells. AgNPs with high and low concentrations and no citrate coating were synthesized by using simple wet chemical method and named as AgNP/HC, AgNP/LC, and AgNP, respectively. Citrate coated particles showed larger zeta potential of -22 mV and AgNp/HC showed the smallest size of 13.2 nm. UV-Visible spectroscopy and dynamic light scattering (DLS) were performed to evaluate particle size and hydrodynamic diameter of NPs in water and cell culture media. Results indicated that higher concentrations of citrate decreased hydrodynamic diameter and NP agglomeration. reactive oxygen species (ROS) production of all AgNPs was similar at 28 ppm although it was significantly higher than control group. Their effects on cell membrane and chromosomal structure were studied using LDH measurement and 4',6-diamidino-2-phenylindole (DAPI) staining, as well. Results demonstrated that AgNP/LC was less toxic to cells owing to higher value of IC50, minimum inhibitory concentration (MIC), and less release of LDH. Cancerous (Human Caucasian neuroblastoma) and immortal cells (Mouse embryonic fibroblast cell line) were about twice more sensitive than HUVECs to toxic effects of AgNPs. DAPI staining results showed that AgNP and AgNP/HC induced highest and lowest breaking of chromosome. Overall results suggest that viability of HUVECs will be higher than 90% when viability of cancerous cells is 50% in AgNPs chemotherapy.


Assuntos
Cromatina/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/citologia , Nanopartículas Metálicas , Organelas/efeitos dos fármacos , Tamanho da Partícula , Prata/química , Prata/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Cromatina/metabolismo , Citratos/química , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , L-Lactato Desidrogenase/metabolismo , Organelas/metabolismo
7.
Mol Neurobiol ; 54(4): 2483-2496, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-26984600

RESUMO

Astroglial scaring and limited neurogenesis are two problematic issues in recovery of spinal cord injury (SCI). In the meantime, it seems that mechanical manipulations of scaffold to inhibit astroglial scarring and improve neurogenesis is worthy of value. In the present investigation, the effect of nanofiber (gel) concentration as a mechanical-stimuli in neurogenesis was investigated. Cell viability, membrane damage, and neural differentiation derived from endometrial stem cells encapsulated into self-assembling peptide nanofiber containing long motif of laminin were assessed. Then, two of their concentrations that had no significant difference of neural differentiation potential were selected for motor neuron investigation in SCI model of rat. MTT assay data showed that nanofibers at the concentrations of 0.125 and 0.25 % w/v induced higher and less cell viability than others, respectively, while cell viability derived from higher concentrations of 0.25 % w/v had ascending trend. Gene expression results showed that noggin along with laminin motif over-expressed TH gene and the absence of noggin or laminin motif did not in all concentrations. Bcl2 over-expression is concomitant with the decrease of nanofiber stiffness, NF+ cells increment, and astrogenesis inhibition and dark neuron decrement in SCI model. It seems that stiffness affects on Bcl2 gene expression and may through ß-Catenin/Wnt signaling pathway and BMP-4 inhibition decreases astrogenesis and improves neurogenesis. However, stiffness had a significant effect on upregulation of GFAP+ cells and motor neuron recovery in in vivo. It might be concluded that eventually there is a critical definitive point concentration that at less or higher than of it changes cell behavior and neural differentiation through different molecular pathways.


Assuntos
Laminina/química , Laminina/farmacologia , Mecanotransdução Celular/efeitos dos fármacos , Nanofibras/química , Neurogênese/efeitos dos fármacos , Peptídeos/farmacologia , Adulto , Motivos de Aminoácidos , Animais , Biomarcadores/metabolismo , Compostos de Bifenilo/química , Nucléolo Celular/efeitos dos fármacos , Nucléolo Celular/metabolismo , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Endométrio/citologia , Feminino , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Imuno-Histoquímica , Filamentos Intermediários/metabolismo , L-Lactato Desidrogenase/metabolismo , Masculino , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Picratos/química , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Células Estromais/citologia , Células Estromais/efeitos dos fármacos , Células Estromais/metabolismo , Regulação para Cima/efeitos dos fármacos
8.
Mol Neurobiol ; 53(8): 5288-99, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-26427854

RESUMO

Spinal cord injury (SCI) in humans stayed a ruining and healless disorder. Since longer laminin motif (CQAASIKVAV (CQIK)) better mimics conformation of native region in active site than isoleucine-lysine-valine-alanine-valine (IKVAV) and resulted in improved cellular response so, for the first time in this study, CQIK bounded with two glycines spacer and (RADA)4 as a self-assembling peptide nanofiber backbone (-CQIK) was used. The purpose of this study was to investigate the role of -CQIK in neural differentiation of human endometrial-derived stromal cells (hEnSCs) in vitro, tubulin polymerization ex vivo, and assess the supportive effect of this hydrogel in an animal model of chronic SCI. Results disclosed that proton concentration has direct effect on hEnSCs membrane damage but not on neuroblastoma cells. However, cell viability of neuroblastoma encapsulated into -CQIK was higher than hEnSCs at the concentration of 0.125 % v/w. Gene expression data confirmed neurogenesis, TH over-expression, and glial fibrillary acidic protein (GFAP) suppression eventually through α6 and ß1 integrin site. However, it revealed higher neurogenesis as compared to bone morrow homing peptides (BMHP). Although, Basso, Beattie, Bresnahan (BBB) score of chronic model of SCI in rat was higher than control and phosphate-buffered saline (PBS) group but significantly was less than BMHP group. However, -CQIK had induced neurite outgrowth and myelination and inhibited astrogliosis. Tubulin polymerization data using UV spectroscopy showed higher degree of polymerization. However, tubulin polymerization was dependent on nanofiber concentration. Based on our results, it might be concluded that peptidic nanofiber containing long motif of laminin holds great promise for spinal cord injury recovery with increment of neurogenesis and astrogliosis decrement.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Laminina/farmacologia , Nanofibras/química , Neurogênese/efeitos dos fármacos , Peptídeos/farmacologia , Polimerização , Tubulina (Proteína)/metabolismo , Adulto , Animais , Biomarcadores/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Endométrio/citologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Ratos Wistar , Ovinos , Células Estromais/citologia , Células Estromais/efeitos dos fármacos , Células Estromais/metabolismo
9.
J Biomed Mater Res A ; 102(12): 4590-7, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24532561

RESUMO

Spinal cord injury (SCI) in humans remains a devastating and incurable disorder. The use of Matrigel, a hydrogel-mimicking extracellular matrix, has been suggested as a scaffold for spinal cord regeneration. Human endometrial-derived stromal cells (hEnSCs) are abundant and available in adult stem cells with low immunological incompatibility, which could be considered for cell replacement therapy. The purpose of this study was to investigate the role of Matrigel in neural differentiation of hEnSCs in vitro and assess the supportive effects of this hydrogel in an animal model of SCI. hEnSCs were isolated and encapsulated into nanofibrous thermogel and cell viability and cell membrane damage were assessed. Encapsulated hEnSCs into Matrigel were treated with neural differentiation medium for 21 days, and then neural genes and protein markers were analyzed using real time-PCR and immunocytochemistry. Matrigel was implanted into rats with SCI and followed for 42 days using a behavioral test. Our study revealed a higher cell viability and neural differentiation in the level of genes and proteins as well as lower cell membrane damage. Substantial recoveries of motor function were observed in animals receiving the Matrigel treatment. The treatment with Matrigel, nanofibrous scaffold, produced beneficial effects on functional recovery following SCI in rats, possibly via assimilation to cytoskeleton fiber, high surface/volume ratio, spatial interconnectivity and containing some adhesive molecules and growth factors, enhancement of anti-inflammation, anti-astrogliosis, neuronal extension, and neuronal regeneration effects.


Assuntos
Células-Tronco Adultas/metabolismo , Diferenciação Celular , Endométrio/metabolismo , Neurônios Motores/metabolismo , Nanofibras/química , Adulto , Células-Tronco Adultas/citologia , Células-Tronco Adultas/transplante , Animais , Endométrio/citologia , Feminino , Xenoenxertos , Humanos , Masculino , Neurônios Motores/citologia , Ratos , Ratos Wistar , Traumatismos da Medula Espinal/terapia , Transplante de Células-Tronco , Células Estromais/citologia , Células Estromais/metabolismo
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