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The irreversible progressive nature of degenerative osteochondral defects, affecting both articular cartilage and the subchondral bone, is a leading cause for joint disease and disability among adults, resulting in severe limitations for people to perform daily activities and represents a heavy economic burden for society. Current treatments do no provide adequate long-term solutions and innovative tissue engineering strategies are regarded as a promising alternative to improve clinical outcomes. A key aspect that needs to be considered is the integration between native tissue and the tissue engineered construct. In this recent article resulting from a collaboration between researchers from iBB and CDRSP-Politécnico de Leiria, published in the Journal Polymers, a new approach to design and manufacture curved scaffolds to mimic osteochondral tissue geometry was presented. The curvature of the sphere was proposed as a template and a procedure was developed for an automated design of scaffolds with explicitly defined curvatures. Fused filament fabrication (3D-Printing) was used for scaffolds’ manufacturing. A Shape fidelity analysis implemented through micro-CT and SEM imaging validated the maximum curvature printability limit predicted from CAD modeling and confirmed the suitability of fused filament fabrication to manufacture curvature-featuring scaffolds. Additionally, a mechanical analysis was conducted with printed scaffolds and through finite element analysis to determine scaffold mechanical properties and identify the regions more susceptible to higher loads.

Diana Marques, a PhD candidate at the Institute for Bioengineering and Biosciences (iBB), was challenged by Fernando Gaspar to answer this question.

In an interview for the podcast "Consórcio do Bem-Estar," Diana discussed the Algae2Fish project's goals with Fernando Gaspar, José Alberto Maia Pereira, and Alvaro Cidrais. Diana is developing research on unique techniques for generating 3D bioprinted fish fillets.

The Algae2Fish project is currently ongoing at iBB and aims to produce seabass fish fillets using 3D bioprinting. These scaffolds will support fish cell growth, provide adequate mechanical and structural properties, and contribute to the final tissue nutritional and organoleptic features.

Extracellular vesicles (EVs) are membrane vesicles fundamental for cellular communication processes. Either by taking advantage of EVs innate properties, or by loading them with therapeutic agents, EVs have emerged as cell-free strategies with diverse clinical applications. Indeed, when compared to mesenchymal stromal cells (MSCs), similar or better therapeutic benefits have been reported for MSC-derived EVs, thus explaining the increased interest in EVs over the years. Despite appealing features, EVs implementation in the clinic is still limited by technical difficulties, like robust isolation methods capable of delivering a high-quality product with high yield. In this work, published in the journal Separation and Purification Technology, we described anion exchange chromatography as a robust and better alternative to already implemented techniques. Following this strategy, more than 85% of injected EVs could be recovered, while complying with the stringencies set by regulatory agencies. This study developed at iBB was led by Dr. Ana Fernandes-Platzgummer and Prof. Ana Azevedo and co-first-authored by PhD students Ricardo Silva and Sara Sousa Rosa.

Human adult stem cells and patient-derived induced pluripotent stem cells represent promising tools to understand human biology, development, and disease. Under a permissive environment, stem cell derivatives can self-organize and reconstruct their native environment, resulting in the creation of organ-like entities known as organoids. Although organoids represent a breakthrough in the stem cell field, there are still considerable shortcomings preventing their widespread use, namely their variability, limited function, and reductionist size. In this recently published review article in Trends in Biotechnology, Miguel Tenreiro (Ph.D. student in Bioengineering), Prof. Margarida Diogo, and co-authors from iBB highlight emerging technologies to fill in the gaps in organoid design and provide insights on how they can be best utilized to fulfill the potential of organoids.

As current treatment options for cartilage degeneration remain ineffective, tissue engineering has emerged as an exciting approach to create cartilage substitutes. In particular, hydrogels seem to be suitable candidates for this purpose due to their high biocompatibility and customizability, being able to be tailored to fit the biophysical properties of native cartilage. Moreover, these hydrogel matrices can be combined with conductive materials in order to simulate the natural electrochemical properties of articular cartilage. In this recent review published in the journal Gels, iBB researchers Filipe Miguel (MSc student in Biotechnology), Frederico Barbosa (PhD student in Bioengineering), Prof. Frederico Ferreira and Dr. João Silva highlight the most common conductive materials combined with hydrogels and their diverse applications, and discuss the current state of research on the development of electrically conductive hydrogels for cartilage tissue engineering.

Understanding the mechano–biological coupling mechanisms of biomaterials for tissue engineering is of major importance to assure proper scaffold performance in situ. Therefore, it is of paramount importance to establish correlations between biomaterials, their processing conditions, and their mechanical behaviour, as well as their biological performance. In a collaborative work between CDRSP-Politécnico de Leiria and SCERG-iBB  (João C. Silva and Frederico Ferreira), it was possible to infer a correlation between the addition of different concentrations of graphene nanoparticles (GPN) in three-dimensional poly(ε-caprolactone) (PCL)-based scaffolds, their extrusion-based processing parameters, and the lamellar crystal orientation observed in the different scaffolds through small-angle X-ray scattering experiments. Moreover, in vitro cell culture studies performed at SCERG-iBB demonstrated the suitability and potential of these novel 3D PCL/GPN scaffolds for tissue engineering applications. The results of this study were just published in Polymers.

The research project “InSilico4OCReg - In silico models guiding in vitro biophysical stimulation of biomimetic hierarchical scaffolds: a computational modelling approach towards functional osteochondral regeneration” was recommended for funding by FCT (250,000 euros in the 2021 Call for SR&TD Project Grants). The project started in January 2022 and aims to develop an innovative combined in silico modelling-in vitro experimental system which, inspired by the properties of the native osteochondral tissue, will be able to optimize tissue engineering strategies towards the production of implants with improved functionality and mechanical properties. The multidisciplinary team of InSilico4OCReg includes researchers from iBB, CDRSP-Politécnico de Leiria, Rensselaer Polytechnic Institute (Troy, NY-USA), and an orthopedic surgeon (Hospital dos Lusíadas). The project, which falls within the scientific area of Mechanical Engineering-Engineering Systems, is headed by João Carlos Silva (PI, SCERG-iBB) and Prof. Paula Pascoal-Faria (co-PI, CDRSP-Politécnico de Leiria).

"This week in the Portuguese newspaper "Público" you will find a summary of the Algae2Fish project, where you can read about the reasons behind the project and its perspectives for the next two years. The team, composed by professor Frederico Ferreira, Dr. Paola Sanjuan Alberte, Dr. Carlos Rodrigues, and Msc Diana Marques, with the support of The Good Food Institute, is currently working to produce the first cell-cultured sea bass fillet!"

Engineering brain organoids from human induced pluripotent stem cells (hiPSCs) is a powerful tool for modeling brain development and neurological disorders. Rett syndrome (RTT), a rare neurodevelopmental disorder, can greatly benefit from this technology, since it affects multiple neuronal subtypes in forebrain sub-regions. SCERG-iBB researchers have recently established dorsal and ventral forebrain organoids from control and RTT patient-specific hiPSCs recapitulating the 3D organization and functional network complexity of this brain region. The data obtained revealed a premature development of the deep-cortical layer, associated to the formation of TBR1 and CTIP2 neurons, and a lower expression of neural progenitor/proliferative cells in RTT dorsal organoids. Moreover, calcium imaging and electrophysiology analysis demonstrated functional defects of RTT neurons. Additionally, assembly of RTT dorsal and ventral organoids revealed impairments of interneuron’s migration. Overall, these models provide a better understanding of RTT during early stages of neural development, demonstrating a great potential for personalized diagnosis and drug screening. The paper was published in Frontiers in Cell Development Biology.

The project "DentalBioMatrix: Exploiting the power of decellularized extracellular matrix to fabricate hierarchical biomimetic scaffolds to regenerate functional periodontal tissues" has been recommended for funding by FCT (2020 Call for SR&TD Project Grants). The goal of DentalBioMatrix is to develop novel bioengineering strategies to regenerate periodontium, by engineering hierarchically designed compartmentalized systems to meet the different characteristics of the tissues involved in periodontal defects. Decellularized extracellular matrix (dECM) will be used to create complex dECM-derived scaffolds to recreate the different niches of periodontium. Collectively, this approach will potentially provide the opportunity to harness the properties of periodontal tissue ECM, recreating a local niche at the tooth interface that can promote periodontal regeneration, especially important for osteoporotic and older patients with compromised ECM. The project, which falls within the scientific area of Medical Biotechnology, is headed by Marta Carvalho from SCERG-iBB.

Today, it is recognized that medicines will eventually be needed during pregnancy, either due to gestation-related medical conditions or pre-existing diseases. Also, the rate of drug prescription to pregnant women has increased over the past few years, in accordance with the increasing trend to postpone childbirth to a later age. However, information regarding teratogenic risk in humans is often missing for most of the purchased drugs. Inter-species differences have limited the suitability of animal models to predict human-specific outcomes, contributing to misidentified human teratogenicity. Therefore, the development of physiologically relevant in vitro humanized models can be the key to surpassing this limitation. In this context, this review describes the pathway towards the introduction of human pluripotent stem cell-derived models in developmental toxicity studies.

As current treatments for periodontitis have failed to promote tissue repair and the coordinated regeneration of the whole periodontium, innovative therapeutic strategies are urgently needed to improve the clinical outcomes. Electrospun nanofibrous scaffolds are particularly promising for applications in periodontal regeneration since they are able to mimic the native tissue extracellular matrix (ECM) features such as the size and alignment of the fibers present in the periodontal ligament (PDL). In this article recently published in the journal Nanomaterials, iBB researchers MSc Mafalda Santos, Dr. Marta Carvalho and Dr. João Silva provided an overview of the current state of the art of the application of electrospun nanofibers in periodontal regeneration, either as guided tissue regeneration (GTR) membranes or as scaffolds for tissue engineering strategies.

Cell and gene therapies (CGT) have reached new therapeutic targets but have noticeably high prices. Solutions to reduce production costs might be found in CGT storage and transportation since they typically involve cryopreservation, which is a heavily burdened process. Encapsulation at hypothermic temperatures (e.g., 2–8 °C) can be a feasible alternative. In this study, we determined the ability of alginate encapsulation to maintain cell viability, identity, and function in the context of mesenchymal stromal cell (MSC)-based therapy manufacturing. MSC encapsulation was proven possible for a remarkable 12 day period, showing its potential to act as a serious competitor to cryopreservation for short to medium time periods. This study developed at iBB was led by Dr. Ana Fernandes-Platzgummer and Prof. Cláudia Lobato da Silva and first-authored by PhD student André Branco.

The embryonic development of the human heart under healthy and pathological conditions is still very poorly understood. In a recently published Nature Communications paper, a iBB team led by Margarida Diogo, in collaboration with Perpetua Pinto do Ó at i3S, engineered a unique organ-like model from stem cells that recreates several stages of embryonic development of the human heart. This humanized model opens now the path to study, in a tissue engineering lab, the development of congenital heart diseases, such as coronary vascular diseases and myocardium non-compaction cardiomyopathies, and can be further applied in drug screening and toxicology assays during embryonic heart development. This last topic is particularly relevant since pregnant women normally do not take part in clinical trials, making this human cell-based in vitro platform the only viable alternative to assess developmental cardiotoxicity. The work was a part of the PhD thesis in Bioengineering – MIT Portugal program of Mariana Branco, the first author of the paper, and was funded by the FCT project “Mini-Hearts - Organoid Engineering for Production of 3D Cardiovascular Microtissues from Human Induced Pluripotent Stem Cells for Cardiotoxicity”.  

Phenylketonuria (PKU) is a recessive genetic disorder of amino-acid metabolism, where impaired phenylalanine hydroxylase function in the liver of patients leads to the accumulation of neurotoxic phenylalanine levels in the brain. Despite the current knowledge, the chronic effect of PKU in the brain is still poorly understood. In a recent publication in Frontiers in Molecular Neurosciences, DBE faculty and SCERG-iBB researcher Tiago Fernandes, discusses the need for better predictive models, able to recapitulate specific mechanisms of this disease. New exciting in vitro platforms to model specific PKU-derived neuronal impairment are presented in a attempt to understand the impact of phenylalanine in the brain of patients, and ultimately contribute to the understanding of this disease.

The project Algae2Fish led by Frederico Ferreira from SCERG-iBB has received funding from the Global Food Institute with the aim of developing a boneless cultivated seabass fillet. 3D printing technology will be used to produce scaffolds that will give the fillet structure, replicating the fibrous texture of fish. The scaffolds will be formed using material from algae and plants, with the algae contributing valuable omega-3 fatty acids such as those found in conventional fish. Read more about the project here.

Endogenous human brain tissue is not easily available for studying neurodevelopment and neurodegenerative diseases. However, human pluripotent stem cells (PSCs) have been used to generate a variety of glial and neuronal cells of the central nervous system. Still, reproducible protocols for generating in vitro models of the human cerebellum are scarce. In this context, Silva et al. describe the scalable production of human PSC-derived cerebellar organoids using single-use vertical-wheel bioreactors. The transcriptomic profile of cerebellar organoids derived under dynamic conditions demonstrates a faster cerebellar differentiation combined with significant enrichment of extracellular matrix and upregulation of transcripts involved in angiogenesis when compared with the static protocol. The authors anticipate that large-scale production of cerebellar organoids may help developing models for drug screening, toxicological tests and studying pathological pathways involved in cerebellar degeneration.

João Carlos Silva and Frederico Castelo Ferreira from SCERG-iBB are guest-editing a special issue for the open-access journal “Polymers” entitled “Advanced Polymeric Scaffolds for Stem Cell Engineering and Regenerative Medicine”. Polymer scaffolds play a crucial role in tissue engineering and regenerative medicine applications since they can closely mimic the architecture of a native extracellular matrix (ECM) and improve the biological performance of cells both in vitro and in vivo. This Special Issue welcomes full research papers, communications and reviews on recent exciting developments of polymeric scaffolds for tissue engineering and regenerative medicine applications.

The project "UROPRINT: Urinary bladder bioprinting for fully autologous transplantation" has been recommended for funding by the EU. The goal of UROPRINT is to laser print fully functional immunocompatible urothelial tissue ex vivo and in vivo for bladder augmentation and replacement. The project is led by the Institute of Biomedical Research of the Academy of Athens and involves iBB-SCERG, Asphalion SL, Institute of Comunication and Computer Systems, Optics11 BV and METATISSUE/JFCC-Biosolutions, LDA.

Photo details: National Institute of Standards and Technology, Public domain, via Wikimedia Commons.