Fiber Alignment Research Articles

Abstract PR018: Inflammation-induced mechanotransduction is necessary and sufficient to create pre-cancerous squamous lung metaplasias and necessary to drive progression to dysplasia

Abstract Chronic exposure to tobacco smoke triggers chronic inflammation, fostering a fibrotic microenvironment and promoting the focal replacement of columnar human bronchial epithelial cells (hBECs) with cells exhibiting squamous morphology, termed metaplasia. How squamous metaplasia progresses to squamous dysplasia and, ultimately, squamous lung carcinoma is under intense investigation. Using human bronchial tissues, we demonstrate that, both in vitro and in vivo, cellular stress associated with chronic inflammation (e.g., oxidative stress, DNA damage) enhances TGF-β signaling and creates a fibroblast state exhibiting upregulated HSP47 expression, driving increased collagen fiber alignment, elevated tissue stiffness, and subsequent activation of YAP-dependent mechanotransduction in adjacent hBECs. These Stress/Tension-producing Instructive Fibroblasts (STIFs) alone were sufficient to induce hBECs to squamous metaplasia, precursors to tumor formation. Furthermore, when tumor suppressor function was compromised in hBECs, STIFs induced dysplastic phenotypes. Mechanistically, STIFs act in a dominant manner modulating epithelial cell identity and malignant phenotypes via extracellular matrix (ECM)-dependent mechanotransduction. Inhibition of HSP47-dependent collagen processing effectively prevented fibroblast-induced squamous metaplasia and dysplasia and was capable of reversing fibroblast-induced metaplasia, restoring bronchial epithelial cell identity. Citation Format: Thea D. Tisty, Deng Pan, Philippe Gascard, Joseph Caruso, Lorenzo Ferri, Chira Chen-Tanyolac. Inflammation-induced mechanotransduction is necessary and sufficient to create pre-cancerous squamous lung metaplasias and necessary to drive progression to dysplasia [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr PR018.

Abstract A033: Integrating spatial transcriptomics and second harmonic generation imaging to identify targets of supermere-associated Discoidin Domain Receptor 1 involved in collagen alignment

Abstract Immune checkpoint blockade has been shown to be an effective treatment for colorectal cancers (CRCs) highly infiltrated by CD8+ T cells. However, there is a paucity of CD8+ T-cell infiltration in the majority of CRCs. We previously identified the collagen-activated receptor Discoidin Domain Receptor 1 (DDR1) as a member of a four-gene signature associated with T-cell exclusion in CRC. The cleaved ectodomain (cECD) of DDR1 induces collagen alignment, leading to T-cell exclusion in a breast cancer model. We found that almost all DDR1 cECD is found in supermeres, a novel secreted nanoparticle identified by our lab, and that DDR1 cECD is required for supermere signaling in recipient cells. We hypothesized that supermere-associated DDR1 cECD signals to CRC stromal cells, leading to collagen alignment. To test this hypothesis, we developed a method to determine which genes are associated with collagen alignment and DDR1 expression. To do this, 30 μm thick CRC sections are imaged using a Nikon AX multiphoton microscope by second harmonic generation imaging, which permits label-free imaging of collagen fibers. Spatial heatmaps of the collagen alignment are generated from the resulting images. Spatial transcriptomics is performed on a serial tumor section using the Visium v2 platform and the gene expression matrix is integrated with the collagen alignment heatmap data. As proof-of-principle, two CRC tumors have been analyzed using this method. Highly aligned regions were found to be associated with expression of genes involved in extracellular matrix organization, keloidal collagen and specific fibroblast subtypes. Genes which may be regulated by DDR1 cECD and be involved in collagen alignment will be confirmed at the protein level by multiplexed immunofluorescence on serial sections and in vitro experiments. This method will provide a rich dataset to elucidate the roles of supermeres and DDR1 cECD in T-cell exclusion and collagen fiber alignment and may support the discovery of new strategies to overcome immune exclusion in CRC. Citation Format: Maxwell S. Hamilton, Oleg Tutanov, Harsimran Kaur, Alan J. Simmons, Clark Massick, Kasey Vickers, Ken S. Lau, Ambra Pozzi, Robert J. Coffey. Integrating spatial transcriptomics and second harmonic generation imaging to identify targets of supermere-associated Discoidin Domain Receptor 1 involved in collagen alignment [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A033.

Effects of Geometry and Supporting Silicone Layers on the Performance of Conductive Composite High-Deflection Strain Gauges

Piezoresistive sensors composed of nickel nanostrands, nickel-coated carbon fibers, and silicone can be used to measure large physical deflections but exhibit viscoelastic properties and creep, leading to a complex and nonlinear electrical response that is difficult to interpret. This study considers the impact of modifying the geometry and architecture of the sensors on their mechanical and electrical performance. Varying the sensor thickness leads to potentially significant differences in conductive fiber alignment, while adding external layers of pure silicone provides elastic support for the sensors, potentially reducing their extreme viscoelastic nature. The impact of such modifications on both mechanical and electrical behavior was assessed by analyzing strain to failure, the magnitude of hysteresis with cycling, the repeatability of the electro-mechanical response, the strain level at which resistance begins to monotonically decrease, and the drift in electrical response with cycling. The results indicate that thicker single-layer sensors have less electrical drift. Sensors with a multilayered architecture exhibit several improvements in behavior, such as increasing the range of the monotonic region by approximately 52%. These improvements become more significant as the thickness of the pure silicone layers increases.

Understanding the interplay between extracellular matrix topology and tumor-immune interactions: Challenges and opportunities.

Modern cancer management comprises a variety of treatment strategies. Immunotherapy, while successful at treating many cancer subtypes, is often hindered by tumor immune evasion and T cell exhaustion as a result of an immunosuppressive tumor microenvironment (TME). In solid malignancies, the extracellular matrix (ECM) embedded within the TME plays a central role in T cell recognition and cancer growth by providing structural support and regulating cell behavior. Relative to healthy tissues, tumor associated ECM signatures include increased fiber density and alignment. These and other differentiating features contributed to variation in clinically observed tumor-specific ECM configurations, collectively referred to as Tumor-Associated Collagen Signatures (TACS) 1-3. TACS is associated with disease progression and immune evasion. This review explores our current understanding of how ECM geometry influences the behaviors of both immune cells and tumor cells, which in turn impacts treatment efficacy and cancer evolutionary progression. We discuss the effects of ECM remodeling on cancer cells and T cell behavior and review recent in silico models of cancer-immune interactions.

Tolerances of the alignment of fibers to photonic integrated circuits

This work presents the solutions to the alignment of two Gaussian beams representing two sides of the optical connection between a fiber and a photonic integrated circuit (PIC). The focus is to use these solutions to derive equations that allow for the straightforward calculation of alignment tolerances for displacement in all three directions, as well as two axes of rotations. These equations can be used to aid in the design of photonic packages by allowing the determination of precision necessary for fabrication of mechanical alignment features, in addition to packaging process development.

Embryo movement is required for limb tendon maturation

IntroductionFollowing early cell specification and tenocyte differentiation at the sites of future tendons, very little is known about how tendon maturation into robust load-bearing tissue is regulated. Between embryonic day (E)16 and E18 in the chick, there is a rapid change in mechanical properties which is dependent on normal embryo movement. However, the tissue, cellular and molecular changes that contribute to this transition are not well defined.MethodsHere we profiled aspects of late tendon development (collagen fibre alignment, cell organisation and Yap pathway activity), describing changes that coincide with tissue maturation. We compared effects of rigid (constant static loading) and flaccid (no loading) immobilisation to gain insight into developmental steps influenced by mechanical cues.ResultsWe show that YAP signalling is active and responsive to movement in late tendon. Collagen fibre alignment increased over time and under static loading. Cells organise into end-to-end stacked columns with increased distance between adjacent columns, where collagen fibres are deposited; this organisation was lost following both types of immobilisation.DiscussionWe conclude that specific aspects of tendon maturation require controlled levels of dynamic muscle-generated stimulation. Such a developmental approach to understanding how tendons are constructed will inform future work to engineer improved tensile load-bearing tissues.

Optimizing aligned steel fiber distribution in UHPC: Unraveling the electromagnetic influence of steel reinforcements

The performance of ultra-high performance concrete (UHPC) can be improved by the magnetic orientation of steel fibers. The reinforcement frame has strong interference with the magnetic field, and this interference may even make the production of oriented steel fibers fail. However, the study of the electromagnetic interference caused by fixed steel reinforcements has not been undertaken. In this paper, the steel fiber distributions under the interference of steel bars were tested, and the impact of different steel reinforcement diameters, lengths, and magnetic flux densities on the magnetic field was studied. The results reveal that stirrups minimally impact the magnetic field, whereas longitudinal reinforcement exerts a substantial influence. Therefore, it is feasible to manufacture an aligned steel fiber UHPC reinforced structure with a steel framework, and only the influence of longitudinal reinforcements needs to be considered. An analytical model based on electromagnetic theory was employed to elucidate the influence of longitudinal reinforcement and analyze the distribution of magnetic lines. Numerical analysis shows that the influenced range of the magnetic lines exhibited a logarithmic relationship with the flux density and a linear relationship with the length and diameter of the reinforcement. Therefore, when accompanied by a judiciously selected magnetic flux density, longer and thinner steel reinforcements are found to be conducive to achieving a comprehensive orientation of steel fibers.

Cocoon microstructures through the lens of topological persistence.

Biological materials display a wide array of functionality, often dictated by complicated microstructures. New geometric and topological strategies allow one to describe the microstructures in a precise and systematic way. This article describes the application of topological persistence and other geometric methods to the microstructural analysis of three-dimensional X-ray micro-computed tomography scans of the Bombyx mori silkworm cocoons. These methods allow conclusions to be drawn about pore space gradients, silk fibre thickness gradients and fibre alignment within the cocoon. The study demonstrates the applicability of these topological and geometric methods to quantify and characterize fibrous materials.

On the Effect of Fibre Orientation and MWCNTs on the Strain‐Sensing Performance of TPU/PANI Electrospun Nanofibres

ABSTRACTThe increasing demand for flexible electromechanical devices with superior stretchability, durability, thermal stability and sensitivity has driven the development of new smart functional materials to replace conventional semiconductors. This study utilises combining electrospinning and in situ polymerisation to fabricate four types of thermoplastic polyurethane/polyaniline (TPU/PANI) nanofibrous membranes with varied fibre orientation and multiwalled carbon nanotubes (MWCNTs) incorporation. The combined effects of material composition and fibre alignment on strain‐sensing performance were systematically explored. All membranes exhibited stable, symmetric ohmic behaviour under strain, with current decreasing as strain increased. Remarkable repeatability and stability were observed in cyclic current–time tests. MWCNTs addition significantly enhanced conductivity, with aligned fibres further boosting performance. Gauge factor (GF) measurements revealed that aligned TPU/PANI mats had GFs of 7 (0%–18% strain) and 14.3 (18%–40%), while aligned TPU/MWCNTs/PANI mats had GFs of 5.2 (0%–13%) and 7.3 (13%–40%). Random TPU/PANI mats showed GFs of 13.9 (0%–11%) and 23.3 (11%–40%) and random TPU/MWCNTs/PANI mats had GFs of 13.9 (0%–19%) and 24.7 (19%–40%). Each configuration demonstrated suitability for specific applications, with random TPU/MWCNT/PANI mats exhibiting the best overall performance. This research provides critical insights into optimising flexible strain sensors by tuning both material composition and fibre orientation.

Confinement Modulates Axial Patterning in Regenerating Hydra

The establishment of the body plan is a major step in animal morphogenesis. The role of mechanical forces and feedback in patterning the body plan remains unclear. Here we explore this question by studying regenerating Hydra tissues confined in narrow cylindrical channels, which constrain their morphology. We find that frustration between the orientation of the channel and the inherited axis in the regenerating tissues can lead to the formation of a multiaxial body plan. The morphological outcome is directly related to the pattern of nematic topological defects that emerges in the organization of the supracellular actomyosin fibers. When the inherited axis, which can be read out from the initial alignment of the supracellular fibers in the confined spheroid, is parallel to the channel's axis, the tissue regenerates normally into animals with a single body axis aligned with the channel. However, regenerating spheroids that are confined in a frustrated perpendicular configuration often develop excess defects (including negatively charged −1/2 defects) and regenerate into multiaxial morphologies. The influence of mechanical constraints on the regenerated body plan argues against an axial patterning mechanism that is based solely on inherited gradients of biochemical morphogens. We further show that the dependence of the regeneration outcomes on the initial tissue orientation can be recapitulated by a biophysical model, which considers the coupled dynamics of the nematic organization of the actomyosin fibers and a morphogen concentration field, incorporating a mechanochemical feedback loop involving strain-dependent morphogen production at defect sites. Published by the American Physical Society 2024

Programmable and flexible wood-based origami electronics

Natural polymer substrates are gaining attention as substitutes for plastic substrates in electronics, aiming to combine high performance, intricate shape deformation, and environmental sustainability. Herein, natural wood veneer is converted into a transparent wood film (TWF) substrate. The combination of 3D printing and origami technique is established to create programmable wood-based origami electronics, which exhibit superior flexibility with high tensile strength (393 MPa) due to the highly aligned cellulose fibers and the formation of numerous intermolecular hydrogen bonds between them. Moreover, the flexible TWF electronics exhibit editable multiplexed configurations and maintain stable conductivity. This is attributed to the strong adhesion between the cellulose-based ink and TWF substrate by non-covalent bonds. Benefiting from its anisotropic structure, the programmability of TWF electronics is achieved through sequentially folding into predesigned shapes. This design not only promotes environmental sustainability but also introduces its customizable shapes with potential applications in sensors, microfluidics, and wearable electronics.

Electrospinning Aligned SF/Magnetic Nanoparticles-Blend Nanofiber Scaffolds for Inducing Skeletal Myoblast Alignment and Differentiation.

In the realm of skeletal muscle tissue engineering, anisotropic materials that emulate natural tissues show substantial promise. Electrospun scaffolds, mimicking the fibrillar structure of the extracellular matrix, are commonly employed but often fall short in achieving optimal alignment and mechanical strength. Silk fibroin has emerged as a versatile material in tissue engineering, valued for its biocompatibility, mechanical robustness, and biodegradability. However, conventional electrospinning methods of SF result in randomly oriented fibers, limiting their efficacy. In this work, we developed a straightforward method to fabricate directional tissue scaffolds using silk fibroin. By integrating a magnetic field collecting device and incorporating Fe3O4 nanoparticles into the spinning solution, we successfully produced well-aligned silk nanofiber scaffolds. These aligned fibers not only improved scaffold orientation and mechanical properties but also exhibited magnetic responsiveness. The aligned SF scaffolds effectively guided the adhesion, proliferation, and differentiation of mesenchymal stem cells along the fiber direction. Cultured on these scaffolds, myoblast C2C12 cells demonstrated oriented growth, highlighting the potential of aligned SF fibers in advancing skeletal muscle engineering for biomedical applications.

An ultrastructure analysis of the developing human anterior cruciate ligament tibial enthesis.

This study aimed to investigate the ultrastructural anatomy of the developing ACL tibial enthesis. We hypothesized that enthesis architecture would progressively mature and remodel, eventually resembling that of the adult by the early postnatal stage. Five fresh-frozen human pediatric cadaveric knees aged 1-36 months underwent anatomical dissection to harvest the ACL insertion and underlying tibial chondroepiphysis. The samples were prepared for scanning electron microscopy (SEM) to examine the ultrastructural anatomy of the enthesis and underwent histological staining for circular polarized light (CPL) and light microscopy imaging. SEM analysis of the 1- and 8-month-old samples revealed a shallow interdigitation between the dense fibrous (ligamentous) tissue and unmineralized chondrogenic tissues, with a minimal transition zone. By 11-month, a more complex transition zone was present. By age 19- and 36-month-old, a progressively more complex and defined fibrocartilage zone was observed. CPL analysis revealed distinct collagen fiber continuity, alignment, and organization changes over time. By 19 and 36 months, the samples exhibited complex fiber arrangements and a progression toward uniform fiber orientation. Similarly, histological analysis demonstrated progressive remodeling of the enthesis with increasing age. Our results suggest that the ACL enthesis of the developing knee begins to mimic that of an adult as early as 19 months of age, as a more complex transition between ligamentous and chondro-epiphyseal tissue can be appreciated. We hypothesize that the observed changes are likely due to mechanical loading of the enthesis with the onset of weightbearing. Future investigations of ACL reconstruction and repair will benefit from improved understanding of the chondro-epiphyseal/ACL regions.

Paste extrusion‐based 3D printing of fiber‐reinforced ultra high‐temperature ceramics

AbstractUltra–high‐temperature ceramics (UHTCs) are valued for their extremely high melting temperatures and resistance to active oxidation. However, their low fracture strengths and the difficulties in shaping them into complex geometries hamper their widespread application. This study aims to fabricate zirconium diboride–silicon carbide (ZrB2‐SiC) composites reinforced with aligned SiC fibers by formulating suspensions containing a preceramic polymer, ZrB2, and SiC fibers. This study assessed the influence of fiber alignment on electrical and thermal conductivities, as well as on mechanical strength. The results revealed a significant enhancement in thermal conductivity, particularly when the fibers were aligned, effectively doubling it compared with the non‐aligned parts. Additionally, increasing the fiber content significantly improved the fracture strength, with composites containing 22.5 vol% fibers reaching fracture strengths over 57 MPa. However, the final values did not meet the theoretical expectations because the porosity in pyrolyzed parts exceeded 10%. Furthermore, the study demonstrated a 10‐fold increase in electrical conductivity with fiber alignment compared to that for non‐aligned composites. These results highlight the capability of paste extrusion‐based additive manufacturing in tailoring ultra–high‐temperature ceramic matrix composites (UHTCMCs) with aligned fibers, realizing their suitability for aerospace applications.

Electric Field-Induced Ordered-Structural Aerogels Enable Superinsulation and Multifunctionality.

1D flexible fibers assembled 3D porous networked ceramic fiber aerogels (CFAs) are developed to overcome the brittleness of traditional ceramic particle aerogels. However, existing CFAs with disordered and quasi-ordered structures fail to balance the relationship between flexibility, robustness, and thermal insulation. Creating novel architectural CFAs with an excellent combination of performances has proven extremely challenging. In this paper, a novel strategy is adopted to fabricate porous mullite fibrous aerogels (MFAs) with ordered structures by combining fiber sedimentation and electric field-induced fiber alignment techniques. For the first time, electric field-induced alignment of ceramic fibers is utilized to prepare bulk aerogels on a large scale. The resulting MFAs exhibit ultra-low high-temperature thermal conductivity of 0.0830 W m-1 K-1 at 1000°C, anisotropic mechanical and sound absorption performances, and multifunctionality in terms of the combination of thermal insulation, sound absorption, and hydrophobicity. The successful synthesis of such fascinating materials may provide new insights into the design and development of multifunctional CFAs for various applications.

Diseased Tendon Models Demonstrate Influence of Extracellular Matrix Alterations on Extracellular Vesicle Profile.

Tendons enable movement through their highly aligned extracellular matrix (ECM), predominantly composed of collagen I. Tendinopathies disrupt the structural integrity of tendons by causing fragmentation of collagen fibers, disorganization of fiber bundles, and an increase in glycosaminoglycans and microvasculature, thereby driving the apparent biomechanical and regenerative capacity in patients. Moreover, the complex cellular communication within the tendon microenvironment ultimately dictates the fate between healthy and diseased tendon, wherein extracellular vesicles (EVs) may facilitate the tendon's fate by transporting biomolecules within the tissue. In this study, we aimed to elucidate how the EV functionality is altered in the context of tendon microenvironments by using polycaprolactone (PCL) electrospun scaffolds mimicking healthy and pathological tendon matrices. Scaffolds were characterized for fiber alignment, mechanical properties, and cellular activity. EVs were isolated and analyzed for concentration, heterogeneity, and protein content. Our results show that our mimicked healthy tendon led to an increase in EV secretion and baseline metabolic activity over the mimicked diseased tendon, where reduced EV secretion and a significant increase in metabolic activity over 5 days were observed. These findings suggest that scaffold mechanics may influence EV functionality, offering insights into tendon homeostasis. Future research should further investigate how EV cargo affects the tendon's microenvironment.

A skin-inspired anisotropic multidimensional sensor based on low hysteresis organohydrogel with linear sensitivity and excellent robustness for directional perception

Human skin has complex sensors to perceive three-dimensional stimuli from the environment. Skin-inspired flexible sensors with multidimensional and directional perception are desirable for applications in intelligent robots and human–machine interaction. Herein, wearable multidimensional sensors have been fabricated by macroscopically assembling 3D-printed microstructured anisotropic organohydrogel sensory units through robust interfacial adhesion. The organohydrogel is crosslinked using microgels and starch microparticles through non-covalent interactions, and composited with short carbon fibers as conductive fillers. The physical network exhibits low hysteresis against 5000 cyclic loadings, which demonstrates excellent robustness in both mechanical properties and sensory performance. The organohydrogel precursor is printed into a cone-shaped sensor with a pressure sensitivity 50 times higher than that of its bulk counterpart. A sea cucumber epidermis-like microstructured sensor with abundant highly sensitive cone array is assembled with 3D-printed anisotropic gels with highly aligned carbon fibers inside, generating a multidimensional sensor capable of perceiving stimuli along X-, Y-, and Z-axis. The assembled sensor can directionally distinguish external forces and identify human motions. This study demonstrates a promising strategy to fabricate well-structured organohydrogel sensors for applications in electronic skin, biomimetic flexible electronics, and artificial intelligence.

Low magnetic field alignment of carbon fibers in a polymer matrix for high-performance thermal interface materials

Carbon-based materials like carbon nanotubes, graphene nanoplatelets, graphite, and carbon fibers (CF) are highly promising fillers for enhancing the thermal conductivity of thermal interface materials (TIMs) due to their high thermal conductivity and low thermal expansion. Aligning these fillers can further improve thermal conductivity, but current alignment methods using high magnetic fields are impractical for industrial use. In this study, we investigated the enhancement of the thermal conductivity of CF–polymer composites by aligning CF fillers with a low magnetic field. We observed up to 17 times enhancement of the thermal conductivity (from 3.1 to 52.8 W/mK) in a CF–graphene–polymer composite film by vertically aligning the CF fillers with a magnetic field of 0.75 T. The analysis of the structural properties of the films using X-ray diffraction and field-emission scanning electron microscopy imaging confirmed that the crystalline c-axis of the graphite plates in the CF was oriented perpendicular to the magnetic field direction. The large anisotropy in the diamagnetic susceptibility of the laminated graphene structure of the CF flakes is at the origin of the filler alignment. Furthermore, the thermal conductivity of the composites showed a strong correlation with the degree of filler alignment, which was dependent on the CF–graphene hybridization and the type of polymer matrix. This study provides a scalable and cost-effective method for enhancing the physical properties of carbon composites by controlling their microarchitecture using a low magnetic field.

Decentralized approach for incorporating waste wind turbine blades into 3D printing filaments using mechanical recycling

AbstractThe increasing awareness in environmental safety has led to rapid development in production of electricity using wind energy with wind turbines. The widespread deployment of wind turbines has outpaced the effective recycling of End‐of‐Life wind turbines. This study explores the potential of mechanically recycling decommissioned wind turbine blades (WTB) as reinforcement material in 3D printing processes. Utilizing mechanical grinding, materials were extracted from the waste blades and subsequently analyzed using Fourier transform infrared spectroscopy, Differential scanning calorimetry, and thermogravimetric analysis to determine optimal processing conditions. The reclaimed materials were then blended with recycled polypropylene through single‐screw extrusion to fabricate tensile test samples via Fused Deposition Modeling. The impact of print orientation on mechanical strength was examined at 0°, 45°, and 90° angles. Morphological analysis was conducted on the fractured specimens to assess the failure characteristics. The findings indicate that samples printed at a 90° orientation exhibited superior mechanical properties, suggesting a viable pathway for incorporating wind turbine waste into sustainable manufacturing cycles.Highlights A decentralized‐mechanical recycling technique to the waste WTB. The necessary material parameters for the operations employed in this study. Reinforced 3D printable filaments from waste WTB. Stronger reinforced filaments obtained from proper fiber alignment.

Full‐field effect of flow‐induced fiber orientation during compression molding of carbon fiber sheet molding compounds

AbstractAn image‐based fiber orientation analysis technique has been employed to study the effect of initial charge coverage and preferential fiber alignment on the flow‐induced fiber orientations in carbon fiber sheet molding compounds (SMCs). The initial charge coverage area exhibited minimal fiber reorientation while significant fiber alignment in the flow direction was observed in the regions of greatest flow. Novel subsurface analysis using the same full‐field orientation analysis technique also showed the presence of a skin‐core structure with greater flow‐induced fiber alignment at the mold surfaces, while the core retained more of the initial fiber orientation distribution. Edge effects were also revealed by this analysis at the mold walls. Tensile testing from two orthogonal directions showed that panels molded from lower charge coverage resulted in better mechanical performance and lower anisotropy. Moreover, by encouraging charge flow in the preform's preferential alignment direction, high tensile stiffness and strength in the preferred direction were achieved (56.7 GPa and 238.9 MPa, respectively) at 30% fiber volume fraction. This provides a practical demonstration of the value in controlling fiber orientation, with respect to charge positioning and mold flow, to maximize and tailor the composite performance of SMCs typically used for automotive applications.Highlights Low‐cost scanning technique is extended to assess internal fiber orientations Full‐field orientations are measured before and after molding Investigation of how charge size and shape can affect orientations Combining effects of flow and initial orientation to tailor SMC performance Circular statistics provides a thorough analysis of orientation distributions