ABSTRACT
Extreme environmental conditions often lead to irreversible structural failure and functional degradation in hydrogels, limiting their service life and applicability. Achieving high toughness, self-healing, and ionic conductivity in cryogenic environments is vital to broaden their applications. Herein, we present a novel approach to simultaneously enhance the toughness, self-healing, and ionic conductivity of hydrogels, via inducing non-freezable water within the zwitterionic cellulose-based hydrogel skeleton. This approach enables resulting hydrogel to achieve an exceptional toughness of 10.8 MJ m-3, rapid self-healing capability (98.9 % in 30 min), and high ionic conductivity (2.9 S m-1), even when subjected to -40 °C, superior to the state-of-the-art hydrogels. Mechanism analyses reveal that a significant amount of non-freezable water with robust electrostatic interactions is formed within zwitterionic cellulose nanofibers-modified polyurethane molecular networks, imparting superior freezing tolerance and versatility to the hydrogel. Importantly, this strategy harnesses the non-freezable water molecular state of the zwitterionic cellulose nanofibers network, eliminating the need for additional antifreeze and organic solvents. Furthermore, the dynamic Zn coordination within these supramolecular molecule chains enhances interfacial interactions, thereby promoting rapid subzero self-healing and exceptional mechanical strength. Demonstrating its potential, this hydrogel can be used in smart laminated materials, such as aircraft windshields.
ABSTRACT
Diabetes is one of the most prevalent diseases worldwide. The tissue regeneration of diabetes patients is known to be rather tricky as the result of vascular dysfunction, and this leads to various clinical complications including diabetic foot ulcers. The vascular endothelial cells, which compactly line the inner surface of blood vessels, are responsible for the growth and maintenance of blood vessels and play an essential role in tissue regeneration. Although the mechanical properties of cells are generally known to be regulated by physiological/pathological conditions, few studies have been performed to investigate vascular endothelial cellular mechanics under hyperglycemia and the biological functions related to tissue regeneration. In this study, we conduct a systematic investigation of this issue. The results suggested that the stiffness of human umbilical vein endothelial cells (HUVECs) can be significantly regulated by the glucose concentration, subsequently, leading to significant alterations in cell migration and proliferation capabilities that are closely related to tissue regeneration. The rearrangement of the cytoskeleton induced by hyperglycemia through Cdc42 was found to be one of the pathways for the alteration of the cell stiffness and the subsequent cell dysfunctions. Therefore, we suggested that the inhibition of Cdc42 might be a promising strategy to facilitate various tissue regeneration for diabetes patients.
ABSTRACT
Poly(lactic acid) (PLA) composites reinforced with cellulose nanocrystals (CNCs) are promising biodegradable materials. However, the poor compatibility and dispersion of CNCs in the PLA matrix remain a significant obstacle to improving the properties of composites. In this study, the modified CNC (CNC-D) was prepared through sulfonation treatment, followed by modification with didecyl dimethyl ammonium chloride (DDAC). Then, CNC-D was mixed with PLA to prepare composite films (PLA-CNC-D). The results revealed that the PLA-CNC-D had higher tensile strength and elongation at break than PLA-CNC at 3 wt% nanofiller content, increasing by 41.53 and 22.18 %, respectively. SEM and DSC analysis indicated that surface modification improved the compatibility and dispersion of CNC-D in the PLA matrix. The sulfonation process increased the anion content on the surface of CNC-D, enabling the CNC-D surface to adsorb more cationic DDAC, consequently sharply reducing the hydrophilicity of CNC-D. Moreover, the PLA-CNC-D exhibited excellent antibacterial activity against S. aureus and E. coli. In summary, this study provides a novel CNC modification approach to enhance the physical properties and antibacterial activity of PLA composite films, enlarging the application of degradable PLA composites.
Subject(s)
Ammonium Compounds , Nanoparticles , Quaternary Ammonium Compounds , Cellulose/chemistry , Polymers/chemistry , Escherichia coli , Staphylococcus aureus , Polyesters/chemistry , Nanoparticles/chemistry , Anti-Bacterial Agents/pharmacologyABSTRACT
BACKGROUND: Titanium (Ti) is now widely used as implant material due to its excellent mechanical properties and superior biocompatibilities, while its inert bioactivities might lead to insufficient osseointegration, and limit its performance in dental applications. METHODS: We introduced a robust and simple approach of modifying titanium surfaces with polysaccharide complexes. Titanium samples were subjected to hydrothermal treatment to create a uniform porous structure on the surface, followed by coating with a bioinspired and self-assembly polydopamine layer. Strontium Eucommia Ulmoides Polysaccharide (EUP-Sr) complexes are then introduced to the polydopamine-coated porous titanium. Multiple morphological and physiochemical characterizations are employed for material evaluation, while cell proliferation and gene expression tests using macrophages, primary alveolar bone osteoblasts, and vascular endothelial cells are used to provide an overall insight into the functions of the product. The significances of statistical differences were analyzed using student's t-test. RESULTS: Microscopic and spectrometric characterizations confirmed that the Ti surface formed a porous structure with an adequate amount of EUP-Sr loading. The attachment was attributed to hydrogen bonding between the ubiquitous glycosidic linkage of the polysaccharide complex and the ring structure of polydopamine, yet the loaded EUP-Sr complex can be gradually released, consequently benefiting the neighboring microenvironment. Cell experiments showed no cytotoxicity of the material, and the product showed promising anti-inflammation, osseointegration, and angiogenesis properties, which were further confirmed by in vivo evaluations. CONCLUSION: We believe the EUP-Sr modified titanium implant is a promising candidate to be used in dental applications with notable osteoimmunomodulation and angiogenesis functions. And the novel technique proposed in this study would benefit the modification of metal/inorganic surfaces with polysaccharides for future research.
ABSTRACT
As a potential anticancer agent, azurin has attracted extensive attraction among chemists, physicists, and material scientists. Its structural and unfolding/folding information has been partially understood, but some detailed information, such as the difference in the unfolding processes between apo-azurin and holo-azurin, the mechanical stability, and the role of the copper cluster in its stability, has not been addressed adequately, especially at the single-molecule level. Here, we employed AFM-based single-molecule force spectroscopy to investigate the unfolding process of azurin in the apo and holo forms under an external force. The results indicated that the unfolding processes of apo-azurin and holo-azurin are different, and holo-azurin requires a stronger force to unfold than does apo-azurin. The copper cluster exhibited a more significant impact on the stability and the folding process of holo-azurin: the copper cluster was completely broken, and the copper ion left the unfolded azurin during the unfolding process of azurin. We suspected that the presence of the disulfide bond in azurin made the unfolding of the copper cluster different from that in pseudoazurin, which is also a type I copper protein like azurin. Rarely reported in previous studies, the mechanical strength of the Cu-N(His) bond of the copper cluster was obtained in this study, which is weaker than that of most metal-S(Cys) bonds but higher than that of the Fe-N(His) bond. Altogether, our results offer a possible new scenario for azurin to widely extend its anticancer activity.
Subject(s)
Azurin , Azurin/metabolism , Copper , Metals , Protein FoldingABSTRACT
Extracellular calcium ion concentration levels increase in human osteoarthritic (OA) joints and contribute to OA pathogenesis. Given the fact that OA is a mechanical problem, the effect of the extracellular calcium level ([Ca2+]) on the mechanical behavior of primary human OA chondrocytes remains to be elucidated. Here, we measured the elastic modulus and cell-ECM adhesion forces of human primary chondrocytes with atomic force microscopy (AFM) at different extracellular calcium ion concentration ([Ca2+]) levels. With the [Ca2+] level increasing from the normal baseline level, the elastic modulus of chondrocytes showed a trend of an increase and a subsequent decrease at the level of [Ca2+], reaching 2.75 mM. The maximum increment of the elastic modulus of chondrocytes is a 37% increase at the peak point. The maximum unbinding force of cell-ECM adhesion increased by up to 72% at the peak point relative to the baseline level. qPCR and immunofluorescence also indicated that dose-dependent changes in the expression of myosin and integrin ß1 due to the elevated [Ca2+] may be responsible for the variations in cell stiffness and cell-ECM adhesion. Scratch assay showed that the chondrocyte migration ability was modulated by cell stiffness and cell-ECM adhesion: as chondrocyte's elastic modulus and cell-ECM adhesion force increased, the migration speed of chondrocytes decreased. Taken together, our results showed that [Ca2+] could regulate chondrocytes stiffness and cell-ECM adhesion, and consequently, influence cell migration, which is critical in cartilage repair.
Subject(s)
Elastic Modulus/physiology , Animals , Calcium/metabolism , Cell Adhesion/physiology , Cell Survival/physiology , Humans , Microscopy, Atomic ForceABSTRACT
Metalloproteins require the corresponding metal cofactors to exert their proper function. The presence of metal cofactors in the metalloprotein makes it more difficult to investigate its folding and unfolding process. In this study, we employed atomic-force-microscopy-based single-molecule force spectroscopy to reveal the unfolding process of pseudoazurin (PAZ) that belongs to blue copper proteins. Our study shows that holo-PAZ requires a higher rupture force for mechanical unfolding comparing with the apo-PAZ. This result demonstrates that the copper atom not only enables PAZ access to transfer electron, but should also have an influence on its stability. The results also suggest that the electronic configuration of the metal cofactors has a striking effect on the strength of the organometallic bonds. Moreover, the results also reveal that there is an intermediate state during the unfolding process of PAZ. This study provides insight into the characteristics of metalloproteins and leads to a better knowledge of their interaction at the individual molecule level.
Subject(s)
Azurin/chemistry , Copper/chemistry , Protein Unfolding , Molecular Dynamics Simulation , Protein ConformationABSTRACT
This study was designed to explore the potential therapeutic effects of Mg-based metal organic frameworks (MOFs) in osteoarthritis (OA) and to broaden the application field of MOFs. Mg/HCOOH-MOF was introduced and characterized by using SEM, XRD, TGA, and FTIR. ICP-MS results proved that Mg/HCOOH-MOF is stable in various physiological media and can maintain the release of Mg2+ ions for a long time. In vitro experiments were carried out to evaluate its potential application in attenuating OA. Mg/HCOOH-MOF was successfully synthesized, and results of MTT assays showed that it is biocompatible and can promote cell proliferation. qPCR results suggested that it can significantly regulate the expression of OCN, Axin2, iNOS and IL-1ß, which indicated its activity in anti-inflammation and bone protection. We believe that Mg/HCOOH-MOF could benefit OA therapy.
Subject(s)
Formates/chemistry , Magnesium/chemistry , Metal-Organic Frameworks/chemistry , Axin Protein/genetics , Axin Protein/metabolism , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Biocompatible Materials/therapeutic use , Cell Differentiation/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Humans , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Metal-Organic Frameworks/pharmacology , Metal-Organic Frameworks/therapeutic use , Osteoarthritis/drug therapy , Osteoarthritis/metabolism , Osteoarthritis/pathology , Osteocalcin/genetics , Osteocalcin/metabolism , Osteogenesis/drug effectsABSTRACT
Leukemia is a commonly seen disease caused by abnormal differentiation of hematopoietic stem cells and blasting in bone marrow. Despite drugs are used to treat the disease clinically, the influence of these drugs on leukemia cells' biomechanical properties, which are closely related to complications like leukostasis or infiltration, is still unclear. Due to non-adherent and viscoelastic nature of leukemia cells, accurate measurement of their elastic modulus is still a challenging issue. In this study, we adopted rate-jump method together with optical tweezers indentation to accurately measure elastic modulus of leukemia cells K562 after phorbol 12-myristate 13-acetate (PMA), all-trans retinoic acid (ATRA), Cytoxan (CTX), and Dexamethasone (DEX) treatment, respectively. We found that compared to control sample, K562â¯cells treated by PMA showed nearly a threefold increase in elastic modulus. Transwell experiment results suggested that the K562â¯cells treated with PMA have the lowest migration capability. Besides, it was shown that the cytoskeleton protein gene α-tubulin and vimentin have a significant increase in expression after PMA treatment by qPCR. The results indicate that PMA has a significant influence on protein expression, stiffness, and migration ability of the leukemia cell K562, and may also play an important role in the leukostasis in leukemia.
Subject(s)
Antineoplastic Agents/pharmacology , Biomechanical Phenomena/drug effects , Cell Movement/drug effects , Leukemia/drug therapy , Cyclophosphamide/pharmacology , Dexamethasone/pharmacology , Elastic Modulus/drug effects , Humans , K562 Cells , Leukemia/pathology , Tetradecanoylphorbol Acetate/pharmacology , Tretinoin/pharmacologyABSTRACT
Magnesium chondroitin sulfate (MgCS) has been fabricated and characterized in this study. We investigated its morphology, composition as well as structure. The results verify that the sodium of sodium chondroitin sulfate (CS) is successfully replaced by magnesium and formed a polysaccharide-metal ion complex. To evaluate the clinical potential of MgCS, cell proliferation and apoptosis test were conducted. The results reveal that MgCS could effectively increase the proliferation and decrease the apoptosis of osteoarthritis (OA) chondrocytes. Moreover, real-time quantitative polymerase chain reaction (RT-qPCR) was conducted to evaluate the gene expression level. RT-qPCR analysis suggests that MgCS could significantly increase the expression of COLII and decrease the expression of IL-1ß and iNOS in OA chondrocytes. Furthermore, significant upregulation of Bcl-2 mRNA expression and downregulation of the expression of apoptosis related gene p53 were observed. Thus, it is indicated that MgCS should have great potentials in OA treatment.
Subject(s)
Apoptosis/drug effects , Chondroitin Sulfates/chemical synthesis , Magnesium/chemistry , Osteoarthritis/metabolism , Apoptosis/physiology , Cells, Cultured , Chondrocytes/drug effects , Chondrocytes/metabolism , Chondrocytes/pathology , Chondroitin Sulfates/pharmacology , Chondroitin Sulfates/therapeutic use , Dose-Response Relationship, Drug , Humans , Magnesium/pharmacology , Magnesium/therapeutic use , Osteoarthritis/drug therapy , Osteoarthritis/pathologyABSTRACT
Chondrocytes culture within three-dimensional (3D) hydrogels is an attractive strategy for expanding their potential in various biomedical applications. Molecular weight (MW) of the polymers used for preparing one-component hydrogels for 3D cell culture can affect the activity of chondrocytes. However, whether MW of double network hydrogels affects cell fate remains to be determined. Here, a ternary hydrogel was employed to encapsulate chondrocytes. By altering MW of alginate, the hybrid hydrogels with different mechanical properties were prepared. The effects of MW on the properties of hydrogels were evaluated in this work. Cell results revealed that chondrocytes encapsulated in low-MW hydrogels exhibited the best results on maintaining cell viability and inhibiting cell death. Additionally, RT-qPCR analysis suggested that the expression of COLII was increased while the expression of IL-1ß and iNOS was decreased in chondrocytes encapsulated in low-MW hydrogels. Hence, it is reasonable to be expected that the developed low-MW hydrogel should be a promising candidate for cartilage tissue engineering.
Subject(s)
Alginates/chemistry , Cell Culture Techniques/methods , Chondrocytes/drug effects , Chondroitin Sulfates/chemistry , Hydrogels/chemistry , Base Sequence , Cell Death/drug effects , Cell Survival/drug effects , Drug Stability , Humans , Hydrogels/chemical synthesis , Molecular Weight , Porosity , RNA, Messenger/metabolism , TemperatureABSTRACT
A composite hydrogel with tunable mechanical properties has been fabricated and characterized in this study. We investigated its swelling degree, morphology, structure and thermal stability. Moreover, the effect of strontium chloride concentration on both the dynamic rheology and nanomechanical properties of the composite hydrogels was confirmed in this work. To eliminate the viscoelastic influences of hydrogels during nanomechanical tests, we first analyzed the elastic modulus of strontium alginate (Alg-Sr) and strontium alginate/chondroitin sulfate (Alg/CS-Sr) hydrogels via atomic force microscopy (AFM) using the rate-jump method. Chondrocytes were cultured with the Alg-Sr and Alg/CS-Sr hydrogels respectively. Cell viability assay reveals that the Alg/CS-Sr hydrogel possesses good cytocompatibility. Flow cytometry, qPCR and western blotting analysis suggest that the Alg/CS-Sr hydrogel exerts a positive effect on the inhibition of apoptosis and may exert anti-inflammatory effects in articular cartilage related applications. Furthermore, the preliminary in vivo study shows that the Alg/CS-Sr composite hydrogel facilitates the repair of cartilage in rabbit cartilage defect. Taken together, it is indicated that the Alg/CS-Sr composite hydrogel might be a promising scaffold to promote the repair of cartilage defects.
Subject(s)
Cartilage/cytology , Cartilage/drug effects , Hydrogels/chemistry , Hydrogels/pharmacology , Mechanical Phenomena , Tissue Engineering/methods , Animals , Apoptosis/drug effects , Cell Proliferation/drug effects , Chondrocytes/cytology , Chondrocytes/drug effects , Rabbits , TemperatureABSTRACT
Collagen, the dominating material in the extracellular matrix, provides the strength, elasticity and mechanical stability to the organisms. The mechanical property of collagen is mainly dominated by its surrounding environments. However, the variation and origin of the mechanics of collagen fibril under different concentrations of calcium ions (χCa) remains unknown. By using the atomic force microscopy based nanoindentation, the mechanics and structure of individual type II collagen fibril were first investigated under different χCa in this study. The results demonstrate that both of the mechanical and structural properties of the collagen fibril show a prominent dependence on χCa. The mechanism of χCa-dependence of the collagen fibril was attributed to the chelation between collagen molecules and the calcium ions. Given the role of calcium in the pathology of osteoarthritis, the current study may cast new light on the understanding of osteoarthritis and other soft tissue hardening related diseases in the future.
Subject(s)
Calcium/metabolism , Collagen Type II/chemistry , Fibrillar Collagens/chemistry , Biomechanical Phenomena , Elastic Modulus , Humans , Ions , Middle AgedABSTRACT
The single-chain mechanics of two similar thermosensitive polymers, poly(N,N-diethylacrylamide) (PDEAM) and poly(N-isopropylacrylamide) (PNIPAM), have been studied by atomic force microscopy-based single-molecule force spectroscopy (SMFS). In a typical nonpolar organic solvent, octane, both of the polymers show the same inherent elasticity, although they have different substitutional groups. However, the mechanics of the two polymers presents large differences in water. The energies needed for the rearrangement of the bound water during elongation at room temperature are estimated by the SMFS method at the single-chain level, which is ~1.13 ± 0.10 and ~5.19 ± 0.10 kJ/mol for PDEAM and PNIPAM, respectively. In addition, PNIPAM shows a temperature-dependent single-chain mechanics when the temperature is increased across the lower critical solution temperature (LCST), while PDEAM does not. These differences observed in aqueous solution originate from the different structures of the two polymers. With a hydrogen bond donor in the amide group, PNIPAM will be more hydrated when T < LCST. When T > LCST, PNIPAM will have larger changes in both conformation and hydration. These findings also suggest that PNIPAM is a good candidate for a thermo-driven single-molecule motor, while PDEAM is not.
Subject(s)
Acrylamides/chemistry , Acrylic Resins/chemistry , Polymers/chemistry , Water/chemistry , Hydrogen Bonding , Microscopy, Atomic Force , Molecular Structure , Octanes/chemistryABSTRACT
We study the single-chain elasticities of three kinds of neutral polymers with a carbon-carbon (C-C) backbone by atomic force microscopy-based single-molecule force spectroscopy in a nonpolar solvent (octane), aiming at measuring the inherent chain elasticity of this very important class of polymers. The finding that the single-chain elasticities of all three polymers in octane are virtually identical in the entire force region implies that the side chains of the polymers have no detectable effects on the single-chain elasticity. By utilizing the single-chain elasticity from quantum mechanics calculations, the freely rotating chain model can provide the best fitting curve when each C-C bond is set to be the rotating unit. Although there are some exceptions when the side chain is very huge, our work provides a general result for the inherent elasticity of single neutral flexible polymer chains with C-C backbones.
Subject(s)
Carbon/chemistry , Polymers/chemistry , Elasticity , Molecular StructureABSTRACT
Radical polymerization from a single initiator molecule in a microenvironment is a nearly ideal system in which bimolecular termination, solution concentration, and viscosity changes could be neglected. In this study, we provide two facile methods of preparing polymers via atom-transfer radical polymerization (ATRP) under single-initiator conditions: tether initiators on planar substrates at superlow density through mixed self-assembled monolayers (SAMs) and encapsulated single initiators in microfluidic droplets. The molecular weight (MW) of the resultant polymers characterized by atomic force microscope-based single-molecule force spectroscopy (AFM-based SMFS) showed that the single-chain ATRP had an extraordinarily faster chain propagation rate (2 unit/s) on planar substrates and gave polymers with much higher MWs (10(5)-10(6) g/mol) than those obtained from traditional ATRP (10(3)-10(5) g/mol). The former method offered a general platform for single-chain polymer synthesis and investigation, and the latter could be amplified to obtain abundant single-chain polymers with ultrahigh molecular weight (UHMW) for commercial applications.
Subject(s)
Polymers/chemical synthesis , Free Radicals/chemical synthesis , Free Radicals/chemistry , Molecular Weight , Polymerization , Polymers/chemistry , Surface PropertiesABSTRACT
Poly(N-isopropyl-acrylamide) (PNIPAM) is a paradigm thermally sensitive polymer, which has a lower critical solution temperature (LCST) of ~32 °C in water. Herein by AFM-based single molecule force spectroscopy (SMFS), we measured the single chain elasticity of PNIPAM across the LCST in water. Below LCST, the force curves obtained at different temperatures have no remarkable difference; while above LCST, an unexpected temperature dependent elasticity is observed, mainly in the middle force regime. We found that 35 °C is a turning point of the variation: from 31 to 35 °C, the middle parts of the force curves drop gradually, whereas from 35 to 40 °C, the middle parts rise gradually. A possible mechanism for the unexpected temperature dependent mechanics is proposed. The single chain contraction against external force upon heating from 35 to 40 °C may cast new light on the design of molecular devices that convert thermal energy to mechanical work.