Matrix Hydrogel Research Articles

Neuroblastoma Invasion Strategies Are Regulated by the Extracellular Matrix.

Simple SummaryPaediatric cancer research in general and neuroblastoma, in particular, has minimal preclinical models of metastasis. As 50% of primary neuroblastomas have already metastasised at the time of diagnosis, it is important to develop models to understand the molecular mechanisms of neuroblastoma metastasis. Here, we describe a novel patient-derived xenograft (PDX)- and cell line-based organoid model. We found that the extracellular matrix (ECM) composition influenced the growth, viability and local invasion of organoids. PDX-derived neuroblastoma organoids displayed four various invasion phenotypes which were dependent on the local microenvironment, while cell lines were more restricted in their invasion strategies. These data support the use of organoid cultures for studying the biology and molecular basis of neuroblastoma invasion into normal tissues.Neuroblastoma is a paediatric malignancy of the developing sympathetic nervous system. About half of the patients have metastatic disease at the time of diagnosis and a survival rate of less than 50%. Our understanding of the cellular processes promoting neuroblastoma metastases will be facilitated by the development of appropriate experimental models. In this study, we aimed to explore the invasion of neuroblastoma cells and organoids from patient-derived xenografts (PDXs) grown embedded in 3D extracellular matrix (ECM) hydrogels by time-lapse microscopy and quantitative image analysis. We found that the ECM composition influenced the growth, viability and local invasion of organoids. The ECM compositions induced distinct cell behaviours, with Matrigel being the preferred substratum for local organoid invasion. Organoid invasion was cell line- and PDX-dependent. We identified six distinct phenotypes in PDX-derived organoids. In contrast, NB cell lines were more phenotypically restricted in their invasion strategies, as organoids isolated from cell line-derived xenografts displayed a broader range of phenotypes compared to clonal cell line clusters. The addition of FBS and bFGF induced more aggressive cell behaviour and a broader range of phenotypes. In contrast, the repression of the prognostic neuroblastoma marker, MYCN, resulted in less aggressive cell behaviour. The combination of PDX organoids, real-time imaging and the novel 3D culture assays developed herein will enable rapid progress in elucidating the molecular mechanisms that control neuroblastoma invasion.

BFGF and collagen matrix hydrogel attenuates burn wound inflammation through activation of ERK and TRK pathway

Burn injuries are most challenging to manage since it causes loss of the integrity of large portions of the skin leading to major disability or even death. Over the years, hydrogels are considered as a significant delivery system for wound treatment because of several advantages over other conventional formulations. We hypothesized that the bFGF-collagen-AgSD incorporated hydrogel formulation can accelerate the rate of burn healing in animal model and would promote fibroblast cell proliferation. Neovascularization and re-epithelialization is a hall mark of burn wound healing. In the present study, histopathological investigation and scanning electron microscopy of skin tissue of Wistar rats showed almost complete epithelialisation after 16 days in the treatment group. The developed hydrogel showed significantly accelerated wound closure compared with a standard and control group. The faster wound closure resulted from increased re-epithelialization and granulation tissue formation because of the presence of collagen and growth factor. Expressions of proteins such as TrkA, p- TrkA, ERK1/2, p-ERK1/2, NF-kβ, and p-NF-kβ involved in nerve growth factor (NGF) signalling pathway were analysed by western blot. All the findings obtained from this study indicated that the hydrogel can be considered as a promising delivery system against second degree burn by faster healing.

In Situ Preconditioning of Human Mesenchymal Stem Cells Elicits Comprehensive Cardiac Repair Following Myocardial Infarction.

Human bone marrow-derived mesenchymal stem cells (BM-MSCs), represented as a population of adult stem cells, have long been considered as one of the most promising sources for cell-based cardiac regenerative therapy. However, their clinical use has been significantly hampered by low survival and poor retention following administration into failing hearts. Here, to improve the therapeutic effectiveness of BM-MSCs, we examined a novel therapeutic platform named in situ preconditioning in a rat myocardial infarction (MI) model. In situ preconditioning was induced by a combinatory treatment of BM-MSCs with genetically engineered hepatocyte growth factor-expressing MSCs (HGF-eMSCs) and heart-derived extracellular matrix (hdECM) hydrogel. Subsequently, our results demonstrated that in situ preconditioning with cell mixture substantially improved the survival/retention of BM-MSCs in the MI-induced rat hearts. Enhanced retention of BM-MSCs ultimately led to a significant cardiac function improvement, which was derived from the protection of myocardium and enhancement of vessel formation in the MI hearts. The results provide compelling evidence that in situ preconditioning devised to improve the therapeutic potential of BM-MSCs can be an effective strategy to achieve cardiac repair of MI hearts.

Development of Decellularized Oviductal Hydrogels as a Support for Rabbit Embryo Culture.

The oviducts (fallopian tubes in mammals) function as the site of fertilization and provide necessary support for early embryonic development, mainly via embryonic exposure to the tubal microenvironment. The main objective of this study was to create an oviduct-specific extracellular matrix (oviECM) hydrogel rich in bioactive components that mimics the native environment, thus optimizing the developmental trajectories of cultured embryos. Rabbit oviducts were decellularized through SDS treatment and enzymatic digestion, and the acellular tissue was converted into oviductal pre-gel extracellular matrix (ECM) solutions. Incubation of these solutions at 37°C resulted in stable hydrogels with a fibrous structure based on scanning electron microscopy. Histological staining, DNA quantification and colorimetric assays confirmed that the decellularized tissue and hydrogels contained no cellular or nuclear components but retained important components of the ECM, e.g. hyaluronic acid, glycoproteins and collagens. To evaluate the ability of oviECM hydrogels to maintain early embryonic development, two-cell rabbit embryos were cultured on oviECM-coated surfaces and compared to those cultured with standard techniques. Embryo development was similar in both conditions, with 95.9% and 98% of the embryos reaching the late morula/early blastocyst stage by 48h under standard culture and oviECM conditions, respectively. Metabolomic analysis of culture media in the presence or absence of embryos, however, revealed that the oviECM coating may include signalling molecules and release compounds beneficial to embryo metabolism.

Tomographic evaluation of direct pulp capping using a novel injectable treated dentin matrix hydrogel: a 2-year randomized controlled clinical trial.

To assess clinically and radiographically the success of pulp capping procedure done in traumatically exposed permanent posterior teeth using a novel injectable treated dentin matrix hydrogel (TDMH), Biodentine, and MTA and to evaluate the formed dentin bridge under the capping materials using CBCT imaging. 45 patients subjected to accidental traumatic pulp exposures by undergraduate dental students are allocated for this study. For each patient, a pulp capping procedure was done. TDMH was formed of TDM powder and sodium alginate to be injected and then hardened in the defect area. Patients were assigned to 3 groups: TDMH, Biodentine, and MTA, respectively, and returned to the clinic after 3, 6, 12, 18, and 24 months for clinical and radiographic examinations. Tomographic data, including thickness and density of formed dentin bridges, were evaluated at the end of the study period compared to the base line. Pulp sensitivity was evaluated throughout the study period using thermal testing and electric pulp tester. During the follow-up period, all patients were asymptomatic with no clinical signs and symptoms and revealed no radiographic signs of pathosis. However, tomographic evaluation showed the tested materials to have different levels of impact on formed dentin bridges with TDMH group resulted in significantly superior dentin bridges of a higher radiodensity and thickness than Biodentine and MTA. TDMH has a greater potential to induce dentin bridge formation than Biodentine and MTA under standardized conditions. Additionally, CBCT imaging was confirmed as a non-invasive and inclusive approach to evaluate the formed dentin bridges after pulp capping procedure. Direct pulp capping can be done successfully with this novel injectable pulp capping material in future clinical applications. PACTR201901866476410.

A three-dimensional in vitro model of the peripheral nervous system

Recent advances in three-dimensional (3D) cell culture models developed on organ-on-a-chip or microfluidic devices have shown their capability to recapitulate the in vivo microenvironment as well as their potential as tools in biomedical research. Here, we present an in vitro model of the peripheral nervous system (PNS) by establishing a coculture model of motor neurons (MNs) and Schwann cells (SCs) in a 3D environment in a microengineered extracellular matrix hydrogel scaffold. The collagen scaffold placed at the center of the microdevice provided a 3D cellular microenvironment where the axons of MNs were allowed to actively interact with SCs during their growth and maturation. By treating the MN–SC coculture model with ascorbic acid, we were able to model the myelination process in the PNS, which was evidenced by the increased expression of myelin markers in SCs. Moreover, we show that this can be reversed by treating myelinated nerve fibers with glial growth factor (neuregulin-1 isoform) to potentially block the formation of the myelin sheath and induce demyelination. Our 3D cell culture model may be used to achieve active control of the myelinating and demyelinating processes in the PNS and thus may offer new opportunities to study pathophysiological processes involved in motor neuron diseases by in vitro modeling.

Near-Infrared Light Induced Dynamic Structural Color Change of Amorphous Photonic Hydrogel

Light-induced photonic systems are rare in nature and usually lack a full color response in artificial systems. Herein, we first report a near-infrared (NIR) light controlled optical hydrogel; its structural color covers the full visible spectrum range. The hydrogel was fabricated by embedding dry thermoresponsive microgels film into a thermoresponsive hydrogel. Under the synergistic effect of the microgels and hydrogel, the hydrogel displayed a dynamic color with temperature changing, and it was reversible. Moreover, the structural colors were angle independent for the amorphous microgel arrays. When carbon nanotubes were integrated into the matrix hydrogel, dynamic structural colors can be displayed under NIR irradiation. Such NIR light-responsive photonic hydrogel film may provide potential applications in artificial skin, the military, and other fields.

3D bioprinting–a step towards heart tissue regeneration

Heart disease and cardiovascular disease is a very serious and growing public health issue. Tissue-engineering has great potential and great strength for regeneration, remolding, and growth. In the case of heart failure, Allografting has been used. 3D bioprinting has a great impact in the field of cardiovascular tissue engineering. It has been observed that 3D Bioprinting is used to construct an artificial heart for transplantation and used to create myocardial cells in case of injury. Recent studies showed that biomaterial used in the treatment of myocardial dysfunction is decellularized cardiac extracellular matrix hydrogel in adults. Collagen, Alginate gelatin, hyaluronic acid, and deECM scaffolds were used as biomaterials in 3D bioprinting. It has been shown that scaffold used with ECM was used to support there generation process A new 3D bioprinting technology was developed in which cells were collected into spheroids and printed on a needle array according to desirable characteristics. Different bio inks such as laser, extrusion, droplet, and stereolithography are used here. Electric stimulation is key to the contractility of cardiomyocytes. A physical cardiac replica was created by image processing software that creates 3D structures. In holographic display 3D, full hearts of patients were printed in flexible material. A process is demonstrated to fabricate robust valves of the heart using the3D bioprinting technique. MRI or CT scans were used to obtained 3D images of the aorta.3D bioprinting plays a huge role in knowing the aortic anatomy involves the aortic valve area and morphology of the root. Recent advances demonstrated that 3D bioprinting can assist in ventricular device placement and perform a specific function in a complex with (CHD) Congenital heart defects. 3D bioprinting holds great prom

Nanofibrous nerve guidance conduits decorated with decellularized matrix hydrogel facilitate peripheral nerve injury repair.

Rationale: Peripheral nerve injury (PNI) is a great challenge for regenerative medicine. Nerve autograft is the gold standard for clinical PNI repair. Due to its significant drawbacks, artificial nerve guidance conduits (NGCs) have drawn much attention as replacement therapies. We developed a combinatorial NGC consisting of longitudinally aligned electrospun nanofibers and porcine decellularized nerve matrix hydrogel (pDNM gel). The in vivo capacity for facilitating nerve tissue regeneration and functional recovery was evaluated in a rat sciatic nerve defect model.Methods: Poly (L-lactic acid) (PLLA) was electrospun into randomly oriented (PLLA-random) and longitudinally aligned (PLLA-aligned) nanofibers. PLLA-aligned were further coated with pDNM gel at concentrations of 0.25% (PLLA-aligned/0.25% pDNM gel) and 1% (PLLA-aligned/1% pDNM gel). Axonal extension and Schwann cells migration were evaluated by immunofluorescence staining of dorsal root ganglia cultured on the scaffolds. To fabricate implantable NGCs, the nanofibrous scaffolds were rolled and covered with an electrospun protection tube. The fabricated NGCs were then implanted into a 5 mm sciatic nerve defect model in adult male Sprague-Dawley rats. Nerves treated with NGCs were compared to contralateral uninjured nerves (control group), injured but untreated nerves (unstitched group), and autografted nerves. Nerve regeneration was monitored by an established set of assays, including T2 values and diffusion tensor imaging (DTI) derived from multiparametric magnetic resonance imaging (MRI), histological assessments, and immunostaining. Nerve functional recovery was evaluated by walking track analysis.Results: PLLA-aligned/0.25% pDNM gel scaffold exhibited the best performance in facilitating directed axonal extension and Schwann cells migration in vitro due to the combined effects of the topological cues provided by the aligned nanofibers and the biochemical cues retained in the pDNM gel. Consistent results were obtained in animal experiments with the fabricated NGCs. Both the T2 and fractional anisotropy values of the PLLA-aligned/0.25% pDNM gel group were the closest to those of the autografted group, and returned to normal much faster than those of the other NGCs groups. Histological assessment indicated that the implanted PLLA-aligned/0.25% pDNM gel NGC resulted in the largest number of axons and the most extensive myelination among all fabricated NGCs. Further, the PLLA-aligned/0.25% pDNM gel group exhibited the highest sciatic nerve function index, which was comparable to that of the autografted group, at 8 weeks post-surgery.Conclusions: NGCs composed of aligned PLLA nanofibers decorated with 0.25% pDNM gel provided both topological and biochemical guidance for directing and promoting axonal extension, nerve fiber myelination, and functional recovery. Moreover, T2-mapping and DTI metrics were found to be useful non-invasive monitoring techniques for PNI treatment.

Pulmonary tissue-mimetic hydrogel niches for small cell lung cancer cell culture.

Although small cell lung cancer (SCLC) is characterized by early metastasis and high resistance to most anti-cancer therapeutics, resulting in poor prognosis, surgical treatment is unavailable for most patients. Instead, clinical treatment for SCLC patients relies largely on chemotherapy. Therefore, an analysis platform supporting research into the physiology of SCLC cells and novel anti-cancer drugs is strongly needed. Decellularized extracellular matrix (dECM) hydrogel is a promising candidate cell-culture system that could provide a tissue-specific environment. However, dECM-based hydrogels have limited property control, poor mechanical properties, and loss of components during decellularization. In this study, porcine decellularized lung tissue and hyaluronic acid (HA) were hybridized via photopolymerization to form a pulmonary tissue-mimetic hydrogel. dECM solution was obtained by decellularization and pepsin digestion. The dECM and HA were then modified with methacrylic moieties, which produced dECM-methacrylate (dECM-MA) and HA methacrylate (HA-MA). dECM-MA/HA-MA hydrogels were fabricated by photopolymerization using a photoinitiator under UV light irradiation. The mechanical properties of the dECM-based hydrogel were compared with those of native tissue. SCLC cells (NCI-H69) were encapsulated in multiple types of dECM-based hydrogels, and they exhibited higher cell proliferation, drug resistance, and CD44 expression in the presence of dECM-MA and HA-MA than in the control condition.

Bi-layer silk fibroin skeleton and bladder acellular matrix hydrogel encapsulating adipose-derived stem cells for bladder reconstruction.

A scaffold, constructed from a bi-layer silk fibroin skeleton (BSFS) and a bladder acellular matrix hydrogel (BAMH) encapsulated with adipose-derived stem cells (ASCs), was developed for bladder augmentation in a rat model. The BSFS, prepared from silk fibroin (SF), had good mechanical properties that allowed it to maintain the scaffold shape and be used for stitching. The prepared BAM was digested by pepsin and the pH was adjusted to harvest the BAMH that provided an extracellular environment for the ASCs. The constructed BSFS-BAMH-ASCs and BSFS-BAMH scaffolds were wrapped in the omentum to promote neovascularization and then used for bladder augmentation; at the same time, a cystotomy was used as the condition for the control group. Histological staining and immunohistochemical analysis confirmed that the omentum incubation could promote scaffold vascularization. Hematoxylin and eosin and Masson's trichrome staining indicated that the BSFS-BAMH-ASCs scaffold regenerated the bladder wall structure. In addition, immunofluorescence analyses confirmed that the ASCs could promote the regeneration of smooth muscle, neurons and blood vessels and the restoration of physiological function. These results demonstrated that the BSFS-BAMH-ASCs may be a promising scaffold for promoting bladder wall regeneration and the restoration of physiological function of the bladder in a rat bladder augmentation model.

Metal-organic framework based mixed matrix hydrogel membranes for highly efficient gas separation

MOF/polymer interface in mixed-matrix membranes (MMMs) has been considered as a crucial issue for achieving highly efficient gas separation performance. In this research, aluminum fumarate framework (A520) based mixed matrix hydrogel self-supported membranes (A520-MMHMs) were prepared by a facile UV photopolymerization. The hydrophilic A520 incorporated into hydrogel polymer effectively avoided the formation of interfacial defects and improved the compatibility between MOF and hydrogel matrix. As a result, the A520-MMHM possessed enhanced CO2 permeability (∼432.87 Barrer) and CO2/CH4 selectivity (∼51.05) in comparison with pure hydrogel membrane. This performance data is very close to the updated McKeown 2019 upper bound, far exceeding the 2008 Robeson upper bound. Therefore, the design of hydrogel membrane incorporated with water-stable MOF may open up a new way for optimizing self-supported hydrogel membrane performance in gas separation.

Prevention of Corneal Myofibroblastic Differentiation In Vitro Using a Biomimetic ECM Hydrogel for Corneal Tissue Regeneration.

Corneal scarring is one of the major causes of blindness, affecting millions worldwide. Despite recent advancements in surgical strategies, there is an unmet need for a clinically feasible material and methods to prevent scarring following corneal injury. In this study, we report the potential utility of a hydrogel derived from cadaveric animal corneas, using a decellularized corneal matrix hydrogel (abbreviated as dCMH), which is prepared by a simple method. This hydrogel is easily injectable, biocompatible, and has the ability to maintain good shape-retention properties at 37 °C, which make it suitable for in vivo applications. Furthermore, our gene expression studies and immunofluorescence studies indicate that dCMH maintains the morphology and function of keratocytes in vitro and prevents their transdifferentiation to myofibroblasts. From the above results, it is evident that dCMH maintains the keratocytes with the ability to regenerate the corneal defect without scar. We thus suggest a simple yet effective approach for corneal tissue decellularization and that dCMH can be a promising material for prophylaxis against blinding scar formation in an injured cornea.

ECM hydrogel improves the delivery of PEG microsphere-encapsulated neural stem cells and endothelial cells into tissue cavities caused by stroke

Intracerebral implantation of neural stem cells (NSCs) to treat stroke remains an inefficient process with <5% of injected cells being retained. To improve the retention and distribution of NSCs after a stroke, we investigated the utility of NSCs’ encapsulation in polyethylene glycol (PEG) microspheres. We first characterized the impact of the physical properties of different syringes and needles, as well as ejection speed, upon delivery of microspheres to the stroke injured rat brain. A 20 G needle size at a 10 μL/min flow rate achieved the most efficient microsphere ejection. Secondly, we optimized the delivery vehicles for in vivo implantation of PEG microspheres. The suspension of microspheres in extracellular matrix (ECM) hydrogel showed superior retention and distribution in a cortical stroke caused by photothrombosis, as well as in a striatal and cortical cavity ensuing middle cerebral artery occlusion (MCAo). Thirdly, NSCs or NSCs + endothelial cells (ECs) encapsulated into biodegradable microspheres were implanted into a large stroke cavity. Cells in microspheres exhibited a high viability, survived freezing and transport. Implantation of 110 cells/microsphere suspended in ECM hydrogel produced a highly efficient delivery that resulted in the widespread distribution of NSCs in the tissue cavity and damaged peri-infarct tissues. Co-delivery of ECs enhanced the in vivo survival and distribution of ∼1.1 million NSCs. The delivery of NSCs and ECs can be dramatically improved using microsphere encapsulation combined with suspension in ECM hydrogel. These biomaterial innovations are essential to advance clinical efforts to improve the treatment of stroke using intracerebral cell therapy.

Characterization of Porcine Urinary Bladder Matrix Hydrogels from Sodium Dodecyl Sulfate Decellularization Method.

Urinary bladder matrix (UBM) is one of the most studied extracellular matrixes (ECM) in the tissue engineering field. Although almost all of the UBM hydrogels were prepared by using peracetic acid (PAA), recent studies indicated that PAA was not a trustworthy way to decellularize UBM. A stronger detergent, such as sodium dodecyl sulfate (SDS), may help tackle this issue; however, its effects on the hydrogels’ characteristics remain unknown. Therefore, the objective of this study was to develop a more reliable protocol to decellularize UBM, using SDS, and to compare the characteristics of hydrogels obtained from this method to the widely employed technique, using PAA. The results indicated that SDS was superior to PAA in decellularization efficacy. Different decellularization methods led to dissimilar gelation kinetics; however, the methods did not affect other hydrogel characteristics in terms of biochemical composition, surface morphology and rheological properties. The SDS-treated hydrogels possessed excellent cytocompatibility in vitro. These results showed that the SDS decellularization method could offer a more stable and safer way to obtain acellular UBM, due to reducing immunogenicity. The hydrogels prepared from this technique had comparable characteristics as those from PAA and could be a potential candidate as a scaffold for tissue remodeling.

Understanding the role of tissue-specific decellularized spinal cord matrix hydrogel for neural stem/progenitor cell microenvironment reconstruction and spinal cord injury

The repair of spinal cord injury (SCI) highly relies on microenvironment remodeling and facilitating the recruitment and neuronal differentiation of endogenous stem/progenitor cells. Decellularized tissue matrices (DTMs) have shown their unique and beneficial characteristics in promoting neural tissue regeneration, especially those derived from the nervous system. Herein, we present a comparative analysis of a DTM hydrogel derived from spinal cord (DSCM-gel) and a decellularized matrix hydrogel derived from peripheral nerves (DNM-gel). The tissue-specificity of DSCM-gel was evaluated both in vitro, using neural stem/progenitor cell (NSPC) culture, and in vivo, using various materials and biological analyses, including transcriptome and proteomics. It was found that DSCM-gel retained an extracellular matrix-like nanofibrous structure but exhibited higher porosity than DNM-gel, which potentiated NSPCs viability, proliferation, and migration in the early stage of 3D culturing, followed by facilitation of the NSPCs differentiation into neurons. Transcriptome analysis indicated that DSCM-gel regulates NSPCs behavior by modulating integrin α2, α9, and β1 expression profiles along with AKT/ERK related signaling pathways. Proteomics analyses suggest that DSCM specific extracellular matrix proteins, such as the tenascin family (TNC) and some soluble growth factor (FGF2) may contribute to these regulations. Furthermore, in vivo assessments confirmed that DSCM-gel provides a suitable microenvironment for endogenous stem/progenitor cell recruitment and axonal regeneration for bridging the lesion site after a completely transected SCI. Thus, this systematic study provides key insights useful for the development of the tissue-specific DTM biomaterials for translational microenvironment replacement therapies and tissue repair.

Decellularized nerve matrix hydrogel scaffolds with longitudinally oriented and size-tunable microchannels for peripheral nerve regeneration

The scaffolding biomaterials and their internal structures are crucial in constructing growth-permissive microenvironment for tissue regeneration. A functional bioscaffold not only requires sufficient extracellular matrix components, but also provides topological guidance by mimicry of the ultrastructure of the native tissue. In our laboratory, a decellularized nerve matrix hydrogel derived from porcine sciatic nerve (pDNM-G) is successfully prepared, which shows great promise for peripheral nerve regeneration. Herein, longitudinally oriented microchannel structures were introduced into pDNM-G bioscaffolds (A-pDNM-G) through controlled unidirectional freeze-drying. The axially aligned microchannels effectively directed and significantly promoted neurite extension and Schwann cell migration, assessed by culturing dorsal root ganglion explants on the longitudinal sections of A-pDNM-G scaffolds. Such regenerative cellular responses can be further optimized by tuning the channel sizes. In vivo studies confirmed that the implanted nerve guidance conduits containing A-pDNM-G scaffolds significantly facilitated axonal extension, myelination, and reached considerable functional recovery in 15-mm rat sciatic nerve defects. The incorporation of nerve growth factor further improved the overall performance in the grafted nerve. The bioactive pDNM-G enables controlled release of neurotrophic factor and easy integration of topological cue provided by the axially aligned microchannels into implantable bioscaffolds, which may serve in future clinical treatments of peripheral nerve injury.

Fabrication of Biomolecule-Loaded Composite Scaffolds Carried by Extracellular Matrix Hydrogel

Fabrication of multifunctional scaffolds with biomimicking physical and biological signals play an important role in enhancing tissue regeneration. Multifunctional features come from the composite scaffold with various bioactive molecules. However, simple, biocompatible, and controllable hybridization strategy is still lacking. In this study, we leverage naturally derived extracellular matrix (ECM) as chemically controllable hydrogel carrier to effectively load functional biomolecules. The use of ECM hydrogel takes advantage of both native functionality of ECM components and tunability of hydrogel in controlling release of loaded molecules. As a proof of concept, porous acellular bone scaffold was selected as the solid pristine scaffold to be composited with BMP-2 and VEGF, which are loaded by spinal cord-derived ECM (SC-ECM) hydrogel. Crosslinking degree of SC-ECM hydrogel is tuned by changing genipin concentration, which renders the control over release kinetics of BMP-2 and VEGF. The mechanical strength of scaffold maintained after hybridization and is not significantly decreased in wet condition. In vitro evaluations of scaffolds cocultured with osteoblasts and mesenchymal stem cells (MSCs) demonstrate the biocompatible and bioactive features resulting from the composite scaffolds. Evidenced by alkaline phosphatase test, immunofluorescence, and real-time polymerase chain reaction, differentiation of MSCs towards osteoblast lineage is significantly enhanced by composite scaffolds. Therefore, our strategy in fabricating composite scaffold enabled by biomolecule-loaded ECM hydrogel holds great promise in regenerating diverse tissue types by appropriate combinations of solid pristine scaffolds, ECM, and bioactive molecules. Impact statement We developed a bioactive molecule (e.g., growth factor, protein) loading method using extracellular matrix hydrogel as a carrier. It brings a new strategy to fabricate composite scaffolds with unique biofunctions.

Facilitate Angiogenesis and Neurogenesis by Growth Factors Integrated Decellularized Matrix Hydrogel.

Neurological functional recovery depends on the synergistic interaction between angiogenesis and neurogenesis after peripheral nerve injury (PNI). Decellularized nerve matrix hydrogels have drawn much attention and been considered as potential therapeutic biomaterials for neurovascularization, due to their intrinsic advantages in construction of a growth-permissive microenvironment, strong affinity to multiple growth factors (GFs), and promotion of neurite outgrowth. In the present study, nerve growth factor (NGF) and vascular endothelial growth factor (VEGF) were incorporated into porcine decellularized nerve matrix hydrogel (pDNM-gel) for PNI treatment. Both GFs bound strongly to pDNM-gel and underwent a controlled release manner, which showed facilitated axonal extension and vascular-like tube formation in vitro. Especially, a companion growth was identified when human umbilical vein endothelial cells and neurons were cocultured on the GFs containing pDNM-gel. In a crushed rat sciatic nerve model, the incorporated NGF and VEGF appeared to contribute for axonal growth and neovascularization correspondingly but separately. Both GFs were equally important in improving nerve functional recovery after in situ administration. These findings indicate that pDNM-gel is not only a bioactive hydrogel-based material that can be used alone, but also serves as suitable carrier of multiple GFs for promoting an effective PNI repair. Impact statement Decellularized matrix hydrogel derived from nerve tissue has demonstrated its effectiveness in promoting nerve reinnervation, remyelination, and functionalization. Meanwhile, angiogenesis is highly desirable for treatment of long-distance peripheral nerve defects. To this end, we incorporated both vascular endothelial growth factor (VEGF) and nerve growth factor (NGF) into porcine decellularized nerve matrix hydrogel (pDNM-gel) to induce neovascularization and neuroregeneration. At the cellular level, the pDNM-gel with both growth factors (GFs) exhibited significant capability in promoting axonal elongation, Schwann cell proliferation and migration, as well as vessel/nerve interaction. In crushed peripheral nerve injury (PNI) rat model, the integrated VEGF was more favorable for angiogenesis, whereas NGF mainly contributed to neurogenesis. However, the combination of both GFs in pDNM-gel highly facilitated motor functional recovery, highlighting the therapeutic promise of decellularized matrix hydrogel for growth factor delivery toward neuroprotection and neuroregeneration after PNI.

Dermal Extracellular Matrix-Derived Hydrogels as an In Vitro Substrate to Study Mast Cell Maturation.

Mast cells (MCs) are pro-inflammatory tissue-resident immune cells that play a key role in inflammation. MCs circulate in peripheral blood as progenitors and undergo terminal differentiation in the tissue microenvironment where they can remain for many years. This in situ maturation results in tissue- and species-specific MC phenotypes, culminating in significant variability in response to environmental stimuli. There are many challenges associated with studying mature tissue-derived MCs, particularly in humans. In cases where cultured MCs are able to differentiate in two-dimensional in vitro cultures, there remains an inability for full maturation. Extracellular matrix (ECM) scaffolds provide for a more physiologically relevant environment for cells in vitro and have been shown to modulate the response of other immune cells such as T cells, monocytes, and macrophages. To improve current in vitro testing platforms of MCs and to assess future use of ECM scaffolds for MC regulation, we studied the in vitro response of human MCs cultured on decellularized porcine dermis hydrogels (dermis extracellular matrix hydrogel [dECM-H]). This study investigated the effect of dECM-H on cellular metabolic activity, cell viability, and receptor expression compared to collagen type I hydrogel (Collagen-H). Human MCs showed different metabolic activity when cultured in the dECM-H and also upregulated immunoglobulin E (IgE) receptors associated with MC maturation/activation compared to collagen type I. These results suggest an overall benefit in the long-term culture of human MCs in the dECM-H compared to Collagen-H providing important steps toward a model that is more representative of in vivo conditions. Graphical abstract [Formula: see text] Impact statement Mast cells (MCs) are difficult to culture in vitro as current culture conditions and substrates fail to promote similar phenotypic features observed in vivo. Extracellular matrix (ECM)-based biomaterials offer three-dimensional, tissue-specific environments that more closely resemble in vivo conditions. Our study explores the use of dermal ECM hydrogels for MC culture and shows significant upregulation of metabolic activity, cell viability, and gene expression of markers associated with MC maturation or activation compared to collagen type I-hydrogel and tissue culture plastic controls at 7 days. These results are among the first to describe MC behavior in response to ECM hydrogels.