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1.
Silane modification is a simple and cost-effective tool to modify existing biomaterials for tissue engineering applications. Aminosilane layer deposition has previously been shown to control NG108-15 neuronal cell and primary Schwann cell adhesion and differentiation by controlling deposition of ─NH2 groups at the submicron scale across the entirety of a surface by varying silane chain length. This is the first study toreport depositing 11-aminoundecyltriethoxysilane (CL11) onto aligned Polycaprolactone (PCL) scaffolds for peripheral nerve regeneration. Fibers are manufactured via electrospinning and characterized using water contact angle measurements, atomic force microscopy (AFM), and X-ray photoelectron spectroscopy (XPS). Confirmed modified fibers are investigated using in vitro cell culture of NG108-15 neuronal cells and primary Schwann cells to determine cell viability, cell differentiation, and phenotype. CL11-modified fibers significantly support NG108-15 neuronal cell and Schwann cell viability. NG108-15 neuronal cell differentiation maintains Schwann cell phenotype compared to unmodified PCL fiber scaffolds. 3D ex vivo culture of Dorsal root ganglion explants (DRGs) confirms further Schwann cell migration and longer neurite outgrowth from DRG explants cultured on CL11 fiber scaffolds compared to unmodified scaffolds. Thus, a reproducible and cost-effective tool is reported to modify biomaterials with functional amine groups that can significantly improve nerve guidance devices and enhance nerve regeneration.  相似文献   

2.
Neural tissue engineering has become a potential technology to restore the functionality of damaged neural tissue with the hope to cure the patients with neural disorder and to improve their quality of life. This paper reports the design and synthesis of polypeptides containing neuron stimulate, glutamic acid, for the fabrication of biomimetic 3D scaffold in neural tissue engineering application. The polypeptides are synthesized by efficient chemical reactions. Monomer γ‐benzyl glutamate‐N‐carboxyanhydride undergoes ring‐opening polymerization to form poly(γ‐benzyl‐l ‐glutamate), then hydrolyzes into poly(γ‐benzyl‐l ‐glutamate)‐r‐poly(glutamic acid) random copolymer. The glutamic acid amount is controlled by hydrolysis time. The obtained polymer molecular weight is in the range of 200 kDa for good quality of fibers. The fibrous 3D scaffolds of polypeptides are fabricated using electrospinning techniques. The scaffolds are biodegradable and biocompatible. The biocompatibility and length of neurite growth are improved with increasing amount of glutamic acid in scaffold. The 3D scaffold fabricated from aligned fibers can guide anisotropic growth of neurite along the fiber and into 3D domain. Furthermore, the length of neurite outgrowth is longer for scaffold made from aligned fibers as compared with that of isotropic fibers. This new polypeptide has potential for the application in the tissue engineering for neural regeneration.  相似文献   

3.
We report the photothermal properties as well as the in vitro cell test results of titanium oxide nanotubes (TiO2 NTs) as a potential therapeutic agent for cancer thermotherapy in combination with near-infrared (NIR) light. TiO2 NTs are found to have a higher photothermal effect upon exposure to NIR laser than Au nanoparticles and single-wall carbon nanotubes, which have also attracted considerable interest as therapeutic agents for cancer thermotherapy. The temperature increase of a TiO2 NT/NaCl suspension during NIR laser exposure is larger than that of a TiO2 NT/D.I. water suspension due to the heat generated by the formation of Na2TiF6. According to the in vitro cell test results the cells exposed to NIR laser without TiO2 NT treatment have a cell viability of 96.4%. Likewise, the cells treated with TiO2 NTs but not with NIR irradiation also have a cell viability of 98.2%. Combination of these two techniques, however, shows a cell viability of 1.35%. Also, the cell deaths are mostly due to necrosis but partly due to late apoptosis. These results suggest that TiO2 NTs can be used effectively as therapeutic agents for cancer thermotherapy due to their excellent photothermal properties and high biocompatibility.  相似文献   

4.
A nano-structured scaffold was designed for bone repair using collagen, hyaluronic acid (HYA) and nano-bioactive glass (NBaG) as its main components. The collagen-HYA/NBaG scaffold was prepared by using a freeze-drying technique and characterized by scanning electron microscopy (SEM). Osteoblastls were seeded on these scaffolds and their proliferation rate, alkaline phosphatase (ALP) activity and ability to form mineralized bone nodules were compared with those osteoblasts grown on cell culture plastic surfaces. The cross-section morphology shows that the collagen-HYA/NBaG scaffold possessed a three-dimensional (3D) interconnected homogenous porous structure. The results obtained from biological assessment show that this scaffold did not negatively affect osteoblasts proliferation rate and improves osteoblasts function as shown by increasing the ALP activity and calcium deposition and formation of mineralized bone nodules. Therefore, the composite scaffolds could provide a favorable environment for initial cell adhesion, maintained cell viability and cell proliferation, and had good in-vitro biocompatibility.  相似文献   

5.
2D in vitro studies have demonstrated that Schwann cells prefer scaffolds with mechanical modulus approximately 10× higher than the modulus preferred by nerves, limiting the ability of many scaffolds to promote both neuron extension and Schwann cell proliferation. Therefore, the goals of this work are to develop and characterize microgel‐based scaffolds that are tuned over the stiffness range relevant to neural tissue engineering and investigate Schwann cell morphology, viability, and proliferation within 3D scaffolds. Using thiol‐ene reaction, microgels with surface thiols are produced and crosslinked into hydrogels using a multiarm vinylsulfone (VS). By varying the concentration of VS, scaffold stiffness ranges from 0.13 to 0.76 kPa. Cell morphology in all groups demonstrates that cells are able to spread and interact with the scaffold through day 5. Although the viability in all groups is high, proliferation of Schwann cells within the scaffold of G* = 0.53 kPa is significantly higher than other groups. This result is ≈5× lower than previously reported optimal stiffnesses on 2D surfaces, demonstrating the need for correlation of 3D cell response to mechanical modulus. As proliferation is the first step in Schwann cell integration into peripheral nerve conduits, these scaffolds demonstrate that the stiffness is a critical parameter to optimizing the regenerative process.

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6.
Polyester‐based scaffolds covalently functionalized with arginine‐glycine‐aspartic acid‐cysteine (RGDC) peptide sequences support the proliferation and osteogenic differentiation of stem cells. The aim is to create an optimized 3D niche to sustain human bone marrow stem cell (hBMSC) viability and osteogenic commitment, without reliance on differentiation media. Scaffolds consisting of poly(lactide‐co‐trimethylene carbonate), poly(LA‐co‐TMC), and functionalized poly(lactide) copolymers with pendant thiol groups are prepared by salt‐leaching technique. The availability of functional groups on scaffold surfaces allows for an easy and straightforward method to covalently attach RGDC peptide motifs without affecting the polymerization degree. The strategy enables the chemical binding of bioactive motifs on the surfaces of 3D scaffolds and avoids conventional methods that require harsh conditions. Gene and protein levels and mineral deposition indicate the osteogenic commitment of hBMSC cultured on the RGDC functionalized surfaces. The osteogenic commitment of hBMSC is enhanced on functionalized surfaces compared with nonfunctionalized surfaces and without supplementing media with osteogenic factors. Poly(LA‐co‐TMC) scaffolds have potential as scaffolds for osteoblast culture and bone grafts. Furthermore, these results contribute to the development of biomimetic materials and allow a deeper comprehension of the importance of RGD peptides on stem cell transition toward osteoblastic lineage.  相似文献   

7.
Ultraviolet-A (365 nm, 120 kJ/m2/h) exposure caused cell death in Pseudomonas aeruginosa at doses at which Escherichia coli cell viability was not affected. We have not found that UVA induced growth delay or any other sublethal effect. Irradiated suspensions of P. aeruginosa showed a marked reduction in membrane-bound succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH) activities. Succinate-driven respiration and several nutrient transport systems were also inhibited. Whereas SDH and LDH activities were independent of the irradiation conditions, cell viability, respiration and transport systems were protected when irradiation was performed in an N2 atmosphere. A similar protective effect was observed when cells were grown in media containing glycerol or when preirradiation bacterial growth was carried out at 30°C (instead of 37°C). Results suggest that UVA induces a differential damaging effect on several biochemical functions of P. aeruginosa. The UVA induced photodamage may fall into two categories: indirect damage mediated by oxygen (cell killing and inhibition of respiration and transport systems) and direct damage to SDH and LDH (apparently not oxygen dependent). These enzymes and leucine transport appear not to be involved in the lethal effect described herein because they were altered despite viability-preserving conditions.  相似文献   

8.
《先进技术聚合物》2018,29(7):2050-2063
Electrospinning has been extensively used to construct tissue‐engineered scaffolds because of its ability to provide the fibrous scaffold with structurally analogous to the naturally occurring protein in the extracellular matrix of native tissues. In addition, the modification of scaffolds with bioactive molecules is beneficial as this can create an environment that consists of biochemical cues to further promote cell adhesion, proliferation, and differentiation. In the present contribution, we prepared and investigated the potential used of aligned electrospun poly(3‐hydroxybutyrate) (PHB) scaffold immobilized with bioactive molecule to serve as nervous scaffold. Laminin was successfully immobilized on the surface using covalent binding between functional groups of modified scaffolds and protein. The ability to use for neural regeneration was evaluated in vitro towards murine neuroblastoma Neuro2a cell line and mouse brain‐derived neural stem cells. The surface modification with laminin immobilized on the PHB fibrous scaffolds supported the attachment and promoted the proliferation of Neuro2a very wells. Despite the good attachment and proliferation of Neuro2a and mouse brain‐derived neural stem cells were not able to proliferate on the neat PHB, hydrolyzed PHB and laminin immobilized on hydrolyzed PHB fibrous scaffold.  相似文献   

9.
A linear Union Carbide PE (UC) has been analyzed by nonstandard calorimetry with a common DSC calorimeter and a Setaram C80 calorimeter. Nonstandard calorimetry entails using a low rate of heating (0.5–1 K/min), a small mass (0.5–1.5 mg), and an open cell (O‐cell) instead of the standard C‐cell. Events in O‐cells overlap less and occur with a faster kinetics than in C‐cells. PE crystals are nascent, solution‐grown (S‐grown), press‐grown (P‐grown), and strained by extrusion. In Part A, the traces show that the phase‐changes in the melt, previously observed in a C80 calorimeter (slow T‐ramp) and characterized by ΔHnetwork, can be observed with a common DSC in nonstandard conditions. In Part B, the difference between the C‐ and O‐cells and the changes in the main peak enthalpy (ΔHortho) are of interest. The main result is that, in O‐cells, the value of ΔHortho around Tortho, exceeds unambiguously in certain conditions ΔHortho found for perfect orthorhombic crystals. The main endotherm contains then another contribution, namely ΔHnetwork. Crystal reorganization during the slow T‐ramp is followed in the C‐ and O‐cells on S‐grown crystals. In O‐cells, lamellar thickening observed in the slow‐ramp is more extensive. The ease of phase‐change depends on the sample history. It is as follows: strained‐part extruded > nascent > S‐grown > P‐grown. Co‐operative chain motions are more hindered in the standard C‐cells than in the O‐cells. In Part C, lower values of m succeed in bringing phase‐changes in P‐grown (O‐cells) samples. The origin of the events is discussed: three processes are thought to contribute to the phase‐changes namely, melting of strained short‐range order, activation of vibrations in the CH2 groups, and fast decay of chain orientation which occurs simultaneously with melting. © 2007 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 1932–1949, 2007  相似文献   

10.
Synthetic hydrogels can be used as scaffolds that not only favor endothelial cells(ECs) proliferation but also manipulate the behaviors and functions of the ECs.In this review paper,the effect of chemical structure,Young’s modulus (E) and zeta potential(ξ) of synthetic hydrogel scaffolds on static cell behaviors,including cell morphology,proliferation, cytoskeleton structure and focal adhesion,and on dynamic cell behaviors,including migration velocity and morphology oscillation,as well as on EC function such as anti-platelet adhesion,are reported.It was found that negatively charged hydrogels,poly(2-acrylamido-2-methylpropanesulfonic sodium)(PNaAMPS) and poly(sodium p-styrene sulphonate) (PNaSS),can directly promote cell proliferation,with no need of surface modification by any cell-adhesive proteins or peptides at the environment of serum-containing medium.In addition,the Young’s modulus(E) and zeta potential(ξ) of hydrogel scaffolds are quantitatively tuned by copolymer hydrogels,poly(NaAMPS-co-DMAAm) and poly(NaSS-co-DMAAm), in which the two kinds of negatively charged monomers NaAMPS and NaSS are copolymerized with neutral monomer,N,N-dimethylacrylamide(DMAAm).It was found that the critical zeta potential of hydrogels manipulating EC morphology,proliferation,and motility isξcritical= -20.83 mV andξcritical= -14.0 mV for poly(NaAMPS-co-DMAAm) and poly(NaSS-co-DMAAm),respectively.The above mentioned EC behaviors well correlate with the adsorption of fibronectin, a kind of cell-adhesive protein,on the hydrogel surfaces.Furthermore,adhered platelets on the EC monolayers cultured on the hydrogel scaffolds obviously decreases with an increase of the Young’s modulus(E) of the hydrogels,especially when E>60 kPa.Glycocalyx assay and gene expression of ECs demonstrate that the anti-platelet adhesion well correlates with the EC-specific glycocalyx.The above investigation suggests that understanding the relationship between physic-chemical properties of synthetic hydrogels and cell responses is essential to design optimal soft and wet scaffolds for tissue engineering.  相似文献   

11.
The highly complex nature of spinal cord injuries (SCIs) requires design of novel biomaterials that can stimulate cellular regeneration and functional recovery. Promising SCI treatments use biomaterial scaffolds, which provide bioactive cues to the cells in order to trigger neural regeneration in the spinal cord. In this work, the use of peptide nanofibers is demonstrated, presenting protein binding and cellular adhesion epitopes in a rat model of SCI. The self‐assembling peptide molecules are designed to form nanofibers, which display heparan sulfate mimetic and laminin mimetic epitopes to the cells in the spinal cord. These neuroactive nanofibers are found to support adhesion and viability of dorsal root ganglion neurons as well as neurite outgrowth in vitro and enhance tissue integrity after 6 weeks of injury in vivo. Treatment with the peptide nanofiber scaffolds also show significant behavioral improvement. These results demonstrate that it is possible to facilitate regeneration especially in the white matter of the spinal cord, which is usually damaged during the accidents using bioactive 3D nanostructures displaying high densities of laminin and heparan sulfate‐mimetic epitopes on their surfaces.  相似文献   

12.
This study was aimed to prepare biodegradable and porous nanocomposite scaffolds with microtubular orientation structure as a model for nerve tissue engineering by thermally induced phase separation (TIPS) method using dioxane as the solvent, crystalline poly (L‐lactic acid) (PLLA) and multi‐walled carbon nanotubes (MWCNTs). In order to overcome dispersion of MWCNTs in the PLLA matrix, heparinization of MWCNTs was performed. Solvent crystallization, oriented structure, the mean pore diameter and porosity percentage of the scaffolds were controlled by fundamental system parameters including temperature‐gradient of the system, polymer solution concentration and carbon nanotube content. Scanning Electron Microscopy (SEM), ImageJ, software and dynamic mechanical thermal analysis (DMTA) were used to investigate the structural and mechanical properties. TEM observation was carried out for characterization of nanotube dispersion in PLLA. It was found that the scaffolds containing heparinized multi‐walled carbon nanotubes (HMWCNTs) exhibited higher storage modulus, better carbon nanotube (CNT) dispersion and tubular orientation structure than those with non heparinized MWCNTs. In‐vitro studies were also conducted by using murine P19 cell line as a suitable model system to analyze neuronal differentiation over a 2‐week period. Immunofluorescence and DAPI staining were used to confirm the cells' attachment and differentiation on the PLLA/HMWCNT nanocomposite scaffolds. Based on the results, we can conclude that the PLLA/HMWCNT scaffolds enhanced the nerve cell differentiation and proliferation, and therefore, acted as a positive cue to support neurite outgrowth. Copyright © 2015 John Wiley & Sons, Ltd.  相似文献   

13.
With the ability to form a submicron-sized fibrous structure with interconnected pores mimicking the extracellular matrix (ECM) for tissue formation, electrospinning was used to fabricate ultra-fine fiber mats of hexanoyl chitosan (H-chitosan) for potential use as skin tissue scaffolds. In the present communication, the in vitro biocompatibility of the electrospun fiber mats was evaluated. Indirect cytotoxicity evaluation of the fiber mats with mouse fibroblasts (L929) revealed that the materials were non-toxic and did not release substances harmful to living cells. The potential for use of the fiber mats as skin tissue scaffolds was further assessed in terms of the attachment and the proliferation of human keratinocytes (HaCaT) and human foreskin fibroblasts (HFF) that were seeded or cultured on the scaffolds at different times. The results showed that the electrospun fibrous scaffolds could support the attachment and the proliferation of both types of cells, especially for HaCaT. In addition, the cells cultured on the fibrous scaffolds exhibited normal cell shapes and integrated well with surrounding fibers. The obtained results confirmed the potential for use of the electrospun H-chitosan fiber mats as scaffolds for skin tissue engineering.  相似文献   

14.
The viability of bacteria plays a critical role in the enhancement of fossil fuels biodesulfurization efficiency since cells are exposed to toxic compounds such as 2-hydroxybiphenyl (2-HBP), the end product of dibenzothiophene (DBT) biodesulfurization. The goal of this work was to study the influence of the carbon source on the resistance of Gordonia alkanivorans strain 1B to 2-HBP. The physiological response of this bacterium, pre-grown in glucose or fructose, to 2-HBP was evaluated using two approaches: a growth inhibition toxicity test and flow cytometry. The results obtained from the growth inhibition bioassays showed that the carbon source has an influence on the sensitivity of strain 1B growing cells to 2-HBP. The highest IC50 value was obtained for the assay using fructose as carbon source in both inoculum growth and test medium (IC50-48 h?=?0.464 mM). Relatively to the evaluation of 2-HBP effect on the physiological state of resting cells by flow cytometry, the results showed that concentrations of 2-HBP >1 mM generated significant loss of cell viability. The higher the 2-HBP concentration, the higher the toxicity effect on cells and the faster the loss of cell viability. In overall, the flow cytometry results highlighted that strain 1B resting cells grown in glucose-SO4 or glucose-DBT are physiologically less resistant to 2-HBP than resting cells grown in fructose-SO4 or fructose-DBT, respectively.  相似文献   

15.
A new cell‐printed scaffold consisting of poly(ϵ‐caprolactone) (PCL) and cell‐embedded alginate struts is designed. The PCL and alginate struts are stacked in an interdigitated pattern in successive layers to acquire a three‐dimensional (3D) shape. The hybrid scaffold exhibits a two‐phase structure consisting of cell (MC3T3‐E1)‐laden alginate struts able to support biological activity and PCL struts able to provide controllable mechanical support of the cell‐laden alginate struts. The hybrid scaffolds exhibit an impressive increase in tensile modulus and maximum strength compared to pure alginate scaffolds. Laden cells are homogeneously distributed throughout the alginate struts and the entire scaffold, resulting in cell viability of approximately 84%.  相似文献   

16.
The high ionic conductive polymer electrolytes were prepared based on poly(vinylidenefluoride) (PVDF) fibers modified via preirradiation grafting poly(methyl methacrylate) (PMMA). In these polymer electrolytes, the PVDF fibers served as the supporting phase providing dimensional stability, and PMMA acted as the gel phase helping for the trapping liquid electrolyte and substituting the nonconductive PVDF phase to provide contact with electrodes well thus increasing conductive area. The modified PVDF fibrous membranes were used as a polymer electrolyte in lithium ion battery after they were activated by uptaking 1 M LiPF6/ethylene carbonate–dimethyl carbonate (1:1 vol) liquid electrolyte, which showed a much higher room-temperature ionic conductivity than the pristine PVDF fibrous membrane. The LiCoO2-mesocarbon microbead coin cells containing the dual-phase fibrous membrane (degree of graft, 111.8%) demonstrated excellent rate performance, and the cell still retained about 86% of discharge capacity at 4C rate, as compared to that at 0.1C rate. The prototype cell showed good cycle performance.  相似文献   

17.
采用磷酸四钙和磷酸氢钙混合粉末制备了nCa/nP比为1.58的非化学计量羟基磷灰石骨水泥(n-HAC)及其多孔支架材料。结果表明:与nCa/nP=1.67的化学计量羟基磷灰石骨水泥(HAC)相比,n-HAC的凝结时间和抗压强度没有明显的区别。XRD和IR显示:n-HAC与HAC都为羟基磷灰石结构,但n-HAC在Tris-HCl缓冲溶液的降解性明显大于HAC。细胞培养结果表明:成骨细胞在n-HAC和HAC两种材料上的粘附和细胞形态没有明显的区别,但细胞在n-HAC上的增殖率明显高于HAC。将多孔n-HAC支架材料植入兔股骨缺损处,观察其修复骨缺损情况,组织学分析结果表明:新生骨在多孔支架的表面形成,并长入其内部;n-HAC在体内的降解比HAC快,能明显地促进新骨生成。  相似文献   

18.
[Cu(pydc)(eim)3]?H2O (1), [Cu(pydc)(4hp)(H2O)] (2), and [Ni(pydc)(3hp)(H2O)2][Cu(pydc)(3hp)(H2O)2]?3H2O (3) (H2pydc = 2,6-pyridinedicarboxylic acid or dipicolinic acid, eim = 2-ethylimidazole, 4hp = 4-hydroxypyridine, 3hp = 3-hydroxypyridine) were synthesized and characterized by elemental analysis, spectroscopic measurements (UV–vis and IR spectra), and single-crystal X-ray diffraction. Crystal analysis revealed that the complexes extended to 3-D supramolecular networks through intermolecular H-bonding and molecular interactions between the ligand moieties and water molecules. The thermal stabilities of complexes are investigated by thermogravimetry, differential thermogravimetry, and differential thermal analysis techniques. The effects of complexes on the proliferation of HT-1080 fibrosarcoma cells were investigated using the quick cell proliferation assay. The cell viability changes were found to depend on the concentrations and type of complex.  相似文献   

19.
One of the major challenges in the processing of hydrogels based on poly(ethylene glycol) (PEG) is to create mechanically robust electrospun hydrogel scaffolds without chemical crosslinking postprocessing. In this study, this is achieved by the introduction of physical crosslinks in the form of supramolecular hydrogen bonding ureido‐pyrimidinone (UPy) moieties, resulting in chain‐extended UPy‐PEG polymers (CE‐UPy‐PEG) that can be electrospun from organic solvent. The resultant fibrous meshes are swollen in contact with water and form mechanically stable, elastic hydrogels, while the fibrous morphology remains intact. Mixing up to 30 wt% gelatin with these CE‐UPy‐PEG polymers introduce bioactivity into these scaffolds, without affecting the mechanical properties. Manipulating the electrospinning parameters results in meshes with either small or large fiber diameters, i.e., 0.63 ± 0.36 and 2.14 ± 0.63 µm, respectively. In that order, these meshes provide support for renal epithelial monolayer formation or a niche for the culture of cardiac progenitor cells.  相似文献   

20.
Sufficient vascularization is critical to sustaining viable tissue‐engineered (TE) constructs after implantation. Despite significant progress, current approaches lack suturability, porosity, and biodegradability, which hinders rapid perfusion and remodeling in vivo. Consequently, TE vascular networks capable of direct anastomosis to host vasculature and immediate perfusion upon implantation still remain elusive. Here, a hybrid fabrication method is presented for micropatterning fibrous scaffolds that are suturable, porous, and biodegradable. Fused deposition modeling offers an inexpensive and automated approach to creating sacrificial templates with vascular‐like branching. By electrospinning around these poly(vinyl alcohol) templates and dissolving them in water, microvascular patterns were transferred to fibrous scaffolds. Results indicated that these scaffolds have sufficient suture retention strength to permit direct anastomosis in future studies. Vascularization of these scaffolds is demonstrated by in vitro endothelialization and perfusion.  相似文献   

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