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1.
Bioact Mater ; 36: 185-202, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38463552

ABSTRACT

Wound healing in cases of excessive inflammation poses a significant challenge due to compromised neovascularization. Here, we propose a multi-functional composite hydrogel engineered to overcome such conditions through recruitment and activation of macrophages with adapted degradation of the hydrogel. The composite hydrogel (G-TSrP) is created by combining gelatin methacryloyl (GelMA) and nanoparticles (TSrP) composed of tannic acid (TA) and Sr2+. These nanoparticles are prepared using a one-step mineralization process assisted by metal-phenolic network formation. G-TSrP exhibits the ability to eliminate reactive oxygen species and direct polarization of macrophages toward M2 phenotype. It has been observed that the liberation of TA and Sr2+ from G-TSrP actively facilitate the recruitment and up-regulation of the expression of extracellular matrix remodeling genes of macrophages, and thereby, coordinate in vivo adapted degradation of the G-TSrP. Most significantly, G-TSrP accelerates angiogenesis despite the TA's inhibitory properties, which are counteracted by the released Sr2+. Moreover, G-TSrP enhances wound closure under inflammation and promotes normal tissue formation with strong vessel growth. Genetic analysis confirms macrophage-mediated wound healing by the composite hydrogel. Collectively, these findings pave the way for the development of biomaterials that promote wound healing by creating regenerative environment.

2.
Small ; 19(25): e2300236, 2023 06.
Article in English | MEDLINE | ID: mdl-36932895

ABSTRACT

The disruption of thyroid hormones because of chemical exposure is a significant societal problem. Chemical evaluations of environmental and human health risks are conventionally based on animal experiments. However, owing to recent breakthroughs in biotechnology, the potential toxicity of chemicals can now be evaluated using 3D cell cultures. In this study, the interactive effects of thyroid-friendly soft (TS) microspheres on thyroid cell aggregates are elucidated and their potential as a reliable toxicity assessment tool is evaluated. Using state-of-the-art characterization methods coupled with cell-based analysis and quadrupole time-of-flight mass spectrometry, it is shown that TS-microsphere-integrated thyroid cell aggregates exhibit improved thyroid function. Specifically, the responses of zebrafish embryos, which are used for thyroid toxicity analysis, and the TS-microsphere-integrated cell aggregates to methimazole (MMI), a known thyroid inhibitor, are compared. The results show that the thyroid hormone disruption response of the TS-microsphere-integrated thyroid cell aggregates to MMI is more sensitive compared with those of the zebrafish embryos and conventionally formed cell aggregates. This proof-of-concept approach can be used to control cellular function in the desired direction and hence evaluate thyroid function. Thus, the proposed TS-microsphere-integrated cell aggregates may yield new fundamental insights for advancing in vitro cell-based research.


Subject(s)
Thyroid Gland , Zebrafish , Animals , Humans , Antithyroid Agents/pharmacology , Thyroid Hormones/pharmacology , Methimazole/toxicity
4.
Small ; 18(22): e2200757, 2022 06.
Article in English | MEDLINE | ID: mdl-35521748

ABSTRACT

Numerous methods have been introduced to produce 3D cell cultures that can reduce the need for animal experimentation. This study presents a unique 3D culture platform that features bioinspired strands of electrospun nanofibers (BSeNs) and aquatic cell lines to compensate for shortcomings in the current cell spheroid generation techniques. The use of BSeNs in 3D zebrafish liver cell cultures is found to improve liver and reproductive functions through spheroid-based in vitro assays such as whole transcriptome sequencing and reproductive toxicity testing, with optimized properties exhibiting results similar to those obtained for fish embryo acute toxicity (FET, OECD TG 236) following exposure to environmental endocrine-disrupting chemicals (17ß-Estradiol (E2), 4-hydroxytamoxifen (4-HT), and bisphenol compounds (bisphenol A (BPA) and 9,9-Bis(4-hydroxyphenyl)fluorene (BPFL)). These findings indicate that the beneficial effects of bioinspired materials that closely mimic ECM environments can yield efficient zebrafish cells with intrinsic functions and xenobiotic metabolism similar to those of zebrafish embryos. As a closer analog for the in vivo conditions that are associated with exposure to potentially hazardous chemicals, the straightforward culture model introduced in this study shows promise as an alternative tool that can be used to further eco-environmental assessment.


Subject(s)
Endocrine Disruptors , Zebrafish , Animals , Endocrine Disruptors/metabolism , Endocrine Disruptors/toxicity , Liver/metabolism , Spheroids, Cellular/metabolism , Toxicity Tests , Zebrafish/metabolism
5.
Bioact Mater ; 11: 118-129, 2022 May.
Article in English | MEDLINE | ID: mdl-34938917

ABSTRACT

Developing a universal culture platform that manipulates cell fate is one of the most important tasks in the investigation of the role of the cellular microenvironment. This study focuses on the application of topographical and electrical field stimuli to human myogenic precursor cell (hMPC) cultures to assess the influences of the adherent direction, proliferation, and differentiation, and induce preconditioning-induced therapeutic benefits. First, a topographical surface of commercially available culture dishes was achieved by femtosecond laser texturing. The detachable biphasic electrical current system was then applied to the hMPCs cultured on laser-textured culture dishes. Laser-textured topographies were remarkably effective in inducing the assembly of hMPC myotubes by enhancing the orientation of adherent hMPCs compared with flat surfaces. Furthermore, electrical field stimulation through laser-textured topographies was found to promote the expression of myogenic regulatory factors compared with nonstimulated cells. As such, we successfully demonstrated that the combined stimulation of topographical and electrical cues could effectively enhance the myogenic maturation of hMPCs in a surface spatial and electrical field-dependent manner, thus providing the basis for therapeutic strategies.

6.
J Mater Chem B ; 9(27): 5560-5571, 2021 07 14.
Article in English | MEDLINE | ID: mdl-34169302

ABSTRACT

The utilization of cell-manipulating techniques reveals information about biological behaviors suited to address a wide range of questions in the field of life sciences. Here, we introduced an on/off switchable physical stimuli technique that offers precise stimuli for reversible cell patterning to allow regulation of the future direction of adherent cellular behavior by leveraging enzymatically degradable alginate hydrogels with defined chemistry and topography. As a proof of concept, targeted muscle cells adherent to TCP exhibited a reshaped structure when the hydrogel-based physical stimuli were applied. This simple tool offers easy manipulation of adherent cells to reshape their morphology and to influence future direction depending on the characteristics of the hydrogel without limitations of time and space. The findings from this study are broadly applicable to investigations into the relationships between cells and physiological extracellular matrix environments as well as has potential to open new horizons for regenerative medicine with manipulated cells.


Subject(s)
Dimethylpolysiloxanes/pharmacology , Extracellular Matrix/chemistry , Hydrogels/pharmacology , Animals , Cell Adhesion/drug effects , Cell Differentiation/drug effects , Cells, Cultured , Dimethylpolysiloxanes/chemical synthesis , Dimethylpolysiloxanes/chemistry , Hydrogels/chemical synthesis , Hydrogels/chemistry , Mice , Particle Size , Surface Properties
7.
Acta Biomater ; 116: 138-148, 2020 10 15.
Article in English | MEDLINE | ID: mdl-32890750

ABSTRACT

Posterior capsular opacification (PCO) is the most common complication of cataract surgery. PCO is due to the proliferation, migration, and epithelial-to-mesenchymal transition of the residual lens epithelial cells (LECs) within the lens capsule. As surface topography influences cellular response, we investigated the effect of modulating the dimensions of periodic nano-textured patterns on the surface of an intraocular lens material to regulate lens epithelial cell functions such as cell adhesion, migration, orientation, and proliferation. Patterned poly(HEMA) samples were prepared by a femtosecond laser microfabrication, and the behaviors of human B-3 LECs were observed on groove/ridge patterns with widths varying from 5 to 40 µm. In the presence of ridge and groove patterns, the adherent cells elongated along the direction of the patterns, and f-actin of the cells was spread to a lesser extent on the nano-textured groove surfaces. Both single and collective cell migrations were significantly inhibited in the perpendicular direction of the patterns on the nano-textured micro-patterned samples. We also fabricated the patterns on the curved surface of a commercially available intraocular lens for in vivo evaluation. In vivo results showed that a patterned IOL could help suppress the progression of PCO by inhibiting cell migration from the edge to the center of the IOL. Our reports demonstrate that nano- and microscale topographical patterns on a biomaterial surface can regulate cellular behavior when it is implanted into animals.


Subject(s)
Capsule Opacification , Lens Capsule, Crystalline , Lenses, Intraocular , Animals , Biocompatible Materials/pharmacology , Cell Movement , Epithelial Cells , Humans , Lasers
8.
Adv Mater ; 32(40): e2004300, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32820574

ABSTRACT

The native extracellular matrix (ECM) can exhibit heterogeneous nano-sequences periodically displaying ligands to regulate complex cell-material interactions in vivo. Herein, an ECM-emulating heterogeneous barcoding system, including ligand-bearing Au and ligand-free Fe nano-segments, is developed to independently present tunable frequency and sequences in nano-segments of cell-adhesive RGD ligand. Specifically, similar exposed surface areas of total Fe and Au nano-segments are designed. Fe segments are used for substrate coupling of nanobarcodes and as ligand-free nano-segments and Au segments for ligand coating while maintaining both nanoscale (local) and macroscale (total) ligand density constant in all groups. Low nano-ligand frequency in the same sequences and terminally sequenced nano-ligands at the same frequency independently facilitate focal adhesion and mechanosensing of stem cells, which are collectively effective both in vitro and in vivo, thereby inducing stem cell differentiation. The Fe/RGD-Au nanobarcode implants exhibit high stability and no local and systemic toxicity in various tissues and organs in vivo. This work sheds novel insight into designing biomaterials with heterogeneous nano-ligand sequences at terminal sides and/or low frequency to facilitate cellular adhesion. Tuning the electrodeposition conditions can allow synthesis of unlimited combinations of ligand nano-sequences and frequencies, magnetic elements, and bioactive ligands to remotely regulate numerous host cells in vivo.


Subject(s)
Cell Adhesion/drug effects , Cell Differentiation/drug effects , Nanotechnology/methods , Stem Cells/cytology , Stem Cells/drug effects , Cell Line , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Gold/chemistry , Humans , Iron/chemistry , Ligands , Oligopeptides/chemistry , Oligopeptides/pharmacology
9.
Adv Sci (Weinh) ; 7(15): 2000800, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32775162

ABSTRACT

Biodegradable metallic materials represent a potential step-change technology that may revolutionize the treatment of broken bones. Implants made with biodegradable metals are significantly stronger than their polymer counterparts and fully biodegradable in vivo, removing the need for secondary surgery or long-term complications. Here, it is shown how clinically approved Mg alloy promotes improved bone repair using an integrated state of the art fetal mouse metatarsal assay coupled with in vivo preclinical studies, second harmonic generation, secretome array analysis, perfusion bioreactor, and high-resolution 3D confocal imaging of vasculature within skeletal tissue, to reveal a vascular-mediated pro-osteogenic mechanism controlling enhanced tissue regeneration. The optimized mechanical properties and corrosion rate of the Mg alloy lead to a controlled release of metallic Mg, Ca, and Zn ions at a rate that facilitates both angiogenesis and coupled osteogenesis for better bone healing, without causing adverse effects at the implantation site. The findings from this study support ongoing development and refinement of biodegradable metal systems to act as crucial portal technologies with significant potential to improve many clinical applications.

10.
Nano Lett ; 20(6): 4188-4196, 2020 06 10.
Article in English | MEDLINE | ID: mdl-32406688

ABSTRACT

Developing materials with remote controllability of macroscale ligand presentation can mimic extracellular matrix (ECM) remodeling to regulate cellular adhesion in vivo. Herein, we designed charged mobile nanoligands with superparamagnetic nanomaterials amine-functionalized and conjugated with polyethylene glycol linker and negatively charged RGD ligand. We coupled negatively a charged nanoligand to a positively charged substrate by optimizing electrostatic interactions to allow reversible planar movement. We demonstrate the imaging of both macroscale and in situ nanoscale nanoligand movement by magnetically attracting charged nanoligand to manipulate macroscale ligand density. We show that in situ magnetic control of attracting charged nanoligand facilitates stem cell adhesion, both in vitro and in vivo, with reversible control. Furthermore, we unravel that in situ magnetic attraction of charged nanoligand stimulates mechanosensing-mediated differentiation of stem cells. This remote controllability of ECM-mimicking reversible ligand variations is promising for regulating diverse reparative cellular processes in vivo.


Subject(s)
Cell Adhesion , Magnetic Phenomena , Oligopeptides , Stem Cells , Cell Differentiation , Extracellular Matrix
11.
Sci Rep ; 10(1): 5969, 2020 04 06.
Article in English | MEDLINE | ID: mdl-32249803

ABSTRACT

Air-pollutants containing toxic particulate matters (PM) deposit in the respiratory tract and increases microbial infections. However, the mechanism by which this occurs is not well understood. This study evaluated the effect of urban particles (UP) on Streptococcus pneumoniae (pneumococcus) in vitro biofilm formation, colonization of human middle ear epithelium cells (HMEECs) as well as mouse nasal cavity and its transition to the middle ear and lungs. The in vitro biofilms and planktonic growth of S. pneumoniae were evaluated in metal ion free medium in the presence of UP. Biofilms were quantified by crystal violet (CV) microplate assay, colony forming unit (cfu) counts and resazurin staining. Biofilm structures were analyzed using a scanning electron microscope (SEM) and confocal microscopy (CM). Gene expressions of biofilms were evaluated using real time RT-PCR. Effects of UP exposure on S. pneumoniae colonization to HMEECs were evaluated using fluorescent in-situ hybridization (FISH), cell viability was detected using the Ezcyto kit, apoptosis in HMEECs were evaluated using Annexin-V/PI based cytometry analysis and reactive oxygen species (ROS) production were evaluated using the Oxiselect kit. Alteration of HMEECs gene expressions on UP exposure or pneumococci colonization was evaluated using microarray. In vivo colonization of pneumococci in the presence of UP and transition to middle ear and lungs were evaluated using an intranasal mice colonization model. The UP exposure significantly increased (*p < 0.05) pneumococcal in vitro biofilms and planktonic growth. In the presence of UP, pneumococci formed organized biofilms with a matrix, while in absence of UP bacteria were unable to form biofilms. The luxS, ply, lytA, comA, comB and ciaR genes involved in bacterial pathogenesis, biofilm formation and quorum sensing were up-regulated in pneumococci biofilms grown in the presence of UP. The HMEECs viability was significantly decreased (p < 0.05) and bacteria colonization was significantly elevated (p < 0.05) in co-treatment (UP + S. pneumoniae) when compared to single treatment. Similarly, increased apoptosis and ROS production were detected in HMEECs treated with UP + pneumococci. The microarray analysis of HMEECs revealed that the genes involve in apoptosis and cell death, inflammation, and immune response, were up-regulated in co-treatment and were unchanged or expressed in less fold in single treatments of UP or S. pneumoniae. The in vivo study showed an increased pneumococcal colonization of the nasopharynx in the presence of UP and a higher transition of bacteria to the middle ear and lungs in the presence of UP. The UP exposure elevated S. pneumoniae in vitro biofilm and colonization of HMEECs, and in vivo mouse nasopharyngeal colonization, and increased dissemination to mouse middle ear and lungs.


Subject(s)
Air Pollutants , Biofilms/drug effects , Ear, Middle/microbiology , Lung/microbiology , Nasopharynx/microbiology , Particulate Matter/administration & dosage , Streptococcus pneumoniae/drug effects , Animals , Gene Expression Regulation, Bacterial/drug effects , Humans , Mice , Quorum Sensing
12.
Sci Rep ; 8(1): 13818, 2018 09 14.
Article in English | MEDLINE | ID: mdl-30218086

ABSTRACT

The cytotoxicity of alloying elements in newly developed biodegradable metals can be assessed through relatively low-cost and rapid in vitro studies using different cell types. However, such approaches have limitations; as such, additional investigations in small mammalian models are required that recapitulate the physiological environment. In this study, we established a zebrafish (Danio rerio) model for cytotoxicity evaluations that combines the physiological aspects of an animal model with the speed and simplicity of a cell-based assay. The model was used to assess the cytotoxicity of five common alloying elements in biodegradable implant materials. Conventional in vitro testing using heart, liver, and endothelial cell lines performed in parallel with zebrafish studies revealed statistically significant differences in toxicity (up to 100-fold), along with distinct changes in the morphology of the heart, liver, and blood vessels that were undetectable in cell cultures. These results indicate that our zebrafish model is a useful alternative to mammalian systems for accurately and rapidly evaluating the in vivo toxicity of newly developed metallic materials.


Subject(s)
Alloys/toxicity , Metals/toxicity , Toxicity Tests/methods , Absorbable Implants , Alloys/metabolism , Animals , Animals, Genetically Modified/metabolism , Embryo, Nonmammalian , Metals/metabolism , Models, Animal , Zebrafish/metabolism
13.
Int J Mol Sci ; 19(3)2018 Mar 06.
Article in English | MEDLINE | ID: mdl-29509688

ABSTRACT

Electrospinning has been used for the fabrication of extracellular matrix (ECM)-mimicking fibrous scaffolds for several decades. Electrospun fibrous scaffolds provide nanoscale/microscale fibrous structures with interconnecting pores, resembling natural ECM in tissues, and showing a high potential to facilitate the formation of artificial functional tissues. In this review, we summarize the fundamental principles of electrospinning processes for generating complex fibrous scaffold geometries that are similar in structural complexity to the ECM of living tissues. Moreover, several approaches for the formation of three-dimensional fibrous scaffolds arranged in hierarchical structures for tissue engineering are also presented.


Subject(s)
Tissue Engineering/methods , Tissue Scaffolds/chemistry , Animals , Extracellular Matrix/chemistry , Extracellular Matrix/metabolism , Humans , Tissue Scaffolds/adverse effects
14.
J Biomed Mater Res A ; 106(6): 1732-1742, 2018 06.
Article in English | MEDLINE | ID: mdl-29468791

ABSTRACT

Biological responses on biomaterials occur either on their surface or at the interface. Therefore, surface characterization is an essential step in the fabrication of ideal biomaterials for achieving effective control of the interaction between the material surface and the biological environment. Herein, we applied femtosecond laser ablation on electrospun fibrous scaffolds to fabricate various hierarchical patterns with a focus on the alignment of cells. We investigated the simultaneously stimulated response of cardiomyoblasts based on multiple topographical cues, including scales, oriented directions, and spatial arrangements, in the fibrous scaffolds. Our results demonstrated a synergistic effect on cell behaviors of one or more structural arrangements in a homogeneous orientation, whereas antagonistic effects were observed for cells arranged on a surface with heterogeneous directions. Taken together, these results indicate that our hierarchically patterned fibrous scaffolds may be useful tools for understanding the cellular behavior on fibrous scaffolds used to mimic an extracellular matrix-like environment. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1732-1742, 2018.


Subject(s)
Biocompatible Materials/chemistry , Myoblasts, Cardiac/cytology , Tissue Scaffolds/chemistry , Animals , Cell Differentiation , Cell Line , Cell Movement , Cell Proliferation , Cell Survival , Electrochemical Techniques , Lasers , Rats , Surface Properties , Tissue Engineering/methods
15.
Adv Healthc Mater ; 6(9)2017 May.
Article in English | MEDLINE | ID: mdl-28230931

ABSTRACT

Although the coculture of multiple cell types has been widely employed in regenerative medicine, in vivo transplantation of cocultured cells while maintaining the hierarchical structure remains challenging. Here, a spatially assembled bilayer cell sheet of human mesenchymal stem cells and human umbilical vein endothelial cells on a thermally expandable hydrogel containing fibronectin is prepared and its effect on in vitro proangiogenic functions and in vivo ischemic injury is investigated. The expansion of hydrogels in response to a temperature change from 37 to 4 °C allows rapid harvest and delivery of the bilayer cell sheet to two different targets (an in vitro model glass surface and in vivo tissue). The in vitro study confirms that the bilayer sheet significantly increases proangiogenic functions such as the release of nitric oxide and expression of vascular endothelial cell genes. In addition, transplantation of the cell sheet from the hydrogels into a hindlimb ischemia mice model demonstrates significant retardation of necrosis particularly in the group transplated with the bilayer sheet. Collectively, the bilayer cell sheet is readily transferrable from the thermally expandable hydrogel and represents an alternative approach for recovery from ischemic injury, potentially via improved cell-cell communication.


Subject(s)
Hydrogels/chemistry , Animals , Cell Communication/drug effects , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Coculture Techniques , Hindlimb/pathology , Human Umbilical Vein Endothelial Cells/cytology , Humans , Hydrogels/pharmacology , Immunohistochemistry , Ischemia/therapy , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Neovascularization, Physiologic/physiology , Temperature , Tissue Engineering/methods
16.
J Mater Chem B ; 5(2): 318-328, 2017 Jan 14.
Article in English | MEDLINE | ID: mdl-32263550

ABSTRACT

A monolayer of endothelial cells (ECs) aligned along the direction of blood flow plays crucial roles in the regulation of anti-thrombogenic and pro-inflammatory reactions in the blood vessel wall. Thus, many researchers have attempted to mimic the aligned structure of ECs in vascular grafts or tissue-engineered blood vessels. In the present study, we fabricated micro-groove patterned nanofibers using a femtosecond laser ablation technique to recapitulate the densely organized anisotropic architecture of the endothelial layer. Femtosecond laser ablation enabled us to generate high-resolution groove patterns (10 µm width) with 20 or 80 µm gaps on randomly oriented electrospun nanofibers. The patterned nanofibers exhibited anisotropic (transverse: 101.1 ± 4.0° and longitudinal: 123.5 ± 9.4°) water contact angles; however, the mechanical properties were consistent in both directions. The micropatterned nanofibers modulated the aligned structure or aspect ratio (20 µm: 0.23 ± 0.11 and 80 µm: 0.42 ± 0.18) of ECs along the pattern direction. In particular, the engineered aligned endothelial layer was effective in eliciting an anti-inflammatory response (approximately 50% greater than that of random or aligned nanofibers), thereby effectively preventing monocyte adhesion following activation by TNF-α treatment. Therefore, micropatterning by laser ablation can be utilized to generate high-resolution microgrooves on various substrates, thereby providing fundamental platforms for vascular tissue engineering.

17.
Adv Healthc Mater ; 5(18): 2396-405, 2016 09.
Article in English | MEDLINE | ID: mdl-27390259

ABSTRACT

Implanted material surfaces make direct contact with body tissues to work on its own purpose. Therefore, studies of the surface properties of implantable materials that determine cell fate are very important for successful implantation. Although numerous studies have addressed the relationship between cells and material surfaces, nonmetallic surfaces and metallic surfaces likely produce different cellular responses because of their intrinsic differences in surface energy, roughness, and chemical composition. Moreover, given the nontransparent property of metal materials, which hampers the real-time imaging of cellular behavior, a detailed cellular-level analysis at the metal-tissue interface has not been performed. In this study, metal-based cell culture platforms (MCPs) with defined microscale topographical patterns are developed using a combination of photolithography and direct current magnetron sputtering techniques. The MCPs allow to observe vascular cells on metals in real time and identify the selective regulation of human aortic smooth muscle cells and Human umbilical vein endothelial cells (HUVECs) by metallic surface topography. Additionally, atomic force microscopy, contact angles, and energy-dispersive X-ray spectroscopy analyses show that the MCPs exhibit nearly identical chemical properties with their bulk counterparts, demonstrating that MCPs can be utilized as an in vitro cell culture platform system for understanding the cellular behavior on metal substrates.


Subject(s)
Aorta/metabolism , Human Umbilical Vein Endothelial Cells/metabolism , Membranes, Artificial , Metals/chemistry , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle/metabolism , Aorta/cytology , Cell Culture Techniques , Human Umbilical Vein Endothelial Cells/cytology , Humans , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/cytology , Surface Properties
18.
ACS Appl Mater Interfaces ; 8(5): 3407-17, 2016 Feb 10.
Article in English | MEDLINE | ID: mdl-26771693

ABSTRACT

Developing an artificial extracellular matrix that closely mimics the native tissue microenvironment is important for use as both a cell culture platform for controlling cell fate and an in vitro model system for investigating the role of the cellular microenvironment. Electrospinning, one of the methods for fabricating structures that mimic the native ECM, is a promising technique for creating fibrous platforms. It is well-known that align or randomly distributed electrospun fibers provide cellular contact guidance in a single pattern. However, native tissues have hierarchical structures, i.e., topographies on the micro- and nanoscales, rather than a single structure. Thus, we fabricated randomly distributed nanofibrous (720 ± 80 nm in diameter) platforms via a conventional electrospinning process, and then we generated microscale grooves using a femtosecond laser ablation process to develop engineered fibrous platforms with patterned hierarchical topographies. The engineered fibrous platforms can regulate cellular adhesive morphology, proliferation, and distinct distribution of focal adhesion proteins. Furthermore, confluent myoblasts cultured on the engineered fibrous platforms revealed that the direction of myotube assembly can be controlled. These results indicate that our engineered fibrous platforms may be useful tools in investigating the roles of nano- and microscale topographies in the communication between cells and ECM.


Subject(s)
Biomimetics , Extracellular Matrix/ultrastructure , Myoblasts/ultrastructure , Tissue Engineering , Cell Adhesion/drug effects , Cell Communication/drug effects , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Cellular Microenvironment , Extracellular Matrix/chemistry , Myoblasts/chemistry
19.
Biomaterials ; 54: 44-54, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25907038

ABSTRACT

Development of stem cell delivery system with ability of control over mutilineage differentiation and improved engraft efficiency is imperative in regenerative medicine. We herein report transfer stamping of human mesenchymal stem cells (hMSCs) patches using thermally expandable hydrogels with tunable cell-adhesive properties. The hydrogels were prepared from functionalized four arm copolymer of Tetronic(®), and the cell adhesion on the hydrogel was modulated by incorporation of fibronectin (FN) or cell-adhesive peptide (RGD). The resulting hydrogels showed spontaneous expansion in size within 10 min in response to the temperature reduction from 37 to 4°C. The adhesion and proliferation of hMSCs on FN-hydrogels were positively tunable in proportion to the amount of FN within hydrogels with complete monolayer of hMSCs (hMSC patch) being successfully achieved. The hMSC patch on the hydrogel was faced to the target substrate, which was then easily detached and re-attached to the target when the temperature was reduced from 37°C up to 4°C. We found that the transfer stamping of cell patch was facilitated at lower temperature of 4°C relative to 25°C, with the use of thinner hydrogels (0.5 mm in thickness relatively to 1.0 or 1.5 mm) and longer transfer time (>15 min). Notably, the hMSC patch was simply transferred from the hydrogel to the subcutaneous mouse skin tissue within 15 min with cold saline solution being dropped to the hydrogel. The hMSC patch following osteogenic or adipogenic commitment was also achieved with long-term culture of hMSCs on the hydrogel, which was successfully detached to the target surface. These results suggest that the hydrogels with thermally expandable and tunable cell-adhesive properties may serve as a universal substrate to harvest hMSC patch in a reliable and effective manner, which could potentially be utilized in many cell-sheet based therapeutic applications.


Subject(s)
Batch Cell Culture Techniques/instrumentation , Hydrogels/chemistry , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/physiology , Molecular Imprinting/methods , Tissue Scaffolds , Batch Cell Culture Techniques/methods , Cell Adhesion/physiology , Cell Proliferation/physiology , Cell Survival/physiology , Cells, Cultured , Equipment Design , Equipment Failure Analysis , Hot Temperature , Humans , Materials Testing , Tissue Engineering/instrumentation , Tissue Engineering/methods
20.
Adv Healthc Mater ; 3(9): 1465-74, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24610737

ABSTRACT

Natural vessel has three types of concentric cell layers that perform their specific functions. Here, the fabrication of vascular structure is reported by transfer printing of three different cell layers using thermosensitive hydrogels. Tetronic-tyramine and RGD peptide are co-crosslinked to prepare cell adhesive and thermosensitive hydrogels. The hydrogel increases its diameter by 1.26 times when the temperature reduces from 37 °C to 4 °C. At optimized seeding density, three types of cells form monolayers on the hydrogel, which is then transferred to the target surface within 3 min. Three monolayers are simultaneously transferred on one substrate with controlled shape and arrangement. The same approach is applied onto nanofiber scaffolds that are cultured for more than 5 d. Every type of monolayer shows proliferation and migration on nanofiber scaffolds, and the formation of robust cell-cell contact is revealed by CD31 staining in endothelial cell layer. A vascular structure with multicellular components is fabricated by transfer of three monolayers on nanofibers that are manually rolled with the diameter and length of the tube being approximately 3 mm and 12 mm, respectively. Collectively, it is concluded that the tissue transfer printing is a useful tool for constructing a vascular structure and mimicking natural structure of different types of tissues.


Subject(s)
Bioprinting/methods , Blood Vessels/cytology , Tissue Engineering/instrumentation , Cell Adhesion , Cell Line , Cell Proliferation , Cells, Cultured , Glass , Human Umbilical Vein Endothelial Cells , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Materials Testing , Models, Biological , Nanofibers/chemistry , Temperature , Tissue Scaffolds/chemistry
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