Organization Of Fibers Research Articles

Exploring natural palm fiber’s mechanical performance using multi-scale fractal structure simulation

Palm fiber, a type of natural multicellular fiber, exhibits distinct mechanical properties, such as excellent elasticity, higher fracture energy, and desirable stretch ability. To reveal the structure-property relationship, a multi-scale layered fractal theoretical model was introduced to investigate the tensile behavior of palm fibers at different scales. A three-circle model was established and used to simulate the hierarchical organization of palm fibers. The palm fibers consisted of cellulose molecular chains, fibril filaments, microfibrils, and cells. Moreover, the characteristics of stress, fracture energy, and Young’s modulus on different scales were calculated and verified by tensile testing and atomic force microscopy (AFM). The results revealed that the fractal model effectively decoupled the contributions of different scales to the tensile properties. In particular, the microfibril mainly influenced the stiffness, whereas the cell determined the toughness of palm fibers. The findings of the current study can be utilized to improve the design and preparation of fiber-based nanomaterials.

Human metaphase chromosome consists of randomly arranged chromatin fibres with up to 30-nm diameter

During cell division, mitotic chromosomes assemble and are equally distributed into two new daughter cells. The chromosome organisation of the two chromatids is essential for even distribution of genetic materials. Although the 11-nm fibre or nucleosome structure is well-understood as a fundamental fibrous structure of chromosomes, the reports on organisation of 30-nm basic chromatin fibres have been controversial, with debates on the contribution of 30-nm or thicker fibres to the higher order inner structure of chromosomes. Here, we used focused ion beam/scanning electron microscopy (FIB/SEM) to show that both 11-nm and 30-nm fibres are present in the human metaphase chromosome, although the higher-order periodical structure could not be detected under the conditions employed. We directly dissected the chromosome every 10-nm and observed 224 cross-section SEM images. We demonstrated that the chromosome consisted of chromatin fibres of an average diameter of 16.9-nm. The majority of the chromatin fibres had diameters between 5 and 25-nm, while those with 30-nm were in the minority. The reduced packaging ratio of the chromatin fibres was detected at axial regions of each chromatid. Our results provide a strong basis for further discussions on the chromosome higher-order structure.

Biomimicry of iridescent, patterned insect cuticles: comparison of biological and synthetic, cholesteric microcells using hyperspectral imaging.

Biological systems inspire the design of multifunctional materials and devices. However, current synthetic replicas rarely capture the range of structural complexity observed in natural materials. Prior to the definition of a biomimetic design, a dual investigation with a common set of criteria for comparing the biological material and the replica is required. Here, we deal with this issue by addressing the non-trivial case of insect cuticles tessellated with polygonal microcells with iridescent colours due to the twisted cholesteric organization of chitin fibres. By using hyperspectral imaging within a common methodology, we compare, at several length scales, the textural, structural and spectral properties of the microcells found in the two-band cuticle of the scarab beetle Chrysina gloriosa with those of the polygonal texture formed in flat films of cholesteric liquid crystal oligomers. The hyperspectral imaging technique offers a unique opportunity to reveal the common features and differences in the spectral-spatial signatures of biological and synthetic samples at a 6-nm spectral resolution over 400 nm-1000 nm and a spatial resolution of 150 nm. The biomimetic design of chiral tessellations is relevant to the field of non-specular properties such as deflection and lensing in geometric phase planar optics.

Sulforaphane protects against skeletal muscle dysfunction in spontaneous type 2 diabetic db/db mice

AimsSkeletal muscle diseases have become to be the most common complication in patients with type 2 diabetic mellitus (T2DM). However, the effective therapies against skeletal muscle diseases are not yet available. Sulforaphane (SFN) is an organic isothiocyanate found in cruciferous plants. Our aim was to explore whether SFN could attenuate the skeletal muscle diseases in spontaneous type 2 diabetic db/db mice. Materials and methodsThe db/m and littermate db/db mice were treated with SFN or dimethyl sulfoxide. The grip strength of mice was measured by a grasping forcing machine. The electron transmission microscopy was used to perform the skeletal muscle. The western blot was used to detect the nuclear factor E2-related factor 2/heme oxygenase 1 (Nrf2/HO-1) signal pathway related proteins, and inflammatory and apoptotic associated proteins. The mRNA levels of anti-inflammatory and anti-oxidative relative genes were detected by RT-QPCR. Key findingsWe found that SFN could significantly increase the grip strength of the db/db mice. The lean mass and gastrocnemius mass were increased in the db/db mice after administration with SFN. Additionally, the db/db mice restored the skeletal muscle fiber organization after SFN treatment. Mechanistically, SFN could activate the Nrf2/HO-1 signal pathway, and downregulate the expression of inflammatory and apoptotic associated proteins. Furthermore, SFN could also regulate the mRNA levels of anti-inflammatory and anti-oxidative related genes. SignificanceOur results demonstrated that SFN can protect against skeletal muscle diseases in db/db type 2 diabetic mice and provide a potential drug to prevent skeletal muscle dysfunction in T2DM patients.

Label-Free Infrared Spectral Histology of Skin Tissue Part I: Impact of Lumican on Extracellular Matrix Integrity.

Proteoglycans (PG) play an important role in maintaining the extracellular matrix (ECM) integrity. Lumican, a small leucine rich PG, is one such actor capable of regulating such properties. In this study, the integrity of the dermis of lumican-deleted Lum–/– vs. wild-type mice was investigated by conventional histology and by infrared spectral histology (IRSH). Infrared spectroscopy is a non-invasive, rapid, label-free and sensitive technique that allows to probe molecular vibrations of biomolecules present in a tissue. Our IRSH results obtained on control (WT, n = 3) and Lum–/– (n = 3) mice showed that different histological structures were identified by using K-means clustering and validated by hematoxylin eosin saffron (HES) staining. Furthermore, an important increase of the dermis thickness was observed in Lum–/– compared to WT mice. In terms of structural information, analysis of the spectral images also revealed an intra-group homogeneity and inter-group heterogeneity. In addition, type I collagen contribution was evaluated by HES and picrosirius red staining as well as with IRSH. Both techniques showed a strong remodeling of the ECM in Lum–/– mice due to the looseness of collagen fibers in the increased dermis space. These results confirmed the impact of lumican on the ECM integrity. The loss of collagen fibers organization due to the absence of lumican can potentially increase the accessibility of anti-cancer drugs to the tumor. These results are qualitatively interesting and would need further structural characterization of type I collagen fibers in terms of size, organization, and orientation.

SUN-217 Skin Glucocorticoid Metabolism in Burn Injury: Towards Novel Treatments That Reduce Scarring

The most common and severe complication of burn injury is the development of excessive scarring/tissue fibrosis. No current treatments reduce scarring after burns. Prolonged exposure to high levels of glucocorticoids (Cushing’s syndrome) detrimentally impacts skin, leading to reduced collagen production and impaired wound healing. Skin can generate active glucocorticoids locally through expression and activity of the 11β-hydroxysteroid dehydrogenase type 1 enzyme (11β-HSD1). We hypothesised that local glucocorticoid activation by 11β-HSD1 is an important regulator of wound healing, fibrosis and scarring after burn injury. We additionally proposed that pharmacological manipulation of this system would improve outcomes of burn wound healing.We examined glucocorticoid metabolism (by RT-PCR, immunohistochemistry and specific enzyme activity assays) in burn and non-burn skin from burn injury patients (n=14) and mouse models of burn injury (1cm2 full thickness burn in C57Bl/6 mice). We utilised mice with genetic or pharmacological deletion of 11β-HSD1 in skin to evaluate the effects of 11β-HSD1 on burn injury healing, wound fibrosis and skin properties (by atomic force microscopy and tensile property testing) after wound healing. We also developed slow release scaffolds containing therapeutic agents including inactive glucocorticoids (prednisone) that are selectively reactivated in skin cells expressing 11β-HSD1.The expression of 11β-HSD1 in human and mouse skin increased substantially after burn injury (7.1+/-1.8 fold increase on day 4–9 post burn compared to non-burn skin, p<0.05). Early after injury, expression was primarily in immune cells but at later stages in fibroblasts. Mice with 11β-HSD1 deletion experienced faster wound healing post burn (17% reduced wound area at day 7 compared to wildtype, p<0.0001) but when healed these wounds had excessive collagen density and skin thickening, and abnormal collagen fibre organisation (assessed by Masson’s Trichrome staining). The post burn scars formed in 11β-HSD1 knockout mice demonstrated different skin elastic properties compared to those formed in wildtype mice. In wildtype mice application of scaffolds loaded with inactive glucocorticoid (prednisone) significantly impacted wound healing demonstrating the feasibility of using enzyme substrates to improve wound outcomes.The findings demonstrate the importance of skin 11β-HSD1 in wound healing and scarring after burn injury and indicate ways in which excessive scarring might be prevented.

The significance of stromal collagen organization in cancer tissue: An in-depth discussion of literature.

It has become clear that carcinogenesis goes beyond tumor cell biology. Cancer research has acknowledged the importance of biological functions of the tumor-microenvironment, wherein not only cellular components seem to hold valuable information but also structural components like collagen fibers. Several studies have focused on the significance of stromal collagen fiber organization and reported on its role in cancer progression, invasiveness and treatment response. In this review, we discuss the different imaging methods for stromal collagen organization, followed by an in-depth discussion of current literature on in-vitro and animal experiments and human studies, highlighting its importance with respect to cancer progression, prognosis and prediction. We can conclude that collagen organization contains valuable information with regard to metastatic potential and clinical outcomes in cancer. However, the significance of an aligned versus disorganized collagen morphology differs between cancer types, implying more research is necessary before steps towards clinical implementation can be made.

Altered organization of collagen fibers in the uninvolved human colon mucosa 10\u2009cm and 20\u2009cm away from the malignant tumor

Remodelling of collagen fibers has been described during every phase of cancer genesis and progression. Changes in morphology and organization of collagen fibers contribute to the formation of microenvironment that favors cancer progression and development of metastasis. However, there are only few data about remodelling of collagen fibers in healthy looking mucosa distant from the cancer. Using SHG imaging, electron microscopy and specialized softwares (CT-FIRE, CurveAlign and FiberFit), we objectively visualized and quantified changes in morphology and organization of collagen fibers and investigated possible causes of collagen remodelling (change in syntheses, degradation and collagen cross-linking) in the colon mucosa 10 cm and 20 cm away from the cancer in comparison with healthy mucosa. We showed that in the lamina propria this far from the colon cancer, there were changes in collagen architecture (width, straightness, alignment of collagen fibers and collagen molecules inside fibers), increased representation of myofibroblasts and increase expression of collagen-remodelling enzymes (LOX and MMP2). Thus, the changes in organization of collagen fibers, which were already described in the cancer microenvironment, also exist in the mucosa far from the cancer, but smaller in magnitude.

PRPS polymerization influences lens fiber organization in zebrafish.

The self-assembly of metabolic enzymes into filaments or foci highlights an intriguing mechanism for the regulation of metabolic activity. Recently, we identified the conserved polymerization of phosphoribosyl pyrophosphate synthetase (PRPS), which catalyzes the first step in purine nucleotide synthesis, in yeast and cultured mammalian cells. While previous work has revealed that loss of PRPS activity regulates retinal development in zebrafish, the extent to which PRPS filament formation affects tissue development remains unknown. By generating novel alleles in the zebrafish PRPS paralogs, prps1a and prps1b, we gained new insight into the role of PRPS filaments during eye development. We found that mutations in prps1a alone are sufficient to generate abnormally small eyes along with defects in head size, pigmentation, and swim bladder inflation. Furthermore, a loss-of-function mutation that truncates the Prps1a protein resulted in the failure of PRPS filament assembly. Lastly, in mutants that fail to assemble PRPS filaments, we observed disorganization of the actin network in the lens fibers. The truncation of Prps1a blocked PRPS filament formation and resulted in a disorganized lens fiber actin network. Altogether, these findings highlight a potential role for PRPS filaments during lens fiber organization in zebrafish.

Macromechanics and polycaprolactone fiber organization drive macrophage polarization and regulate inflammatory activation of tendon in vitro and in vivo

Appropriate macrophage response to an implanted biomaterial is crucial for successful tissue healing outcomes. In this work we investigated how intrinsic topological cues from electrospun biomaterials and extrinsic mechanical loads cooperate to guide macrophage activation and macrophage-tendon fibroblast cross-talk. We performed a series of in vitro and in vivo experiments using aligned or randomly oriented polycaprolactone nanofiber substrates in both mechanically loaded and unloaded conditions. Across all experiments a disorganized biomaterial fiber topography was alone sufficient to promote a pro-inflammatory signature in macrophages, tendon fibroblasts, and tendon tissue. Extrinsic mechanical loading was found to strongly regulate the character of this signature by reducing pro-inflammatory markers both in vitro and in vivo. We observed that macrophages generally displayed a stronger response to biophysical cues than tendon fibroblasts, with dominant effects of cross-talk between these cell types observed in mechanical co-culture models. Collectively our data suggest that macrophages play a potentially important role as mechanosensory cells in tendon repair, and provide insight into how biological response might be therapeutically modulated by rational biomaterial designs that address the biomechanical niche of recruited cells.

Mode of Ossification and Extracellular Fiber Characterization of Osteoderm Matrix in the American Alligator (Alligator mississippiensis) and Comparisons to Inherited and Acquired Heterotopic Ossification Disorders

Osteoderms are bones that form in the dermis of several reptilian species and have been a source of scientific curiosity since at least the 1800s. Though there are several hypotheses for their function, a consensus has not yet emerged. The function of osteoderms is further obscured, since there is little histomorphological information on these structures. Indeed, understanding the cellular and molecular events that initiate and drive osteoderm development may clarify the functional significance of these structures. Dubansky &amp; Dubansky (2018) conducted the first histological description of the development of the osteoderm in the American alligator, and identified discrete developmental stages based on the organization of collagen fibers and contributing cell mediators, as well as the degree of mineralization of the matrix. However, questions remain regarding the type of collagen and the identity of the cell precursors that are involved at different stages, as well as the existence of a hyaline cartilage model. In this study, differential staining techniques characterized changes in the fibrous component of the developing matrix of the osteoderm, and investigated whether ossification proceeds via an endochondral mechanism. Early stage osteoderms exhibit an extensive reticulin (type III collagen) fiber scaffold, which is replaced by disorganized, thick type I collagen fibers in mineralizing maturing matrix, and highly organized, thin type I collagen at the proliferating edge of the matrix. Despite morphological similarities to cartilage in some areas, staining showed that hyaline cartilage was absent and, therefore, the mode of ossification more resembled intramembranous ossification, than endochondral ossification. Osteoderm development resembles Heterotopic Ossification (HO) disorders in humans and is being used as a model to study several forms of this pathology. Based on this study, osteoderm histomorphology most closely resembles the inherited disorder Progressive Osseous Heteroplasia, in which pathological bone formation is restricted to the dermis and superficial fascia and proceeds via intramembranous ossification. Documenting similarities and differences among the various types of HO lesions and osteoderms may provide insights into the disease progression and future therapeutic strategies.Support or Funding InformationThis research was supported by a Faculty‐Student Research and Creative Activity Internal Grant from Tarleton State University, Office of Research and Innovation.

Design and application of (Fe3O4)-GOTfOH based AgNPs doped starch/PEG-poly (acrylic acid) nanocomposite as the magnetic nanocatalyst and the wound dress

As a novel, recyclable nanocatalyst, (Fe3O4)-GOTfOH based Ag nanoparticles doped Starch/PEG-poly (acrylic acid) nanocomposite (Fe3O4@GOTfOH/Ag/St-PEG-AcA) was applied for one-pot synthesis of 2,4,6-triarylpyridine derivatives under water solvent conditions. The prepared nanocomposite was also evaluated in terms of biocompatibility for wound healing. Fe3O4@GOTfOH/Ag/St-PEG-AcA could be easily removed from the mixture of the reaction by an external magnet and recycled without a considerable decrease of activity even after 10 runs. The new nanocatalyst offered better efficiencies than other commercially available sulfonic acid catalysts. In terms of the bioactivity of nanocatalyst, good antimicrobial efficiency was confirmed on E. coli bacteria. Besides, histology of repaired wounds in Fe3O4@GOTfOH/Ag/St-PEG-AcA for a healed group of mice showed better fibroblast distribution and more compact collagen fiber organization compared to wounds in the control group.

Factors affecting optic nerve head biomechanics in a rat model of glaucoma.

Glaucoma is the leading cause of irreversible blindness and is characterized by the death of retinal ganglion cells, which carry vision information from the retina to the brain. Although it is well accepted that biomechanics is an important part of the glaucomatous disease process, the mechanisms by which biomechanical insult, usually due to elevated intraocular pressure (IOP), leads to retinal ganglion cell death are not understood. Rat models of glaucoma afford an opportunity for learning more about these mechanisms, but the biomechanics of the rat optic nerve head (ONH), a primary region of damage in glaucoma, are only just beginning to be characterized. In a previous study, we built finite-element models with individual-specific rat ONH geometries. Here, we developed a parametrized model of the rat ONH and used it to perform a sensitivity study to determine the influence that six geometric parameters and 13 tissue material properties have on rat optic nerve biomechanical strains due to IOP elevation. Strain magnitudes and patterns in the parametrized model generally matched those from individual-specific models, suggesting that the parametrized model sufficiently approximated rat ONH anatomy. Similar to previous studies in human eyes, we found that scleral properties were highly influential: the six parameters with highest influence on optic nerve strains were optic nerve stiffness, IOP, scleral thickness, the degree of alignment of scleral collagen fibres, scleral ground substance stiffness and the scleral collagen fibre uncrimping coefficient. We conclude that a parametrized modelling strategy is an efficient approach that allows insight into rat ONH biomechanics. Further, scleral properties are important influences on rat ONH biomechanics, and additional efforts should be made to better characterize rat scleral collagen fibre organization.

Identification of Key Factors in Cardiomyocyte Development by Single‐Cell Transcriptome Analysis

BackgroundA promising intervention for heart diseases is the application of cardiomyocytes differentiated from pluripotent stem cells. However, the driving factors of the differentiation process still remain unclear. This study aims to map the transcriptional landscapes of directional differentiation of pluripotent stem cells at multiple stages, using single‐cell sequencing data, in order to identify the key drivers of the differentiation process.MethodSingle‐cell sequencing data from pluripotent stem cell‐derived cardiomyocytes at age day 0, day 2, day 5, day 15 and day 30 were included and normalized. UMAP algorithm was then used to reduce the dimensionality of the transcriptome profiles, followed by community detection to group the cells and cell type identification using marker genes. Graph‐autocorrelation analysis was implemented to identify differentially expressed genes among cell groups with Gene Ontology term enrichment. Next, we calculated the pseudotimes of each cardiomyocyte to construct the pseudotime trajectories. Finally, knowledge based dynamic changes in gene expression were analyzed to identify the key factors driving the cardiomyocyte development.ResultWe identified 9 different cell types in the process of cardiomyocyte development, including cardiomyocytes, induced pluripotent stem cells, cardiogenic mesoderm cells, cardiovascular progenitor cells, embryonic stem cells, endoderm cells, epithelial progenitor cells, cardiomyocyte progenitor cells and multipotent progenitor cells. Identification of cardiomyocyte was confirmed with specific gene expression profiles such as significant enrichment in the biological processes of cell adhesion and supramolecular fiber organization. The expression of NKX2‐5, which encodes a homeobox‐containing transcription factor, was found to be at a lower level in the early stages and continually increased as cardiomyocyte develop, suggesting that NKX2‐5 was one of the driving factors of cardiac development, acting through promoting myocardial maturation and development.ConclusionSingle cell sequencing data revealed the dynamic changes of gene expression during cardiomyocyte development differentiation. NKX2‐5 was identified as a key factor in cardiac development. Our finding contributes to the understanding of the mechanism behind the cardiomyocyte development, and may serve as the basis for stem cell based therapies.Support or Funding InformationThis work was supported by National Natural Science Foundation of China 81774152 (to RZ) and 81770571 (to LZ).

Corneal Collagen Ordering After In Vivo Rose Bengal and Riboflavin Cross-Linking

PurposePhotoactivated cornea collagen cross-linking (CXL) increases corneal stiffness by initiating formation of covalent bonds between stromal proteins. Because CXL depends on diffusion to distribute the photoinitiator, a gradient of CXL efficiency with depth is expected that may affect the degree of stromal collagen organization. We used second harmonic generation (SHG) microscopy to investigate the differences in stromal collagen organization in rabbit eyes after corneal CXL in vivo as a function of depth and time after surgery.MethodsRabbit corneas were treated in vivo with either riboflavin/UV radiation (UVX) or Rose Bengal/green light (RGX) and evaluated 1 and 2 months after CXL. Collagen fibers were imaged with a custom-built SHG scanning microscope through the central cornea (350 µm depth, 225 × 225 µm en face images). The order coefficient (OC), a metric for collagen organization, and total SHG signal were computed for each depth and compared between treatments.ResultsOC values of CXL-treated corneas were larger than untreated corneas by 27% and 20% after 1 month and 38% and 33% after 2 months for the RGX and UVX, respectively. RGX OC values were larger than UVX OC values by 3% and 5% at 1 and 2 months. The SHG signal was higher in CXL corneas than untreated corneas, both at 1 and 2 months after surgery, by 18% and 26% and 1% and 10% for RGX and UVX, respectively.ConclusionsIncreased OC corresponded with increased collagen fiber organization in CXL corneas. Changes in collagen organization parallel reported temporal changes in cornea stiffness after CXL and also, surprisingly, are detected deeper in the stroma than the regions stiffened by collagen cross-links.

Core Matrisome Protein Signature During Periodontal Ligament Maturation From Pre-occlusal Eruption to Occlusal Function.

The pre-occlusal eruption brings the molars into functional occlusion and initiates tensional strains during mastication. We hypothesized that upon establishment of occlusal contact, the periodontal ligament (PDL) undergoes cell and extracellular matrix maturation to adapt to this mechanical function. The PDL of 12 Wistar male rats were laser microdissected to observe the proteomic changes between stages of pre-occlusal eruption, initial occlusal contact and 1-week after occlusion. The proteome was screened by mass spectrometry and confirmed by immunofluorescence. The PDL underwent maturation upon establishment of occlusion. Downregulation of alpha-fetoprotein stem cell marker and protein synthesis markers indicate cell differentiation. Upregulated proteins were components of the extracellular matrix (ECM) and were characterized with the matrisome project database. In particular, periostin, a major protein of the PDL, was induced following occlusal contact and localized around collagen α-1 (III) bundles. This co-localization coincided with organization of collagen fibers in direction of the occlusal forces. Establishment of occlusion coincides with cellular differentiation and the maturation of the PDL. Co-localization of periostin and collagen with subsequent fiber organization may help counteract tensional forces and reinforce the ECM structure. This may be a key mechanism of the PDL to adapt to occlusal forces and maintain structural integrity.

Myocardial deformation imaging in anesthesia and perioperative medicine: a non systematic review

Measuring the systolic function of the left ventricle (LV) is essential in clinical practice. However, the complex organization of the myocardial fibers whose contraction results in the ejection of the stroke volume renders this assessment challenging. The ejection fraction of the left ventricle (LVEF) has long been the most popular measure of the systolic function of the left ventricle despite its numerous technical and non- technical limitations. More recently, the development of speckle-tracking echocardiography allowed the widespread adoption of myocardial deformation imaging indices such as the strain and the strain rate. Strain, and in particular, global longitudinal strain (GLS) has quickly gained popularity as an additional measure of the systolic function of the left ventricle. In comparison with the ejection fraction, GLS is easier to use, more reproducible, and more sensitive to mild changes in myocardial contractility. Strain is an interesting tool for diagnosis and prognostic stratification in both surgical and non-surgical patients. The purpose of this review is to describe the principles of strain use and to review its main applications, while focusing on the aspects relevant to the practice of anesthesia and intensive care medicine.

Spatially Controlled Supramolecular Polymerization of Peptide Nanotubes by Microfluidics.

Despite the importance of spatially resolved self-assembly for molecular machines, the spatial control of supramolecular polymerization with synthetic monomers had not been experimentally established. Now, a microfluidic-regulated tandem process of supramolecular polymerization and droplet encapsulation is used to control the position of self-assembled microfibrillar bundles of cyclic peptide nanotubes in water droplets. This method allows the precise preferential localization of fibers either at the interface or into the core of the droplets. UV absorbance, circular dichroism and fluorescence microscopy indicated that the microfluidic control of the stimuli (changes in pH or ionic strength) can be employed to adjust the packing degree and the spatial position of microfibrillar bundles of cyclic peptide nanotubes. Additionally, this spatially organized supramolecular polymerization of peptide nanotubes was applied in the assembly of highly ordered two-dimensional droplet networks.

Single-Cell Response to the Rigidity of Semiconductor Nanomembranes on Compliant Substrates.

Single-crystalline semiconductor nanomembranes (NMs) bonded to compliant substrates are increasingly used for biomedical research and in health care. Nevertheless, there is a limited understanding of how individual cells sense the unique mechanical properties of these substrates and adjust their behavior in response to them. In this work, we performed proliferation assays, cytoskeleton analysis, and focal adhesion (FA) studies for NIH-3T3 fibroblasts on 220 and 20 nm single-crystalline Si on polydimethylsiloxane (PDMS) substrates with an elastic modulus of ∼31 kPa. We also characterized cell response on bulk Si as a reference. Our in vitro studies show that varying the thickness of the NM between 20 and 220 nm affects the proliferation rate of the cells, their cytoskeleton, fiber organization, spread area, and degree of FA. For example, cultured cells on 220 nm Si/PMDS exhibit the same response as on bulk Si, that is, they are well-spread with a pentagonal (or dendritic) shape and show a good organization of stress fibers and FAs. On the other hand, the cells on 20 nm Si/PDMS are spherical, with fiber organization and FAs in undetectable levels. We explained the results of our in vitro studies through a shear-lag mechanical model. The calculated FA-substrate contact stiffnesses for fibroblasts on bulk Si and 220 nm Si/PDMS closely match, and they are significantly higher than the stiffness of the integrin clutches and the plaque. Conversely, focal contacts with 20 nm Si/PDMS have comparable lateral compliance to adhesion-mediating intracellular organisms. In conclusion, our work relies on recent advances in NM technology to fill a critical knowledge gap about how individual cells sense and react to the mechanical properties of NM-based substrates. Our findings will have a major impact on the design of flexible electronic materials for applications in biomedical science and health care.

Interaction between Autonomous and Microtubule Guidance Systems Controls Cellulose Synthase Trajectories.

The organization of cellulose microfibrils is critical for the strength and growth of plant cell walls. Microtubules have been shown to play a key role in controlling microfibril organization by guiding cellulose synthase complexes [1-4]. However, cellulose synthase trajectories can be maintained when microtubules are removed by drugs, suggesting a separate guidance mechanism is also at play [1, 5, 6]. By slowing down microtubule dynamics, we reveal such a mechanism by showing that cellulose synthase complexes can interact with the trails left by other complexes, causing them to follow the trails or disappear. The stability of the trails, together with the sensitivity of their directions to cellulase treatment, indicates they most likely reflect nascent cellulose microfibrils. Over many hours, this autonomous mechanism alone can lead to a change in the dominant orientation of cellulose synthase trajectories. However, the mechanism can be overridden by the microtubule guidance system. Our findings suggest a dual guidance model, in which an autonomous system, involving interaction between cellulose synthases and microfibrils, can maintain aligned cellulose synthase trajectories, while a microtubule guidance system allows alignments to be steered by environmental and developmental cues.