RESUMEN
Localization of tumors during laparoscopic surgery is generally performed by locally injecting India ink into the submucosal layer of the gastrointestinal tract using endoscopy. However, the location of the tumor is obscured because of the black-stained surgical field and the blurring caused by India ink. To solve this problem, in this study, we developed a tissue-adhesive porphyrin with polycations consisting of quaternary ammonium salt groups. To evaluate the ability of tissue-adhesive porphyrin in vivo, low-molecular-weight hematoporphyrin and tissue-adhesive porphyrin were injected into the anterior wall of the exposed stomach in rats. Local injection of low-molecular-weight hematoporphyrin into the anterior wall of the stomach was not visible even after 1 day because of its rapid diffusion. In contrast, the red fluorescence of the tissue-adhesive porphyrin was visible even after 7 days due to the electrostatic interactions between the positively-charged moieties of the polycation in the tissue-adhesive porphyrin and the negatively-charged molecules in the tissue. In addition, intraperitoneal injection of tissue-adhesive porphyrin in rats did not cause adverse effects such as weight loss, hepatic or renal dysfunction, or organ adhesion in the abdominal cavity. These results indicate that tissue-adhesive porphyrin is a promising fluorescent tissue-marking agent.
Asunto(s)
Porfirinas , Adhesivos Tisulares , Animales , Colorantes , Hematoporfirinas , Polielectrolitos , Compuestos de Amonio Cuaternario , RatasRESUMEN
Combination of different therapies is an attractive approach for cancer therapy. However, it is a challenge to synchronize different therapies for maximization of therapeutic effects. In this work, a smart composite scaffold that could synchronize magnetic hyperthermia and chemotherapy was prepared by hybridization of magnetic Fe3O4 nanoparticles and doxorubicin (Dox)-loaded thermosensitive liposomes with biodegradable polymers. Irradiation of alternating magnetic field (AMF) could not only increase the scaffold temperature for magnetic hyperthermia but also trigger the release of Dox for chemotherapy. The two functions of magnetic hyperthermia and chemotherapy were synchronized by switching AMF on and off. The synergistic anticancer effects of the composite scaffold were confirmed by in vitro cell culture and in vivo animal experiments. The composite scaffold could efficiently eliminate breast cancer cells under AMF irradiation. Moreover, the scaffold could support proliferation and adipogenic differentiation of mesenchymal stem cells for adipose tissue reconstruction after anticancer treatment. In vivo regeneration experiments showed that the composite scaffolds could effectively maintain their structural integrity and facilitate the infiltration and proliferation of normal cells within the scaffolds. The composite scaffold possesses multi-functions and is attractive as a novel platform for efficient breast cancer therapy.
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Doxorrubicina/análogos & derivados , Hipertermia Inducida , Neoplasias , Animales , Línea Celular Tumoral , Doxorrubicina/farmacología , Doxorrubicina/uso terapéutico , Hipertermia , Fenómenos Magnéticos , PolietilenglicolesRESUMEN
The treatment of melanoma requires not only the elimination of skin cancer cells but also skin regeneration to heal defects. To achieve this goal, a bifunctional composite scaffold of poly(DL-lactic-co-glycolic acid) (PLGA), collagen and black phosphorus nanosheets (BPNSs) was prepared by hybridizing a BPNS-embedded collagen sponge with a PLGA knitted mesh. The composite mesh increased the temperature under near-infrared laser irradiation. The incorporation of BPNSs provided the PLGA-collagen-BPNS composite mesh with excellent photothermal properties for the photothermal ablation of melanoma cells both in vitro and in vivo. The PLGA-collagen-BPNS composite mesh had high mechanical strength for easy handling. The PLGA-collagen-BPNS composite mesh facilitated the proliferation of fibroblasts, promoted the expression of angiogenesis-related genes and the genes of components of the extracellular matrix for skin tissue regeneration. The high mechanical strength, photothermal ablation capability and skin tissue regeneration effects demonstrate that the bifunctional PLGA-collagen-BPNS composite mesh is a versatile and effective platform for the treatment of melanoma and the regeneration of skin defects.
Asunto(s)
Antineoplásicos/uso terapéutico , Melanoma/tratamiento farmacológico , Fósforo/uso terapéutico , Regeneración/efectos de los fármacos , Fenómenos Fisiológicos de la Piel/efectos de los fármacos , Andamios del Tejido/química , Animales , Antineoplásicos/química , Antineoplásicos/efectos de la radiación , Línea Celular Tumoral , Colágeno/química , Femenino , Humanos , Rayos Infrarrojos , Ratones Endogámicos BALB C , Ratones Desnudos , Nanoestructuras/química , Nanoestructuras/efectos de la radiación , Nanoestructuras/uso terapéutico , Fósforo/química , Fósforo/efectos de la radiación , Terapia Fototérmica/métodos , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Ingeniería de Tejidos/métodosRESUMEN
Synergistic therapy, especially the combination of photothermal therapy and chemotherapy, has been proposed as an effective therapeutic approach for breast cancer treatment. In this study, a smart platform for synergistic photothermal therapy and chemotherapy was developed by hybridizing doxorubicin-encapsulated thermosensitive liposomes and gold nanorods into porous scaffolds of gelatin and polyglutamic acid (Dox-lipo/AuNR/Gel/PGA). The Dox-lipo/AuNR/Gel/PGA composite scaffolds had good photothermal conversion and temperature-dependent doxorubicin release properties. Under near-infrared laser irradiation, the composite scaffolds increased the local temperature to not only kill the breast cancer cells in the scaffolds but also accelerate the release of doxorubicin to eliminate the breast cancer cells surrounding the scaffolds. In vitro cell culture and in vivo mouse experiments demonstrated that the synergistic effects of photothermal ablation combined with doxorubicin-induced inhibition of the breast cancer cells in and surrounding the composite scaffolds under near-infrared laser irradiation. Moreover, after drug release was complete, the composite scaffolds fostered human bone marrow-derived mesenchymal stem cell proliferation. These results suggested that the composite scaffolds provided synergistic photothermal therapy and chemotherapy for breast cancer cell elimination at the early stage and promoted stem cell activities at the late stage. Therefore, this composite scaffold holds great potential as a synergistic therapy platform for breast cancer treatment.
Asunto(s)
Neoplasias de la Mama , Liposomas , Animales , Femenino , Humanos , Ratones , Neoplasias de la Mama/tratamiento farmacológico , Línea Celular Tumoral , Doxorrubicina , Terapia FototérmicaRESUMEN
Micropatterned surfaces are very useful to control cell microenvironment and investigate the physical effects on cell function. In this study, poly(vinyl alcohol) (PVA) micropatterns on polystyrene cell-culture plates were prepared using UV photolithography. Cell adhesive polystyrene geometries of triangle, square, pentagon, hexagon, and circle were surrounded by cell nonadhesive PVA to manipulate cell shapes. These different geometries had the same small surface areas for cell spreading. Human mesenchymal stem cells (MSCs) were cultured on the micropatterned surface, and the effect of cell geometry on adipogenic differentiation was investigated. MSCs adhered to the geometric micropatterns and formed arrays of single cell with different shapes. The distribution patterns of actin filaments were similar among these cell shapes and remolded during adipogenesis. The adipogenic differentiation potential of MSCs was similar on the small size triangular, square, pentagonal, hexagonal, and circular geometries according to lipid vacuoles staining. This simple micropatterning technique using photoreactive molecules will be useful for creating micropatterns of arbitrary design on an organic surface, and cell functions can be directly and systematically compared on a single surface without external factors resulting from separate cell culture and coating method.
Asunto(s)
Adipogénesis/efectos de los fármacos , Técnicas de Cultivo de Célula/métodos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Microtecnología/métodos , Alcohol Polivinílico/química , Alcohol Polivinílico/farmacología , Adipogénesis/efectos de la radiación , Forma de la Célula , Humanos , Células Madre Mesenquimatosas/efectos de la radiación , Poliestirenos/química , Impresión , Propiedades de Superficie , Rayos UltravioletaRESUMEN
Interconnected pore structures of scaffolds are important to control the cell functions for cartilage tissue engineering. In this study, collagen scaffolds with interconnected pore structures were prepared using poly(D,L-lactide-co-glycolide) (PLGA) sponges as sacrificial templates. Six types of PLGA sponges of different pore sizes and porosities were prepared by the solvent casting/particulate leaching method and used to regulate the interconnectivity of the collagen scaffolds. The integral and continuous templating structure of PLGA sponges generated well-interconnected pore structures in the collagen scaffolds. Bovine articular chondrocytes cultured in collagen scaffolds showed homogenous distribution, fast proliferation, high expression of cartilaginous genes and high secretion of cartilaginous extracellular matrix. In particular, the collagen scaffold templated by the PLGA sacrificial sponge that was prepared with a high weight ratio of PLGA and large salt particulates showed the most promotive effect on cartilage tissue formation. The interconnected pore structure facilitated cell distribution, cell-cell interaction and cartilage tissue regeneration.
Asunto(s)
Cartílago/fisiología , Colágeno/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Ingeniería de Tejidos/métodos , Andamios del Tejido , Animales , Materiales Biocompatibles , Bovinos , Adhesión Celular , Supervivencia Celular , Condrocitos/fisiologíaRESUMEN
PURPOSE: We studied the regenerated cartilage in tracheal defect repair and compared the bio-materials used versus native trachea using basic fibroblast growth factor (bFGF)-impregnated gelatin hydrogel. MATERIALS AND METHODS: A full-thickness anterior defect was created in the cervical trachea of 15 experimental rabbits. The defect was implanted with a hybrid scaffold of poly(lactic-co-glycolic acid) (PLGA) knitted mesh and collagen sponge. The implanted trachea was reinforced with a copolymer stent of polycaprolactone and poly(lactic acid) coarse fiber mesh. A gelatin hydrogel was used for providing a sustained release of bFGF. The reconstructed tracheas were divided into three groups with wrapped materials; without gelatin hydrogel (control group, n = 5), a gelatin hydrogel with saline (gelatin group, n = 5), and a gelatin hydrogel with 100 microg of bFGF (bFGF group, n = 5). One of the five rabbits in each group at 1 month after operation, one at 3 months, and three at 6 months were killed and the engineered tracheas were evaluated histologically. Biomechanical properties were evaluated on samples at 6 months postoperatively. RESULTS: The rigid support in the defect portion was maintained during 6 months postoperatively. The newly regenerated cartilages were recognized between the host cartilage stumps at 3 months postoperatively in the bFGF group, and limited new cartilage growth and epithelialization were observed at 6 months postoperatively. CONCLUSIONS: The experiment shows that using bFGF, better mechanical strength was obtained but with poor cartilage growth.
Asunto(s)
Cartílago/fisiología , Regeneración , Ingeniería de Tejidos , Tráquea/cirugía , Heridas y Lesiones/cirugía , Animales , Materiales Biocompatibles , Fenómenos Biomecánicos , Colágeno , Modelos Animales de Enfermedad , Factor 2 de Crecimiento de Fibroblastos/farmacología , Gelatina , Hidrogel de Polietilenoglicol-Dimetacrilato , Implantes Experimentales , Ácido Láctico , Masculino , Ácido Poliglicólico , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Polímeros , Conejos , Regeneración/efectos de los fármacos , Stents , Mallas Quirúrgicas , Andamios del Tejido , Tráquea/lesionesRESUMEN
Extracellular matrices (ECMs) are dynamically altered and remodeled during tissue development. How the dynamic remodeling of ECM affects stem cell functions remains poorly understood due to the difficulty of obtaining biomimetic ECMs. In this study, stepwise osteogenesis-mimicking ECM-deposited hybrid meshes were prepared by culturing human mesenchymal stem cells (hMSCs) in poly (lactic-co-glycolic acid) (PLGA)-collagen hybrid meshes and controlling the stages of the osteogenesis of hMSCs. Three types of hybrid mesh mimicking the ECMs that were secreted from stem cell stage of hMSCs (SC-ECM), early stage (EO-ECM) and late stage (LO-ECM) osteogenesis of hMSCs were prepared. The stepwise osteogenesis-mimicking ECM deposited PLGA-collagen hybrid meshes showed different ECM compositions associated with the stage of osteogenesis. Their effects on the osteogenic differentiation of hMSCs differed. EO-ECM scaffold increased and LO-ECM scaffold moderately promoted the osteogenic differentiation of hMSCs. However, SC-ECM scaffold inhibited the osteogenic differentiation of hMSCs. The novel PLGA-collagen-ECM hybrid meshes will provide useful tools for stem cell culture and tissue engineering.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Diferenciación Celular , Colágeno/química , Matriz Extracelular/química , Osteogénesis , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Adhesión Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Humanos , Células Madre Mesenquimatosas/citología , Osteoblastos/citología , Osteoblastos/metabolismo , Osteogénesis/efectos de los fármacos , Ingeniería de TejidosRESUMEN
Three kinds of photoreactive polyelectrolytes of polyallylamine (PAAm), poly(acrylic acid) (PAAc), and poly(vinyl alcohol) (PVA) were synthesized by the introduction of azidophenyl groups in the respective polymers. The photoreactive PAAm, PAAc, and PVA were micropatterned on polystyrene surfaces by photolithography. Observation with optical microscopy and scanning probe microscopy demonstrated the formation of a striped pattern of polyelectrolytes with a width of 200 microm. The micropatterned polyelectrolytes swelled in water. The micropatterned surfaces were used for cell culture of mesenchymal stem cells (MSCs) and their effects on adipogenic differentiation were investigated. The MSCs adhered to and proliferated evenly on the PAAm- and PAAc-patterned surfaces while they formed a cell pattern on the PVA-patterned surface. The PAAm-, PAAc-grafted, and polystyrene surfaces supported cell adhesion while the PVA-grafted surface did not. When cultured in adipogenic differentiation medium, the adipogenic differentiation of MSCs on the polyelectrolyte-patterned surfaces was demonstrated by the formation of lipid vacuoles and gene expression analysis. Oil Red-O-positive cells showed an even distribution on the PAAm- and PAAc-patterned surfaces, while they showed a pattern on the PVA-patterned surface. The fraction of Oil RedO-positive cells increased with culture time. The MSCs cultured on the PAAm-, PAAc-grafted, and polystyrene surfaces in adipogenic differentiation medium expressed the adipogenesis marker genes of peroxisome proliferator-activated receptor gamma2 (PPARgamma2), lipoprotein lipase (LPL), and fatty acid binding protein 4 (FABP4). These results indicate that the PAAm-, and PAAc-grafted, and polystyrene surfaces supported the adipogenesis of MSCs while a PVA-grafted surface did not.
Asunto(s)
Adipocitos/citología , Adipocitos/fisiología , Materiales Biocompatibles/química , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/fisiología , Polímeros/química , Ingeniería de Tejidos/métodos , Técnicas de Cultivo de Célula/métodos , Diferenciación Celular/fisiología , Células Cultivadas , Electrólitos , Humanos , Ensayo de MaterialesRESUMEN
Cell delivery in cell therapy is typically challenged by the low cell survival rate and immunological rejection during cells injection and circulation. Encapsulation of cells with semipermeable hydrogels or membranes can improve cell viability by resisting high shear force and inhibit immune response with the physical isolation effect. Herein, the individual HeLa cells and human mesenchymal stem cells (hMSCs) were encapsulated with enzyme responsive polymer nanoshell. The encapsulation shell was prepared via the Layer-by-Layer (LbL) assembly of functionalized gelatin and click chemistry of peptide linker and gelatin. The encapsulated cells showed high cell viability and could resist the physical stress. Moreover, the encapsulation shell had a prolonged encapsulation sustaining period and could effectively prevent the invasion of external entities. In addition, on-site cell release was realized via enzymolysis of the encapsulation shell by human matrix metalloproteinase-7 (MMP-7), an overexpressed enzyme on tumor area. The finding of this study proved a potential approach in cell therapy, especially for cell-based cancer therapy.
Asunto(s)
Células Madre Mesenquimatosas/citología , Nanocáscaras/química , Polímeros/química , Biocatálisis , Supervivencia Celular , Células Inmovilizadas/citología , Células HeLa , Humanos , Metaloproteinasa 7 de la Matriz/químicaRESUMEN
A shift in osteogenesis toward adipogenesis of human bone marrow-derived mesenchymal stem cells (hMSCs) is a crucial pathological factor in the progression of osteoporosis. The development of an in vitro three-dimensional (3D) model that reflects the dynamic remodeling of extracellular matrices (ECMs) during simultaneous osteogenesis and adipogenesis of hMSCs can provide a useful tool to mimic the process and to investigate how 3D ECMs balance the osteogenic and adipogenic differentiation of hMSCs. In this study, ECMs secreted by hMSCs during their stepwise osteogenesis-co-adipogenesis were deposited on hybrid meshes of poly(lactide-co-glycolide) (PLGA) and collagen to prepare biomimetic PLGA-collagen-ECM hybrid scaffolds. Four types of stepwise differentiation ECMs were prepared: ECMs secreted by hMSCs at early stages of osteogenesis and adipogenesis (EOEA-ECMs), hMSCs at early stages of osteogenesis and late stages of adipogenesis (EOLA-ECMs), hMSCs at late stages of osteogenesis and early stages of adipogenesis (LOEA-ECMs) and hMSCs at late stages of osteogenesis and late stages of adipogenesis (LOLA-ECMs). The deposited ECMs had different compositions that were dependent on the different stages of osteogenesis and adipogenesis. They also showed different effects on balancing the adipogenic and osteogenic differentiation of hMSCs. The EOEA-ECM scaffold had a promotive effect on adipogenesis and a suppressive effect on the osteogenesis of hMSCs. The LOEA-ECM and LOLA-ECM scaffolds showed a promotive effect on osteogenesis and a moderate effect on the adipogenic differentiation of hMSCs. The EOLA-ECM scaffold exhibited a suppressive effect on both osteogenesis and adipogenesis of hMSCs. However, the EOLA-ECM scaffold promoted hMSC proliferation more strongly than the other ECM scaffolds. The results indicated that dynamically remodeling ECM scaffolds could affect the osteogenic and adipogenic differentiation of hMSCs and should provide a useful 3D cell culture model for the investigation of ECM-cell interactions.
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Materiales Biomiméticos/química , Colágeno/química , Matriz Extracelular/química , Células Madre Mesenquimatosas/química , Poliglactina 910/química , Adipogénesis , Técnicas de Cultivo de Célula , Células Cultivadas , Humanos , Células Madre Mesenquimatosas/citología , Osteogénesis , Tamaño de la Partícula , Propiedades de Superficie , Andamios del TejidoRESUMEN
Osteogenic differentiation and commitment of mesenchymal stem cells (MSCs) is a complex process that is induced and regulated by various biological factors and biophysical cues. Although cell spreading area, as a biophysical cue, has been demonstrated to play a critical role in the regulation of osteogenic differentiation of MSCs, it is unclear how it affects the maintenance of the committed phenotype after osteogenic differentiation of MSCs. In this study, poly (vinyl alcohol) was micropatterned on a tissue culture polystyrene surface, and the micropatterns were used to culture MSCs to control their cell spreading area. The influence of cell spreading area on osteogenic differentiation and maintenance of the differentiated phenotype of MSCs was investigated. MSCs with a larger spreading area showed a higher degree of osteogenic differentiation, slower loss of differentiated phenotype and slower re-expression of stem cell markers compared with MSCs with a smaller spreading area. A large cell spreading area was beneficial for osteogenic differentiation of MSCs and maintenance of their differentiated phenotype.
Asunto(s)
Células Madre Mesenquimatosas/citología , Osteogénesis , Fenotipo , Fosfatasa Alcalina/metabolismo , Calcio/metabolismo , Técnicas de Cultivo de Célula , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Endoglina/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Células Madre Mesenquimatosas/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Alcohol Polivinílico/farmacologíaRESUMEN
Evaluation of the degradability of porous scaffolds is very important for tissue engineering. A protocol in which the condition is close to the in vivo pH environment was established for in vitro evaluation of biodegradable porous scaffolds. Degradation of PLGA sponges in phosphate-buffered solution (PBS) was evaluated with the protocol. The PLGA sponges degraded with incubation time. For the first 12 weeks, the weight loss increased gradually and then remarkably after 12 weeks. In contrast, the number-average molecular weight (Mn) decreased dramatically for the first 12 weeks and then less markedly after 12 weeks. Thermal analysis showed that the glass transition temperatures (Tg) decreased rapidly for the first 12 weeks, and the change became less evident after 12 weeks. These results suggest that the degradation mechanism of PLGA sponges was dominated by autocatalyzed bulk degradation for the first 12 weeks and then by surface degradation after 12 weeks. Physical aging was observed during incubation at 37 degrees C. The heterogeneous structure caused by physical aging might be one of the driving forces that induced autocatalyzed bulk degradation. The degradation mechanism was further supported by the data of pH change and the morphology of the degraded PLGA sponges. The autocatalyzed acidic products flooded out after 8 weeks, the pH dropped, and the walls of the sponges became more porous. The increase of the pore surface area facilitated surface degradation after 12 weeks. The pH was in the range between 7.43 and 7.24 during the entire incubation time. The protocol suppressed extreme changes of the pH and will be useful in the biodegradation evaluation of porous scaffolds for tissue engineering.
Asunto(s)
Materiales Biocompatibles/química , Ácido Láctico/química , Ácido Poliglicólico/química , Ensayo de Materiales/métodos , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ingeniería de TejidosRESUMEN
Human mesenchymal stem cells (MSCs) were cultured on polystyrene surfaces modified with photoreactive azidophenyl-derivatives of three different chargeable polymers, poly(acrylic acid) (PAAc), polyallylamine (PAAm), and poly(ethylene glycol) (PEG). The MSCs adhered and spread both on a PAAm-modified surface and on PAAc-modified and polystyrene (control) surfaces. However, the cells adhered more easily to the PAAm-modified surface. The MSCs did not attach to the PEG-modified surface and aggregated to form pellets immediately after cell seeding. The cells proliferated on the PAAc-, PAAm-modified and control surfaces with culture time, formed a monolayer, and aggregated to form pellets. The cells in the pellets that formed on the PAAm- and PEG-modified surfaces after 2 weeks culture had a round morphology and the extracellular matrices were positively stained by safranin O and toluidine blue, while those that formed on the PAAc-modified and control surfaces had a spindle, fibroblast-like morphology and were not positively stained by safranin O and toluidine blue. The pellets that formed on the PAAm- and PEG-modified surfaces contained significantly higher levels of sulfated glycosaminoglycans than did those that formed on the PAAc-modified and control surfaces. Type II collagen and cartilage proteoglycan were immunohistologically detected in the pellets that formed on PAAm- and PEG-modified surfaces, but not those that formed on the PAAc-modified and control surfaces. The MSCs cultured on the PAAm- and PEG-modified surfaces expressed a high level of cartilaginous genes encoding type II collagen and aggrecan, while the MSCs cultured on the PAAc-modified and control surfaces did not express these genes. These results suggest that the PAAm-modified surface supported cell adhesion and proliferation and also promoted chondrogenic differentiation of the MSCs. The PAAc-modified and polystyrene surfaces supported cell adhesion and proliferation, but not chondrogenic differentiation. The PEG-modified surfaces did not support cell adhesion, but did promote chondrogenic differentiation. The adhesion, proliferation, and differentiation of the MSCs could be controlled by surface chemistry.
Asunto(s)
Diferenciación Celular , Condrocitos/citología , Células Madre Mesenquimatosas/citología , Polímeros/química , Adhesión Celular , Proliferación Celular , Células Cultivadas , Condrocitos/metabolismo , Colágeno/metabolismo , Regulación de la Expresión Génica , Humanos , Inmunohistoquímica , Estructura Molecular , Fotoquímica , Proteoglicanos/metabolismo , ARN Mensajero/genética , Propiedades de SuperficieRESUMEN
Three-dimensional porous scaffolds play an important role in cartilage and bone tissue engineering as temporary templates for transplanted cells to control their adhesion and proliferation to guide the formation of the new tissues. The scaffolds should be biodegradable, biocompatible, mechanically strong, and capable of being formed into desired shapes. Biodegradable synthetic polymers and naturally derived collagen have their respective advantages and drawbacks. Hybridization of the two kinds of polymers has been carried out to combine their respective advantages. This review will summarize some of the recently developed porous scaffolds having hybrid, biphasic and leakproof structures, and their application to bone and cartilage tissue engineering.
Asunto(s)
Materiales Biocompatibles , Huesos , Cartílago , Medicina Regenerativa , Ingeniería de Tejidos , Andamios del Tejido , Animales , Regeneración Ósea , Condrocitos , Colágeno , Humanos , Ácido Láctico , Ácido Poliglicólico , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Polímeros , PorosidadRESUMEN
Nanoencapsulation of individual mammalian cells has great potential in biomedical, biotechnological and bioelectronic applications. However, existing techniques for cell nanoencapsulation generally yield short sustaining period and loose structure of encapsulation shell, which fails to meet the long-term cytoprotection and immunosuppression requirements. Here, we report a mild method to realize the nanoencapsulation of individual mammalian cells by layer-by-layer (LbL) assembly of gelatin inner layer and cross-linking of poly(ethylene glycol) (PEG) outer layer through thiol-click chemistry. With the present method, the encapsulated individual HeLa cells showed a high viability, long persistence period and effective resistance against macro external entities and high physical stress. Moreover, on-demand cell release could also be achieved by selective cleavage of succinimide thioether linkage in the outer PEG layer. The approach presented here may provide a new and versatile method for the cleavable nanoencapsulation of individual mammalian cells.
Asunto(s)
Química Clic/métodos , Polietilenglicoles/química , Polímeros/química , Supervivencia Celular/efectos de los fármacos , Portadores de Fármacos/efectos adversos , Portadores de Fármacos/química , Células HeLa , Humanos , Polímeros/efectos adversosRESUMEN
Chondrocytes are important for cartilage tissue engineering. However, dedifferentiation during chondrocyte subculture prevents the application of cartilage tissue engineering. Therefore, prevention of this dedifferentiation is required. Here, the possibility of poly(2-methoxyethyl acrylate) (PMEA) and its analogous polymers, poly(tetrahydrofurfuryl acrylate) (PTHFA) and poly(2-(2-methoxyethoxy) ethyl acrylate-co-butyl acrylate) (PMe2A), for chondrocyte subculture without dedifferentiation is examined. Chondrocytes spread on PTHFA and polyethylene terephthalate (PET), whereas their spreading is delayed on PMEA and PMe2A. When primary chondrocytes are subcultured on these polymers, the expression levels of cartilaginous genes are higher on PMEA and PMe2A than on PET and PTHFA. Integrin contribution to the initial cell adhesion is lower on PMEA and PMe2A than on PTHFA and PET. This low level of integrin contribution to cell adhesion may cause a delay in cell spreading and the maintenance of cartilaginous gene expression. These results indicate that PMEA and PMe2A may be favorable substrates for chondrocyte subculture and cartilage tissue engineering.
Asunto(s)
Acrilatos/química , Cartílago Articular/citología , Condrocitos/fisiología , Regulación de la Expresión Génica , Polímeros/química , Animales , Bovinos , Adhesión Celular , Técnicas de Cultivo de Célula/instrumentación , Técnicas de Cultivo de Célula/métodos , Proliferación Celular , Células Cultivadas , Condrocitos/citologíaRESUMEN
Mouse embryonic stem (ES) cells were cultured on four types of polymer with different surface properties. The polymers were poly(acrylic acid), polyallylamine, gelatin, and poly(2-methacryloyloxyethyl phosphorylcholine-co-methacrylic acid) (PMAc50), and were coupled with azidophenyl groups and photoimmobilized on conventional polystyrene cell-culture dishes. Mouse ES cells were cultured on the immobilized polymer surfaces, and cell morphology, cell growth, staining for alkaline phosphatase, activation of the transcription factor stat3, and expression of the octamer-binding protein 3/4 (Oct3/4) transcription factor and the zinc finger-containing transcription factor (GATA4) were observed. Morphology and growth rate were significantly affected by the polymer surface properties. The ES cells attached to gelatin or polyallylamine surfaces; however, colonies formed on the former but not the latter. In addition, significant enhancement of growth was observed on the gelatin surface. In contrast, ES cells aggregated to form an embryoid body on the photoimmobilized poly(acrylic acid) surface and the PMAc50 surface, although cell growth was reduced. Significant enhancement of aggregation of ES cells on the PMAc50 surface was observed in morphology and gene expression analyses.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Células Madre Embrionarias/citología , Células Madre Embrionarias/fisiología , Ácidos Polimetacrílicos/química , Ingeniería de Tejidos/métodos , Animales , Técnicas de Cultivo de Célula/instrumentación , Diferenciación Celular , Línea Celular , Proliferación Celular , Luz , Ratones , Fotoquímica/métodos , Ácidos Polimetacrílicos/efectos de la radiación , Ingeniería de Tejidos/instrumentaciónRESUMEN
Nanoparticles (NPs) have shown great potential for biomedical applications because of their unique physical and structural properties. A critical aspect for their clinical applications is cellular uptake that depends on both particle properties and the cell mechanical state. Despite the numerous studies trying to disclose the influencing factors, the role of cell size on cellular uptake remains unclear. In this study, poly(vinyl alcohol) was micropatterned on tissue culture polystyrene surfaces using UV photolithography to control the cell size, and the influence of cell size on the cellular uptake of gold NPs was investigated. Cells with a large size had a high total cellular uptake, but showed a low average uptake per unit area of cells. Cells with a small size showed opposite behaviors. The results were related to both cell/NP contacting area and membrane tension. A large cell size was beneficial for a high total cellular uptake due to the large contact area with the NPs. On the other hand, the large cell size resulted in high membrane tension that required high wrapping energy for engulfing of NPs and thus reduced the uptake. The two oppositely working effects decided the cellular uptake of NPs. The results would shed light on the influence of the cellular microenvironment on cellular uptake behavior.
Asunto(s)
Tamaño de la Célula , Endocitosis , Células Madre Mesenquimatosas/metabolismo , Nanopartículas/química , Nanopartículas/metabolismo , Membrana Celular/metabolismo , Proliferación Celular , Técnica del Anticuerpo Fluorescente , Oro/química , Oro/metabolismo , Humanos , Células Madre Mesenquimatosas/citología , Tamaño de la Partícula , Polietilenglicoles/química , Propiedades de SuperficieRESUMEN
Adhesion and spreading are essential processes of anchorage dependent cells involved in regulation of cell functions. Cells interact with their extracellular matrix (ECM) resulting in different degree of adhesion and spreading. However, it is not clear whether cell adhesion or cell spreading is more important for cell functions. In this study, 10 types of isotropical micropatterns that were composed of 2 µm microdots were prepared to precisely control the adhesion area and spreading area of human mesenchymal stem cells (MSCs). The respective influence of adhesion and spreading areas on stem cell functions was investigated. Adhesion area showed more significant influences on the focal adhesion formation, binding of myosin to actin fibers, cytoskeletal organization, cellular Young's modulus, accumulation of YAP/TAZ in nuclei, osteogenic and adipogenic differentiation of MSCs than did the spreading area. The results indicated that adhesion area rather than spreading area played more important roles in regulating cell functions. This study should provide new insight of the influence of cell adhesion and spreading on cell functions and inspire the design of biomaterials to process in an effective manner for manipulation of cell functions.