RESUMEN

Last twenties, tissue engineering has rapidly advanced to address the shortage of organ donors. Decellularization techniques have been developed to mitigate immune rejection and alloresponse in transplantation. However, a clear definition of effective decellularization remains elusive. This study compares various decellularization protocols using the human fascia lata model. Morphological, structural and cytotoxicity/viability analyses indicated that all the five tested protocols were equivalent and met Crapo's criteria for successful decellularization. Interestingly, only the in vivo immunization test on rats revealed differences. Only one protocol exhibited Human Leucocyte Antigen (HLA) content below 1% residual threshold, the only criterion preventing rat immunization with an absence of rat anti-human IgG switch after one month (N=4 donors for each of the 7 groups, added by negative and positive controls, n=28). By respecting a refined set of criteria, i.e. lack of visible nuclear material, <50ng DNA/mg dry weight of extracellular matrix, and <1% residual HLA content, the potential for adverse host reactions can be drastically reduced. In conclusion, this study emphasizes the importance of considering not only nuclear components but also major histocompatibility complex in decellularization protocols and proposes new guidelines to promote safer clinical development and use of bioengineered scaffolds.

Asunto(s)

Fascia Lata , Antígenos HLA , Ingeniería de Tejidos , Humanos , Animales , Ingeniería de Tejidos/métodos , Antígenos HLA/inmunología , Ratas , Andamios del Tejido/química , Materiales Biocompatibles/química , Masculino , Matriz Extracelular Descelularizada/química , Matriz Extracelular/química , Matriz Extracelular/metabolismo

RESUMEN

Improper management of diabetic wound effusion and disruption of the endogenous electric field can lead to passive healing of damaged tissue, affecting the process of tissue cascade repair. This study developed an extracellular matrix sponge scaffold (K1P6@Mxene) by incorporating Mxene into an acellular dermal stroma-hydroxypropyl chitosan interpenetrating network structure. This scaffold is designed to couple with the endogenous electric field and promote precise tissue remodelling in diabetic wounds. The fibrous structure of the sponge closely resembles that of a natural extracellular matrix, providing a conducive microenvironment for cells to adhere grow, and exchange oxygen. Additionally, the inclusion of Mxene enhances antibacterial activity(98.89%) and electrical conductivity within the scaffold. Simultaneously, K1P6@Mxene exhibits excellent water absorption (39 times) and porosity (91%). It actively interacts with the endogenous electric field to guide cell migration and growth on the wound surface upon absorbing wound exudate. In in vivo experiments, the K1P6@Mxene sponge reduced the inflammatory response in diabetic wounds, increased collagen deposition and arrangement, promoted microvascular regeneration, Facilitate expedited re-epithelialization of wounds, minimize scar formation, and accelerate the healing process of diabetic wounds by 7 days. Therefore, this extracellular matrix sponge scaffold, combined with an endogenous electric field, presents an appealing approach for the comprehensive repair of diabetic wounds.

Asunto(s)

Antibacterianos , Cicatrización de Heridas , Cicatrización de Heridas/efectos de los fármacos , Animales , Antibacterianos/farmacología , Antibacterianos/química , Masculino , Matriz Extracelular/química , Hemostáticos/farmacología , Hemostáticos/química , Andamios del Tejido/química , Diabetes Mellitus Experimental/complicaciones , Ratones , Quitosano/química , Ratas , Humanos , Conductividad Eléctrica , Ratas Sprague-Dawley

RESUMEN

The field of bone tissue engineering (BTE) has witnessed a revolutionary breakthrough with the advent of three-dimensional (3D) bioprinting technology, which is considered an ideal choice for constructing scaffolds for bone regeneration. The key to realizing scaffold biofunctions is the selection and design of an appropriate bioink, and existing bioinks have significant limitations. In this study, a composite bioink based on natural polymers (gelatin and alginate) and liver decellularized extracellular matrix (LdECM) was developed and used to fabricate scaffolds for BTE using 3D bioprinting. Through in vitro studies, the concentration of LdECM incorporated into the bioink was optimized to achieve printability and stability and to improve the proliferation and osteogenic differentiation of loaded rat bone mesenchymal stem cells (rBMSCs). Furthermore, in vivo experiments were conducted using a Sprague Dawley rat model of critical-sized calvarial defects. The proposed rBMSC-laden LdECM-gelatin-alginate scaffold, bioprinted layer-by-layer, was implanted in the rat calvarial defect and the development of new bone growth was studied for four weeks. The findings showed that the proposed bioactive scaffolds facilitated angiogenesis and osteogenesis at the defect site. The findings of this study suggest that the developed rBMSC-laden LdECM-gelatin-alginate bioink has great potential for clinical translation and application in solving bone regeneration problems.

Asunto(s)

Bioimpresión , Hígado , Células Madre Mesenquimatosas , Osteogénesis , Ratas Sprague-Dawley , Ingeniería de Tejidos , Andamios del Tejido , Animales , Andamios del Tejido/química , Ingeniería de Tejidos/métodos , Bioimpresión/métodos , Ratas , Células Madre Mesenquimatosas/citología , Osteogénesis/fisiología , Hígado/citología , Impresión Tridimensional , Matriz Extracelular Descelularizada/química , Regeneración Ósea/fisiología , Gelatina/química , Diferenciación Celular , Alginatos/química , Proliferación Celular , Matriz Extracelular/química , Huesos/fisiología , Tinta

RESUMEN

Covalent adaptable networks (CANs) are polymeric networks with cross-links that can break and reform in response to external stimuli, including pH, shear, and temperature, making them potential materials for use as injectable cell delivery vehicles. In the native niche, cells rearrange the extracellular matrix (ECM) to undergo basic functions including migration, spreading, and proliferation. Bond rearrangement enables these hydrogels to mimic viscoelastic properties of the native ECM which promote migration and delivery from the material to the native tissue. In this work, we characterize thioester CANs to inform their design as effective cell delivery vehicles. Using bulk rheology, we characterize the rearrangement of these networks when they are subjected to strain, which mimics the strain applied by a syringe, and using multiple particle tracking microrheology (MPT) we measure cell-mediated remodeling of the pericellular region. Thioester networks are formed by photopolymerizing 8-arm poly(ethylene glycol) (PEG)-thiol and PEG-thioester norbornene. Bulk rheology measures scaffold properties during low and high strain and demonstrates that thioester scaffolds can recover rheological properties after high strain is applied. We then 3D encapsulated human mesenchymal stem cells (hMSCs) in thioester scaffolds. Using MPT, we characterize degradation in the pericellular region. Encapsulated hMSCs degrade these scaffolds within ≈4 days post-encapsulation. We hypothesize that this degradation is mainly due to cytoskeletal tension that cells apply to the matrix, causing adaptable thioester bonds to rearrange, leading to degradation. To verify this, we inhibited cytoskeletal tension using blebbistatin, a myosin-II inhibitor. Blebbistatin-treated cells can degrade these networks only by secreting enzymes including esterases. Esterases hydrolyze thioester bonds, which generate free thiols, leading to bond exchange. Around treated cells, we measure a decrease in the extent of pericellular degradation. We also compare cell area, eccentricity, and speed of untreated and treated cells. Inhibiting cytoskeletal tension results in significantly smaller cell area, more rounded cells, and lower cell speeds when compared to untreated cells. Overall, this work shows that cytoskeletal tension plays a major role in hMSC-mediated degradation of thioester networks. Cytoskeletal tension is also important for the spreading and motility of hMSCs in these networks. This work informs the design of thioester scaffolds for tissue regeneration and cell delivery.

Asunto(s)

Hidrogeles , Células Madre Mesenquimatosas , Reología , Compuestos de Sulfhidrilo , Hidrogeles/química , Humanos , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/citología , Compuestos de Sulfhidrilo/química , Polietilenglicoles/química , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Ésteres/química , Andamios del Tejido/química

RESUMEN

The incidence of liver diseases is high worldwide. Many factors can cause liver fibrosis, which in turn can lead to liver cirrhosis and even liver cancer. Due to the shortage of donor organs, immunosuppression, and other factors, only a few patients are able to undergo liver transplantation. Therefore, how to construct a bioartificial liver that can be transplanted has become a global research hotspot. With the rapid development of three-dimensional (3D) bioprinting in the field of tissue engineering and regenerative medicine, researchers have tried to use various 3D bioprinting technologies to construct bioartificial livers in vitro. In terms of the choice of bioinks, liver decellularized extracellular matrix (dECM) has many advantages over other materials for cell-laden hydrogel in 3D bioprinting. This review mainly summarizes the acquisition of liver dECM and its application in liver 3D bioprinting as a bioink with respect to availability, printability, and biocompatibility in many aspects and puts forward the current challenges and prospects.

Asunto(s)

Bioimpresión , Matriz Extracelular Descelularizada , Hígado , Impresión Tridimensional , Ingeniería de Tejidos , Humanos , Bioimpresión/métodos , Hígado/metabolismo , Hígado/citología , Ingeniería de Tejidos/métodos , Animales , Matriz Extracelular Descelularizada/química , Matriz Extracelular Descelularizada/metabolismo , Andamios del Tejido/química , Hidrogeles/química , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Materiales Biocompatibles/química

RESUMEN

Human space activities have been continuously increasing. Astronauts experiencing spaceflight are faced with health problems caused by special space environments such as microgravity, and the investigation of cell injury is fundamental. The development of a platform capable of cell culture and injury detection is the prerequisite for the investigation. Constructing a platform suitable for special conditions in space life science research is the key issue. The ground-based investigation is an indispensable part of the research. Accordingly, a simulated microgravity (SMG)-oriented integrated chip platform capable of 3D cell culture and in situ visual detection of superoxide anion radical (O2•-) is developed. SMG can cause oxidative stress in human cells, and O2•- is one of the signaling molecules. Thus, a O2•--responsive aggregation-induced emission (AIE) probe is designed, which shows high selectivity and sensitivity to O2•-. Moreover, the probe exhibits abilities of long-term and wash-free staining to cells due to the AIE behavior, which is precious for space cell imaging. Meanwhile, a chip with a high-aspect-ratio chamber for adequate medium storage for the lack of the perfusion system during the SMG experiment and a cell culture chamber which can integrate the extracellular matrix (ECM) hydrogel for the bioinspired 3D cell culture is fabricated. In addition, a porous membrane is introduced between the chambers to prevent the hydrogel from separating during the SMG experiment. The afforded AIE probe-ECM hydrogel-integrated chip can achieve 3D culturing of U87-MG cells and in situ fluorescent detection of endogenous O2•- in the cells after long-term staining under SMG. The chip provides a powerful and potential platform for ground-based investigation in space life science and biomedical research.

Asunto(s)

Técnicas Biosensibles , Hidrogeles , Superóxidos , Humanos , Superóxidos/análisis , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Hidrogeles/química , Matriz Extracelular/química , Técnicas de Cultivo de Célula/instrumentación , Simulación de Ingravidez , Diseño de Equipo , Colorantes Fluorescentes/química , Dispositivos Laboratorio en un Chip , Ingravidez , Estrés Oxidativo

RESUMEN

Injectable extracellular matrix (iECM) is a versatile biological material with beneficial properties such as good degradability, promotion of cell survival, immunomodulation, and facilitation of vascular formation. However, intravenous injection of iECM faces challenges like a short retention time in vivo and low concentration at the lesion site. To address these issues, we prepared a composite hydrogel composed of sodium alginate and iECM and administered it via intrapericardial injection, forming a structure akin to cardiac patches within the pericardium. Compared with intramyocardial injection, intrapericardial injection avoids direct myocardial injury and ectopic tumor formation, offering less invasiveness and better biocompatibility. This study demonstrates that the sodium alginate/infusible extracellular matrix (SA/iECM) composite hydrogel can effectively prolong the local retention time of iECM in the heart, enhance electrical conduction between cardiomyocytes, promote angiogenesis at ischemic myocardial sites, inhibit apoptosis in the infarcted region, mitigate left ventricular remodeling postmyocardial infarction (MI), and improve cardiac function after infarction. Precise coordination of cardiomyocyte contraction and relaxation depends on the rhythmic occurrence of calcium-dependent action potentials. Cardiac dysfunction is partially attributed to the disruption of the excitation-contraction coupling (ECC) mechanism, which is associated with prolonged intracellular Ca2+ transients and alterations in contraction and relaxation Ca2+ levels. Our results show that the SA/iECM composite hydrogel improves electrical conduction, as evidenced by increased Cx43 expression and enhanced intercellular electrical connectivity. This research establishes that intrapericardial injection of a SA/iECM composite hydrogel is a safe and effective treatment modality, providing a theoretical basis for the use of biomaterials in MI therapy.

Asunto(s)

Alginatos , Matriz Extracelular , Hidrogeles , Infarto del Miocardio , Neovascularización Fisiológica , Pericardio , Alginatos/química , Alginatos/farmacología , Animales , Infarto del Miocardio/tratamiento farmacológico , Infarto del Miocardio/patología , Infarto del Miocardio/fisiopatología , Hidrogeles/química , Hidrogeles/farmacología , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Pericardio/efectos de los fármacos , Ratones , Neovascularización Fisiológica/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Masculino , Ratas , Angiogénesis

RESUMEN

Traditional hepatocellular carcinoma-chip models lack the cell structure and microenvironments necessary for high pathophysiological correlation, leading to low accuracy in predicting drug efficacy and high production costs. This study proposed a decellularized hepatocellular carcinoma-on-a-chip model to screen anti-tumor nanomedicine. In this model, human hepatocellular carcinoma (HepG2) and human normal liver cells (L02) were co-cultured on a three-dimensional (3D) decellularized extracellular matrix (dECM) in vitro to mimic the tumor microenvironments of human hepatocellular carcinoma in vivo. Additionally, a smart nanomedicine was developed by encapsulating doxorubicin (DOX) into the ferric oxide (Fe3O4)-incorporated liposome nanovesicle (NLV/Fe+DOX). NLV/Fe+DOX selectively killed 78.59% ± 6.78% of HepG2 cells through targeted delivery and synergistic chemo-chemodynamic-photothermal therapies, while the viability of surrounding L02 cells on the chip model retained high, at over 90.0%. The drug efficacy tested using this unique chip model correlated well with the results of cellular and animal experiments. In summary, our proposed hepatocellular carcinoma-chip model is a low-cost yet accurate drug-testing platform with significant potential for drug screening.

Asunto(s)

Carcinoma Hepatocelular , Doxorrubicina , Dispositivos Laboratorio en un Chip , Neoplasias Hepáticas , Nanomedicina , Humanos , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/patología , Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/patología , Carcinoma Hepatocelular/terapia , Doxorrubicina/farmacología , Doxorrubicina/química , Doxorrubicina/uso terapéutico , Células Hep G2 , Nanomedicina/métodos , Animales , Liposomas/química , Matriz Extracelular/química , Matriz Extracelular/efectos de los fármacos , Compuestos Férricos/química , Técnicas Biosensibles/métodos , Microambiente Tumoral/efectos de los fármacos , Antineoplásicos/farmacología , Antineoplásicos/química , Antineoplásicos/uso terapéutico

RESUMEN

Cells can sense the physical properties of the extracellular matrices (ECMs), such as stiffness and ligand density, through cell adhesions to actively regulate their behaviors. Recent studies have shown that varying ligand spacing of ECMs can influence adhesion size, cell spreading, and even stem cell differentiation, indicating that cells have the spatial sensing ability of ECM ligands. However, the mechanism of the cells' spatial sensing remains unclear. In this study, we have developed a lattice-spring motor-clutch model by integrating cell membrane deformation, the talin unfolding mechanism, and the lattice spring for substrate ligand distribution to explore how the spatial distribution of integrin ligands and substrate stiffness influence cell spreading and adhesion dynamics. By applying the Gillespie algorithm, we found that large ligand spacing reduces the superposition effect of the substrate's displacement fields generated by pulling force from motor-clutch units, increasing the effective stiffness probed by the force-sensitive receptors; this finding explains a series of previous experiments. Furthermore, using the mean-field theory, we obtain the effective stiffness sensed by bound clutches analytically; our analysis shows that the bound clutch number and ligand spacing are the two key factors that affect the superposition effects of deformation fields and, hence, the effective stiffness. Overall, our study reveals the mechanism of cells' spatial sensing, i.e., ligand spacing changes the effective stiffness sensed by cells due to the superposition effect of deformation fields, which provides a physical clue for designing and developing biological materials that effectively control cell behavior and function.

Asunto(s)

Adhesión Celular , Matriz Extracelular , Ligandos , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Modelos Biológicos , Integrinas/metabolismo , Integrinas/química , Membrana Celular/metabolismo , Membrana Celular/química , Talina/metabolismo , Talina/química

RESUMEN

High-molecular-weight (HMW) hyaluronic acid (HA) is a highly abundant natural polysaccharide and a fundamental component of the extracellular matrix (ECM). Its size and concentration regulate tissues' macro- and microenvironments, and its upregulation is a hallmark feature of certain tumors. Yet, the conformational dynamics of HMW-HA and how it engages with the components of the ECM microenvironment remain poorly understood at the molecular level. Probing the molecular structure and dynamics of HMW polysaccharides in a hydrated, physiological-like environment is crucial and also technically challenging. Here, we deploy advanced magic-angle spinning (MAS) solid-state NMR spectroscopy in combination with isotopic enrichment to enable an in-depth study of HMW-HA to address this challenge. This approach resolves multiple coexisting HA conformations and dynamics as a function of environmental conditions. By combining 13C-labeled HA with unlabeled ECM components, we detect by MAS NMR HA-specific changes in global and local conformational dynamics as a consequence of hydration and ECM interactions. These measurements reveal atom-specific variations in the dynamics and structure of the N-acetylglucosamine moiety of HA. We discuss possible implications for interactions that stabilize the structure of HMW-HA and facilitate its recognition by HA-binding proteins. The described methods apply similarly to the studies of the molecular structure and dynamics of HA in tumor contexts and in other biological tissues as well as HMW-HA hydrogels and nanoparticles used for biomedical and/or pharmaceutical applications.

Asunto(s)

Ácido Hialurónico , Espectroscopía de Resonancia Magnética , Peso Molecular , Ácido Hialurónico/química , Humanos , Matriz Extracelular/química , Matriz Extracelular/metabolismo

RESUMEN

Liver sinusoidal endothelial cells (LSECs) are key targets for addressing metabolic dysfunction-associated steatotic liver disease (MASLD). However, isolating and culturing primary LSECs is challenging due to rapid dedifferentiation, resulting in loss of function. The extracellular matrix (ECM) likely plays a crucial role in maintaining the fate and function of LSECs. In this study, we explored the influence of liver-ECM (L-ECM) on liver cells and developed culture conditions that maintain the differentiated function of liver cells in vitro for prolonged periods. Porcine liver-derived L-ECM, containing 34.9 % protein, 0.045 % glycosaminoglycans, and negligible residual DNA (41.2 ng/mg), was utilized to culture primary rat liver cells in generated hydrogels. Proteomic analyses and molecular weight distribution of proteins of solubilized L-ECM revealed the typical diverse ECM core matrisome, with abundant collagens. L-ECM hydrogels showed suitable stiffness and stress relaxation properties. Furthermore, we demonstrated that collagen-rich L-ECM hydrogels enhanced LSECs' and hepatocytes' viability, and reduced the dedifferentiation rate of LSECs. In addition, hepatocyte function was maintained longer by culture on L-ECM hydrogels compared to traditional culturing. These beneficial effects are likely attributed to the bioactive macromolecules including collagens, and mechanical and microarchitectural properties of the L-ECM hydrogels.

Asunto(s)

Supervivencia Celular , Colágeno , Células Endoteliales , Matriz Extracelular , Hepatocitos , Hidrogeles , Hígado , Animales , Hidrogeles/química , Hidrogeles/farmacología , Hepatocitos/metabolismo , Hepatocitos/efectos de los fármacos , Hepatocitos/citología , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Ratas , Células Endoteliales/metabolismo , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Colágeno/química , Colágeno/farmacología , Colágeno/metabolismo , Hígado/metabolismo , Hígado/citología , Porcinos , Células Cultivadas , Masculino

RESUMEN

Extracellular matrix sponge plays a positive role in the wound healing process, but requires proper structural strength and biological properties. In order to solve the problem of lyophilized dissolution of placenta-derived sponge, glutaraldehyde was selected for use in the lyophilized crosslinking process to improve the necessary mechanical properties of the placental decellularization matrix sponge. In this work, the effects of three cross-linking methods of glutaraldehyde (Fumigation/Slurry/Soak) on the physical and biological characteristics of lyophilised sponges derived from placental acellular matrix was investigated. The results revealed that the sponges prepared by all three cross-linking methods exhibited excellent blood coagulation ability and stability. The fumigation cross-linked sponges had good mechanical properties of soft and elastic, and safe cytotoxicity, which were more compatible with the requirements of wound dressing. The slurry cross-linking process was uneven due to the stacked matrix materials, resulting in obvious cracks and easy to break when stretching. The soak crosslinking can obtain a higher degree of crosslinking, which leads to the poor antibacterial performance and the harder sponge scaffold with larger elastic modulus and smaller tensile ratio. In general, fumigation cross-linking is more suitable for the preparation of acellular sponge derived from placenta materials which can maintain basic mechanical properties and biological validity.

Asunto(s)

Reactivos de Enlaces Cruzados , Glutaral , Placenta , Glutaral/química , Placenta/citología , Femenino , Reactivos de Enlaces Cruzados/química , Embarazo , Animales , Matriz Extracelular Descelularizada/química , Matriz Extracelular Descelularizada/farmacología , Humanos , Andamios del Tejido/química , Liofilización/métodos , Coagulación Sanguínea/efectos de los fármacos , Resistencia a la Tracción , Matriz Extracelular/química , Ensayo de Materiales

RESUMEN

Decellularized extracellular matrix (dECM) hydrogels are engineered constructs that are widely-used in the field of regenerative medicine. However, the development of ECM-based hydrogels for bone tissue engineering requires enhancement in its osteogenic properties. For this purpose, we initially employed bone-derived dECM hydrogel (dECM-Hy) in combination with calcium phosphate cement (CPC) paste to improve the biological and structural properties of the dECM hydrogel. A decellularization protocol for bovine bone was developed to prepare dECM-Hy, and the mechanically-tuned dECM/CPC-Hy was built based on both rheological and mechanical characteristics. The dECM/CPC-Hy displayed a double swelling ratio and compressive strength. An interconnected structure with distinct hydroxyapatite crystals was evident in dECM/CPC-Hy. The expression levels of Alp, Runx2 and Ocn genes were upregulated in dECM/CPC-Hy compared to the dECM-Hy. A 14-day follow-up of the rats receiving subcutaneous implanted dECM-Hy, dECM/CPC-Hy and mesenchymal stem cells (MSCs)-embedded (dECM/CPC/MSCs-Hy) showed no toxicity, inflammatory factor expression or pathological changes. Radiography and computed tomography (CT) of the calvarial defects revealed new bone formation and elevated number of osteoblasts-osteocytes and osteons in dECM/CPC-Hy and dECM/CPC/MSCs-Hy compared to the control groups. These findings indicate that the dECM/CPC-Hy has substantial potential for bone tissue engineering.

Asunto(s)

Cementos para Huesos , Regeneración Ósea , Fosfatos de Calcio , Células Madre Mesenquimatosas , Animales , Fosfatos de Calcio/química , Regeneración Ósea/efectos de los fármacos , Bovinos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Ratas , Cementos para Huesos/química , Cementos para Huesos/farmacología , Matriz Extracelular Descelularizada/química , Matriz Extracelular Descelularizada/farmacología , Hidrogeles/química , Hidrogeles/farmacología , Osteogénesis/efectos de los fármacos , Ratas Sprague-Dawley , Ingeniería de Tejidos , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Matriz Extracelular/química , Matriz Extracelular/metabolismo

RESUMEN

The extracellular matrix (ECM) has evolved around complex covalent and non-covalent interactions to create impressive function-from cellular signaling to constant remodeling. A major challenge in the biomedical field is the de novo design and control of synthetic ECMs for applications ranging from tissue engineering to neuromodulation to bioelectronics. As we move towards recreating the ECM's complexity in hydrogels, the field has taken several approaches to recapitulate the main important features of the native ECM (i.e. mechanical, bioactive and dynamic properties). In this review, we first describe the wide variety of hydrogel systems that are currently used, ranging from fully natural to completely synthetic to hybrid versions, highlighting the advantages and limitations of each class. Then, we shift towards supramolecular hydrogels that show great potential for their use as ECM mimics due to their biomimetic hierarchical structure, inherent (controllable) dynamic properties and their modular design, allowing for precise control over their mechanical and biochemical properties. In order to make the next step in the complexity of synthetic ECM-mimetic hydrogels, we must leverage the supramolecular self-assembly seen in the native ECM; we therefore propose to use supramolecular monomers to create larger, hierarchical, co-assembled hydrogels with complex and synergistic mechanical, bioactive and dynamic features.

Asunto(s)

Materiales Biocompatibles , Materiales Biomiméticos , Matriz Extracelular , Hidrogeles , Matriz Extracelular/química , Matriz Extracelular/metabolismo , Hidrogeles/química , Materiales Biomiméticos/química , Materiales Biocompatibles/química , Humanos , Ingeniería de Tejidos , Animales

RESUMEN

Bone extracellular matrix (ECM) has been shown to mimic aspects of the tissue's complex microenvironment, suggesting its potential role in promoting bone repair. However, current ECM-based therapies suffer from limitations such as inefficient scale-up, lack of mechanical integrity, and sub-optimal efficacy. Here, we fabricated hydrogels from decellularized ECM (dECM) from wild type (WT) and thrombospondin-2 knock-out (TSP2KO) mouse bones. TSP2KO bone ECM hydrogel was found to have distinct mechanical properties and collagen fibril assembly from WT. Furthermore, TSP2KO hydrogel promoted mesenchymal stem cell (MSC) attachment, spreading, and invasion in vitro. Similarly, it promoted formation of tube-like structures by human umbilical vein endothelial cells (HUVECs). When applied to a murine calvarial defect model, TSP2KO hydrogel enhanced repair, in part, due to increased angiogenesis. Our study suggests the pro-angiogenic therapeutic potential of TSP2KO bone ECM hydrogel in bone repair. STATEMENT OF SIGNIFICANCE: The study describes the first successful preparation of a novel hydrogel made from decellularized bones from wild-type mice and mice lacking thrombospondin-2 (TSP2). Hydrogels from TSP2 knock-out (TSP2KO) bones have unique characteristics in structure and biomechanics. These gels interacted well with cells in vitro and helped repair damaged bone in a mouse model. Therefore, TSP2KO bone-derived hydrogel has translational potential for accelerating repair of bone defects that are otherwise difficult to heal. This study not only creates a new material with promise for accelerated healing, but also validates tunability of native biomaterials by genetic engineering.

Asunto(s)

Matriz Extracelular , Células Endoteliales de la Vena Umbilical Humana , Hidrogeles , Ratones Noqueados , Trombospondinas , Animales , Trombospondinas/metabolismo , Trombospondinas/genética , Hidrogeles/química , Humanos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Ratones , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/citología , Neovascularización Fisiológica/efectos de los fármacos , Regeneración Ósea/efectos de los fármacos , Matriz Extracelular Descelularizada/química , Matriz Extracelular Descelularizada/farmacología , Huesos/efectos de los fármacos

RESUMEN

Extracellular matrix (ECM) elasticity remains a crucial parameter to determine cell-material interactions (viz. adhesion, growth, and differentiation), cellular communication, and migration that are essential to tissue repair and regeneration. Supramolecular peptide hydrogels with their 3-dimensional porous network and tuneable mechanical properties have emerged as an excellent class of ECM-mimetic biomaterials with relevant dynamic attributes and bioactivity. Here, we demonstrate the design of minimalist amyloid-inspired peptide amphiphiles, CnPA (n = 6, 8, 10, 12) with tuneable peptide nanostructures that are efficiently biomineralized and cross-linked using bioactive silicates. Such hydrogel composites, CnBG exhibit excellent mechanical attributes and possess excellent self-healing abilities and collagen-like strain-stiffening ability as desired for bone ECM mimetic scaffold. The composites exhibited the formation of a hydroxyapatite mineral phase upon incubation in a simulated body fluid that rendered mechanical stiffness akin to the hydroxyapatite-bridged collagen fibers to match the bone tissue elasticity eventually. In a nutshell, peptide nanostructure-guided temporal effects and mechanical attributes demonstrate C8BG to be an optimal composite. Finally, such constructs feature the potential for adhesion, proliferation of U2OS cells, high alkaline phosphatase activity, and osteoconductivity.

Asunto(s)

Matriz Extracelular , Nanofibras , Péptidos , Nanofibras/química , Péptidos/química , Péptidos/farmacología , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Humanos , Materiales Biomiméticos/química , Hidrogeles/química , Huesos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología

RESUMEN

Extracellular matrix (ECM) is essential for tissue development, providing structural support and a microenvironment that is necessary for cells. As tissue engineering advances, there is a growing demand for ECM mimics. Polycaprolactone (PCL) is a commonly used synthetic polymer for ECM mimic materials. However, its biologically inactive surface limits its direct application in tissue engineering. Our study aimed to improve the biocompatibility of PCL by incorporating hemoglobin nanofibrils (HbFs) into PCL using an electrospinning technique. HbFs were formed from bovine hemoglobin (Hb) extracted from industrial byproducts and designed to offer PCL an improved cell adhesion property. The fabricated HbFs@PCL electrospun scaffold exhibits improved fibroblast adherence, proliferation, and deeper fibroblast infiltration into the scaffold compared with the pure PCL scaffold, indicating its potential to be an ECM mimic. This study represents the pioneering utilization of Hb-sourced nanofibrils in the electrospun PCL scaffolds for tissue engineering applications.

Asunto(s)

Materiales Biocompatibles , Matriz Extracelular , Hemoglobinas , Ensayo de Materiales , Nanofibras , Poliésteres , Ingeniería de Tejidos , Hemoglobinas/química , Animales , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Bovinos , Nanofibras/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Poliésteres/química , Proliferación Celular/efectos de los fármacos , Adhesión Celular/efectos de los fármacos , Andamios del Tejido/química , Tamaño de la Partícula , Ratones , Fibroblastos

RESUMEN

Hyalocytes, which are considered to originate from the monocyte/macrophage lineage, play active roles in vitreous collagen and hyaluronic acid synthesis. Obtaining a hyalocyte-compatible bioink during the 3D bioprinting of eye models is challenging. In this study, we investigated the suitability of a cartilage-decellularized extracellular matrix (dECM)-based bioink for printing a vitreous body model. Given that achieving a 3D structure and environment identical to those of the vitreous body necessitates good printability and biocompatibility, we examined the mechanical and biological properties of the developed dECM-based bioink. Furthermore, we proposed a 3D bioprinting strategy for volumetric vitreous body fabrication that supports cell viability, transparency, and self-sustainability. The construction of a 3D structure composed of bioink microfibers resulted in improved transparency and hyalocyte-like macrophage activity in volumetric vitreous mimetics, mimicking real vitreous bodies. The results indicate that our 3D structure could serve as a platform for drug testing in disease models and demonstrate that the proposed printing technology, utilizing a dECM-based bioink and volumetric vitreous body, has the potential to facilitate the development of advanced eye models for future studies on floater formation and visual disorders.

Asunto(s)

Bioimpresión , Matriz Extracelular , Tinta , Impresión Tridimensional , Cuerpo Vítreo , Cuerpo Vítreo/metabolismo , Cuerpo Vítreo/citología , Matriz Extracelular/química , Matriz Extracelular/metabolismo , Animales , Bioimpresión/métodos , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Humanos , Cartílago/citología , Cartílago/química , Cartílago/metabolismo , Supervivencia Celular , Macrófagos/metabolismo , Macrófagos/citología

RESUMEN

Regenerating periodontal defects in osteoporosis patients presents a significant clinical challenge. Unlike the relatively straightforward regeneration of homogeneous bone tissue, periodontal regeneration requires the intricate reconstruction of the cementum-periodontal ligament-alveolar bone interface. Strontium (Sr)-doped biomaterials have been extensively utilized in bone tissue engineering due to their remarkable pro-osteogenic attributes. However, their application in periodontal tissue regeneration has been scarcely explored. In this study, we synthesized an innovative injectable Sr-BGN/GNM scaffold by integrating Sr-doped bioactive glass nanospheres (Sr-BGNs) into the nanofiber architecture of gelatin nanofiber microspheres (GNMs). This design, mimicking the natural bone extracellular matrix (ECM), enhanced the scaffold's mechanical properties and effectively controlled the sustained release of Sr ions (Sr2+), thereby promoting the proliferation, osteogenic differentiation, and ECM secretion of PDLSCs and BMSCs, as well as enhancing vascularization in endothelial cells. In vivo experiments further indicated that the Sr-BGNs/GNMs significantly promoted osteogenesis and angiogenesis. Moreover, the scaffold's tunable degradation kinetics optimized the prolonged release and pro-regenerative effects of Sr2+ in vivo, matching the pace of periodontal regeneration and thereby facilitating the regeneration of functional periodontal tissues under osteoporotic conditions. Therefore, Sr-BGNs/GNMs emerge as a promising candidate for advancing periodontal regeneration strategies.

Asunto(s)

Matriz Extracelular , Microesferas , Nanofibras , Osteoporosis , Estroncio , Estroncio/química , Estroncio/farmacología , Nanofibras/química , Osteoporosis/tratamiento farmacológico , Humanos , Matriz Extracelular/química , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Animales , Osteogénesis/efectos de los fármacos , Andamios del Tejido/química , Diferenciación Celular/efectos de los fármacos , Ingeniería de Tejidos , Proliferación Celular/efectos de los fármacos , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/citología , Regeneración/efectos de los fármacos

RESUMEN

Articular cartilage defects can lead to pain and even disability in patients and have significant socioeconomic loss. Repairing articular cartilage defects remains a long-term challenge in medicine owing to the limited ability of cartilage to regenerate. At present, the treatment methods adopted in clinical practice have many limitations, thereby necessitating the rapid development of biomaterials. Among them, decellularized biomaterials have been particularly prominent, with numerous breakthroughs in research progress and translational applications. Although many studies show that decellularized cartilage biomaterials promote tissue regeneration, any differences in cellular morphology, dynamics, and functionality among various biomaterials upon comparison have not been reported. In this study, we prepared cartilage-derived extracellular matrix (cdECM) biomaterials with different bioactive contents and various physical properties to compare their effects on the morphology, dynamics and functionality of chondrocytes. This cellular multimodal analysis of the characteristics of cdECM biomaterials provided a theoretical basis for understanding the interactions between biomaterials and cells, thus laying an experimental foundation for the translation and application of decellularized cartilage biomaterials in the treatment of cartilage defects.

Asunto(s)

Materiales Biocompatibles , Condrocitos , Matriz Extracelular , Animales , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Condrocitos/citología , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Cartílago Articular , Células Cultivadas , Humanos , Proliferación Celular/efectos de los fármacos , Tamaño de la Partícula , Andamios del Tejido/química