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1.
Artigo em Inglês | MEDLINE | ID: mdl-32208666

RESUMO

Surface-mediated gene delivery attracts more and more attentions in biomedical researches and applications due to its characteristics of low toxicity and localized delivery. Herein, a novel visible-light regulated surface-mediated gene delivery platform is exhibited, arising from the photo-induced surface charge accumulation on silicon. Silicon with a pn junction is used, and tested subsequently for the behavior of surface-mediated gene delivery under visible-light illumination. It is found that positive charge accumulation under light illumination changes the surface potential, and then facilitates the delivery of gene-loaded carriers. As a result, the gene expression efficiency shows a significant improvement from 6% to 28% under a 10 min visible-light illumination. Such improvement is ascribed to that the increased surface potential caused by light illumination, which promotes both the release of gene-loaded carriers and the cellular uptake. This work suggests that silicon with photovoltaic effect could offer a new strategy for surface-mediated gene delivery related biomedical researches and applications.

2.
Stem Cells ; 38(3): 395-409, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31721356

RESUMO

Mesenchymal stem cells (MSCs), which are undifferentiated stem cells with the property of stemness and the potential to differentiate into multiple lineages, including osteoblasts, have attracted a great deal of attention in bone tissue engineering. Consistent with the heterogeneity of MSCs, various surface markers have been used. However, it is still unclear which markers of MSCs are best for cell amplification in vitro and later bone regeneration in vivo. Krüppel-like Factor 2 (KLF2) is an important indicator of the stemness of human MSCs (hMSCs) and as early vascularization is also critical for bone regeneration, we used KLF2 as a novel in vitro marker for MSCs and investigated the angiogenesis and osteogenesis between KLF2+ MSCs and endothelial cells (ECs). We found a synergistic interaction between hMSCs and human umbilical vein ECs (HUVECs) in that KLF2+ stemness-maintained hMSCs initially promoted the angiogenesis of HUVECs, which in turn more efficiently stimulated the osteogenesis of hMSCs. In fact, KLF2+ hMSCs secreted angiogenic factors initially, with some of the cells then differentiating into pericytes through the PDGF-BB/PDGFR-ß signaling pathway, which improved blood vessel formation. The matured HUVECs in turn synergistically enhanced the osteogenesis of KLF2+ hMSCs through upregulated vascular endothelial growth factor. A three-dimensional coculture model using cell-laden gelatin methacrylate (GelMA) hydrogel further confirmed these results. This study provides insight into the stemness-directed synergistic interaction between hMSCs and HUVECs, and our results will have a profound impact on further strategies involving the application of KLF2+ hMSC/HUVEC-laden GelMA hydrogel in vascular network bioengineering and bone regeneration.

3.
ACS Omega ; 4(21): 19350-19357, 2019 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-31763559

RESUMO

Naringin, a Chinese herbal medicine, has been demonstrated to concentration-dependently promote osteogenic differentiation of mesenchymal stem cells (MSCs). However, it remains a challenge to load naringin on coatings for osteogenesis and further control the release kinetics. Here, we demonstrated that the release behavior of naringin on rutile nanorod films could be controlled by either mixing naringin with gelatin methacryloyl (GelMA) before spinning onto the films or soaking the obtained GelMA-incorporated films with the naringin solution to achieve the distinct degradation-type release and diffusion-type release, respectively. We further revealed that the naringin-loaded coatings facilitated adhesion, proliferation and late differentiation, and mineralization of MSCs. Our findings provided a novel strategy to engineer the coatings with controlled release of naringin and emphasized the bioactivity of naringin for the osteogenic differentiation of MSCs.

4.
ACS Appl Mater Interfaces ; 11(47): 43857-43864, 2019 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-31692325

RESUMO

Graphene (Gr) presents promising applications in regulating the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Light illumination is regarded as a spatiotemporally controllable, easily applicable, and noninvasive mean to modulate material responses. Herein, Gr-transferred silicon (Gr/Si) with a Schottky junction is utilized to evaluate the visible-light-promoted osteogenic differentiation of BMSCs. Under light illumination, light-induced charges, owing to the formation of the Schottky junction at the interface of Gr and Si, accumulated on the surface and then changed the surface potential of Gr/Si. The Schottky junction and surface potential at the interface of Gr and Si was measured by photovoltaic test and scanning Kelvin probe microscopy. Alkaline phosphatase (ALP) activity and quantitative real-time polymerase chain reaction (PCR) measurement showed that such variations of surface improved the osteogenic differentiation of BMSCs, and the activation of the voltage-gated calcium channels through surface potential and accumulation of cytosolic Ca2+ could be the reason. Moreover, X-ray photoelectron spectroscopy characterization showed that surface charge could also affect BMSCs differentiation through the promotion or inhibition of the adsorption of osteogenic growth factors. Such light-promoted osteogenic differentiation of BMSCs on Gr/Si may have huge potential for biomedical materials or devices for bone regeneration application.

5.
J Nanobiotechnology ; 17(1): 102, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31581945

RESUMO

Natural extracellular matrices (ECMs) are three-dimensional (3D) and multi-scale hierarchical structure. However, coatings used as ECM-mimicking structures for osteogenesis are typically two-dimensional or single-scaled. Here, we design a distinct quasi-three-dimensional hierarchical topography integrated of density-controlled titania nanodots and nanorods. We find cellular pseudopods preferred to anchor deeply across the distinct 3D topography, dependently of the relative density of nanorods, which promote the osteogenic differentiation of osteoblast but not the viability of fibroblast. The in vivo experimental results further indicate that the new bone formation, the relative bone-implant contact as well as the push-put strength, are significantly enhanced on the 3D hierarchical topography. We also show that the exposures of HFN7.1 and mAb1937 critical functional motifs of fibronectin for cellular anchorage are up-regulated on the 3D hierarchical topography, which might synergistically promote the osteogenesis. Our findings suggest the multi-dimensions and multi-scales as vital characteristic of cell-ECM interactions and as an important design parameter for bone implant coatings.

6.
Biomaterials ; 222: 119456, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31476662

RESUMO

Geometry sensing of cells inevitably involves cytoskeletal remodeling and the activation of biochemical signaling, which control multiple aspects of cell behaviors, such as proliferation, differentiation and migration. A variety of size-, shape- and geometry-dependent cell behaviors have been revealed, but the role of geometric chirality in regulating cellular behaviors and the underlying biophysical mechanisms remain elusive. Here, we report an intriguing mechanotransduction of stem cells on chiral geometries that human mesenchymal stem cells (hMSCs) prefer to migrate towards dextral geometry with nearly 30% relative advantage in migration speed, referred to as "chirotaxis". We also found that cell adhesion, proliferation, and differentiation of hMSCs are greatly enhanced for cells cultured on dextral geometry than those on sinistral geometry, by triggering transcription factor AP-1 complex through p38/MAPK signaling that regulates hMSCs fate and activity. We demonstrated that the cytoskeletal network consisting of transverse and radial stress fibers exhibits a strengthening/offsetting effect on dextral/sinistral geometry through focal adhesion sites, and consequently, cell's cytoskeletal contractility on the dextral geometry is nearly 80% higher. These findings highlight the importance of geometric chirality as an extracellular cue in regulating stem cell's behaviors through cell-material interactions.

7.
ACS Appl Mater Interfaces ; 11(28): 25457-25464, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31282143

RESUMO

A facile strategy is needed for accurate time-space supply of suitable growth factors or drugs. Polypyrrole (PPy) was able to carry almost all kinds of negatively charged biomolecules through anodizing method, which made it an appropriate way for codeposition of multiple molecules. The difference in the conjugation between different molecules and PPy makes it possible for selective release when the redox state of PPy changes. In this work, bovine serum albumin (BSA) and heparin (Hep) were chosen to be the model molecules in view of their differences in the level of electronegativity and molecular weight. Double-layer deposition method was used to improve the biocompatibility of PPy/BSA/Hep film. It was found the content of BSA and Hep in the film can be controlled by regulating deposition current and time. BSA release was facilitated under positive voltage and then promote the proliferation of preosteoblasts, while Hep release was promoted under negative voltage and enhance cell differentiation. Our work provides a dual-molecule model in PPy for selective release and further explores the mechanism of release selectivity, this discovery has potential applications in tissue engineering and regenerative medicine.


Assuntos
Adesão Celular , Condutividade Elétrica , Heparina/química , Membranas Artificiais , Osteoblastos/metabolismo , Polímeros/química , Pirróis/química , Soroalbumina Bovina/química , Animais , Bovinos , Linhagem Celular , Camundongos , Osteoblastos/citologia
8.
ACS Appl Mater Interfaces ; 11(25): 22218-22227, 2019 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-31199127

RESUMO

The surface electric potential of biomaterials has been extensively proven to play a critical role in stem cells' fate. However, there are ambiguous reports on the relation of stem cells' osteogenic capacity to surface potential characteristics (potential polarity and intensity). To address this, we adopted a surface with a wide potential range and both positive/negative polarity in a comprehensive view to get insight into surface potential-regulating cellular osteogenic differentiation. Tb xDy1- xFe2 alloy/poly(vinylidene fluoride-trifluoroethylene) magnetoelectric films were prepared, and the film could provide controllable surface potential characteristics with positive or negative polarity and potential (ϕME) intensity variation from 0 to ±120 mV as well as keep the surface chemical composition and microstructure unchanged. Cell culture results showed that osteogenic differentiation of mesenchymal stem cells on both positive and negative potential films was obviously upregulated when the /ϕME/ intensities were set from 0-55 mV. Differently, the highest upregulated osteogenic differentiation on the positive potential films corresponded to the /ϕME/ intensity from 35-55 mV and was better than that on the negative potential films whereas the highest on the negative potential films corresponded to the /ϕME/ intensity from 0-35 mV and was better than that on the positive potential films. This fact could illustrate why previous reports appeared ambiguously; i.e., the comparative result in osteogenic differentiation between the positive and negative potential films strongly depends on the selection of surface potential intensity. On the basis of assaying of the exposed functional sites (RGD and PHSRN) of the adsorbed fibronectin (FN) and the expression of cellular integrin α5 and ß1 subunits, the difference in the behavior between the positive and negative potential films was attributed to the distinct conformation of adsorbed fibronectin (FN) and the opposite changing trend with /ϕME/ for the two films, which triggers the osteogenesis-related FAK/ERK signaling pathway to a different extent. This study could provide new cognition for the in-depth understanding of the regulation mechanism underlying surface potential characteristics in cell behaviors.


Assuntos
Fibronectinas/metabolismo , Adesão Celular/fisiologia , Proliferação de Células/fisiologia , Humanos , Integrina alfa5/metabolismo , Integrina beta1/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Osteogênese/fisiologia , Espectroscopia Fotoeletrônica , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
9.
Materials (Basel) ; 12(5)2019 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-30857349

RESUMO

It is well known that three-dimensional (3D) printing is an emerging technology used to produce customized implants and surface characteristics of implants, strongly deciding their osseointegration ability. In this study, Ti alloy microspheres were printed under selected rational printing parameters in order to tailor the surface micro-characteristics of the printed implants during additive manufacturing by an in situ, controlled way. The laser path and hatching space were responsible for the appearance of the stripy structure (S), while the bulbous structure (B) and bulbous⁻stripy composite surface (BS) were determined by contour scanning. A nano-sized structure could be superposed by hydrothermal treatment. The cytocompatibility was evaluated by culturing Mouse calvaria-derived preosteoblastic cells (MC3T3-E1). The results showed that three typical microstructured surfaces, S, B, and BS, could be achieved by varying the 3D printing parameters. Moreover, the osteogenic differentiation potential of the S, B, and BS surfaces could be significantly enhanced, and the addition of nano-sized structures could be further improved. The BS surface with nano-sized structure demonstrated the optimum osteogenic differentiation potential. The present research demonstrated an in situ, controlled way to tailor and optimize the surface structures in micro-size during the 3D printing process for an implant with higher osseointegration ability.

10.
Colloids Surf B Biointerfaces ; 172: 355-361, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30189386

RESUMO

This study presents a convenient and versatile way to prepare functionalized composite polydopamine/titanium dioxide (PDA/TiO2) film on polystyrene (PS). First, polystyrene substrate was immersed in dopamine chloride solution, and then collosol containing TiO2 and water was spun on it, to produce uniform, continuous PDA/TiO2 composite films. The thickness of film was controllable by adjustment of the spin speed. It was found that the films were strongly adhered on the PS substrate, with peel strength and shear strength of 2.78 MPa and 37.78 MPa, respectively. After 20 min of ultraviolet (365 nm) illumination, over 90% of fibroblasts and 77% of osteoblasts detached from the PDA/TiO2 composite film. Additionally, the detached cells showed good viability, allowing further culture and applications. This preparation method could be widely applied for cell and cell sheet harvesting directly from PS-based culture wares.


Assuntos
Técnicas de Cultura de Células/métodos , Indóis/síntese química , Polímeros/síntese química , Titânio/química , Animais , Sobrevivência Celular , Camundongos , Células NIH 3T3 , Análise Espectral Raman
11.
J Mater Sci Mater Med ; 29(9): 136, 2018 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-30120587

RESUMO

Surface nanotopography and bioactive ions have been considered to play critical roles on the interactions of biomaterials with cells. In this study, a TiO2 nanorod film incorporated with Zn-containing bioactive glass (TiO2/Zn-BG) was prepared on tantalum substrate, trying to evaluate the synergistic effects of nanotopograpgy and bioactive ions to promote cellular osteogenic differentiation activity. The expression of osteogenic-related genes, ALP as well as the ECM mineralization on TiO2/Zn-BG film were significantly upregulated compared to that of the film without TiO2 nanorod nanostructure (Zn-BG) or without Zn (TiO2/BG). Moreover, a much low Zn2+ release level on TiO2/Zn-BG film was beneficial to promote the osteogenesis, which could be ascribed to that a semi-closed space established by TiO2 nanorods with adhered cells provided an appropriate micro-environment that facilitated Zn2+ adsorption.


Assuntos
Cerâmica , Nanopartículas Metálicas/química , Nanotubos/química , Osteogênese , Titânio/química , Zinco/química , Células 3T3 , Adsorção , Fosfatase Alcalina/metabolismo , Animais , Materiais Biocompatíveis/química , Adesão Celular , Diferenciação Celular , Matriz Extracelular/metabolismo , Íons , Camundongos , Nanoestruturas/química , Osteocalcina/metabolismo , Propriedades de Superfície , Tantálio/química
12.
Acta Biomater ; 74: 291-301, 2018 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-29729416

RESUMO

Surface potential of biomaterials can dramatically influence cellular osteogenic differentiation. In this work, a wide range of surface potential on ferroelectric polyvinylidene fluoride trifluoroethylene (P(VDF-TrFE)) films was designed to get insight into the interfacial interaction of cell-charged surface. The P(VDF-TrFE) films poled by contact electric poling at various electric fields obtained well stabilized surface potential, with wide range from -3 to 915 mV. The osteogenic differentiation level of cells cultured on the films was strongly dependent on surface potential and reached the optimum at 391 mV in this system. Binding specificity assay indicated that surface potential could effectively govern the binding state of the adsorbed fibronectin (FN) with integrin. Molecular dynamic (MD) simulation further revealed that surface potential brought a significant difference in the relative distance between RGD and synergy PHSRN sites of adsorbed FN, resulting in a distinct integrin-FN binding state. These results suggest that the full binding of integrin α5ß1 with both RGD and PHSRN sites of FN possesses a strong ability to activate osteogenic signaling pathway. This work sheds light on the underlying mechanism of osteogenic differentiation behavior on charged material surfaces, and also provides a guidance for designing a reasonable charged surface to enhance osteogenic differentiation. STATEMENT OF SIGNIFICANCE: The ferroelectric P(VDF-TrFE) films with steady and a wide range of surface potential were designed to understand underlying mechanism of cell-charged surface interaction. The results showed that the charged surface well favored upregulation of osteogenic differentiation of MC3T3-E1 cells, and more importantly, a highest level occurred on the film with a moderate surface potential. Experiments and molecular dynamics simulation demonstrated that the surface potential could govern fibronectin conformation and then the integrin-fibronectin binding. We propose that a full binding state of integrin α5ß1 with fibronectin induces effective activation of integrin-mediated FAK/ERK signaling pathway to upregulate cellular osteogenic differentiation. This work provides a guidance for designing a reasonable charged surface to enhance osteogenic differentiation.


Assuntos
Diferenciação Celular , Condutividade Elétrica , Membranas Artificiais , Osteoblastos/metabolismo , Osteogênese , Polivinil/química , Animais , Linhagem Celular , Humanos , Camundongos , Osteoblastos/citologia , Propriedades de Superfície
13.
Colloids Surf B Biointerfaces ; 167: 213-219, 2018 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-29656204

RESUMO

Titanium and tantalum have been widely used for orthopedic and dental implant applications. However, how their inherent surface features regulate cellular osteogeneses still remains elusive. In this study, we engineered two distinct TiO2 and Ta2O5 nanorod films as the two model oxidized surfaces to investigate their intrinsic osteogenic behaviors. The results indicated that the distinctive gradient on zeta potential against pH, corresponding to the deprotonation rate, but not the hydroxyl amount or hydroxylation polarity played a critical role on the cellular osteogenic performance. TiO2 nanorod film with a higher deprotonation rate significantly upregulated the expression of osteogeneses-related gene and protein, comparing to that of Ta2O5 nanorod film. These results might be attributed to that surface with higher deprotonation rateprovided more Bronsted acid-base surface sites to react with protein residues, leading to a mild change in conformation of the absorbed proteins, and subsequently facilitating to trigger the integrin-focal adhesion cytoskeleton actin transduction pathway. This study, therefore, provides a new insight into the understanding the role of material surface hydroxylation on cellular osteogenic responses.


Assuntos
Nanotubos/química , Osteogênese/efeitos dos fármacos , Titânio/química , Células 3T3 , Animais , Células Cultivadas , Hidroxilação , Camundongos , Tamanho da Partícula , Propriedades de Superfície
14.
Acta Biomater ; 71: 49-60, 2018 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-29550443

RESUMO

Mechanical stimuli at the bone-implant interface are considered to activate the mechanotransduction pathway of the cell to improve the initial osseointegration establishment and to guarantee clinical success of the implant. However, control of the mechanical stimuli at the bone-implant interface still remains a challenge. In this study, we have designed a strategy of a magnetically responsive coating on which the mechanical stimuli is controlled because of coating deformation under static magnetic field (SMF). The iron oxide nanoparticle/mineralized collagen (IOP-MC) coatings were electrochemically codeposited on titanium substrates in different quantities of IOPs and distributions; the resulting coatings were verified to possess swelling behavior with flexibility same as that of hydrogel. The relative quantity of IOP to collagen and the IOP distribution in the coatings were demonstrated to play a critical role in mediating cell behavior. The cells present on the outer layer of the distributed IOP-MC (O-IOP-MC) coating with a mass ratio of 0.67 revealed the most distinct osteogenic differentiation activity being promoted, which could be attributed to the maximized mechanical stimuli with exposure to SMF. Furthermore, the enhanced osteogenic differentiation of the stimulated MC3T3-E1 cells originated from magnetically actuated mechanotransduction signaling pathway, embodying the upregulated expression of osteogenic-related and mechanotransduction-related genes. This work therefore provides a promising strategy for implementing mechanical stimuli to activate mechanotransduction on the bone-implant interface and thus to promote osseointegration. STATEMENT OF SIGNIFICANCE: The magnetically actuated coating is designed to produce mechanical stimuli to cells for promoting osteogenic differentiation based on the coating deformation. Iron oxide nanoparticles (IOPs) were incorporated into the mineralized collagen coatings (MC) forming the composite coatings (IOP-MC) with spatially distributed IOPs, and the IOP-MC coatings with outer distributed IOPs (O-IOPs-MC) shows the maximized mechanical stimuli to cells with enhanced osteogenic differentiation under static magnetic field. The upregulated expression of the associated genes reveals that the enabled mechanotransduction signaling pathway is responsible for the promoted cellular osteogenic differentiation. This work therefore provides a promising strategy for implementing mechanical stimuli to activate mechanotransduction on the bone-implant interface to promote osseointegration.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Materiais Revestidos Biocompatíveis , Colágeno , Óxido Ferroso-Férrico , Nanopartículas de Magnetita/química , Osteogênese/efeitos dos fármacos , Animais , Linhagem Celular , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Colágeno/química , Colágeno/farmacologia , Óxido Ferroso-Férrico/química , Óxido Ferroso-Férrico/farmacologia , Campos Magnéticos , Camundongos
15.
ACS Appl Mater Interfaces ; 10(14): 11508-11518, 2018 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-29564888

RESUMO

Extracellular matrix (ECM) provides a dynamic and complex environment to determine the fate of stem cells. In this work, light harvested cell sheets were treated with paraformaldehyde or ethanol, which eventually become ECM. Such ECM was then immobilized on titanium substrates via polydopamine chemistry. Their effects on bone marrow mesenchymal stromal cells (BMSCs) behaviors were investigated. It was found that paraformaldehyde-treated ECM coating (PT-ECM) showed a well-maintained microstructure, whereas that of ethanol-treated (ET-ECM) was completely changed. As a result, different amide structures and distributions of ECM components, such as laminin and collagen I, were exhibited. Alkaline phosphatase activity, osteocalcin secretion, related gene expression, and mineral deposition were evaluated for BMSCs cultured on both ECM coatings. PT-ECM was demonstrated to promote osteogenic differentiation much more efficiently than that of ET-ECM. That is ascribed to the preservation of native ECM milieu of PT-ECM. Such ECM acquirement and immobilization method could establish surfaces being able to direct stem cell responses on various materials. That shows promising potential in bone tissue engineering and other related biomedical applications.


Assuntos
Matriz Extracelular , Células da Medula Óssea , Diferenciação Celular , Células Cultivadas , Células-Tronco Mesenquimais , Osteogênese
16.
ACS Appl Mater Interfaces ; 10(9): 7841-7851, 2018 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-29412633

RESUMO

The binding of cell integrins to proteins adsorbed on the material surface is a highly dynamic process critical for guiding cellular responses. However, temporal dynamic regulation of adsorbed proteins to meet the spatial conformation requirement of integrins for a certain cellular response remains a great challenge. Here, an active CoFe2O4/poly(vinylidene fluoride-trifluoroethylene) nanocomposite film, which was demonstrated to be an obvious surface potential variation (Δ V ≈ 93 mV) in response to the applied magnetic field intensity (0-3000 Oe), was designed to harness the dynamic binding of integrin-adsorbed proteins by in situ controlling of the conformation of adsorbed proteins. Experimental investigation and molecular dynamics simulation confirmed the surface potential-induced conformational change in the adsorbed proteins. Cells cultured on nanocomposite films indicated that cellular responses in different time periods (adhesion, proliferation, and differentiation) required distinct magnetic field intensity, and synthetically programming the preferred magnetic field intensity of each time period could further enhance the osteogenic differentiation through the FAK/ERK signaling pathway. This work therefore provides a distinct concept that dynamically controllable modulation of the material surface property fitting the binding requirement of different cell time periods would be more conducive to achieving the desired osteogenic differentiation.


Assuntos
Nanocompostos , Diferenciação Celular , Células Cultivadas , Osteogênese , Células-Tronco
17.
J Biomed Mater Res B Appl Biomater ; 106(5): 2037-2045, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29098765

RESUMO

The combination of bone marrow-derived mesenchymal stem cells (BMSCs) and biological scaffolds has been demonstrated to be a promising strategy for bone regeneration. However, this method does not result in satisfactory bone regeneration, because the BMSCs are dispersed in the biological scaffolds. The current study developed a new bone regeneration system, which combines synthetic porous three-dimensional scaffolds of ß-TCP/COL-I composite with cultured osteogenic sheets of BMSCs. Activity of alkaline phosphatase (ALP), a marker of bone regeneration, was assayed in vitro using enzyme-linked immunosorbent assays and quantitative real-time polymerase chain reaction. In vivo bone regeneration was assayed in male nude mice. The study samples were BMSC sheet, scaffold/scattered BMSCs, scaffold/BMSC sheet, and scaffold alone. The samples were implanted dorsally in the mice. In vitro analysis showed that ß-TCP/COL-I scaffold combined with BMSC sheets significantly upregulated both gene expression and protein levels of ALP, osteocalcin, and osteopontin. Histological and micro-computed tomography showed that the only implants that demonstrated new bone formation after 4 weeks were scaffold/BMSC sheet implants. These results underscore the crucial requirement of a synergistic effect of ß-TCP/COL-I scaffolds and BMSC sheets. This could be a promising novel strategy for bone tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2037-2045, 2018.


Assuntos
Regeneração Óssea , Fosfatos de Cálcio/química , Células Imobilizadas/metabolismo , Células Imobilizadas/transplante , Colágeno Tipo I/química , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Tecidos Suporte/química , Animais , Células Imobilizadas/patologia , Xenoenxertos , Masculino , Células-Tronco Mesenquimais/patologia , Camundongos , Camundongos Nus , Ratos , Ratos Sprague-Dawley
18.
Sci Rep ; 7(1): 17926, 2017 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-29263335

RESUMO

In electrical stimulation (ES), daily stimulation time means the interacting duration with cells per day, and is a vital factor for mediating cellular function. In the present study, the effect of stimulation time on osteogenic differentiation of MC3T3-E1 cells was investigated under ES on polypyrrole (Ppy) planar interdigitated electrodes (IDE). The results demonstrated that only a suitable daily stimulation time supported to obviously upregulate the expression of ALP protein and osteogenesis-related genes (ALP, Col-I, Runx2 and OCN), while a short or long daily stimulation time showed no significant outcomes. These might be attributed to the mechanism that an ES induced transient change in intracellular calcium ion concentration, which was responsible for activating calcium ion signaling pathway to enhance cellular osteogenic differentiation. A shorter daily time could lead to insufficient duration for the transient change in intracellular calcium ion concentration, and a longer daily time could give rise to cellular fatigue with no transient change. This work therefore provides new insights into the fundamental understanding of cell responses to ES and will have an impact on further designing materials to mediate cell behaviors.


Assuntos
Diferenciação Celular , Estimulação Elétrica , Osteoblastos/citologia , Osteogênese , Polímeros/química , Pirróis/química , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Cálcio/metabolismo , Proliferação de Células , Células Cultivadas , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Eletrodos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Osteoblastos/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Crânio/citologia , Crânio/metabolismo
19.
ACS Appl Mater Interfaces ; 9(42): 36513-36524, 2017 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-28984126

RESUMO

Well-organized orientation of cells and anisotropic extracellular matrix (ECM) are crucial in engineering biomimetic tissues, such as muscles, arteries, and nervous system, and so on. This strategy, however, is only beginning to be explored. Here, we demonstrated a light-induced cell alignment and harvest for anisotropic cell sheets (ACS) technology using light-responsive TiO2 nanodots film (TNF) and photo-cross-linkable gelatin methacrylate (GelMA). Cell initial behaviors on TNF might be controlled by micropatterns of light-induced distinct surface hydroxyl features, owing to a sensing mechanism of myosin II-driven retraction of lamellipodia. Further light treatment allowed ACS detachment from TNF surface while simultaneously solidified the GelMA, realizing the automatic transference of ACS. Moreover, two detached ACS were successfully stacked into a 3D bilayer construct with controllable orientation of individual layer and maintained cell alignment for more than 7 days. Interestingly, the anisotropic HFF-1 cell sheets could further induce the HUVECs to form anisotropic capillary-like networks via upregulating VEGFA and ANGPT1 and producing anisotropic ECM. This developed integrated-functional ACS technology therefore provides a novel route to produce complex tissue constructs with well-defined orientations and may have a profound impact on regenerative medicine.


Assuntos
Anisotropia , Matriz Extracelular , Gelatina , Medicina Regenerativa , Engenharia Tecidual
20.
ACS Appl Mater Interfaces ; 9(34): 28250-28259, 2017 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-28795563

RESUMO

Effective regulation of cell-surface interactions is critical for regenerative medicine and other cell-based therapies. Herein, visible-light-induced cell sheet harvesting based on silicon wafers with a p/n junction [Si(p/n)] is introduced. Cell sheets could quickly detach from the Si(p/n) surface after 10 min of visible-light illumination with maintained cell viability and functions. It is found that preadsorbed proteins on the Si(p/n) surface like BSA and collagen-I show light-induced desorption behaviors. Molecular dynamics simulation also indicates that long-range force caused by the photovoltaic effect of Si(p/n) under visible-light illumination plays a key role in triggering the release of the preadsorbed protein. It is suggested that such protein desorption behavior mediated by the photovoltaic effect is responsible for cell release. This work not only shows promising potential for cell sheet harvesting, but also provides new insights into protein-material interactions.


Assuntos
Luz , Animais , Sobrevivência Celular , Colágeno Tipo I , Soroalbumina Bovina , Silício
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