Clara R. Correia

• Doctor of Tissue Engineering Regenerative Medicine and Stem Cells
• Senior Scientist at Stemmatters SA

Background Surface topography has been shown to influence cell behavior and direct stromal cell differentiation into distinct lineages. Whereas this phenomenon has been verified in two-dimensional cultures, there is an urgent need for a thorough investigation of topography’s role within a three-dimensional (3D) environment, as it better replicates...

Background Surface topography has been shown to influence cell behavior and direct stromal cell differentiation into distinct lineages. Whereas this phenomenon has been verified in two-dimensional cultures, there is an urgent need for a thorough investigation of topography’s role within a three-dimensional (3D) environment, as it better replicates...

It is essential to design a multifunctional well‐controlled platform to transfer mechanical cues to the cells in different magnitudes. This study introduces a platform, a miniaturized bioreactor, which enables to study the effect of shear stress in microsized compartmentalized structures. In this system, the well‐established cell encapsulation syst...

In order to solve the clinical challenges related to bone grafting, several tissue engineering (TE) strategies have been proposed to repair critical-sized defects. Generally, the classical TE approaches are designed to promote bone repair via intramembranous ossification. Although promising, strategies that direct the osteogenic differentiation of...

Extraordinary capabilities underlie the potential use of immune cells, particularly macrophages, in bone tissue engineering. Indeed, the depletion of macrophages during bone repair often culminates in disease scenarios. Inspired by the native dynamics between immune and skeletal systems, this work proposes a straightforward in vitro method to bioen...

The well-known synergetic interplay between the skeletal and immune systems has changed the design of advanced bone tissue engineering strategies. The immune system is essential during the bone lifetime, with macrophages playing multiple roles in bone healing and biomaterial integration. If in the past, the most valuable aspect of implants was to a...

Recreating the biological complexity of living bone marrow (BM) in a single in vitro strategy has faced many challenges. Most bioengineered strategies propose the co-culture of BM cellular components entrapped in different matrices limiting their migration and self-organization capacity or in open scaffolds enabling their escaping. We propose a met...

Structure and organisation are key aspects of the native tissue environment, which ultimately condition cell fate via a myriad of processes, including the activation of mechanotransduction pathways. By modulating the formation of integrin-mediated adhesions and consequently impacting cell contractility, engineered geometrical and topographical cues...

Replacement orthopedic surgeries are among the most common surgeries worldwide, but clinically used passive implants cannot prevent failure rates and inherent revision arthroplasties. Optimized non-instrumented implants, resorting to preclinically tested bioactive coatings, improve initial osseointegration but lack long-term personalized actuation...

Dynamic soft materials that have the ability to expand and contract, change stiffness, self-heal or dissolve in response to environmental changes, are of great interest in applications ranging from biosensing and drug delivery to soft robotics and tissue engineering. This book covers the state-of-the-art and current trends in the very active and ex...

The therapeutic effectiveness and biological relevance of technologies based on adherent cells depend on platforms that enable long‐term culture in controlled environments. Liquid‐core capsules have been suggested as semipermeable moieties with spatial homogeneity due to the high mobility of all components in their core. The lack of cell‐adhesive s...

From an “over‐engineering” era in which biomaterials played a central role, now it is observed to the emergence of “developmental” tissue engineering (TE) strategies which rely on an integrative cell‐material perspective that paves the way for cell self‐organization. The current challenge is to engineer the microenvironment without hampering the sp...

The repair process of bone fractures is a complex biological mechanism requiring the recruitment and in situ functionality of stem/stromal cells from the bone marrow (BM). While BM mesenchymal stem/stromal cells have been widely explored in multiple bone tissue engineering applications, the use of hematopoietic stem cells (HSCs) has been poorly exp...

A critical determinant of successful clinical outcomes is the host's response to the biomaterial. Therefore, the prediction of the immunomodulatory bioperformance of biomedical devices following implantation is of utmost importance. Herein, liquefied capsules are proposed as immunomodulatory miniaturized 3D platforms for the high‐content combinator...

Manufacturing macroscale cell-laden architectures is one of the biggest challenges faced nowadays in the domain of tissue engineering. Such living constructs, in fact, pose strict requirements for nutrients and oxygen supply that can hardly be addressed through simple diffusion in vitro or without a functional vasculature in vivo. In this context,...

An old but still hot topic in tissue engineering (TE) is the establishment of efficient vascularization networks proving fine, controlled, and long-term distribution of oxygen and nutrients. Combining elegant three-dimensional (3D) fabrication techniques with unconventional living microorganisms, namely photosynthetic species, complex 3D-printed TE...

In article number 2000127, Clara R. Correia, João F. Mano, and co‐workers describe the high‐throughput production of alginate‐based microgels with defined geometries using an in situ crosslinking procedure. This encapsulation system is surrounded by a permselective polymeric membrane that wraps cells and microparticles in a liquefied environment. T...

In the human body, highly organized tissues rely on the compartmentalization effect of basement membranes (BMs) that separate different types of cells. We recently reported an artificial basement membrane (A-BM) composed of type-IV collagen and laminin (Col-IV/LM), which are the main components of natural BMs, for cell compartmentalization in three...

A plethora of bioinspired cell‐laden hydrogels are being explored as building blocks that once assembled are able to create complex and highly hierarchical structures recapitulating the heterogeneity of living tissues. Yet, the resulting 3D bioengineered systems still present key limitations, mainly related with limited diffusion of essential molec...

Grooved topography and inherent cell contact guidance has shown promising results regarding cell proliferation, morphology, and lineage‐specific differentiation. Yet these approaches are limited to 2D applications. Sandwich‐culture conditions are developed to bridge the gap between 2D and 3D culture, enabling both ventral and dorsal cell surface st...

In article number 1907434, Michiya Matsusaki and co‐workers introduce an artificial basement membrane based on collagen type IV and laminin by a layer‐by‐layer assembly technique. Benefited by the efficient cell compartmentalization property of multilayered nanofilms, patterned co‐culture between fibroblasts and endothelial cells will contribute to...

In recent decades, tissue engineering techniques have attracted much attention in the construction of 3D tissues or organs. However, even though precise control of cell locations in 3D has been achieved, the organized cell locations are easily destroyed because of the cell migration during the cell culture period. In human body, basement membranes...

In the primordial cell encapsulation systems, the main goal is to treat endocrine diseases avoiding the action of the immune system. Although lessons afforded by such systems are of outmost importance for the demands of tissue engineering and regenerative medicine, the paradigm has recently completely changed. If before the most important feature w...

Cells with differentiation potential into mesodermal types are the focus of emerging bone tissue engineering (TE) strategies as an alternative autologous source. When the source of cells is extremely limited or not readily accessible, such as in severe injuries, a tissue biopsy may not yield the required number of viable cells. In line, adipose-der...

Cell encapsulation systems must ensure the diffusion of molecules to avoid the formation of necrotic cores. The architectural design of hydrogels, the gold standard tissue engineering strategy, is thus limited to a microsize range. To overcome such a limitation, liquefied microcapsules encapsulating cells and microparticles are proposed. Microcapsu...

Grooved topographical features have effectively modulated cell differentiation on two-dimensional substrates. To transpose patterning into a 3D context, nanogrooved microdiscs, “topodiscs”, are produced as cell-carriers for bottom-up cell-mediated assembly. While...

Cell encapsulation is a widely used technique in the field of Tissue Engineering and Regenerative Medicine (TERM). However, for the particular case of liquefied compartmentalised systems, only a limited number of studies have been reported in the literature. We have been exploring a unique cell encapsulation system composed by liquefied and multila...

Significance Honey bees present a peculiar hairy structure that covers the surface of their body, which is able to transport large quantities of pollen particles for pollination purposes. Inspired by this natural phenomenon and to overcome the current problems associated with drug patches with passive delivery, we propose the concept of a micropatt...

In the last few years, several strategies have been proposed to fabricate scaffolds for tissue engineering (TE) applications; however, they are based on harsh and time‐consuming techniques. The choice for natural polymers such as cashew gum (CG) allows to circumvent the demands of biocompatibility and degradability of TE systems. In this work, CG,...

Cell encapsulation systems are being increasingly applied as multifunctional strategies to regenerate tissues. Lessons afforded with encapsulation systems aiming to treat endocrine diseases seem to be highly valuable for the tissue engineering and regenerative medicine (TERM) systems of today, in which tissue regeneration and biomaterial integratio...

Statement of significance: The diffusion efficiency of essential molecules for cell survival is a main issue in cell encapsulation. Former studies reported the superior biological outcome of encapsulated cells within liquified systems. However, most cells used in TE are anchorage-dependent, requiring a solid substrate to perform main cellular proc...

Macrophages play a crucial role in the biological performance of biomaterials as key factors in defining the optimal inflammation-healing balance towards tissue regeneration and implant integration. Here, we investigate how different surface modifications performed on poly(L-lactic acid) (PLLA) films would influence the differentiation of human mon...

TGF-β3 is enzymatically immobilized by transglutaminase-2 action to poly(l-lactic acid) microparticles coated with collagen II. Microparticles are then encapsulated with stem cells inside liquified spherical compartments enfolded with a permselective shell through layer-by-layer adsorption. Magnetic nanoparticles are electrostatically bound to the...

A new concept of semipermeable reservoirs containing co-cultures of cells and supporting microparticles is presented, inspired by the multi-phenotypic cellular environment of bone. Based on the deconstruction of the “stem cell niche”, the developed capsules are designed to drive a self-regulated osteogenesis. PLLA microparticles functionalized with...

Cartilage tissue is a complex nonlinear, viscoelastic, anisotropic, and multiphasic material with a very low coefficient of friction, which allows to withstand millions of cycles of joint loading over decades of wear. Upon damage, cartilage tissue has a low self-reparative capacity due to the lack of neural connections, vascularization, and a laten...

Osteogenic Differentiation of Adipose Stem Cells by Endothelial Cells Co-culture Within Liquified Capsules Clara R. Correia1,2, Rogério P. Pirraco1,2, Mariana T. Cerqueira1,2, Alexandra P. Marques1,2, Rui L. Reis1,2, João F. Mano1,2 1 3B’s Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the Europ...

Chondrogenic Differentiation Within Magnetic-multilayered Liquified Capsules Containing Collagen II/TGF-β3 Microparticles Clara R. Correia,1,2 Sara Gil,1,2 Rui L. Reis,1,2 and João F. Mano1,2 1 3B’s Research Group - Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue En...

Stable liquid marbles (LM) are produced by coating liquid droplets with a hydrophobic powder. The used hydrophobic powder is produced by fluorosi­lanization of diatomaceous earth, used before to produce superhydrophobic structures. Here, the use of LM is proposed for high-throughput drug screening on anchorage-dependent cells. To provide the requir...

We propose a novel hanging spherical drop system for anchoring arrays of droplets of cell suspension based on the use of biomimetic superhydrophobic flat substrates, with controlled positional adhesion and minimum contact with a solid substrate. By facing down the platform, it was possible to generate independent spheroid bodies in a high throughpu...

Magnetically targeted cells with internalized magnetic nanoparticles (MNPs) could allow the success of cell transplantation and cell-based therapies, overcoming low cell retention that occurs when delivering cells by intravenous or local injection. Upon magnetization, these cells could then accumulate and stimulate the regeneration of the tissue in...

Polymeric multilayered capsules (PMCs) have found great applicability in bioencapsulation, an evolving branch of tissue engineering and regenerative medicine. Here, we describe the production of hierarchical PMCs composed by an external multilayered membrane by layer-by-layer assembly of poly(L-lysine), algi-nate, and chitosan. The core of the PMCs...

Self-supporting, millimeter length 3D constructs consisting of individualized liquefied compartments, were produced using cell encapsulated hydrogel beads as building blocks. A perfusion-based layer-by-layer approach was used that allowed bioencapsulated beads to assemble, pattern, hold and attach to produce non-liquefied 3D constructs with control...

The development of high-throughput and combinatorial technologies is helping to speed up research that is applicable in many areas of chemistry, engineering, and biology. A new model is proposed for flat devices for the high-throughput screening of accelerated evaluations of multiplexed processes and reactions taking place in aqueous-based environm...

In a marine environment, specific proteins are secreted by mussels and used as a bioglue to stick to a surface. These mussel proteins present an unusual amino acid 3,4-dihydroxyphenylalanine (known as DOPA). The outstanding adhesive properties of these materials in the sea harsh conditions have been attributed to the presence of the catechol groups...

Magnetically targeted cells with internalized magnetic nanoparticles (MNPs) could allow the success of cell transplantation and cell-based therapies, overcoming low cell retention that occurs when delivering cells by intravenous or local injection. Upon magnetization, these cells could then accumulate and stimulate the regeneration of the tissue in...

We suggest the use of biomimetic superhydrophobic patterned chips produced by a bench-top methodology as low-cost and waste-free platforms for the production of arrays of cell spheroids/microtissues by the hanging drop methodology. Cell spheroids have a wide range of applications in biotechnology fields. For drug screening, they allow studying 3D m...

Chitosan is soluble in acidic media, which makes it incompatible for the encapsulation of cells and pH-sensitive molecules. In this work, a mild chitosan-based system with two sequential gelation steps is proposed, where the model drug dexamethasone and L929 cells are immobilized inside hydrogel beads. Superhydrophobic surfaces were used to produce...

Medical adhesives and sealants often require that long-term adhesiveness is achieved. In this work, nanostructured coatings consisting of chitosan and the adhesive bacterial exopolysaccharide levan are fabricated using layer-by-layer (LbL) assembly. Taking advantage of the electrostatic self-assembly mechanism of LbL, the charges of both chitosan a...

We report the development of liquified multilayer hierarchical capsules capable of providing cell adhesion sites to the encapsulated cells. The proof of principle is demonstrated with the example of a chitosan–alginate shell via layer-by-layer assembly, encapsulating cells adhered to the functionalized surface of poly(L-lactic acid) microparticles.

Liquified capsules featuring (i) an external shell by layer-by-layer assembly of poly(L-lysine), alginate and chitosan, and encapsulating (ii) surface functionalized poly(L-lactic acid) (PLLA) microparticles were developed. We hypothesize that, while the liquified environment enhances the diffusion of essential molecules for cell survival, micropar...

Scaffolds derived from natural polysaccharides are very promising in tissue engineering applications and regenerative medicine, as they resemble glycosaminoglycans in the extracellular matrix (ECM). In this study, we have prepared freeze-dried composite scaffolds of chitosan (CHT) and hyaluronic acid (HA) in different weight ratios containing eithe...

Scaffolds derived from natural polysaccharides are very promising in tissue engineering applications and regenerative medicine, as they resemble glycosaminoglycans in the extracellular matrix. In this study we have prepared freeze-dried composite scaffolds of chitosan (CHT) and hyaluronic acid (HA), in different weight ratios containing either no H...