RESUMO
A dome-shaped elastic poly(l-lactide-co-caprolactone) (PLCL) scaffold with a channel and pore structure was fabricated by a combinative method of 3D printing technology and the gel pressing method (13 mm in diameter and 6.5 mm in thickness) for patient-specific regeneration. The PLCL scaffold was combined with adipose decellularized extracellular matrix (adECM) and heart decellularized extracellular matrix (hdECM) hydrogels and human adipose-derived stem cells (hADSCs) to promote adipogenesis and angiogenesis. These scaffolds had mechanical properties similar to those of native adipose tissue for improved tissue regeneration. The results of the in vitro real-time PCR showed that the dECM hydrogel mixture induces adipogenesis. In addition, the in vivo study at 12 weeks demonstrated that the tissue-engineered PLCL scaffolds containing the hydrogel mixture (hdECM/adECM (80:20)) and hADSCs promoted angiogenesis and adipose tissue formation, and suppressed apoptosis. Therefore, we expect that our constructs will be clinically applicable as material for the regeneration of patient-specific large-sized adipose tissue.
Assuntos
Adipogenia/efeitos dos fármacos , Tecido Adiposo/crescimento & desenvolvimento , Neovascularização Fisiológica/efeitos dos fármacos , Regeneração/genética , Tecido Adiposo/transplante , Animais , Apoptose/efeitos dos fármacos , Matriz Extracelular Descelularizada/farmacologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Humanos , Hidrogéis/química , Hidrogéis/farmacologia , Células-Tronco Mesenquimais/citologia , Camundongos , Miocárdio/citologia , Miocárdio/metabolismo , Neovascularização Fisiológica/genética , Poliésteres/farmacologia , Impressão Tridimensional , Regeneração/efeitos dos fármacosRESUMO
Signal transducer and activator of transcription 3 (STAT3) protein signaling is crucial for the survival, invasion, and growth of human cancer cells; thus, STAT3 protein is an ideal target for a new drug screening system. Herein, we developed a label-free sensor for anticancer drug-discovery based on the localized surface plasmon resonance (LSPR) shift response by tracking of STAT3 signaling including phosphorylation and dimerization. This enables ultrasensitive monitoring of the molecular interactions that occur on the surface of single gold nanoparticles. The red shift of the LSPR λmax was observed as 3.46 nm and 9.00 nm, respectively, indicating phosphorylation and dimerization of the STAT3 signaling pathway. In screening of anticancer candidates, the system worked well in the presence of STA-21 which inhibits STAT3 dimerization. The LSPR λmax shift in the inhibition condition is three times lower than that in the absence of an inhibitor. Interestingly, the system reveals high specificity, reproducibility and compatibility with real samples (MCF-7 cell line). Therefore, these results demonstrated that this system has strong potential to be an accurate and effective sensor for tracking of signaling pathways and drug screening of anticancer candidates for anticancer therapy.
Assuntos
Antineoplásicos/farmacologia , Descoberta de Drogas/métodos , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Ressonância de Plasmônio de Superfície/métodos , Humanos , Células MCF-7 , Fosforilação/efeitos dos fármacos , Compostos Policíclicos/farmacologia , Multimerização Proteica/efeitos dos fármacos , Estrutura Quaternária de Proteína , Fator de Transcrição STAT3/químicaRESUMO
Three-dimensional (3D) bioprinter including screw extruder was developed, and the polycaprolactone (PCL) grafts fabricated by screw-type and pneumatic pressure-type bioprinters were comparatively evaluated. The density and tensile strength of the single layers printed by the screw-type were 14.07% and 34.76% higher, respectively, than those of the single layers produced by the pneumatic pressure-type. The adhesive force, tensile strength, and bending strength of the PCL grafts printed by the screw-type bioprinter were 2.72 times, 29.89%, and 67.76% higher, respectively, than those of the PCL grafts prepared by the pneumatic pressure-type bioprinter. By evaluating the consistency with the original image of the PCL grafts, we found that it had a value of about 98.35%. The layer width of the printing structure was 485.2 ± 0.004919 µm, which was 99.5% to 101.8% compared to the set value (500 µm), indicating high accuracy and uniformity. The printed graft had no cytotoxicity, and there were no impurities in the extract test. In the in vivo studies, the tensile strength of the sample 12 months after implantation was reduced by 50.37% and 85.43% compared to the initial point of the sample printed by the screw-type and the pneumatic pressure-type, respectively. Through observing the fractures of the samples at 9- and 12-month samples, we found that the PCL grafts prepared by the screw-type had better in vivo stability. Therefore, the printing system developed in this study can be used as a treatment for regenerative medicine.
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Currently, dermal fillers are largely based on commercialized cross-linked hyaluronic acid (HA) injections, which require a large injection force. Additionally, HA can be easily decomposed by enzymes, and HA-treated tissues present a risk of developing granuloma. In this study, a chitosan-based dermal filler is presented that operates on a liquid-to-gel transition and allows the injection force to be kept ≈4.7 times lower than that required for HA injections. Evaluation of the physical properties of the chitosan filler indicates high viscoelasticity and recovery rate after gelation at 37 °C. Furthermore, in an in vivo evaluation, the liquid injection-type chitosan filler transitions to a gel state within 5 min after injection into the body, and exhibits a compressive strength that is ≈2.4 times higher than that of cross-linked HA. The filler also exhibits higher moldability and maintains a constant volume in the skin for a longer time than the commercial HA filler. Therefore, it is expected that the chitosan filler will be clinically applicable as a novel material for dermal tissue restoration and supplementation.
Assuntos
Quitosana , Preenchedores Dérmicos , Materiais Biocompatíveis , Elasticidade , Ácido HialurônicoRESUMO
BACKGROUND: Critical bone defects remain challenges for clinicians, which cannot heal spontaneously and require medical intervention. Following the development of three-dimensional (3D) printing technology is widely used in bone tissue engineering for its outstanding customizability. The 3D printed scaffolds were usually accompanied with growth factors, such as bone morphometric protein 2 (BMP-2), whose effects have been widely investigated on bone regeneration. We previously fabricated and investigated the effect of a polylactic acid (PLA) cage/Biogel scaffold as a carrier of BMP-2. In this study, we furtherly investigated the effect of another shape of PLA cage/Biogel scaffold as a carrier of BMP-2 in a rat calvaria defect model and an ectopic ossification (EO) model. METHOD: The PLA scaffold was printed with a basic commercial 3D printer, and the PLA scaffold was combined with gelatin and alginate-based Biogel and BMP-2 to induce bone regeneration. The experimental groups were divided into PLA scaffold, PLA scaffold with Biogel, PLA scaffold filled with BMP-2, and PLA scaffold with Biogel and BMP-2 and were tested both in vitro and in vivo. One-way ANOVA with Bonferroni post-hoc analysis was used to determine whether statistically significant difference exists between groups. RESULT: The in vitro results showed the cage/Biogel scaffold released BMP-2 with an initial burst release and followed by a sustained slow-release pattern. The released BMP-2 maintained its osteoinductivity for at least 14 days. The in vivo results showed the cage/Biogel/BMP-2 group had the highest bone regeneration in the rat calvarial defect model and EO model. Especially, the bone regenerated more regularly in the EO model at the implanted sites, which indicated the cage/Biogel had an outstanding ability to control the shape of regenerated bone. CONCLUSION: In conclusion, the 3D printed PLA cage/Biogel scaffold system was proved to be a proper carrier for BMP-2 that induced significant bone regeneration and induced bone formation following the designed shape.
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BACKGROUND: Three-dimensional (3D) in vitro cultures recapitulate the physiological microenvironment and exhibit high concordance with in vivo conditions. Improving co-culture models with different kind of cell types cultured on a 3D scaffold can closely mimic the in vivo environment. In this study, we examined the osteogenic response of pre-osteoblast MC3T3-E1 cells and Raw264.7 mouse monocytes in a 3D-encapsulated co-culture environment composed of the Cellrix® 3D culture system, which provides a physiologically relevant environment. METHODS: The Cellrix® 3D Bio-Gel scaffolds were used to individually culture or co-culture two type cells in 3D microenvironment. Under 3D culture conditions, osteoblastic behavior was evaluated with an ALP assay and staining. ACP assay and TRAP staining were used as osteoclastic behavior indicator. RESULTS: Treatment with osteoblastic induction factors (+3F) and RANKL had on positively effect on alkaline phosphatase activity but significantly inhibited to acid phosphatase activity during osteoclastic differentiation in 3D co-culture. Interestingly, alkaline phosphatase activity or acid phosphatase activity in 3D co-culture was stimulated with opposite differentiation factors at an early stage of differentiation. We guess that these effects may be related to RANK-RANKL signaling, which is important in osteoblast regulation of osteoclasts. CONCLUSION: In this study, the osteogenic response of 3D encapsulated pre-osteoblast MC3T3-E1 cells and mouse monocyte Raw264.7 cells was successfully demonstrated. Our 3D culture conditions will be able to provide a foundation for developing a high-throughput in vitro bone model to study the effects of various drugs and other agents on molecular pathways.
Assuntos
Osteoblastos , Osteogênese , Animais , Diferenciação Celular , Técnicas de Cocultura , Camundongos , OsteoclastosRESUMO
It has been known that single-strand DNA wraps around a single-walled carbon nanotube (SWNT) by pi-stacking. In this paper it is demonstrated that such DNA is dissociated from the SWNT by Watson-Crick base-pairing with a complementary sequence. Measurement of field effect transistor characteristics indicates a shift of the electrical properties as a result of this "unwrapping" event. We further confirm the suggested process through Raman spectroscopy and gel electrophoresis. Experimental results are verified in view of atomistic mechanisms with molecular dynamics simulations and binding energy analyses.
Assuntos
Pareamento de Bases , DNA de Cadeia Simples/química , Nanotubos de Carbono/química , Elétrons , Modelos Moleculares , Transistores EletrônicosRESUMO
A field effect transistor (FET) measurement of a single-walled carbon nanotube (SWNT) shows a transition from a metallic one to a p-type semiconductor after helical wrapping of DNA. Water is found to be critical to activate this metal-semiconductor transition in the ssDNA-SWNT hybrid. Raman spectroscopy confirms the same change in electrical behaviors. According to our ab initio calculations, a band gap can open up in a metallic SWNT with wrapped ssDNA in the presence of water molecules due to charge transfer.
Assuntos
DNA de Cadeia Simples/química , Metais/química , Nanotubos de Carbono , Semicondutores , Transferência de Energia , Fluorescência , Microscopia Eletrônica de Varredura , Análise Espectral RamanRESUMO
3D printing technology has various advantages, and the incorporation of bioactive substances into the 3D printed scaffold provides the biological and architectural characteristics of the scaffolds, which is very important for obtaining a good osseointegration effect. In this relation, this study prepared a novel porous hollow cage poly(lactic acid) (PLA) 3D printed scaffold and combined recombinant human bone morphogenetic protein-2 (rhBMP-2) and/or mesenchymal stem cells (MSCs) with Biogel composed of gelatin and alginate. Then, the scaffolds were used to evaluate the resulting bone regeneration through both in vitro and in vivo tests. The experimental group was divided into four groups as follows: only PLA scaffold (PLA); PLA scaffold filled with BMP-2 loaded on Biogel (P-BG-B2); PLA scaffold filled with MSCs encapsulated Biogel (P-BG-M); PLA scaffold filled with both BMP-2 and MSCs loaded on Biogel (P-BG-B2-M). Then in vitro results showed that the PLA-Biogel-based scaffold increased cell proliferation, and the P-BG-B2-M group showed a higher alkaline phosphatase activity and bone-related gene expression than was seen with the P-BG-M group at all the time points. It was shown that four weeks post-operative micro-CT analysis showed that within the defect site the P-BG-B2 group had a significantly higher percent bone volume (BV/TV) than the PLA group and P-BG-M group. And, out of the defect site, the P-BG-B2-M group BV/TV was shown significantly higher than the PLA group (p < 0.05). Histologically, defects in the P-BG-B2-M group showed a homogeneous new bone distribution, however the P-BG-B2 group and P-BG-M group presented a notably higher bone formation in the internal region than in the proximal region of the bone defect site. In conclusion, the 3D PLA-Biogel-based scaffold adapted rhBMP-2 and MSCs with carrier PLA showed good biocompatibility and high possibility as an effective and satisfactory bone graft material.
Assuntos
Proteína Morfogenética Óssea 2/administração & dosagem , Regeneração Óssea , Células-Tronco Mesenquimais/citologia , Alicerces Teciduais/química , Fator de Crescimento Transformador beta/administração & dosagem , Animais , Materiais Biocompatíveis/química , Regeneração Óssea/efeitos dos fármacos , Regeneração Óssea/genética , Regeneração Óssea/fisiologia , Proliferação de Células , Células Cultivadas , Géis , Humanos , Técnicas In Vitro , Masculino , Teste de Materiais , Células-Tronco Mesenquimais/fisiologia , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Osteogênese/fisiologia , Poliésteres/química , Porosidade , Impressão Tridimensional , Coelhos , Proteínas Recombinantes/administração & dosagem , Tíbia/efeitos dos fármacos , Tíbia/lesões , Tíbia/fisiologia , Engenharia Tecidual/métodos , Microtomografia por Raio-XRESUMO
We present a thin membrane transducer (TMT) that can detect nucleic acid based biomolecular reactions including DNA hybridization and protein recognition by aptamers. Specific molecular interactions on an extremely thin and flexible membrane surface cause the deflection of the membrane due to surface stress change which can be measured by a compact capacitive circuit. A gold-coated thin PDMS membrane assembled with metal patterned glass substrate is used to realize the capacitive detection. It is demonstrated that perfect match and mismatch hybridizations can be sharply discriminated with a 16-mer DNA oligonucleotide immobilized on the gold-coated surface. While the mismatched sample caused little capacitance change, the perfectly matched sample caused a well-defined capacitance decrease vs. time due to an upward deformation of the membrane by a compressive surface stress. Additionally, the TMT demonstrated the single nucleotide polymorphism (SNP) capabilities which enabled a detection of mismatching base pairs in the middle of the sequence. It is intriguing that the increase of capacitance, therefore a downward deflection due to tensile stress, was observed with the internal double mismatch hybridization. We further present the detection of thrombin protein through ligand-receptor type recognition with 15-mer thrombin aptamer as a receptor. Key aspects of this detection such as the effect of concentration variation are investigated. This capacitive thin membrane transducer presents a completely new approach for detecting biomolecular reactions with high sensitivity and specificity without molecular labelling and optical measurement.
Assuntos
Aptâmeros de Nucleotídeos/análise , Técnicas Biossensoriais/métodos , DNA/análise , Membranas Artificiais , Proteínas/análise , Trombina/análise , Aptâmeros de Nucleotídeos/química , Pareamento Incorreto de Bases/genética , Pareamento Incorreto de Bases/fisiologia , Sequência de Bases , Técnicas Biossensoriais/instrumentação , DNA/química , Vidro/química , Ouro/química , Hibridização de Ácido Nucleico , Polimorfismo de Nucleotídeo Único , Proteínas/química , Trombina/química , Fatores de Tempo , TransdutoresRESUMO
A new bacterial strain isolated from activated sludge, identified as Pseudomonas aeruginosa EMS1, produced a biosurfactant when grown on acidified soybean oil as the sole carbon source. An optimum biosurfactant production of 5 g/L was obtained with the following medium composition: 2% acidified soybean oil, 0.3% NH4NO3, 0.03% KH2PO4, 0.03% K2HPO4, 0.02% MgSO4.7H2O and 0.025% CaCl2.2H2O, with shaking at 200 rpm for an incubation period of 100 h at 30 degrees C. The production of the biosurfactant was found to be a function of cell growth, with maximum production occurring during the exponential phase. Hemolysis of erythrocytes and thin-layer chromatography studies revealed that the secreted biosurfactant was rhamnolipid. To overcome the complex environmental regulation with respect to rhamnolipid biosynthesis, and to replace the opportunistic pathogen P. aeruginosa with a safe industrial strain, attempts were made to achieve rhamnolipid production in a heterologous host, Pseudomonas putida, using molecular cloning of rhlAB rhamnosyltransferase genes with the rhlRI quorum sensing system, assuming that a functional rhamnosyltransferase would catalyze the formation of rhamnosyl-6-hydroxydecanoyl-6-hydroxydecanoate (mono-rhamnolipid) in P. putida. It was shown that rhamnolipid can be produced in the heterologous strain, P. putida, when provided with the rhamnosyltransferase genes.
Assuntos
Pseudomonas aeruginosa/metabolismo , Pseudomonas putida/metabolismo , Tensoativos/metabolismo , Sequência de Bases , Primers do DNA , Tensão SuperficialRESUMO
BACKGROUND: Recent evidence from in vitro and in vivo studies indicates that bisphosphonates may promote osteoblastic bone formation and potently inhibit osteoclast activity. However, little is known about the potential effect of bisphosphonates on the recruitment of osteoblastic precursors from patient-derived bone marrow stromal cells due to difficulties in accessing human bone marrow from healthy and disease subjects. METHODS: In this study, we evaluated the potential of using FDA-approved and clinically utilized bisphosphonates such as alendronate, ibandronate, and zoledronate to enhance the development of bone forming osteoblasts from osteoporosis patient- and healthy-person derived hBMSCs (op-MSCs and hp-MSCs, respectively). hBMSCs were obtained from postmenopausal women without endocrine diseases or receiving hormone replacement therapy. Cells were treated with or without a bisphosphonate (alendronate, ibandronate, and zoledronate) and analyzed over 21 days of culture. RESULTS: hBMSC from osteoporosis-patient with bisphosphonates treatment demonstrated a significant increase in Alizarin red staining after 7 days compared to that from healthy-person. Calcium contents and alkaline phosphatase (ALP) enzyme activity also demonstrated an increased propensity in hMSCs from osteoporosis-patient compared to those from healthy-person, although there were inter-individual variations. Gene expression levels varied among different donors. There were no significant differences in the effect on the osteoblastic differentiation of hBMSCs among alendronate, ibandronate, and zoledronate. Statistical significance in the osteoblastic differentiation of hBMSCs between the positive control group cultured in osteogenic medium alone and groups cultured in osteogenic medium supplemented with bisphosphonate was not shown either. These results might be due to various cell types of hBMSCs from individual clinical patients and concentrations of bisphosphonate used. CONCLUSION: Our study using a clinically relevant in vitro model suggests that bisphosphonate treatment is more effective for patients with osteoporosis than its preventive effect for healthy person. In addition, patient-specific responses to bisphosphonates should be considered rather than bisphosphonate type prior to prescription. Further investigations are needed to determine how bisphosphonates influence hBMSCs function to mediate bone quality and turnover in osteoporotic patients. Such studies can generate novel approaches to treat age-related osteoporotic bone loss.
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This paper presents a hybrid micropump actuated by the up-down motion of a dome shaped cell-polymer membrane composite. The contractile force induced from self-beating cardiomyocytes cultured on the membrane causes shrinkage and relaxation of a microchamber, leading to a flow in a microchannel. Flow direction is controlled by the geometry of diffuser/nozzle in the microchannel. The fabrication process is noninvasive to cells, thus, cardiomyocytes can robustly maintain their activity for a long time. The fluid motion in the microchannel was monitored by tracking 2 microm polystyrene beads. A net flow rate of 0.226 nl min(-1) was obtained in our microscale device. Our device demonstrates a unique performance of a cell-microdevice hybrid lab-on-a-chip that does not require any external power source, preventing electrical or heat shock to analytes.
Assuntos
Biomimética/instrumentação , Técnicas de Cultura de Células/instrumentação , Dimetilpolisiloxanos/química , Técnicas Analíticas Microfluídicas/instrumentação , Contração Miocárdica/fisiologia , Miócitos Cardíacos/fisiologia , Polímeros/química , Silicones/química , Animais , Animais Recém-Nascidos , Biomimética/métodos , Técnicas de Cultura de Células/métodos , Células Cultivadas , Desenho de Equipamento , Análise de Falha de Equipamento , Técnicas Analíticas Microfluídicas/métodos , Miócitos Cardíacos/citologia , Pressão , Ratos , Ratos Sprague-Dawley , Estresse MecânicoRESUMO
Carbon nanotubes (CNTs) have emerged as a leading nanomaterial for biomedical applications because of their extraordinary properties, which make them useful as delivery vehicles for drugs, proteins, and DNA into cells. However, the numerous applications of carbon nanotubes inevitably increase the potential risk of this nanomaterial. To address this issue, it is necessary to develop protocols for the effective and safe degradation of CNTs. In this study, we demonstrate a self-degradation route for single-wall carbon nanotubes mediated by the built-in peroxidase-like activity of bacterial magnetic nanoparticles (BMPs). Biocompatible BMPs which originated from Magnetospirillum sp. AMB-1 were directly conjugated through covalent bonding to functionalized SWNTs (f-SWNTs) without any additional functionalization processes. This SWNT-BMP hybrid was proven to exhibit highly synergetic peroxidase-like activity, and BMPs act as a highly effective intrinsic peroxidase for the self-degradation of BMP-decorated SWNTs. Moreover, it was shown to be an inhibitor that reduces the formation of ß-amyloid (Aß) fibrils, which are considered a key element in Alzheimer's disease. Thereby the SWNT-BMP hybrid exerts neuroprotective effects against ß-amyloid (Aß) fibrillation-induced neurotoxicity in SH-SY5Y human neuroblastoma cells. These results suggest that the SWNT-BMP hybrid could offer a new approach for treating or preventing neurodegenerative diseases.
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To develop environment-friendly biofertilizer solubilizing insoluble phosphates, salt- and pH-tolerant, insoluble inorganic phosphate-solubilizing bacterium was isolated from soybean rhizosphere. On the basis of its physiological characteristics and Vitek analysis, this bacterium was identified as Pantoea agglomerans. The optimal medium composition and cultural conditions for the solubilization of insoluble phosphate by P. agglomerans R-42 were 3% (w/v) of glucose, 0.1% (w/v) of NH4NO3, 0.02% (w/v) of MgSO4 x 7H2O, and 0.06% (w/v) of CaCl2 x 2H2O along with initial pH 7.5 at 30 degree C. The soluble phosphate production under optimal condition was around 900 mg/l, which was approximately 4.6-fold higher than the yield in the MPVK medium. The solubilization of insoluble phosphate was associated with a drop in the pH of the culture medium. P. agglomerans R-42 showed resistance against different environmental stresses like 5-45 degrees C temperature, 1-5% salt concentration and 3-11 pH range. Insoluble phosphate solubilization was highest from CaHPO4 (1367 mg/l), hydroxyapatite (1357 mg/l) and Ca3(PO4)2 (1312 mg/l). However, the strain produced soluble phosphate to the culture broth with the concentrations of 28 mg/l against FePO4, and 19 mg/l against AlPO4, respectively.
Assuntos
Glycine max/microbiologia , Pantoea/isolamento & purificação , Pantoea/metabolismo , Fosfatos/metabolismo , Raízes de Plantas/microbiologia , Cloreto de Sódio/farmacologia , Concentração de Íons de Hidrogênio , Pantoea/química , Pantoea/efeitos dos fármacos , SolubilidadeRESUMO
Long-range chromosomal travel is a phenomenon unique to cell division. Methods for non-invasive, artificial manipulation of chromosomes, such as optical or magnetic tweezers, have difficulty in producing the motion of whole chromosomes in live cells. Here, we report the spatial control of chromosomes over 10 µm in a live mouse oocyte using magnetic particles driven by an external magnetic field. Selective capture of the chromosomes was achieved using antibodies specific for histone H1 in the chromosome that were conjugated to magnetic particles (H1-BMPs). When an external magnetic field was applied, the chromosomes captured by the H1-BMPs traveled through the cytosol and accumulated near the cell membrane though the movement of the chromosomes captured by H1-BMPs was strongly disturbed by the distribution of the cytoskeleton (e.g. actin filaments). Being non-invasive in nature, our approach will enable new opportunities in the remote manipulation of subcellular elements.
Assuntos
Cromossomos de Mamíferos/metabolismo , Campos Magnéticos , Nanopartículas/química , Oócitos/metabolismo , Animais , Feminino , Histonas/metabolismo , Camundongos , Oócitos/citologiaRESUMO
An ideally designed scaffold for tissue engineering must be able to provide an environment that recapitulates the physiological conditions to control stem cell function. Here, we compared vertically aligned single-crystal apatite nanowires sheathed in graphitic layers (SANGs) with single-crystal apatite nanowires (SANs), which had the same geometric properties as--but differing nanotopographic surface chemistry than--SANGs, in order to evaluate the effect of the graphitic layer on the behavior of human mesenchymal stem cells (hMSCs). The difference in nanotopographic surface chemistry did not affect hMSC adhesion, growth, or morphology. However, hMSCs were more effectively differentiated into bone cells on SANGs through interaction with graphitic layers, which later degraded and thereby allowed the cells to continue differentiation on the bare apatite nanowires. Thus, SANGs provide an excellent microenvironment for the osteogenic differentiation of hMCS.
Assuntos
Apatitas/química , Técnicas de Cultura de Células/instrumentação , Células-Tronco Mesenquimais/citologia , Nanofios/química , Osteogênese , Cálcio/metabolismo , Adesão Celular , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Grafite/química , Humanos , Sialoproteína de Ligação à Integrina/genética , Sialoproteína de Ligação à Integrina/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Propriedades de Superfície , Engenharia TecidualRESUMO
Specific targeting of cells to sites of tissue damage and delivery of high numbers of transplanted cells to lesion tissue in vivo are critical parameters for the success of cell-based therapies. Here, we report a promising in vitro model system for studying the homing of transplanted cells, which may eventually be applicable for targeted regeneration of damaged neurons in spinal cord injury. In this model system, neurospheres derived from human neuroblastoma SH-SY5Y cells labeled with bacterial magnetic nanoparticles were guided by a magnetic field and successfully accumulated near the focus site of the magnetic field. Our results demonstrate the effectiveness of using an in vitro model for testing bacterial magnetic nanoparticles to develop successful stem cell targeting strategies during fluid flow, which may ultimately be translated into in vivo targeted delivery of cells through circulation in various tissue-repair models.
Assuntos
Nanopartículas de Magnetita/uso terapêutico , Neurônios/transplante , Traumatismos da Medula Espinal/terapia , Diferenciação Celular/fisiologia , Linhagem Celular Tumoral , Sobrevivência Celular/fisiologia , Humanos , Magnetismo/métodos , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Neurônios/ultraestruturaRESUMO
Vertically aligned one-dimensional hybrid structures, which are composed of apatite and graphitic structures, can be beneficial for orthopedic applications. However, they are difficult to generate using the current method. Here, we report the first synthesis of a single-crystal apatite nanowire encapsulated in graphitic shells by a one-step chemical vapor deposition. Incipient nucleation of apatite and its subsequent transformation to an oriented crystal are directed by derived gaseous phosphorine. Longitudinal growth of the oriented apatite crystal is achieved by a vapor-solid growth mechanism, whereas lateral growth is suppressed by the graphitic layers formed through arrangement of the derived aromatic hydrocarbon molecules. We show that this unusual combination of the apatite crystal and the graphitic shells can lead to an excellent osteogenic differentiation and bony fusion through a programmed smart behavior. For instance, the graphitic shells are degraded after the initial cell growth promoted by the graphitic nanostructures, and the cells continue proliferation on the bare apatite nanowires. Furthermore, a bending experiment indicates that such core-shell nanowires exhibited a superior bending stiffness compared to single-crystal apatite nanowires without graphitic shells. The results suggest a new strategy and direction for bone grafting materials with a highly controllable morphology and material conditions that can best stimulate bone cell differentiation and growth.
Assuntos
Apatitas/química , Substitutos Ósseos/síntese química , Transplante Ósseo/instrumentação , Grafite/química , Células-Tronco Mesenquimais/citologia , Nanofios/química , Osteoblastos/citologia , Diferenciação Celular , Células Cultivadas , Cristalização/métodos , Módulo de Elasticidade , Desenho de Equipamento , Análise de Falha de Equipamento , Humanos , Substâncias Macromoleculares/química , Teste de Materiais , Conformação Molecular , Nanofios/ultraestrutura , Osteogênese/fisiologia , Propriedades de Superfície , Engenharia Tecidual/instrumentação , Alicerces TeciduaisRESUMO
We first demonstrate the effects of magnetic trapping of mitochondria using aptamer conjugated to bacterial magnetic nanoparticles that allowed targeting of the mitochondrial cytochrome c in the treatment of cancer cells. Our findings offer a new approach for targeted cell therapy, with the advantage of remote control over subcellular elements.