Porous polylactic acid-silica hybrids: preparation, characterization, and study of mesenchymal stem cell osteogenic differentiation

A novel approach to reinforce polymer porous membranes is presented. In the prepared hybrid materials, the inorganic phase of silica is synthesized in-situ and inside the pores of aminolyzed polylactic acid (PLA) membranes by sol-gel reactions using tetraethylorthosilicate (TEOS) and glycidoxypropyltrimethoxysilane (GPTMS) as precursors. The hybrid materials present a porous structure with a silica layer covering the walls of the pores while GPTMS serves also as coupling agent between the organic and inorganic phase. The adjustment of silica precursors ratio allows the modulation of the thermomechanical properties. Culture of mesenchymal stem cells on these supports in osteogenic medium shows the expression of characteristic osteoblastic markers and the mineralization of the extracellular matrix.The research project is implemented within the framework of the Action "Supporting Postdoctoral Researchers" of the Operational Program "Education and Lifelong Learning'' (Action's Beneficiary: General Secretariat for Research and Technology), and is co-financed by the European Social Fund (ESF) and the Greek State, Grant No.: NARGEL-PE5(2551). J.R.R. acknowledges funding of his PhD by the Generalitat Valenciana through VALi+d grant (ACIF/2010/238). J.F.M. thanks the Portuguese Foundation for Science and Technology (FCT) for financial support through the PTDC/FIS/115048/2009 project. J.L.G.R. acknowledges the support of the Ministerio de Economia y Competitividad, MINECO, through theMAT2013-46467-C4-1-R project. CIBER-BBN is an initiative funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with the assistance from the European Regional Development Fund

Macroporous thin membranes for cell transplant in regenerative medicine

The aim of this paper is to present a method to produce macroporous thin membranes made of poly (ethyl acrylate-co-hydroxyethyl acrylate) copolymer network with varying cross-linking density for cell transplantation and prosthesis fabrication. The manufacture process is based on template techniques and anisotropic pore collapse. Pore collapse was produced by swelling the membrane in acetone and subsequently drying and changing the solvent by water to produce 100 microns thick porous membranes. These very thin membranes are porous enough to hold cells to be transplanted to the organism or to be colonized by ingrowth from neighboring tissues in the organism, and they present sufficient tearing stress to be sutured with surgical thread. The obtained pore morphology was observed by Scanning Electron Microscope, and confocal laser microscopy. Mechanical properties were characterized by stress-strain experiments in tension and tearing strength measurements. Morphology and mechanical properties were related to the different initial thickness of the scaffold and the cross-linking density of the polymer network. Seeding efficiency and proliferation of mesenchymal stem cells inside the pore structure were determined at 2 hours, 1, 7, 14 and 21 days from seeding.This work was funded by the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2013-46467-C4-1-R (including the FEDER financial support). J.R.R. acknowledges funding of his PhD by the Generalitat Valenciana through VALi+d grant (ACIF/2010/238). CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. The authors acknowledge too the advice of Dr. Daniel Kelly, Dr. Conor Buckley and Dr. Yurong Liu about the isolation and expansion of porcine MSCs. The authors acknowledge the assistance and advice of Electron Microscopy Service of the UPV.Antolinos Turpín, CM.; Morales Román, RM.; Ródenas Rochina, J.; Gómez Ribelles, JL.; Gómez-Tejedor, JA. (2015). Macroporous thin membranes for cell transplant in regenerative medicine. Biomaterials. 67:254-263. https://doi.org/10.1016/j.biomaterials.2015.07.032S2542636

Influence of oxygen levels on chondrogenesis of porcine mesenchymal stem cells cultured in polycaprolactone scaffolds

[EN] Chondrogenesis of mesenchymal stem cells (MSCs) is known to be regulated by a number of environmental factors, including local oxygen levels. The hypothesis of this study is that the response of MSCs to hypoxia is dependent on the physical and chemical characteristics of the substrate used. The objective of this study was to explore how different modifications to polycaprolactone (PCL) scaffolds influenced the response of MSCs to hypoxia. PCL, PCL-hyaluronic acid (HA), and PCL-Bioglass (R) (BG) scaffolds were seeded with MSCs derived from bone marrow and cultured for 35 days under normoxic or low oxygen conditions, and the resulting biochemical properties of the MSC laden construct were assessed. Low oxygen tension has a positive effect over cell proliferation and macromolecules biosynthesis. Furthermore, hypoxia enhanced the distribution of collagen and glycosaminoglycans (GAGs) deposition through the scaffold. On the other hand, MSCs displayed certain material dependent responses to hypoxia. Low oxygen tension had a positive effect on cell proliferation in BG and HA scaffolds, but only a positive effect on GAGs synthesis in PCL and HA scaffolds. In conclusion, hypoxia increased cell viability and expression of chondrogenic markers but the cell response was modulated by the type of scaffold used.Contract grant sponsors: VI National R&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions, Instituto de Salud Carlos III, and European Regional Development FundRódenas Rochina, J.; Kelly, DJ.; Gómez Ribelles, JL.; Lebourg, MM. (2017). Influence of oxygen levels on chondrogenesis of porcine mesenchymal stem cells cultured in polycaprolactone scaffolds. Journal of Biomedical Materials Research Part A. 105(6):1684-1691. https://doi.org/10.1002/jbm.a.36043S16841691105

An experimental fatigue study of a porous scaffold for the regeneration of articular cartilage

The aim of this experimental study is to predict the long-term mechanical behavior of a porous scaffold implanted in a cartilage defect for tissue engineering purpose. Fatigue studies were performed by up to 100,000 unconfined compression cycles in a polycaprolactone (PCL) scaffold with highly interconnected pores architecture. The scaffold compliance, stress strain response and hysteresis energy have been measured after different number of fatigue cycles, while the morphology has been observed by scanning electron microscopy at the same fatigue times. To simulate the growing tissue in the scaffold/tissue construct, the scaffold was filled with an aqueous solution of polyvinyl alcohol (PVA) and subjected to repeating cycles of freezing and thawing that increase the hydrogel stiffness. Fatigue studies show that the mechanical loading provokes failure of the dry scaffold at a smaller number of deformation cycles than when it is immersed in water, and also that 100,000 compressive dynamic cycles do not affect the scaffold/gel construct. This shows the stability of the scaffold implanted in a chondral defect and gives a realistic simulation of the mechanical performance from implantation of the empty scaffold to regeneration of the new tissue inside the scaffold's pores.This work was funded by the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2013-46467-C4-1-R (including the FEDER financial support). CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. The authors acknowledge the assistance and advice of Electron Microscopy Service of the UPVVikingsson, LKA.; Gómez-Tejedor, JA.; Gallego Ferrer, G.; Gómez Ribelles, JL. (2015). An experimental fatigue study of a porous scaffold for the regeneration of articular cartilage. Journal of Biomechanics. 48(7):1310-1317. https://doi.org/10.1016/j.jbiomech.2015.02.013S1310131748

Prediction of the in vivo mechanical behavior of biointegrable acrylic macroporous scaffolds

[EN] This study examines a biocompatible scaffold series of random copolymer networks P(EA-HEA) made of Ethyl Acrylate, EA, and 2-Hydroxyl Ethyl Acrylate, HEA. The P(EA-HEA) scaffolds have been synthesized with varying crosslinking density and filled with a Poly(Vinyl Alcohol), PVA, to mimic the growing cartilaginous tissue during tissue repair. In cartilage regeneration the scaffold needs to have sufficient mechanical properties to sustain the compression in the joint and, at the same time, transmit mechanical signals to the cells for chondrogenic differentiation. Mechanical tests show that the elastic modulus increases with increasing crosslinking density of P(EA-HEA) scaffolds. The water plays an important role in the mechanical behavior of the scaffold, but highly depends on the crosslinking density of the proper polymer. Furthermore, when the scaffold with hydrogel is tested it can be seen that the modulus increases with increasing hydrogel density. Even so, the mechanical properties are inferior than those of the scaffolds with water filling the pores. The hydrogel inside the pores of the scaffolds facilitates the expulsion of water during compression and lowers the mechanical modulus of the scaffold. The P(EA-HEA) with PVA shows to be a good artificial cartilage model with mechanical properties close to native articular cartilage.This work was funded by the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2013-46467-C4-1-R (including the FEDER financial support). CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program. CIBER actions are financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. The authors acknowledge the assistance and advice of Electron Microscopy Service of the UPV.Vikingsson, L.; Antolinos Turpín, CM.; Gómez-Tejedor, JA.; Gallego Ferrer, G.; Gómez Ribelles, JL. (2016). Prediction of the in vivo mechanical behavior of biointegrable acrylic macroporous scaffolds. Materials Science and Engineering: C. 61:651-658. https://doi.org/10.1016/j.msec.2015.12.068S6516586

Biomedical applications of natural-based polymers combined with bioactive glass nanoparticles

In recent years, the combination of natural polymers with nanoparticles has permitted the development of sophisticated and efficient bioinspired constructs. In this regard, the incorporation of bioactive glass nanoparticles (BGNPs) confers a bioactive nature to these constructs, which can then induce the formation of a bone-like apatite layer upon immersion in a physiological environment. Moreover, the incorporation of bioactive glass nanoparticles has been found to be beneficial; the constructs proved to be biocompatible, promote cell adhesion and spreading, and regulate osteogenic commitment. This review provides a summary and discussion of the composition, design, and applications of bioinspired nanocomposite constructs based on BGNPs. Examples of nanocomposite systems will be highlighted with relevance to biomedical applications. It is expected that understanding the principles and the stateof-the-art of natural nanocomposites may lead to breakthroughs in many research areas, including tissue engineering and orthopaedic devices. The challenges regarding the future translation of these nanostructured composites into clinical use are also summarized.A´lvaro J. Leite acknowledges the Portuguese Foundation for Science and Technology (FCT) for his doctoral grant (SFRH/BD/73174/2010).info:eu-repo/semantics/publishedVersio

Designing microenvironments for optimal outcomes in tissue engineering and regenerative medicine: From biopolymers to culturing conditions

Bone marrow mesenchymal stem cells have been extensively used for tissue engineering and regenerative medicine applications due to their ease of isolation and expansion and their ability to differentiate towards various lineages of mesodermal origin. Despite these properties, their clinical potential is often hampered by the simplicity of the in vitro environment and its inability to resemble the complex in vivo niche. Herein, different microenvironmental cues (e.g. surface topography, substrate stiffness, mechanical stimulation, oxygen tension and co-culture systems) that have been utilised to enhance the therapeutic efficacy of bone marrow mesenchymal stem cells are discussed.The authors would like to acknowledge the following entities for financial support: H2020, Marie Skłodowska-Curie Actions, Innovative Training Networks 2015 Tendon Therapy Train project (Grant No. 676338); Science Foundation Ireland (SFI) / European Regional Development Fund (Grant Number 13/RC/2073); and SFI Career Development Award (Grant Number 15/CDA/3629)

Hybrid polycaprolactone/silica porous membranes produced by sol-gel

Hybrid porous membranes of polycaprolactone (PCL) and silica were obtained by a two stage procedure. First a porous PCL membrane was produced by freeze extraction technique and then a sol-gel reaction was used to form a uniform silica coating on the pore walls. Sol-gel reaction was performed at low pH with tetraethylorthosilicate (TEOS) as the precursor of silica. In this work we analyze the influence of several clue parameters on the silica film formation and in turn on the properties of the hybrid material. Water: TEOS molar ratio and drying protocol were considered in this study.Pandis, C.; Trujillo, S.; Roganowicz, M.; Gómez Ribelles, JL. (2014). Hybrid polycaprolactone/silica porous membranes produced by sol-gel. Macromolecular Symposia. 341(1):34-44. doi:10.1002/masy.201300155S3444341