Percorrer por autor "Rosadas, Marta"
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- Beyond DNA removal: assessing the immunological response to decellularized rabbit dermal matricesPublication . Rosadas, Marta; Sánchez Espinel, Christian; Peleteiro, Mercedes; Sousa, Alda; Ribeiro, Viviana P.; González-Fernández, África; Oliveira, Ana L.Introduction: Decellularized matrices have attracted considerable attention in tissue engineering and regenerative medicine due to their ability to preserve the biochemical composition and microarchitecture of native tissues. Although DNA removal is commonly used as the main indicator of decellularization efficiency, other factors—such as endotoxin contamination, damage-associated molecular patterns, and residual reagents—can significantly influence the host immune response. In this study, decellularized rabbit dermal matrices (dRDM) produced using two different protocols— resulting in DNA levels below or above the proposed safety threshold of 50 ng/mg dry tissue—were evaluated for their immunological performance. Conclusions: Two decellularization protocols produced dRDMs with distinct biological profiles. Although the SDC 3H protocol achieved DNA levels below the recommended threshold, it was associated with greater collagen degradation, whereas the SDC 30 min protocol better preserved native dermal collagen. Both matrices were sterile, endotoxin-free, and did not induce ROS production in PBMCs. However, dry dRDMs and SDC 3H-treated matrices promoted complement activation and increased PBMC necrosis. Overall, despite not meeting the recommended DNA threshold, the SDC 30 min protocol demonstrated superior immunocompatibility and collagen preservation, highlighting that DNA content alone is not a reliable predictor of decellularization efficiency. Instead, immunocompatibility, collagen integrity, physical cell–matrix interactions, and potential detergent residues play a critical role in defining matrix performance.
- Biohybrid dressings: integrating silk fibroin textiles with decellularized biological tissue for wound healingPublication . Sousa, Teresa; Vale, Inês; Rosadas, Marta; Silva, Inês V.; Ribeiro, Viviana P.; Oliveira, Ana L.Burns affect 11 million people globally each year with 180,000 associated fatalities. This study proposes a multilayer hybrid dressing combining a silk fibroin (SF) textile with decellularized porcine small intestine (dPSI) capable of simultaneously provide wound protection and tissue regeneration. A silk sericin (SS) hydrogel was embedded into the decellularized tissue ensuring the integration with the textile layer while promoting anti-inflammatory benefits, enhancing the hybrid dressing’s biological performance.
- Biohybrid solutions for burn care: merging silk medical textiles with decellularized matricesPublication . Sousa, Teresa; Vale, Inês; Rosadas, Marta; Silva, Inês V.; Ribeiro, Viviana P.; Oliveira, Ana L.
- Biohybrid solutions for burn care: merging silk medical textiles with decellularized matricesPublication . Sousa, Teresa; Vale, Inês; Rosadas, Marta; Silva, Inês V.; Ribeiro, Viviana P.; Oliveira, Ana L.
- Cutting-edge hybrid dressings: combining silk medical textiles and decellularized biological tissue for advanced burn wound carePublication . Sousa, Teresa; Vale, Inês; Rosadas, Marta; Silva, Inês V.; Ribeiro, Viviana P.; Oliveira, Ana L.Aim: Burns affect 11 million people globally each year, with 180,000 fatalities [1]. This study investigates a multilayer burn dressing combining silk fibroin (SF) fabric with highly- preserved decellularized porcine small intestine (dPSI) to support tissue regeneration and wound integration [2]. A silk sericin (SS) hydrogel is included as interface for structural integrity and anti-inflammatory benefits, enhancing the hybrid dressing’s biological performance. Methods: An innovative decellularization protocol was proposed to obtain dPSI, maintaining submucosa, serosa, and muscle layers, using cycles of decellularizing agents (SDS, SDC, DMSO), washing agents (upH₂O, PBS), and sterilization (PAA/ethanol). The serosa and submucosa of dPSI were integrated with SF-based textiles using HRP- crosslinked SS hydrogels. Decellularization and integrity were assessed via DNA quantification and histology, while hybrid dressings’ morphology and mechanics were evaluated by SEM and tensile tests. Degradation profile was tested in simulated wound fluid, and biological performance was assessed by culturing human dermal fibroblasts (hDFs) on the submucosa layer up to 10 days. Results: The dPSI was successfully achieved (<50 ng/mg dsDNA). SEM images confirmed the full integration of the dPSI with SF-based textiles, especially when serosa faced the textile. The presence of the textile structure resulted in an enhancement of the mechanical strength. dPSI was able to degrade first in the multilayer dressing, and hDFs adhered and proliferated on the submucosa over 10 days, supporting hybrid structural integrity. Conclusions: This study is pioneer in confirming promising results for the first multilayer hybrid dressing combining medical textiles and dPSI for burn wound applications.
- Decellularized dermal matrices: unleashing the potential in tissue engineering and regenerative medicinePublication . Rosadas, Marta; Silva, Inês V.; Costa, João B.; Ribeiro, Viviana P.; Oliveira, Ana L.Decellularized dermal matrices (dDMs) have emerged as effective biomaterials that can revolutionize regenerative medicine, particularly in the field of wound healing and tissue regeneration. Derived from animal or human skin, dDMs offer great biocompatibility, remarkable biochemistry, and a macromolecular architecture equivalent to the native tissue. Notably, among the biomimetic extracellular matrix (ECM)-based scaffolds, dDMs stand out due to their inherent dermal microenvironment, holding high value for skin regeneration and reconstructive surgery. The integration of dDMs as a biomaterial base for bioinks in advanced manufacturing technologies opens promising avenues for crafting precise, biomimetic tissue engineering (TE) constructs with optimized recellularization properties. This mini review outlines the main sources, differential decellularization techniques applied to dDMs, and their significance intissue engineering and regenerative medicine. It subsequently delves into the different categories of decellularized materials obtained, their unique physical and biochemical attributes, as well as their applications to promote wound healing and regenerating skin and soft tissues. Additionally, the currently available market products based on dDMs are examined and the main outcomes are compared. Finally, the article highlights current barriers in the field and anticipates the future challenges and applications of dDMs-based therapies.
- Decellularized dermal matrix-based hydrogels and their potential as immunomodulatory biomaterialsPublication . Pereira, Ana Beatriz V.; Rosadas, Marta; Gomes, Patricia; Sousa, Alda; Oliveira, Ana Leite; Ribeiro, Viviana P.Decellularization is a process that aims to remove cellular and nuclear components from tissues, while preserving the bioactivity of the extracellular matrix (ECM) and minimizing the immunogenicity. Decellularized ECMs positively influence cell adhesion, proliferation, and differentiation, establishing them as promising biomaterials for tissue regeneration. Recent research has highlighted their potential immunomodulatory effect, including their ability to regulate the crosstalk between macrophages and T cells and influence the polarization of macrophages, which leads to an anti-inflammatory and pro-remodelling response. For example, hydrogels derived from decellularized dermal matrix (dDM) have reported potential immunomodulatory effect, namely the regulation of macrophage phenotypes. Using techniques involving enzymatic digestion and collagen crosslinking, dDMs can be converted into hydrogels that preserve growth factors, bioactive binding sites, and the fundamental structural and functional proteins of the ECM. Moreover, hydrogels have the benefit of being injectable and can be used in defect areas with irregular shapes when compared to three-dimensional porous scaffolds. The aim of this study is to develop an hydrogel matrix derived from decellularized rabbit dermal matrix dRDMs and assess its immunomodulatory capabilities for applications in tissue engineering and regenerative medicine. These dermal matrices (dDMs), mostly consisting of collagen, elastin, fibronectin, and laminin, offer advantages over other decellularized tissues regarding availability and adaptability. In addition, by combining the innate immunomodulatory characteristics of dDMs with the adjustable physical properties of hydrogels, we hypothesize that the developed hydrogel will promote a favorable immune response, potentially enabling an immunomodulatory environment. dRDM, obtained through a chemical decellularization, will be lyophilized and subsequently processed into hydrogels through a pepsin-mediated digestion, followed by pH neutralization and warming to 37 ºC to induce gelation. Since the immunomodulatory response may be conditioned by the hydrogel synthesis process, different collagen crosslinking strategies will be evaluated, including photo, thermal, and chemical approaches. Hydrogel formulations will be characterized in terms of rheological properties, microstructure, biocompatibility, and immune response. Immunological assessment will include analysis of cytokine profiles, macrophages polarization markers and immune-related genes.
- Decellularized dermal matrix-based hydrogels and their potential as immunomodulatory biomaterialsPublication . Pereira, Ana Beatriz V.; Rosadas, Marta; Gomes, Patricia; Sousa, Alda; Oliveira, Ana Leite; Ribeiro, Viviana P.
- Decellularized small intestine for burn wound treatment: a tissue engineering paradigm shift?Publication . Silva, Inês V.; Rosadas, Marta; Rodrigues, Ilda; Sousa, Clara; Ribeiro, Viviana; Costa, Raquel; Moroni, Lorenzo; Oliveira, Ana L.Introduction: Burn injuries are a significant global health issue, causing approximately 11 million injuries and 180,000 fatalities each year (1). Beyond physical trauma, burn injuries lead to complications such as infections and sepsis. Burn scars can also diminish quality of life by affecting joint mobility and daily activities (2,3). Conventional dressings and autografts have limitations, necessitating novel treatment strategies (4). Decellularized xenografts, particularly from porcine small intestine (SI), offer a promising alternative due to their content of growth factors and structural proteins essential for wound healing (5,6). Preserving these bioactive molecules while ensuring cost-effectiveness requires carefully designed decellularization processes. This study investigates a new decellularization protocol aimed at creating a safe and highly preserved extracellular matrix (ECM) from porcine SI for optimal functional wound dressing. Conclusion: Our results indicate that the protocol implemented effectively preserves essential ECM components and structure while removing cellular contaminants. The material demonstrates anisotropic preserved mechanical properties, adequate swelling capacity, and WTVR similar to skin. The treated samples present biocompatibility, as they do not hinder human fibroblast metabolic activity. This innovative strategy presents a promising approach to produce preserved ECM that could be further process to become a solution for wound healing and tissue regeneration, particularly in challenging cases like burns. Future research will focus on enhancing its antibacterial and anti-inflammatory properties to further improve its efficacy as a dressing for challenging wounds.
- Development of a decellularized extracelular matrix from porcine aorta for heart valve applications in the Ross procedurePublication . Reis, Mariana S.; Rosadas, Marta; Ho, Chou I.; Costa, João; Vervenne, Thibault; Oliveira, Ana L.; Ribeiro, Viviana P.; Mignon, ArnCardiovascular diseases are the leading cause of adult mortality worldwide, according to the WHO [1]. An important surgical approach for treating diseased aortic valves is the Ross procedure, in which the affected aortic valve is replaced with an autograft from the patient’s own pulmonary valve. The main concern about this procedure is linked to wall dilatation, which can lead to valve leakage and reoperation. Dilatation occurs due to the fivefold increase in blood pressure when transitioning from pulmonary to aortic conditions. Current solutions rely on permanet and stiff synthetic materials to provide structural support, however, these lack biological functionality. Our approach aim to incorporate a decellularized extracellular matrix (dECM) in a semi-permanent textile wrapped around the autograft promoting benign biological adaptation. The decellularization process by removing cellular components, reduces the risk of inflammatory responses and immune rejection. Moreover, essential ECM components that regulate cellular behavior are mantained, which is crucial for effective decellularization outcomes [2]. This study is focused in the development and optimization of an efficient decellularization protocol for obtaining dECM from porcine aorta with mild effects on ECM components preservation. Decellularization of the porcine aortic tissue was performed using a detergent and enzymatic-based protocol combined with supercritical CO₂ (scCO₂). Additional steps of sonication, agitation, washing and freeze-thaw were performed to enhance decellularization efficiency. Graphic A shows a significant decrease in the DNA content after optimized decellularization processing, indicating the removal of approximatly 85% of the DNA from the native tissue, and suggest an effective reduction of potential immunogenic components. Further characterization of ECM components (i.e. glycosaminoglycans, total collagen and elastin) is required to evaluate its preservation and the biological potencial of the dECM when integrated in the Ross processure. To do so, aorta dECM powder will be combined with polycaprolactone and elastin and processed through electrospinning used to create a tubular device to enhance bio-mechanocompatibility, further improving current external supports for the Ross procedure and enhancing its general clinical outcomes. Since this support is biodegradable, it allows the new valve to take over the required strength. The dECM specifically offers essential biochemical cues to promote tissue integration and enable long-term functional repair after the Ross procedure.