RESUMO
Photo-functionalization of titanium orthopedic/prosthetic implants using ultraviolet illumination is known to improve osteogenesis. Therefore, in this study, we aimed to examine the influence of vacuum ultraviolet (VUV)-treated titanium surfaces on osteoblast cell adhesion, activity, and differentiation. Osteoblastic cells were cultured on titanium substrates treated with various VUV treatment conditions (0, 6.2, 18.7, and 37.4 J/cm2) and their behavior was evaluated. The results revealed that cell adhesion was increased whereas cell activity and differentiation ability were decreased upon cell culture on VUV-treated substrates. In particular, cell activity and differentiation ability were dramatically suppressed with 18.7 J/cm2 VUV irradiation. Within the limitations of this cell-based experiment, we clarified the VUV treatment conditions in which cell adhesion was improved but cell activity and differentiation ability were suppressed. These results indicate that VUV-treatment can be used to influence cell growth properties and can be used to accelerate or suppress cell differentiation on implant substrates.
Assuntos
Diferenciação Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Osteogênese/efeitos dos fármacos , Titânio/farmacologia , Adesão Celular/efeitos dos fármacos , Adesão Celular/efeitos da radiação , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Humanos , Osteoblastos/efeitos dos fármacos , Osteoblastos/efeitos da radiação , Osteogênese/genética , Osteogênese/efeitos da radiação , Especificidade por Substrato , Propriedades de Superfície/efeitos dos fármacos , Propriedades de Superfície/efeitos da radiação , Raios Ultravioleta/efeitos adversos , VácuoRESUMO
Intensive adjuvant radiotherapy (RT) is a standard treatment for glioblastoma multiforme (GBM) patients; however, its effect on the normal brain tissue remains unclear. Here, we investigated the short-term effects of multiple irradiation on the cellular and extracellular glycosylated components of normal brain tissue and their functional significance. Triple irradiation (7 Gy*3 days) of C57Bl/6 mouse brain inhibited the viability, proliferation and biosynthetic activity of normal glial cells, resulting in a fast brain-zone-dependent deregulation of the expression of proteoglycans (PGs) (decorin, biglycan, versican, brevican and CD44). Complex time-point-specific (24-72 h) changes in decorin and brevican protein and chondroitin sulfate (CS) and heparan sulfate (HS) content suggested deterioration of the PGs glycosylation in irradiated brain tissue, while the transcriptional activity of HS-biosynthetic system remained unchanged. The primary glial cultures and organotypic slices from triple-irradiated brain tissue were more susceptible to GBM U87 cells' adhesion and proliferation in co-culture systems in vitro and ex vivo. In summary, multiple irradiation affects glycosylated components of normal brain extracellular matrix (ECM) through inhibition of the functional activity of normal glial cells. The changed content and pattern of PGs and GAGs in irradiated brain tissues are accompanied by the increased adhesion and proliferation of GBM cells, suggesting a novel molecular mechanism of negative side-effects of anti-GBM radiotherapy.
Assuntos
Neoplasias Encefálicas , Encéfalo , Proliferação de Células/efeitos da radiação , Raios gama , Glioblastoma , Neoplasias Experimentais , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/radioterapia , Adesão Celular/efeitos da radiação , Proteínas da Matriz Extracelular/metabolismo , Glioblastoma/metabolismo , Glioblastoma/patologia , Glioblastoma/radioterapia , Masculino , Camundongos , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Neoplasias Experimentais/radioterapia , Proteoglicanas/metabolismoRESUMO
Anti-inflammatory low-dose therapy is well established, whereas the immunomodulatory impact of doses below 0.1 Gy is much less clear. In this study, we investigated dose, dose rate and time-dependent effects in a dose range of 0.005 to 2 Gy on immune parameters after whole body irradiation (IR) using a pro-inflammatory (ApoE-/-) and a wild type mouse model. Long-term effects on spleen function (proliferation, monocyte expression) were analyzed 3 months, and short-term effects on immune plasma parameters (IL6, IL10, IL12p70, KC, MCP1, INFγ, TGFß, fibrinogen, sICAM, sVCAM, sE-selectin/CD62) were analyzed 1, 7 and 28 days after Co60 γ-irradiation (IR) at low dose rate (LDR, 0.001 Gy/day) and at high dose rate (HDR). In vitro measurements of murine monocyte (WEHI-274.1) adhesion and cytokine release (KC, MCP1, IL6, TGFß) after low-dose IR (150 kV X-ray unit) of murine endothelial cell (EC) lines (H5V, mlEND1, bEND3) supplement the data. RT-PCR revealed significant reduction of Ki67 and CD68 expression in the spleen of ApoE-/- mice after 0.025 to 2 Gy exposure at HDR, but only after 2 Gy at LDR. Plasma levels in wild type mice, showed non-linear time-dependent induction of proinflammatory cytokines and reduction of TGFß at doses as low as 0.005 Gy at both dose rates, whereas sICAM and fibrinogen levels changed in a dose rate-specific manner. In ApoE-/- mice, levels of sICAM increased and fibrinogen decreased at both dose rates, whereas TGFß increased mainly at HDR. Non-irradiated plasma samples revealed significant age-related enhancement of cytokines and adhesion molecules except for sICAM. In vitro data indicate that endothelial cells may contribute to systemic IR effects and confirm changes of adhesion properties suggested by altered sICAM plasma levels. The differential immunomodulatory effects shown here provide insights in inflammatory changes occurring at doses far below standard anti-inflammatory therapy and are of particular importance after diagnostic and chronic environmental exposures.
Assuntos
Apolipoproteínas E/deficiência , Inflamação/patologia , Radiação Ionizante , Envelhecimento/sangue , Animais , Adesão Celular/efeitos da radiação , Linhagem Celular , Citocinas/metabolismo , Relação Dose-Resposta à Radiação , Células Endoteliais/efeitos da radiação , Feminino , Inflamação/sangue , Interleucina-6/metabolismo , Camundongos Endogâmicos C57BL , Monócitos/efeitos da radiação , Baço/efeitos da radiação , Fatores de TempoRESUMO
Peri-implantitis is an unsolved but critical problem with dental implants. It is postulated that creating a seal of gingival soft tissue around the implant neck is key to preventing peri-implantitis. The objective of this study was to determine the effect of UV surface treatment of titanium disks on the adhesion strength and retention time of oral connective tissues as well as on the adherence of mucosal fibroblasts. Titanium disks with a smooth machined surface were prepared and treated with UV light for 15 min. Keratinized mucosal tissue sections (3 × 3 mm) from rat palates were incubated for 24 h on the titanium disks. The adhered tissue sections were then mechanically detached by agitating the culture dishes. The tissue sections remained adherent for significantly longer (15.5 h) on the UV-treated disks than on the untreated control disks (7.5 h). A total of 94% of the tissue sections were adherent for 5 h or longer on the UV-treated disks, whereas only 50% of the sections remained on the control disks for 5 h. The adhesion strength of the tissue sections to the titanium disks, as measured by tensile testing, was six times greater after UV treatment. In the culture studies, mucosal fibroblasts extracted from rat palates were attached to titanium disks by incubating for 24, 48, or 96 h. The number of attached cells was consistently 15-30% greater on the UV-treated disks than on the control disks. The cells were then subjected to mechanical or chemical (trypsinization) detachment. After mechanical detachment, the residual cell rates on the UV-treated surfaces after 24 and 48 h of incubation were 35% and 25% higher, respectively, than those on the control surfaces. The remaining rate after chemical detachment was 74% on the control surface and 88% on the UV-treated surface for the cells cultured for 48 h. These trends were also confirmed in mouse embryonic fibroblasts, with an intense expression of vinculin, a focal adhesion protein, on the UV-treated disks even after detachment. The UV-treated titanium was superhydrophilic, whereas the control titanium was hydrophobic. X-ray photoelectron spectroscopy (XPS) chemical analysis revealed that the amount of carbon at the surface was significantly reduced after UV treatment, while the amount of TiOH molecules was increased. These ex vivo and in vitro results indicate that the UV treatment of titanium increases the adhesion and retention of oral mucosa connective tissue as a result of increased resistance of constituent fibroblasts against exogenous detachment, both mechanically and chemically, as well as UV-induced physicochemical changes of the titanium surface.
Assuntos
Adesão Celular/efeitos da radiação , Tecido Conjuntivo/metabolismo , Fibroblastos/metabolismo , Mucosa Bucal/metabolismo , Titânio/metabolismo , Titânio/efeitos da radiação , Raios Ultravioleta , Animais , Carbono/metabolismo , Implantes Dentários , Adesões Focais/metabolismo , Gengiva/citologia , Gengiva/metabolismo , Masculino , Camundongos , Células NIH 3T3 , Espectroscopia Fotoeletrônica/métodos , Ratos , Ratos Sprague-Dawley , Propriedades de Superfície/efeitos da radiação , Resistência à Tração , Vinculina/metabolismoRESUMO
Although anti-cancer properties of the natural compound curcumin have been reported, low absorption and rapid metabolisation limit clinical use. The present study investigated whether irradiation with visible light may enhance the inhibitory effects of low-dosed curcumin on prostate cancer cell growth, proliferation, and metastasis in vitro. DU145 and PC3 cells were incubated with low-dosed curcumin (0.1-0.4 µg/mL) and subsequently irradiated with 1.65 J/cm2 visible light for 5 min. Controls remained untreated and/or non-irradiated. Cell growth, proliferation, apoptosis, adhesion, and chemotaxis were evaluated, as was cell cycle regulating protein expression (CDK, Cyclins), and integrins of the α- and ß-family. Curcumin or light alone did not cause any significant effects on tumor growth, proliferation, or metastasis. However, curcumin combined with light irradiation significantly suppressed tumor growth, adhesion, and migration. Phosphorylation of CDK1 decreased and expression of the counter-receptors cyclin A and B was diminished. Integrin α and ß subtypes were also reduced, compared to controls. Irradiation distinctly enhances the anti-tumor potential of curcumin in vitro and may hold promise in treating prostate cancer.
Assuntos
Curcumina/farmacologia , Luz , Neoplasias da Próstata/patologia , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Adesão Celular/efeitos dos fármacos , Adesão Celular/efeitos da radiação , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/efeitos da radiação , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Movimento Celular/efeitos da radiação , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Células Clonais , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Integrinas/metabolismo , Masculino , Metástase NeoplásicaRESUMO
Regulating stem cell functions by precisely controlling the nanoscale presentation of bioactive ligands has a substantial impact on tissue engineering and regenerative medicine but remains a major challenge. Here it is shown that bioactive ligands can become mechanically "invisible" by increasing their tether lengths to the substrate beyond a critical length, providing a way to regulate mechanotransduction without changing the biochemical conditions. Building on this finding, light switchable tethers are rationally designed, whose lengths can be modulated reversibly by switching a light-responsive protein, pdDronpa, in between monomer and dimer states. This allows the regulation of the adhesion, spreading, and differentiation of stem cells by light on substrates of well-defined biochemical and physical properties. Spatiotemporal regulation of differential cell fates on the same substrate is further demonstrated, which may represent an important step toward constructing complex organoids or mini tissues by spatially defining the mechanical cues of the cellular microenvironment with light.
Assuntos
Ligantes , Luz , Mecanotransdução Celular/fisiologia , Engenharia de Proteínas , Adesão Celular/efeitos da radiação , Diferenciação Celular/efeitos da radiação , Dimerização , Elastina/química , Elastina/metabolismo , Humanos , Integrinas/química , Integrinas/metabolismo , Proteínas Luminescentes/química , Proteínas Luminescentes/metabolismo , Mecanotransdução Celular/efeitos da radiação , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Microscopia de Força Atômica , Oligopeptídeos/química , Oligopeptídeos/metabolismoRESUMO
Astrocytes act as neural stem cells (NSCs) that have the potential to self-renew and differentiate into other neuronal cells. The protein expression of these astrocytes depends on the stage of differentiation, showing sequential expression of multiple proteins such as octamer-binding transcription factor 4 (Oct4), nestin, glial fibrillary acidic protein (GFAP), and aldehyde dehydrogenase 1 family member L1 (aldh1L1). Photobiomodulation (PBM) affects cell apoptosis, proliferation, migration, and adhesion. We hypothesized that astrocyte proliferation and differentiation would be modulated by PBM. We used an optimized astrocyte culture method and a 660-nanometer light-emitting diode (LED) to enhance the biological actions of many kinds of cells. We determined that the 660-nanometer LED promoted the biological actions of cultured astrocytes by increasing the reactive oxygen species levels. The overall viability of the cultured cells, which included various cells other than astrocytes, did not change after LED exposure; however, astrocyte-specific proliferation was observed by the increased co-expression of GFAP and bromodeoxyuridine (BrdU)/Ki67. Furthermore, the 660-nanometer LED provides evidence of differentiation, as shown by the decreased Oct4 and GFAP co-expression and increased nestin and aldh1L1 expression. These results demonstrate that a 660-nanometer LED can modify astrocyte proliferation, which suggests the efficacy of the therapeutic application of LED in various pathological states of the central nervous system.
Assuntos
Astrócitos/efeitos da radiação , Proliferação de Células/efeitos da radiação , Expressão Gênica/efeitos da radiação , Neurônios/efeitos da radiação , Animais , Apoptose/genética , Apoptose/efeitos da radiação , Astrócitos/citologia , Astrócitos/metabolismo , Encéfalo/citologia , Encéfalo/metabolismo , Adesão Celular/efeitos da radiação , Diferenciação Celular/efeitos da radiação , Movimento Celular/efeitos da radiação , Técnicas de Cocultura , Embrião de Mamíferos , Proteína Glial Fibrilar Ácida/genética , Proteína Glial Fibrilar Ácida/metabolismo , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Lasers Semicondutores , Luz , Nestina/genética , Nestina/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Fator 3 de Transcrição de Octâmero/genética , Fator 3 de Transcrição de Octâmero/metabolismo , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Cultura Primária de Células , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/agonistas , Espécies Reativas de Oxigênio/metabolismoRESUMO
The mammalian skin is essential to protect the organism from external damage while at the same time enabling communication with the environment. Aging compromises skin function and regeneration, which is further exacerbated by external influences, such as UVR from the sun. Aging and UVR are also major risk factors contributing to the development of skin cancer. Whereas aging research traditionally has focused on the role of DNA damage and metabolic and stress pathways, less is known about how aging affects tissue architecture and cell dynamics in skin homeostasis and regeneration and whether changes in these processes promote skin cancer. This review highlights how key regulators of cell polarity and adhesion affect epidermal mechanics, tissue architecture, and stem cell dynamics in skin aging and cancer.
Assuntos
Polaridade Celular/genética , Epiderme/patologia , Envelhecimento da Pele/genética , Neoplasias Cutâneas/patologia , Animais , Adesão Celular/genética , Adesão Celular/efeitos da radiação , Polaridade Celular/efeitos da radiação , Dano ao DNA/efeitos da radiação , Modelos Animais de Doenças , Epiderme/efeitos da radiação , Humanos , Camundongos , Regeneração/genética , Regeneração/efeitos da radiação , Envelhecimento da Pele/efeitos da radiação , Neoplasias Cutâneas/etiologia , Células-Tronco , Raios Ultravioleta/efeitos adversosRESUMO
This study examined the effects of gold nanoparticles (AuNPs) and/or ionizing radiation (IR) on the viability and motility of human primary colon epithelial (CCD841) and colorectal adenocarcinoma (SW48) cells as well as human primary epidermal melanocytes (HEM) and melanoma (MM418-C1) cells. AuNPs up to 4 mM had no effect on the viability of these cell lines. The viability of the cancer cells was ~60% following exposure to 5 Gy. Exposure to 5 Gy X-rays or 1 mM AuNPs showed the migration of the cancer cells ~85% that of untreated controls, while co-treatment with AuNPs and IR decreased migration to ~60%. In the non-cancerous cell lines gap closure was enhanced by ~15% following 1 mM AuNPs or 5 Gy treatment, while for co-treatment it was ~22% greater than that for the untreated controls. AuNPs had no effect on cell re-adhesion, while IR enhanced only the re-adhesion of the cancer cell lines but not their non-cancerous counterparts. The addition of AuNPs did not enhance cell adherence. This different reaction to AuNPs and IR in the cancer and normal cells can be attributed to radiation-induced adhesiveness and metabolic differences between tumour cells and their non-cancerous counterparts.
Assuntos
Movimento Celular/efeitos dos fármacos , Ouro/química , Nanopartículas Metálicas/uso terapêutico , Adesão Celular/efeitos dos fármacos , Adesão Celular/efeitos da radiação , Linhagem Celular Tumoral , Movimento Celular/efeitos da radiação , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/efeitos da radiação , Humanos , Melanócitos/efeitos dos fármacos , Melanócitos/efeitos da radiação , Radiação Ionizante , Raios XRESUMO
DUSP3 is a phosphatase expressed and active in several tissues that dephosphorylates tyrosine residues in many regulatory proteins of cellular activities such as proliferation, survival, and cell death. Recently, two new independent functions were assigned to this enzyme: dephosphorylation of focal adhesion kinase (FAK) and regulation of nucleotide-excision repair (NER) pathway. Genotoxic stress by UV radiation is known to affect cell morphology, adhesion, and migration for affecting, for example, the Rho GTPases that regulate actin cytoskeleton. This work investigated the intersection of DUSP3 function, XPA protein activity, and UV toxicity by examining cell migration, FAK, and SRC kinase phosphorylation status, in addition to cell morphology, in fibroblast cells proficient (MRC-5) or deficient (XPA) of the NER pathway. DUSP3 loss reduced cell migration of normal cells, which was stimulated by the genotoxic stress, effects evidenced in presence of serum mitogenic stimulus. However, NER-deficient cells migration response was the opposite since DUSP3 loss increased migration, especially after cells being exposed to UV stress. The levels of pFAK(Y397) peaked 15 min and 1 h after UV radiation in normal cells, but only slightly increased in repair-deficient cells. However, the DUSP3 knockdown strongly raised pFAK(Y397) levels in both cells, but especially in XPA cells as supported by the higher SRC activity. These effects impacted on the dynamics of actin-based structures formation, such as stress fibres, apparently dependent on DUSP3 and DNA-repair (NER) proficiency of the cells. Altogether our findings suggest this dual-phosphatase is bridging gaps between the complex regulation of cell morphology, motility, and genomic stability.
Assuntos
Movimento Celular/efeitos da radiação , Fosfatase 3 de Especificidade Dupla/metabolismo , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Raios Ultravioleta , Adesão Celular/efeitos da radiação , Linhagem Celular , Reparo do DNA/efeitos da radiação , Fosfatase 3 de Especificidade Dupla/antagonistas & inibidores , Fosfatase 3 de Especificidade Dupla/genética , Proteína-Tirosina Quinases de Adesão Focal/genética , Humanos , Fosforilação/efeitos da radiação , Interferência de RNA , RNA Interferente Pequeno/metabolismoRESUMO
Anti-inflammatory effects of low-dose irradiation often follow a non-linear dose-effect relationship. These characteristics were also described for the modulation of leukocyte adhesion to endothelial cells. Previous results further revealed a contribution of reactive oxygen species (ROS) and anti-oxidative factors to a reduced leukocyte adhesion. Here, we evaluated the expression of anti-oxidative enzymes and the transcription factor Nrf2 (Nuclear factor-erythroid-2-related factor 2), intracellular ROS content, and leukocyte adhesion in primary human microvascular endothelial cells (HMVEC) upon low-dose irradiation under physiological laminar shear stress or static conditions after irradiation with X-ray or Carbon (C)-ions (0-2 Gy). Laminar conditions contributed to increased mRNA expression of anti-oxidative factors and reduced ROS in HMVEC following a 0.1 Gy X-ray and 0.5 Gy C-ion exposure, corresponding to reduced leukocyte adhesion and expression of adhesion molecules. By contrast, mRNA expression of anti-oxidative markers and adhesion molecules, ROS, and leukocyte adhesion were not altered by irradiation under static conditions. In conclusion, irradiation of endothelial cells with low doses under physiological laminar conditions modulates the mRNA expression of key factors of the anti-oxidative system, the intracellular ROS contents of which contribute at least in part to leucocyte adhesion, dependent on the radiation source.
Assuntos
Células Endoteliais/citologia , Leucócitos/citologia , Microvasos/citologia , Espécies Reativas de Oxigênio/metabolismo , Carbono , Adesão Celular/efeitos da radiação , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Relação Dose-Resposta à Radiação , Células Endoteliais/efeitos da radiação , Regulação da Expressão Gênica/efeitos da radiação , Humanos , Leucócitos/efeitos da radiação , Modelos Biológicos , Fator 2 Relacionado a NF-E2/metabolismo , Oxirredução , Estresse Oxidativo/efeitos da radiação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Raios XRESUMO
Radiation-induced multiorgan dysfunction is thought to result primarily from damage to the endothelial system, leading to a systemic inflammatory response that is mediated by the recruitment of leukocytes. The Eph-ephrin signaling pathway in the vascular system participates in various disease developmental processes, including cancer and inflammation. In this study, we demonstrate that radiation exposure increased intestinal inflammation via endothelial dysfunction, caused by the radiation-induced activation of EphA2, an Eph receptor tyrosine kinase, and its ligand ephrinA1. Barrier dysfunction in endothelial and epithelial cells was aggravated by vascular endothelial-cadherin disruption and leukocyte adhesion in radiation-induced inflammation both in vitro and in vivo. Among all Eph receptors and their ligands, EphA2 and ephrinA1 were required for barrier destabilization and leukocyte adhesion. Knockdown of EphA2 in endothelial cells reduced radiation-induced endothelial dysfunction. Furthermore, pharmacological inhibition of EphA2-ephrinA1 by the tyrosine kinase inhibitor dasatinib attenuated the loss of vascular integrity and leukocyte adhesion in vitro. Mice administered dasatinib exhibited resistance to radiation injury characterized by reduced barrier leakage and decreased leukocyte infiltration into the intestine. Taken together, these data suggest that dasatinib therapy represents a potential approach for the protection of radiation-mediated intestinal damage by targeting the EphA2-ephrinA1 complex.
Assuntos
Dasatinibe/uso terapêutico , Intestinos/lesões , Intestinos/efeitos da radiação , Lesões por Radiação/tratamento farmacológico , Receptor EphA2/antagonistas & inibidores , Animais , Adesão Celular/efeitos dos fármacos , Adesão Celular/efeitos da radiação , Permeabilidade da Membrana Celular/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos da radiação , Dasatinibe/farmacologia , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/efeitos da radiação , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/patologia , Endotélio Vascular/efeitos da radiação , Efrina-A1/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos da radiação , Humanos , Intestinos/efeitos dos fármacos , Intestinos/patologia , Leucócitos/efeitos dos fármacos , Leucócitos/efeitos da radiação , Ligantes , Masculino , Camundongos Endogâmicos C57BL , Fosforilação/efeitos dos fármacos , Fosforilação/efeitos da radiação , Radiação Ionizante , Receptor EphA2/metabolismoRESUMO
Ultraviolet (UV) light and non-thermal plasma (NTP) are promising chair-side surface treatment methods to overcome the time-dependent aging of dental implant surfaces. After showing the efficiency of UV light and NTP treatment in restoring the biological activity of titanium and zirconia surfaces in vitro, the objective of this study was to define appropriate processing times for clinical use. Titanium and zirconia disks were treated by UV light and non-thermal oxygen plasma with increasing duration. Non-treated disks were set as controls. Murine osteoblast-like cells (MC3T3-E1) were seeded onto the treated or non-treated disks. After 2 and 24 h of incubation, the viability of cells on surfaces was assessed using an MTS assay. mRNA expression of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) were assessed using real-time reverse transcription polymerase chain reaction analysis. Cellular morphology and attachment were observed using confocal microscopy. The viability of MC3T3-E1 was significantly increased in 12 min UV-light treated and 1 min oxygen NTP treated groups. VEGF relative expression reached the highest levels on 12 min UV-light and 1 min NTP treated surfaces of both disks. The highest levels of HGF relative expression were reached on 12 min UV light treated zirconia surfaces. However, cells on 12 and 16 min UV-light and NTP treated surfaces of both materials had a more widely spread cytoskeleton compared to control groups. Twelve min UV-light and one min non-thermal oxygen plasma treatment on titanium and zirconia may be the favored times in terms of increasing the viability, mRNA expression of growth factors and cellular attachment in MC3T3-E1 cells.
Assuntos
Regulação da Expressão Gênica , Peptídeos e Proteínas de Sinalização Intercelular/biossíntese , Osteoblastos/metabolismo , Oxigênio/farmacologia , Gases em Plasma/farmacologia , RNA Mensageiro/sangue , Titânio/química , Raios Ultravioleta , Zircônio/química , Animais , Adesão Celular/efeitos dos fármacos , Adesão Celular/efeitos da radiação , Linhagem Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos da radiação , Camundongos , Osteoblastos/citologia , Propriedades de SuperfícieRESUMO
The self-assembly of different cell types into multicellular structures and their organization into spatiotemporally controlled patterns are both challenging and extremely powerful to understand how cells function within tissues and for bottom-up tissue engineering. Here, we not only independently control the self-assembly of two cell types into multicellular architectures with blue and red light, but also achieve their self-sorting into distinct assemblies. This required developing two cell types that form selective and homophilic cell-cell interactions either under blue or red light using photoswitchable proteins as artificial adhesion molecules. The interactions were individually triggerable with different colors of light, reversible in the dark, and provide noninvasive and temporal control over the cell-cell adhesions. In mixtures of the two cells, each cell type self-assembled independently upon orthogonal photoactivation, and cells sorted out into separate assemblies based on specific self-recognition. These self-sorted multicellular architectures provide us with a powerful tool for producing tissue-like structures from multiple cell types and investigate principles that govern them.
Assuntos
Adesão Celular/efeitos da radiação , Luz , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Linhagem Celular Tumoral , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Humanos , Cinética , Optogenética , Fotorreceptores Microbianos/genética , Fotorreceptores Microbianos/metabolismo , Proteínas Quinases/genética , Proteínas Quinases/metabolismoRESUMO
Zirconia has been regarded as a promising material for dental implants, and Nd:YAG laser treatment has been proposed as a potential strategy to improve its bioactivity. The main aim of the present study was to evaluate the in vitro behavior of human fetal osteoblasts in contact with laser-textured zirconia implant surfaces assessing the effect of different texture patterns, spacing between laser passes and number of laser passes. Zirconia discs were produced and treated with Nd:YAG laser according to test group variables: texture (microgrooves and micropillar array), distance between surface features (25 µm, 30 µm and 35 µm), and laser passes [1, 2, 4, and 8]. Untextured sandblasted and acid-etched zirconia discs (SBAE) were used as controls. Human osteoblasts (hFOB 1.19) were cultured for 14 days on test and control samples. Morphology and cellular adhesion were observed using scanning electron microscopy (SEM). Cell viability and proliferation were evaluated at 1, 3, 7, and 14 days using a commercial resazurin-based method. Collagen type I was evaluated at 3 days using ELISA. Alkaline phosphatase (ALP) activity was evaluated at 7 days using a colorimetric enzymatic technique. Group comparisons were tested using ANOVA or Mann-Whitney test (Tukey's post hoc) using statistical software, and significance was set at p < 0.05. Cell viability and proliferation increased over time for all groups with statistically higher values for laser-textured groups when compared with control at 7 and 14 days in culture (p < 0.05). Collagen type I levels were higher for study groups (p < 0.05) when compared with control group. No statistically differences were detected for ALP activity levels between texture and control groups (p > 0.05). The results suggest that laser-machined zirconia implant surfaces may benefit biological osteoblast response. However, the type of texture, spacing at the range of 25-35 µm, and number of laser passes did not seem to be relevant variables.
Assuntos
Lasers de Estado Sólido , Osteoblastos/efeitos da radiação , Próteses e Implantes , Zircônio/farmacologia , Adesão Celular/efeitos dos fármacos , Adesão Celular/efeitos da radiação , Forma Celular/efeitos dos fármacos , Forma Celular/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Células Cultivadas , Humanos , Osteoblastos/citologia , Osteoblastos/ultraestrutura , Propriedades de SuperfícieRESUMO
Increasing studies demonstrated that photobiomodulation (PBM) influenced specific biological effects in cells, tissues and organs, and these effects rely on the production of light irradiation. In this study, we aimed to precisely manipulate the spatial arrangement of adhesion cells in a traditional culture condition with 450 nm low intensity laser. Through 450 nm laser PBM, the adhesion of the cultured cells was significantly improved and resisted the digestion of 0.1% trypsin. Combined with a computer aided design system (CAD) and computer numerical control (CNC) system, the designed laser irradiation pattern induced the specific cell micropattern in the culture dish. RNA sequencing and biochemical experiments confirmed that the 450 nm laser prompted low-density lipoprotein (LDL) bonding to the cell surface and induced lipid peroxidation, which crosslinked and modified the protein molecules on the irradiated cell surface. In this way, the peroxidation product-modified proteins resisted trypsin proteolysis, ultimately leading to a differential detachment between the irradiated and non-irradiated cells under trypsin treatment. This convenient method did not require special biomaterial processing, has no impact on cell viability and functions, and required no changes to the conventional cell culture conditions. The photo-induced cell capturing is a great complement to existing tools by providing spatial resolution.
Assuntos
Terapia com Luz de Baixa Intensidade/métodos , Animais , Adesão Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Perfilação da Expressão Gênica , Peroxidação de Lipídeos/efeitos da radiação , Camundongos , Células NIH 3T3 , ProteóliseRESUMO
Two-dimensional (2D) molybdenum disulfide (MoS2) nanomaterials are an emerging class of biomaterials that are photoresponsive at near-infrared wavelengths (NIR). Here, we demonstrate the ability of 2D MoS2 to modulate cellular functions of human stem cells through photothermal mechanisms. The interaction of MoS2 and NIR stimulation of MoS2 with human stem cells is investigated using whole-transcriptome sequencing (RNA-seq). Global gene expression profile of stem cells reveals significant influence of MoS2 and NIR stimulation of MoS2 on integrins, cellular migration, and wound healing. The combination of MoS2 and NIR light may provide new approaches to regulate and direct these cellular functions for the purposes of regenerative medicine as well as cancer therapy.
Assuntos
Dissulfetos/efeitos da radiação , Células-Tronco Mesenquimais/efeitos da radiação , Molibdênio/efeitos da radiação , Nanoestruturas/efeitos da radiação , Adesão Celular/efeitos da radiação , Movimento Celular/efeitos da radiação , Sobrevivência Celular , Dissulfetos/química , Dissulfetos/metabolismo , Perfilação da Expressão Gênica , Humanos , Raios Infravermelhos , Integrinas/genética , Integrinas/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Molibdênio/química , Molibdênio/metabolismo , Nanoestruturas/química , Fármacos Fotossensibilizantes , Transdução de Sinais/efeitos da radiaçãoRESUMO
Spatiotemporal control of integrin-mediated cell adhesions to extracellular matrix regulates cell behavior with has numerous implications for biotechnological applications. In this work, two approaches for regulating cell adhesions in space and time with high precision are reported, both of which utilize green light. In the first design, CarH, which is a tetramer in the dark, is used to mask cRGD adhesion-peptides on a surface. Upon green light illumination, the CarH tetramer dissociates into its monomers, revealing the adhesion peptide so that cells can adhere. In the second design, the RGD motif is incorporated into the CarH protein tetramer such that cells can adhere to surfaces functionalized with this protein. The cell adhesions can be disrupted with green light, due to the disassembly of the CarH-RGD protein. Both designs allow for photoregulation with noninvasive visible light and open new possibilities to investigate the dynamical regulation of cell adhesions in cell biology.
Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/efeitos da radiação , Adesão Celular/efeitos da radiação , Luz , Oligopeptídeos/metabolismo , Proteínas de Bactérias/química , Integrinas/química , Integrinas/metabolismo , Análise Espaço-Temporal , Thermus thermophilusRESUMO
BACKGROUND: Changes in the cellular behavior depend on environmental and intracellular interactions. Cancer treatments force the changes, first on the molecular level, but the main visible changes are macroscopic. During radiotherapy, cancer cell's adhesion, proliferation and migration should be well monitored. In over 60% of diagnosed cancers cases, patients are given treatments with different protocols of radiotherapy, which result in possible metastasis and acute whole body response to toxic radiation. OBJECTIVE: Effectiveness of the therapy used depends on the sensitivity/resistance of irradiated cancer cells. Cellular mechanisms of cancer protection, such as the activation of DNA damage and repair pathways, antioxidants production and oxidative stress suppression during treatments are not desirable. Cancer cells monitoring require the development of novel techniques, and the best techniques are non-invasive and long-term live observation methods, which are shown in this study. METHODS: In cancers, invasive and metastatic phenotypes could be enhanced by stimulation of proliferation rate, decreased adhesion with simultaneous increase of motility and migration potential. For such reasons, the Ionizing Radiation (IR) stimulated proliferation; migration with lowered adhesiveness of cancer Me45 and normal fibroblasts NHDF were studied. Using impedance measurements technique for live cells, the adhesion of cells after IR exposition was assessed. Additionally proliferation and migration potential, based on standard Wound Healing assay were evaluated by timelapse microscopic observations. RESULTS: We found simulative IR dose-ranges (0.2-2 Gy) for Me45 and NHDF cells, with higher proliferation and adhesion rates. On the other hand, lethal impact of IR (10-12 Gy) on both the cell lines was indicated. CONCLUSION: Over-confluence cell populations, characterized with high crowd and contact inhibition could modulate invasiveness of individual cells, convert them to display migration phenotype and advance motility, especially after radiotherapy treatments.
Assuntos
Adesão Celular/efeitos da radiação , Movimento Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Impedância Elétrica , Radiação Ionizante , Linhagem Celular Tumoral , Proliferação de Células/genética , Fibroblastos/citologia , Fibroblastos/efeitos da radiação , Humanos , Melanoma/patologia , Microscopia de Contraste de Fase , Tolerância a Radiação , Imagem com Lapso de TempoRESUMO
Cross-linked albumin films, which are non-cell adhesive, were altered to be cell-adhesive by the combination of varied concentrations of fluorescein isothiocyanate and blue light irradiation. In this system, cell patterning was developed with two different cell lines by sequential seeding.Abbreviations: BSA: bovine serum albumin; EGDE: ethylene glycol diglycidyl ether; PBS: Dulbecco's phosphate buffered saline.