Sandra Camarero-Espinosa
@polymat.eu
University of The Basque Country
POLYMAT
Scopus Publications
Scholar Citations
Scholar h-index
Scholar i10-index
Scopus Publications
Neda Kathami, Carolina Moreno-Vicente, Pablo Martín, Jhonatan A. Vergara-Arce, Raquel Ruiz-Hernández, Daniela Gerovska, Ana M. Aransay, Marcos J. Araúzo-Bravo, Sandra Camarero-Espinosa, and Ander Abarrategi
Springer Science and Business Media LLC
Abstract Background Recombinant human bone morphogenetic protein 2 (rhBMP-2) and human bone marrow mesenchymal stromal cells (hBM-MSCs) have been thoroughly studied for research and translational bone regeneration purposes. rhBMP-2 induces bone formation in vivo, and hBM-MSCs are its target, bone-forming cells. In this article, we studied how rhBMP-2 drives the multilineage differentiation of hBM-MSCs both in vivo and in vitro. Methods rhBMP-2 and hBM-MSCs were tested in an in vivo subcutaneous implantation model to assess their ability to form mature bone and undergo multilineage differentiation. Then, the hBM-MSCs were treated in vitro with rhBMP-2 for short-term or long-term cell-culture periods, alone or in combination with osteogenic, adipogenic or chondrogenic media, aiming to determine the role of rhBMP-2 in these differentiation processes. Results The data indicate that hBM-MSCs respond to rhBMP-2 in the short term but fail to differentiate in long-term culture conditions; these cells overexpress the rhBMP-2 target genes DKK1, HEY-1 and SOST osteogenesis inhibitors. However, in combination with other differentiation signals, rhBMP-2 acts as a potentiator of multilineage differentiation, not only of osteogenesis but also of adipogenesis and chondrogenesis, both in vitro and in vivo. Conclusions Altogether, our data indicate that rhBMP-2 alone is unable to induce in vitro osteogenic terminal differentiation of hBM-MSCs, but synergizes with other signals to potentiate multiple differentiation phenotypes. Therefore, rhBMP-2 triggers on hBM-MSCs different specific phenotype differentiation depending on the signalling environment.
Mahtab Asadian, Clarissa Tomasina, Yuliia Onyshchenko, Ke Vin Chan, Mohammad Norouzi, Jip Zonderland, Sandra Camarero‐Espinosa, Rino Morent, Nathalie De Geyter, and Lorenzo Moroni
Wiley
AbstractBone marrow‐derived mesenchymal stromal cells (BMSCs) are extensively being utilized for cartilage regeneration owing to their excellent differentiation potential and availability. However, controlled differentiation of BMSCs towards cartilaginous phenotypes to heal full‐thickness cartilage defects remains challenging. This study investigates how different surface properties induced by either coating deposition or biomolecules immobilization onto nanofibers (NFs) could affect BMSCs chondro‐inductive behavior. Accordingly, electrospun poly(ε‐caprolactone) (PCL) NFs were exposed to two surface modification strategies based on medium‐pressure plasma technology. The first strategy is plasma polymerization, in which cyclopropylamine (CPA) or acrylic acid (AcAc) monomers were plasma polymerized to obtain amine‐ or carboxylic acid‐rich NFs, respectively. The second strategy uses a combination of CPA plasma polymerization and a post‐chemical technique to immobilize chondroitin sulfate (CS) onto the NFs. These modifications could affect surface roughness, hydrophilicity, and chemical composition while preserving the NFs' nano‐morphology. The results of long‐term BMSCs culture in both basic and chondrogenic media proved that the surface modifications modulated BMSCs chondrogenic differentiation. Indeed, the incorporation of polar groups by different modification strategies had a positive impact on the cell proliferation rate, production of the glycosaminoglycan matrix, and expression of extracellular matrix proteins (collagen I and collagen II). The chondro‐inductive behavior of the samples was highly dependent on the nature of the introduced polar functional groups. Among all samples, carboxylic acid‐rich NFs promoted chondrogenesis by higher expression of aggrecan, Sox9, and collagen II with downregulation of hypertrophic markers. Hence, this approach showed an intrinsic potential to have a non‐hypertrophic chondrogenic cell phenotype.
Neda Khatami, Pedro Guerrero, Pablo Martín, Eztizen Quintela, Viviana Ramos, Laura Saa, Aitziber L. Cortajarena, Koro de la Caba, Sandra Camarero-Espinosa, and Ander Abarrategi
Elsevier BV
Sandra Camarero‐Espinosa, Ivo Beeren, Hong Liu, David B. Gomes, Jip Zonderland, Ana Filipa H Lourenço, Denis van Beurden, Marloes Peters, David Koper, Pieter Emans,et al.
Wiley
AbstractThe regeneration of the osteochondral unit represents a challenge due to the distinct cartilage and bone phases. Current strategies focus on the development of multiphasic scaffolds that recapitulate features of this complex unit and promote the differentiation of implanted bone‐marrow derived stem cells (BMSCs). In doing so, challenges remain from the loss of stemness during in vitro expansion of the cells and the low control over stem cell activity at the interface with scaffolds in vitro and in vivo. Here, this work scaffolds inspired by the bone marrow niche that can recapitulate the natural healing process after injury. The construct comprises an internal depot of quiescent BMSCs, mimicking the bone marrow cavity, and an electrospun (ESP) capsule that “activates” the cells to migrate into an outer “differentiation‐inducing” 3D printed unit functionalized with TGF‐β and BMP‐2 peptides. In vitro, niche‐inspired scaffolds retained a depot of nonproliferative cells capable of migrating and proliferating through the ESP capsule. Invasion of the 3D printed cavity results in location‐specific cell differentiation, mineralization, secretion of alkaline phosphatase (ALP) and glycosaminoglycans (GAGs), and genetic upregulation of collagen II and collagen I. In vivo, niche‐inspired scaffolds are biocompatible, promoted tissue formation in rat subcutaneous models, and regeneration of the osteochondral unit in rabbit models.
R. Olmos-Juste, G. Larrañaga-Jaurrieta, I. Larraza, S. Ramos-Diez, S. Camarero-Espinosa, N. Gabilondo, and A. Eceiza
Elsevier BV
Sandra Camarero‐Espinosa, Huipin Yuan, Pieter J. Emans, and Lorenzo Moroni
Wiley
Anterior cruciate ligament (ACL) is the connective tissue providing mechanical stability to the knee joint. ACL reconstruction upon rupture remains a clinical challenge due to the high mechanical properties required for proper functioning. ACL owes its outstanding mechanical properties to the arrangement of the ECM and to the cells with distinct phenotypes present along the length of the tissue. Tissue regeneration appears as an ideal alternative. In this study, we developed a tri-phasic fibrous scaffold that mimics the structure of collagen in the native ECM, presenting a wavy intermediate zone and two aligned uncurled extremes. The mechanical properties of the wavy scaffolds presented a toe region, characteristic of the native ACL, and an extended yield and ultimate strain as compared to traditional aligned scaffolds. The presentation of a wavy fiber arrangement affected the cell organization and the deposition of a specific ECM characteristic of fibrocartilage. Cells cultured in wavy scaffolds grew in aggregates, deposited an abundant ECM rich in fibronectin and collagen II, and expressed higher amounts of collagen II, X and tenomodulin as compared to aligned scaffolds. In-vivo implantation in rabbits showed a high cellular infiltration and the formation of an oriented ECM, as compared to traditional aligned scaffolds. This article is protected by copyright. All rights reserved.
Sandra Ramos-Díez, Garazi Larrañaga-Jaurrieta, Leire Iturriaga, Ander Abarrategi, and Sandra Camarero-Espinosa
Royal Society of Chemistry (RSC)
A library of low molecular weight biocompatible inks has been developed to be used in DLP printing. The resulting inks present low viscosity and are printable without diluents or solvents, resulting in structures with high shape fidelity.
Clarissa Tomasina, Giorgia Montalbano, Sonia Fiorilli, Paulo Quadros, António Azevedo, Catarina Coelho, Chiara Vitale-Brovarone, Sandra Camarero-Espinosa, and Lorenzo Moroni
Elsevier BV
Ivo A O Beeren, Pieter J Dijkstra, Ana Filipa H Lourenço, Ravi Sinha, David B Gomes, Hong Liu, Nicole Bouvy, Matthew B Baker, Sandra Camarero-Espinosa, and Lorenzo Moroni
IOP Publishing
Abstract Melt extrusion-based additive manufacturing (AM) is often used to fabricate scaffolds for osteochondral (OC) regeneration. However, there are two shortcomings associated with this scaffold manufacturing technique for engineering of tissue interfaces: (a) most polymers used in the processing are bioinert, and (b) AM scaffolds often contain discrete (material) gradients accompanied with mechanically weak interfaces. The inability to mimic the gradual transition from cartilage to bone in OC tissue leads to poor scaffold performance and even failure. We hypothesized that introducing peptide gradients on the surface could gradually guide human mesenchymal stromal cell (hMSC) differentiation, from a chondrogenic towards on osteogenic phenotype. To work towards this goal, we initially manufactured poly(ϵ-caprolactone)-azide (PCLA) and PCL-maleimide (PCLM) scaffolds. The surface exposed click-type functional groups, with a surface concentration in the 102pmol cm−2 regime, were used to introduce bone morphogenic protein-2 or transforming growth factor-beta binding peptide sequences to drive hMSC differentiation towards osteogenic or chondrogenic phenotypes, respectively. After 3 weeks of culture in chondrogenic medium, we observed differentiation towards hypertrophic chondrogenic phenotypes with expression of characteristic markers such as collagen X. In osteogenic medium, we observed the upregulation of mineralization markers. In basic media, the chondro-peptide displayed a minor effect on chondrogenesis, whereas the osteo-peptide did not affect osteogenesis. In a subcutaneous rat model, we observed a minimal foreign body response to the constructs, indicating biocompatibility. As proof-of-concept, we finally used a novel AM technology to showcase its potential to create continuous polymer gradients (PCLA and PCLM) across scaffolds. These scaffolds did not display delamination and were mechanically stronger compared to discrete gradient scaffolds. Due to the versatility of the orthogonal chemistry applied, this approach provides a general strategy for the field; we could anchor other tissue specific cues on the clickable groups, making these gradient scaffolds interesting for multiple interfacial tissue applications.
Ivo A.O. Beeren, Pieter J. Dijkstra, Philippe Massonnet, Sandra Camarero-Espinosa, Matthew B. Baker, and Lorenzo Moroni
Elsevier BV
Maria Soledad Orellano, Oihane Sanz, Sandra Camarero-Espinosa, Ana Beloqui, and Marcelo Calderón
Future Medicine Ltd
Sandra Camarero‐Espinosa, Maria Carlos‐Oliveira, Hong Liu, João F. Mano, Nicole Bouvy, and Lorenzo Moroni
Wiley
Tissue regeneration evolves towards the biofabrication of sophisticated 3D scaffolds. However, the success of these will be contingent to their capability to integrate within the host. The control of the mechanical or topographical properties of the implant, appear as ideal methods to modulate the immune response. However, the interplay between these properties is yet not clear. We created dual-porosity scaffolds with varying mechanical and topographical features and evaluated their immunomodulatory properties in rat alveolar macrophages in vitro, and in vivo in a rat subcutaneous model. Scaffolds were fabricated via additive-manufacturing and thermally induced phase separation (TIPS) methods from two copolymers with virtually identical chemistries, but different stiffness. The introduction of porosity enabled the modulation of macrophages towards anti-inflammatory phenotypes, with secretion of IL-10 and TGF-β. Soft scaffolds (< 5 kPa) resulted in a pro-inflammatory phenotype in contrast to stiffer (> 40 kPa) scaffolds of comparable porosities supporting a pro-healing phenotype, which appeared to be related to the surface spread area of cells. In vivo, stiff scaffolds integrated, while softer scaffolds appeared encapsulated after 3 weeks of implantation, resulting on chronic inflammation after 6 weeks. Our results demonstrate the importance of evaluating the interplay between topography and stiffness of candidate scaffolds. This article is protected by copyright. All rights reserved.
Leire Iturriaga, Kyle D. Van Gordon, Garazi Larrañaga-Jaurrieta, and Sandra Camarero‐Espinosa
Wiley
Regeneration of tissues represents a current challenge and a need to treat damage and diseased organs, and is becoming determinant to alleviate the burden of healthcare systems with the increasing age of the population. Although many strategies are being developed and some scaffolds are already reaching the market, issues of formation of well‐structured and functional tissues are still prevalent. Additive manufacturing and particularly 3D printing have emerged as ideal technologies to fabricate patient‐specific scaffolds and allow for the easy modification of multiple structural parameters. Yet, these are generally composed of smooth fibers that are not able to drive, by themselves, the formation of well‐structured tissues. The use of physical cues to modulate cellular processes such as migration, proliferation, differentiation, and matrix synthesis has been proven effective in 2D. Thus, the extrapolation of these physical cues to 3D‐printed scaffolds appears as a tempting approach to promote the formation of functional tissues and thus, many strategies are being developed to this end. Herein, an overview of developed techniques to introduce topography and porosity to 3D‐printed scaffolds and their effect on the cell response and tissue formation is provided.
Sandra Camarero-Espinosa and Lorenzo Moroni
Springer Science and Business Media LLC
AbstractThe application of physical stimuli to cell cultures has shown potential to modulate multiple cellular functions including migration, differentiation and survival. However, the relevance of these in vitro models to future potential extrapolation in vivo depends on whether stimuli can be applied “externally”, without invasive procedures. Here, we report on the fabrication and exploitation of dynamic additive-manufactured Janus scaffolds that are activated on-command via external application of ultrasounds, resulting in a mechanical nanovibration that is transmitted to the surrounding cells. Janus scaffolds were spontaneously formed via phase-segregation of biodegradable polycaprolactone (PCL) and polylactide (PLA) blends during the manufacturing process and behave as ultrasound transducers (acoustic to mechanical) where the PLA and PCL phases represent the active and backing materials, respectively. Remote stimulation of Janus scaffolds led to enhanced cell proliferation, matrix deposition and osteogenic differentiation of seeded human bone marrow derived stromal cells (hBMSCs) via formation and activation of voltage-gated calcium ion channels.
Giorgia Montalbano, Clarissa Tomasina, Sonia Fiorilli, Sandra Camarero-Espinosa, Chiara Vitale-Brovarone, and Lorenzo Moroni
MDPI AG
The use of biomaterials and scaffolds to boost bone regeneration is increasingly gaining interest as a complementary method to the standard surgical and pharmacological treatments in case of severe injuries and pathological conditions. In this frame, the selection of biomaterials and the accurate assessment of the manufacturing procedures are considered key factors in the design of constructs able to resemble the features of the native tissue and effectively induce specific cell responses. Accordingly, composite scaffolds based on type-I-collagen can mimic the composition of bone extracellular matrix (ECM), while electrospinning technologies can be exploited to produce nanofibrous matrices to resemble its architectural organization. However, the combination of collagen and electrospinning reported several complications due to the frequent denaturation of the protein and the variability of results according to collagen origin, concentration, and solvent. In this context, the strategies optimized in this study enabled the preparation of collagen-based electrospun scaffolds characterized by about 100 nm fibers, preserving the physico-chemical properties of the protein thanks to the use of an acetic acid-based solvent. Moreover, nanoparticles of mesoporous bioactive glasses were combined with the optimized collagen formulation, proving the successful design of composite scaffolds resembling the morphological features of bone ECM at the nanoscale.
Tobias Kuhnt and Sandra Camarero-Espinosa
Elsevier BV
Cellulose nanomaterials (CNMs) have attracted great attention in the last decades due to the abundance of the biopolymer, the biorenewable character and the outstanding mechanical properties they account for. These, together with their biocompatibility makes them ideal candidates for tissue engineering (TE) applications. Additive manufacturing is an ideal biofabrication approach for TE, providing rapid and reliable technologies to produce scaffolds aimed for the guidance of host or implanted cells to form functional tissues. However, the control of parameters at the nanoscale that regulate cellular functions such as proliferation and differentiation remain challenging. This review article presents the latest advances in the use of CNMs as platforms to guide cellular functions in additive manufactured scaffolds. Special attention is given to functionalization routes, methods to exploit them as topographical cues and to improve the local mechanical properties together with the resulting cell-CNM interactions.
Carlos Mota, Sandra Camarero-Espinosa, Matthew B. Baker, Paul Wieringa, and Lorenzo Moroni
American Chemical Society (ACS)
Bioprinting techniques have been flourishing in the field of biofabrication with pronounced and exponential developments in the past years. Novel biomaterial inks used for the formation of bioinks have been developed, allowing the manufacturing of in vitro models and implants tested preclinically with a certain degree of success. Furthermore, incredible advances in cell biology, namely, in pluripotent stem cells, have also contributed to the latest milestones where more relevant tissues or organ-like constructs with a certain degree of functionality can already be obtained. These incredible strides have been possible with a multitude of multidisciplinary teams around the world, working to make bioprinted tissues and organs more relevant and functional. Yet, there is still a long way to go until these biofabricated constructs will be able to reach the clinics. In this review, we summarize the main bioprinting activities linking them to tissue and organ development and physiology. Most bioprinting approaches focus on mimicking fully matured tissues. Future bioprinting strategies might pursue earlier developmental stages of tissues and organs. The continuous convergence of the experts in the fields of material sciences, cell biology, engineering, and many other disciplines will gradually allow us to overcome the barriers identified on the demanding path toward manufacturing and adoption of tissue and organ replacements.
Jip Zonderland, David B. Gomes, Yves Pallada, Ivan L. Moldero, Sandra Camarero-Espinosa, and Lorenzo Moroni
Wiley
Stanniocalcin‐1 (STC1) secreted by mesenchymal stromal cells (MSCs) has anti‐inflammatory functions, reduces apoptosis, and aids in angiogenesis, both in vitro and in vivo. However, little is known about the molecular mechanisms of its regulation. Here, we show that STC1 secretion is increased only under specific cell‐stress conditions. We find that this is due to a change in actin stress fibers and actin‐myosin tension. Abolishment of stress fibers by blebbistatin and knockdown of the focal adhesion protein zyxin leads to an increase in STC1 secretion. To also study this connection in 3D, where few focal adhesions and actin stress fibers are present, STC1 expression was analyzed in 3D alginate hydrogels and 3D electrospun scaffolds. Indeed, STC1 secretion was increased in these low cellular tension 3D environments. Together, our data show that STC1 does not directly respond to cell stress, but that it is regulated through mechanotransduction. This research takes a step forward in the fundamental understanding of STC1 regulation and can have implications for cell‐based regenerative medicine, where cell survival, anti‐inflammatory factors, and angiogenesis are critical.
Ravi Sinha, Fergal J. O'Brien, and Sandra Camarero-Espinosa
Frontiers Media SA
S. Camarero-Espinosa, C. Tomasina, A. Calore, and L. Moroni
Elsevier BV
Articular cartilage was thought to be one of the first tissues to be successfully engineered. Despite the avascular and non-innervated nature of the tissue, the cells within articular cartilage – chondrocytes – account for a complex phenotype that is difficult to be maintained in vitro. The use of bone marrow–derived stromal cells (BMSCs) has emerged as a potential solution to this issue. Differentiation of BMSCs toward stable and non-hypertrophic chondrogenic phenotypes has also proved to be challenging. Moreover, hyaline cartilage presents a set of mechanical properties – relatively high Young's modulus, elasticity, and resilience – that are difficult to reproduce. Here, we report on the use of additive manufactured biodegradable poly(ester)urethane (PEU) scaffolds of two different structures (500 μm pore size and 90° or 60° deposition angle) that can support the loads applied onto the knee while being highly resilient, with a permanent deformation lower than 1% after 10 compression-relaxation cycles. Moreover, these scaffolds appear to promote BMSC differentiation, as shown by the deposition of glycosaminoglycans and collagens (in particular collagen II). At gene level, BMSCs showed an upregulation of chondrogenic markers, such as collagen II and the Sox trio, to higher or similar levels than that of traditional pellet cultures, with a collagen II/collagen I relative expression of 2–3, depending on the structure of the scaffold. Moreover, scaffolds with different pore architectures influenced the differentiation process and the final BMSC phenotype. These data suggest that additive manufactured PEU scaffolds could be good candidates for cartilage tissue regeneration in combination with microfracture interventions.
Sandra Camarero-Espinosa, Andrea Calore, Arnold Wilbers, Jules Harings, and Lorenzo Moroni
Elsevier BV
Although a growing knowledge on the field of tissue engineering of articular cartilage exists, reconstruction or in-vitro growth of functional hyaline tissue still represents an unmet challenge. Despite the simplicity of the tissue in terms of cell population and absence of innervation and vascularization, the outstanding mechanical properties of articular cartilage, which are the result of the specificity of its extra cellular matrix (ECM), are difficult to mimic. Most importantly, controlling the differentiation state or phenotype of chondrocytes, which are responsible of the deposition of this specialized ECM, represents a milestone in the regeneration of native articular cartilage. In this study, we fabricated fused deposition modelled (FDM) scaffolds with different pore sizes and architectures from an elastic and biodegradable poly(ester)urethane (PEU) with mechanical properties that can be modulated by design, and that ranged the elasticity of articular cartilage. Cell culture in additive manufactured 3D scaffolds exceeded the chondrogenic potential of the gold-standard pellet culture. In-vitro cell culture studies demonstrated the intrinsic potential of elastic (PEU) to drive the re-differentiation of de-differentiated chondrocytes when cultured in-vitro, in differentiation or basal media, better than pellet cultures. The formation of neo-tissue was assessed as a high deposition of GAGs and fibrillar collagen II, and a high expression of typical chondrogenic markers. Moreover, the collagen II / collagen I ratio commonly used to evaluate the differentiation state of chondrocytes (ratio > 1 being chondrocytes and, ratio < 0 being de-differentiated chondrocytes) was higher than 5.
Tobias Kuhnt, Ramiro Marroquín García, Sandra Camarero-Espinosa, Aylvin Dias, A. Tessa ten Cate, Clemens A. van Blitterswijk, Lorenzo Moroni, and Matthew B. Baker
Royal Society of Chemistry (RSC)
Biodegradable polyester urethane acrylate oligomers were created to address the scarcity of resins for Digital Light Processing (DLP). Cell adhesion was heavily influenced by copolymer composition.
Sandra Camarero-Espinosa and Justin J. Cooper-White
Elsevier BV
Human articular cartilage is a complex multi-zonal tissue in which cells displaying three chondrocyte phenotypes (persistent, transient and hypertrophic) are supported and maintained by distinctly different (zonal) combinations of extracellular matrix (ECM) molecules. Articular cartilage has limited regenerative capacity, even though adjacent to the medullary cavity, an easily accessible reservoir of multipotent progenitor cells capable of eliciting repair, (human) mesenchymal stromal/stem cells (hMSCs). A greater understanding of the impacts of the extracellular cues provided in each zone of articular cartilage on hMSCs thus offers the potential to develop new scaffolds that can effect multi-zonal cartilage generation. In this work, we have systematically surveyed combinatorial mixtures of peptide sequences derived from ECM and cell adhesion molecules (CAMs) found to be present in cartilage and bone tissues, at a range of concentrations and ratios, to assess their ability to modulate hMSC fate. We show that directed differentiation of hMSCs towards persistent, transient and hypertrophic chondrogenic phenotypes is possible via the controlled presentation of specific peptide combinations on self-assembled polymeric coatings displaying hexagonally-packed nanodomains. These biomimetic substrates highlight that a high level of spatial and compositional control over biochemical cues is required by hMSCs in order to specify different cellular sub-phenotypes.
Brody Frost, Sandra Camarero-Espinosa, and E. Foster
MDPI AG
Disc degeneration affects 12% to 35% of a given population, based on genetics, age, gender, and other environmental factors, and usually occurs in the lumbar spine due to heavier loads and more strenuous motions. Degeneration of the extracellular matrix (ECM) within reduces mechanical integrity, shock absorption, and swelling capabilities of the intervertebral disc. When severe enough, the disc can bulge and eventually herniate, leading to pressure build up on the spinal cord. This can cause immense lower back pain in individuals, leading to total medical costs exceeding $100 billion. Current treatment options include both invasive and noninvasive methods, with spinal fusion surgery and total disc replacement (TDR) being the most common invasive procedures. Although these treatments cause pain relief for the majority of patients, multiple challenges arise for each. Therefore, newer tissue engineering methods are being researched to solve the ever-growing problem. This review spans the anatomy of the spine, with an emphasis on the functions and biological aspects of the intervertebral discs, as well as the problems, associated solutions, and future research in the field.