Fibrous Viscoelastic Extracellular Matrix Assists Precise Neuronal Connectivity

Synapse formation in complex neuronal network is a pivotal process for normal functioning of nervous system. Although intense research has been conducted, how neurons and axons are guided toward the target remains largely unclear. In traditional opinions, axons are directed through chemotaxis, while recently mechanotaxis has been brought up as a potential complementary mechanism, as it can provide delicately controlled signals in addition to the random diffusive chemical cues. To further explore the path-finding mechanism, a quasi-3D in vitro model for neuronal cells is constructed by integrating hydrogel collagen I as extracellular matrix (ECM), and primary mouse cortical neurons and PC12 cells are tested. It is strikingly found out that axons and neuronal cells can be precisely guided toward target neurites via ECM. By developing a label-free traction force microscopy technique, the force networks among neurons are presented, validating that the fibrous matrix-transmitted paratensile signals can assist the axon pathfinding. This precise axon guidance is related to the activation of mechanosensitive ion channels, calcium signaling, and probably the following F-actin assembly. This mechanism can potentially assist developing clinical applications and designing biomaterials in near future.     

Gelatin-Hyaluronan Click-Crosslinked Cryogels Elucidate Human Macrophage Invasion Behavior

Hydrogel models of metastasis traditionally focus on the invasion of cancer cells; however, other cells in the tumor microenvironment that are associated with metastasis also have the ability to migrate. Macrophage phenotype plays a key role in the tumor microenvironment, yet understanding their migration within tunable 3D in vitro models has been limited. To gain a greater understanding of macrophage invasive behavior, stable and transparent oxime-crosslinked cryogels comprised of click-crosslinked gelatin-oxyamine and hyaluronan-aldehyde (GELox-HAa) are synthesized. Fibronectin-derived, oxyamine-modified PHSRN-RGDSP peptides are incorporated to further mimic the tumor extracellular matrix without impacting cryogel mechanics. It is found that primary human macrophages migrate to a greater depth in cryogels with greater porosity and pore size. To better understand the mechanism of migration, cells are treated with either inhibitors of matrix metalloproteinases (MMPs) or rho-associated protein kinase (ROCK) and a predominantly MMP-mediated mechanism of invasion is found. Macrophage polarization studies reveal that anti-inflammatory, interleukin-4/13 (IL4/IL13)-treated macrophages migrate through cryogels to a greater extent than pro-inflammatory, interferon-gamma/lipopolysaccharide (IFNγ/LPS)-treated cells. Interestingly, polarized macrophages move through cryogels using a combination of amoeboid and mesenchymal migration. These findings of macrophage invasion in this cryogel platform set the stage for their further study in a biomimetic tumor microenvironment.     

Injectable Scaffolds for In Vivo Programmed Macrophages Manufacture and Postoperative Cancer Immunotherapy

Cancer recurrence and metastasis after surgical resection is a vital reason of treatment failure. The modification of immune cells through implanted biomaterials is a promising postoperative immunotherapy. Herein, an injectable hydrogel scaffold loaded with engineered exosome mimetics that in vivo recruits and programs endogenous macrophages into M1 binding with anti-CD47 antibody (M1-aCD47 macrophages) for postoperative cancer immunotherapy is developed. Briefly, M1 macrophages-derived exosome mimetics co-modified with vesicular stomatitis virus glycoprotein (VSV-G) and aCD47 (V-M1EM-aCD47) are encapsulated in injectable chitosan hydrogel. Such hydrogel recruits inherent macrophages in situ and releases V-M1EM-aCD47 that programs M2 to M1-aCD47 macrophages. M1-aCD47 macrophages own dual-functions of tumor-homing and enhanced phagocytosis. They can actively target to tumor cells for delivery of aCD47 that blocks the “don't eat me” signal, thereby promoting phagocytosis of macrophages to cancer cells. Furthermore, V-M1EM-aCD47 hydrogel implanted into resection site of 4T1 breast tumor inhibits tumor recurrence and metastasis by phagocytosis of M1-aCD47 macrophages and T cell-mediated immune responses. The findings demonstrate that biomaterials can be designed in vivo to program inherent macrophages, thereby activating the innate and adaptive immune systems for prevention of postoperative tumor recurrence and metastasis.     

Macrophage-Mediated Tumor Cell Phagocytosis: Opportunity for Nanomedicine Intervention

Macrophages are one of the most abundant non-malignant cells in the tumor microenvironment, playing critical roles in mediating tumor immunity. As important innate immune cells, macrophages possess the potential to engulf tumor cells and present tumor-specific antigens for adaptive antitumor immunity induction, leading to growing interest in targeting macrophage phagocytosis for cancer immunotherapy. Nevertheless, live tumor cells have evolved to evade phagocytosis by macrophages via the extensive expression of anti-phagocytic molecules, such as CD47. In addition, macrophages also rapidly recognize and engulf apoptotic cells (efferocytosis) in the tumor microenvironment, which inhibits inflammatory responses and facilitates immune escape of tumor cells. Thus, intervention of macrophage phagocytosis by blocking anti-phagocytic signals on live tumor cells or inhibiting tumor efferocytosis presents a promising strategy for the development of cancer immunotherapies. Here, the regulation of macrophage-mediated tumor cell phagocytosis is first summarized, followed by an overview of strategies targeting macrophage phagocytosis for the development of antitumor therapies. Given the potential off-target effects associated with the administration of traditional therapeutics (for example, monoclonal antibodies and small molecule inhibitors), the opportunity for nanomedicine in macrophage phagocytosis intervention is highlighted.     

A Nano “Immune-Guide” Recruiting Lymphocytes and Modulating the Ratio of Macrophages from Different Origins to Enhance Cancer Immunotherapy

Artificially modulating the type, density, and location of immune cells within the tumor microenvironment can suppress tumor growth and efficiently promote current immunotherapy. In this study, a magnetite nanoparticle-based “immune-guide” is developed by the functionalization of magnetite nanoparticles with hyaluronic acid (HA). HA, an extracellular matrix component, can target various CD44-overexpressing tumors and mediate the adhesion and migration of multiple types of immune cells. Thus, HA-functionalized magnetite nanoparticles (HA-PDA@Fe₃O₄) can highly efficiently accumulate in breast cancer and penetrate deep into the tumor parenchyma. Consequently, high intratumoral concentration of HA, serving as a “guidepost,” can directly recruit lymphocytes and elicit more chemokine production through cascading amplification effects, turning the immune “cold” tumor into a “hot” one. More importantly, HA-PDA@Fe₃O₄ can effectively remodel the diversity, origin, and activation of tumor-associated macrophages by recruiting and activating infiltrating macrophages, while simultaneously reducing the M2-maintained tissue-resident macrophages. Thus, HA-PDA@Fe₃O₄ synergistically improves T cell- and macrophage-based immunotherapies as well as interferes with the formation of premetastatic niches in the lung. By redistributing the localization of HA in tumors by using magnetite nanoparticles, this study provides a unique strategy to modulate the tumor immune microenvironment and potentiate tumor immunotherapies by using biocompatible nanomaterials without any therapeutic drug.     

Engineered 3D Silk‐Based Metastasis Models: Interactions Between Human Breast Adenocarcinoma,Mesenchymal Stem Cells and Osteoblast‐Like Cells

Bone metastasis occurs in 70% of breast cancer patients and is a frequent cause of morbidity in cancer patients. A delicate balance exists in the bone microenvironment, but the functional dynamics underlying the tumor cell‐microenvironment interactions remain poorly understood. 3D in vitro model systems of metastasis can throw new light on this phenomenon. Silk protein fibroin scaffolds, are cytocompatible for 3D cancer cell culture. They are structurally more resistant to protease degradation than other native biomaterials making these matrices suitable for cancer modeling. In this report, human breast adenocarcinoma cells, human osteoblast like cells and mesenchymal stem cells are co‐cultered. Cancer cells and osteoblast‐like cells are found to interact through secreted products. Decreased population of osteoblast‐like cells and mineralization of extracellular matrix are observed as a result of co‐culture. Significantly increased migration of breast cancer cells is observed in the bone‐like constructs than in non‐seeded scaffolds. The co‐culture constructs show significant increase in drug resistance, invasiveness and angiogenicity. Co‐culture of breast cancer cells with osteoblast like cells and mesenchymal stem cells also indicate that the interaction of cancer cells with bone microenvironment varies with spatial organization, presence of osteogenic factors as well as stromal cell type. Here, results show that 3D in vitro co‐culture models is possibly a better system to study and target cancer progression.     

Designer Biomaterials to Model Cancer Cell Invasion In Vitro: Predictive Tools or Just Pretty Pictures?

Metastasis is the leading cause of mortality in cancer patients. Underlying this process is the invasion and colonization of cancer cells into healthy tissues. Engineered hydrogel models of tumor microenvironments present an opportunity to understand the microenvironmental determinants of cellular invasion. The biochemical and mechanical cues, presented in the form of adhesion sites, degradable cues, matrix stiffness, and architecture, have significant effects on the extent of cancer cell migration, and the mechanisms employed by these cells to move through their matrix. Coculture with stromal cells such as cancer associated fibroblasts, endothelial cells, and immune cells that are associated with poor prognosis demonstrate that these cells exacerbate cancer cell invasion. With these models, researchers aim not only to recapitulate known cancer cell behaviors in a dish, but also to uncover new insights into mechanisms underlying these phenomena, paving the way for novel treatment strategies. In this perspective, the design of engineered models that are used to study cancer cell invasion and metastasis in vitro is discussed. To this end, the authors seek to understand and put into perspective: do these models reveal relevant mechanisms of cancer cell migration, or are they simply pretty pictures with little biological translatability?     

Inhibition of Cancer Cell Migration by Gold Nanorods: Molecular Mechanisms and Implications for Cancer Therapy

Gold nanorods have received much attention because of their distinct physicochemical properties and promising applications in bioimaging, biosensing, drug delivery, photothermal therapy, and optoelectronic devices. However, little is known regarding their effect on tumor metastasis. In the present investigation, serum protein‐coated gold nanorods (AuNRs) at low concentrations is shown to exhibit no apparent effects on the viability and proliferation of three different metastatic cancer cell lines, that is, MDA‐MB‐231 human breast cancer cells, PC3 human prostate cancer cells, and B16F10 mouse melanoma cells, but effectively inhibit their migration and invasion in vitro. Quantitative proteomics and real‐time PCR array analyses indicate that exposure of cells to AuNRs can down‐regulate the expression of diverse energy generation‐related genes, which accounts for their inhibition of mitochondrial oxidative phosphorylation (OXPHOS) and glycolysis. The impairment of OXPHOS and glycolysis results in a distinctive reduction of ATP production and subsequent inhibition of F‐actin cytoskeletal assembly, which is crucial for the migration and invasion of cancer cells. The inhibitory effect of AuNRs on cancer cell migration is also confirmed in vivo. Taken together, the unique mechanism in inhibiting cancer cell migration by AuNRs might provide a new approach to specific cancer therapeutic treatment.     

Magnetic Electrospun Fibers for Cancer Therapy

Iron oxide nanoparticles (IONPs) for magnetic hyperthermia in cancer treatment have recently gained substantial interest. Unfortunately, the use of free IONPs still faces major challenges such as poor tumor targetability, high variability in the amount of IONPs taken up by the tumor and the IONP leakage from dead cancer cells into the surrounding healthy tissues. The present work reports on electrospun fiber webs, heavily loaded with 50 nm sized IONPs. The high loading capacity of the fibers enables significant heating of the environment upon applying an alternating magnetic field. Furthermore, magnetic fibers can be repeatedly heated without loss of heating capacity or release of IONPs. Upon functionalization of the fiber surface with collagen, human SKOV‐3 ovarian cancer cells attached well to the fibers. Applying an alternating magnetic field during 10 minutes to the fiber webs killed all fiber‐associated cancer cells. Killing the cells using this method seemed more efficient compared to the use of a warm water bath. As the fiber webs can be i) loaded with a well‐controlled amount of IONPs and ii) localized in the body by Magnetic Resonance Imaging, magnetic electrospun fibers may become promising materials for a highly reproducible (repeated) heating of cancer tissues in vivo.     

An Interplay between Matrix Anisotropy and Actomyosin Contractility Regulates 3D‐Directed Cell Migration

Directed cell migration is essential for many biological processes, such as embryonic development or cancer progression. Cell contractility and adhesion to the extracellular matrix are known to regulate cell locomotion machinery. However, the cross‐talk between extrinsic and intrinsic factors at the molecular level on the biophysical mechanism of three dimensional (3D)‐directed cell migration is still unclear. In this work, a novel physiologically relevant in vitro model of the extracellular microenvironment is used to reveal how the topological anisotropy of the extracellular matrix synergizes with actomyosin contractility to modulate directional cell migration morphodynamics. This study shows that cells seeded on polarized 3D matrices display asymmetric protrusion morphodynamics and in‐vivo‐like phenotypes. It is found that matrix anisotropy significantly enhances cell directionality, but strikingly, not the invasion distance of cells. In Rho‐inhibited cells, matrix anisotropy counteracts the lack of actomyosin‐driven forces to stabilize cell directionality suggesting a myosin‐II‐independent mechanism for cell guidance. Finally, this study shows that on isotropic 3D environments, cell directionality is independent of actomyosin contractility. Altogether, this study provides novel quantitative data on the biomechanical regulation of directional cell motion and shows the important regulatory role of matrix anisotropy and actomyosin forces to guide cell migration in 3D microenvironments.     

Dual Targeted Immunotherapy via In Vivo Delivery of Biohybrid RNAi‐Peptide Nanoparticles to Tumor‐Associated Macrophages and Cancer Cells

Lung cancer is associated with very poor prognosis and considered one of the leading causes of death worldwide. Here, highly potent and selective biohybrid RNA interference (RNAi)‐peptide nanoparticles (NPs) are presented that can induce specific and long‐lasting gene therapy in inflammatory tumor associated macrophages (TAMs), via an immune modulation of the tumor milieu combined with tumor suppressor effects. The data here prove that passive gene silencing can be achieved in cancer cells using regular RNAi NPs. When combined with M2 peptide–based targeted immunotherapy that immuno‐modulates TAMs cell population, a synergistic effect and long‐lived tumor eradication can be observed along with increased mice survival. Treatment with low doses of siRNA (ED₅₀ 0.0025–0.01 mg kg^?1) in a multi and long‐term dosing system substantially reduces the recruitment of inflammatory TAMs in lung tumor tissue, reduces tumor size (≈95%), and increases animal survival (≈75%) in mice. The results here suggest that it is likely that the combination of silencing important genes in tumor cells and in their supporting immune cells in the tumor microenvironment, such as TAMs, will greatly improve cancer clinical outcomes.     

Double Charge Transfer Dominates in Carrier Localization in Low Bandgap Sites of Heterogeneous Lead Halide Perovskites

Heterogeneous organic-inorganic halide perovskites possess inherent non-uniformities in bandgap that are sometimes engineered and exploited on purpose, like in quasi-2D perovskites. In these systems, charge carrier and excitation energy migration to lower-bandgap sites are key processes governing luminescence. The question, which of them dominates in particular materials and under specific experimental conditions, still remains unanswered, especially when charge carriers comprise excitons. In this study transient absorption (TA) and transient photoluminescence (PL) techniques are combined to address the excited state dynamics in quasi-2D and other heterogeneous perovskite structures in broad temperature range, from room temperature down to 15 K. The data provide clear evidence that charge carrier transfer rather than energy migration dominates in heterogeneous quasi-2D perovskite films.     

A Non‐Mulberry Silk Fibroin Protein Based 3D In Vitro Tumor Model for Evaluation of Anticancer Drug Activity

The limitations of clinical chemotherapy are credited primarily to drug resistance. Effective development and screening of new drugs require appropriate in vitro tumor models that resemble the in vivo situation to evaluate drug efficiency and to decrease the use of experimental animals. 3D in vitro model systems that are able to mimic in vivo microenvironments are now highly sought after in cancer research. Here, the characteristics of breast cancer cell line MDA‐MB‐231 cells on 3D, and 2D Antheraea mylitta silk matrices and tissue culture plates are compared. After long term culture of breast cancer cells in the silk scaffold, the engineered tumor construct shows different zones of cell proliferation, such as an avascular tumor. Silk fibroin matrix 3D tumor models are studied for the evaluation of various anticancer drugs. The cytotoxic effects of three different drugs (Paclitaxel, Celecoxib, and ZD6474) at different concentrations are evaluated for MDA‐MB‐231 grown on 2D films as well as on a 3D fibroin scaffold. Higher drug concentrations are required to achieve a comparable reduction in cell viability and invasive potential in 3D culture. Combinatorial treatment of drugs at IC₅₀ concentrations result in up to 84% death of cancer cells. The results indicate that 3D in vitro tumor models may be better systems to evaluate cancer treatment strategies.     

Domain Distribution Management of Quasi-2D Perovskites toward High-Performance Blue Light-Emitting Diodes

Quasi-2D perovskites, as one of the promising materials applied in perovskite light-emitting diodes (PeLEDs), have attracted great attention for their superior semiconductor properties. The inherent multiquantum well structure can induce a strong confinement effect, which is especially suitable for blue emission. However, compared to their green counterparts, blue emitters constructed from quasi-2D perovskites are more sensitive to n domain distribution (where n represents the number of PbX₆ inorganic layers). Suffering from inefficient domain distribution management, blue PeLEDs now face a variety of negative issues, including color instability, multipeak emission, and poor fluorescence yield. In this review, the development of blue PeLEDs and the optical properties of quasi-2D perovskites are overviewed. Then, a classification and summary of strategies for domain distribution management are proposed. Finally, the challenges and potential directions of domain distribution management in quasi-2D perovskites are summarized. This review is expected to provide a comprehensive perspective and reference on domain distribution management toward efficient blue quasi-2D PeLEDs.     

The Antitumor Efficacy of CpG Oligonucleotides is Improved by Encapsulation in Plant Virus‐Like Particles

Oligodeoxynucleotides (ODNs) with CpG motifs have potent immunostimulatory effects on many subsets of immune cells. For example, Class B CpG‐ODNs, such as ODN1826 induce the phagocytic activity of macrophages by activating the Toll‐like receptor 9 signaling pathway. Systemic ODN delivery results in unfavorable pharmacokinetic profiles and can trigger adverse effects. To address this issue, plant virus‐like particles (VLPs) are developed for the targeted delivery of ODN1826 to tumor‐associated macrophages (TAMs). ODN1826 is encapsulated by the in vitro disassembly and reassembly of Cowpea chlorotic mottle virus (CCMV), producing VLPs that are structurally analogous to the native virus. The encapsulation of ODN1826 in CCMV‐derived VLPs promotes ODN uptake by TAMs ex vivo and significantly enhance their phagocytic activity. The antitumor activity of the VLPs in vivo is also evaluated, revealing that the direct injection of ODN1826 VLPs into established tumors induces a robust antitumor response by increasing the phagocytic activity of TAMs in the tumor microenvironment. CCMV encapsulation significantly enhances the efficacy of ODN1826 compared to the free drug, slowing tumor growth and prolonging survival in mouse models of colon cancer and melanoma.     

Smart Injectable Hydrogels for Cancer Immunotherapy

Hydrogels, a class of materials with a 3D network structure, are widely used in various fields especially in biomedicine. Injectable hydrogels could facilitate the encapsulation and controlled release of small molecular drugs, macromolecular therapeutics, and even cells. With the rapid development of cancer immunotherapy, such injectable hydrogels have attracted wide attention for local immunomodulation to boost systemic anticancer immune responses, realizing more effective immunotherapy at lower doses. The latest progresses in the development of various smart injectable hydrogels for cancer immunotherapy are summarized here. Although applied locally, such injectable hydrogels can activate systemic antitumor immune responses, safely and effectively inhibiting the tumor metastasis and recurrence. Moreover, it is discussed how injectable hydrogel‐based cancer immunotherapy would contribute to the development of next generation of cancer treatment together with their potential for clinical translation.     

Engineered Oxygen Factories Synergize with STING Agonist to Remodel Tumor Microenvironment for Cancer Immunotherapy

Immunotherapy is a revolutionary achievement in cancer treatment. However, inadequate immune cells infiltration in tumor microenvironment (TME) always leads to treatment failure. Moreover, hypoxic TME hampers normal functions of immune cells. Here, it is found that hypoxia suppresses the STING signaling and immune cells activation in the work. Remodeling tumor immune microenvironment and relieving hypoxia are thus essential for enhancing immunotherapy efficiency. Herein, a spirulina platensis (SP)-based magnetic biohybrid system is constructed as an oxygen factory and loaded with stimulator of interferon genes (STING) agonist ADU-S100 (ADU@Fe-SP) for tumor immunotherapy. Magnet-guided biohybrid SP can actively target tumor tissues and produce oxygen in situ through photosynthesis, which reverses the hypoxic TME and facilitates the function of immune cells. Besides, the targeted delivery of ADU-S100 can activate the STING/TBK1/IRF3 signaling and boost cytokines production in tumor and innate immune cells. The ADU@Fe-SP system thus induces efficient immune cells infiltration in TME, which efficiently inhibits tumor progression and significantly enhances anti-PD-1 therapy efficiency in SCC VII-bearing tumor xenograft. ADU@Fe-SP exerts antitumor effect in a STING-dependent manner by in vivo STING-knockout mice model. The efficiency of this immunotherapy strategy is also demonstrated in patient-derived xenograft model originating from oral cancer, showing great clinical potential.     

Tumor‐Penetrating Nanotherapeutics Loading a Near‐Infrared Probe Inhibit Growth and Metastasis of Breast Cancer

The tumor growth and metastasis is the leading reason for the high mortality of breast cancer. Herein, it is first reported a deep tumor‐penetrating photothermal nanotherapeutics loading a near‐infrared (NIR) probe for potential photothermal therapy (PTT) of tumor growth and metastasis of breast cancer. The NIR probe of 1,1‐dioctadecyl‐3,3,3,3‐tetramethylindotricarbocyanine iodide (DiR), a lipophilicfluorescent carbocyanine dye with strong light‐absorbing capability, is entrapped into the photothermal nanotherapeutics for PTT application. The DiR‐loaded photothermal nanotherapeutics (DPN) is homogeneous nanometer‐sized particles with the mean diameter of 24.5 ± 4.1 nm. Upon 808 nm laser irradiation, DPN presents superior production of thermal energy than free DiR both in vitro and in vivo. The cell proliferation and migration activities of metastatic 4T1 breast cancer cells are obviously inhibited by DPN in combination with NIR irradiation. Moreover, DPN can induce a higher accumulation in tumor and penetrate into the deep interior of tumor tissues. The in vivo PTT measurements indicate that the growth and metastasis of breast cancer are entirely inhibited by a single treatment of DPN with NIR irradiation. Therefore, the deep tumor‐penetrating DPN can provide a promising strategy for PTT of tumor progression and metastasis of breast cancer.     

Efficient and Stable Quasi-2D Perovskite Solar Cells Enabled by Thermal-Aged Precursor Solution

Quasi-2D perovskites have received wide attention in photovoltaics owing to their excellent materials robustness and merits in the device stability. However, the highest power conversion efficiency (PCE) reported on quasi-2D perovskite solar cells (PSCs) still lags those of the 3D counterparts, mainly caused by the relatively high voltage loss. Here, a study is presented on the mitigation of voltage loss in quasi-2D PSCs via usage of thermal-aged precursor solutions (TAPSs). Based on the (AA)₂MA₄Pb₅I₁₆ (n = 5) quasi-2D perovskite absorber with a bandgap of ≈1.60 eV, a record-high open-circuit voltage of 1.24 V is obtained, resulting in boosting the PCE to 18.68%. The enhanced photovoltaic performance afforded by TAPS is attributed to the thermal-aged solution processing that triggers colloidal aggregations to reduce the nucleation sites inside the solution. As a result, formation of high-quality perovskite films featuring compact morphology, preferential crystal orientation, and lowered trap density is allowed. Of importance, with the improved film quality, the corrosion of Ag electrode induced by ion migrations is effectively restrained, which leads to a satisfactory storage stability with <2% degradation after 1200 h under nitrogen environment without encapsulation.     

Highly Aligned Nanofibrous Scaffold Derived from Decellularized Human Fibroblasts

Native tissues are endowed with a highly organized nanofibrous extracellular matrix (ECM) that directs cellular distribution and function. The objective of this study is to create a purely natural, uniform, and highly aligned nano­fibrous ECM scaffold for potential tissue engineering applications. Synthetic nanogratings (130 nm in depth) are used to direct the growth of human dermal fibroblasts for up to 8 weeks, resulting in a uniform 70 μm‐thick fibroblast cell sheet with highly aligned cells and ECM nanofibers. A natural ECM scaffold with uniformly aligned nanofibers of 78 ± 9 nm in diameter is generated after removing the cellular components from the fibroblast sheet. The elastic modulus of the scaffold is well maintained after the decellularization process because of the preservation of elastin fibers. Reseeding human mesenchymal stem cells (hMSCs) shows the excellent capacity of the scaffold in directing and supporting cell alignment and proliferation along the underlying fibers. The scaffold's biocompatibility is further examined by an in vitro inflammation assay with seeded macrophages. The aligned ECM scaffold induces a significantly lower immune response compared to its unaligned counterpart, as detected by the pro‐inflammatory cytokines secreted from macrophages. The aligned nanofibrous ECM scaffold holds great potential in engineering organized tissues.