Jorge L. Escobar Ivirico

Chitosan microparticles as injectable scaffolds for tissue engineering

The use of chitosan microparticles as injectable carriers for cell transplantation represents a promising alternative to avoid the drawbacks of the implantation of other forms of three‐dimensional (3D) scaffolds seeded with cells. In this study, a 3D construct is obtained in vitro by combining chitosan microparticles crosslinked with genipin and goat bone marrow stromal cells (GBMCs). Cell viability and the morphology of GBMCs were evaluated after culture for 7 and 14 days. Our results show the feasibility of chitosan microparticles as potential injectable scaffolds for tissue engineering and regenerative medicine. Copyright

Bioactive organic inorganic poly(CLMA-co-HEA)/silica nanocomposites

A series of novel poly(CLMA-co-HEA)/silica nanocomposites is synthesized from caprolactone 2-(methacryloyloxy)ethyl ester (CLMA) and 2-hydroxyethyl acrylate (HEA) as organic comonomers and the simultaneous sol-gel polymerization of tetraethyloxysilane (TEOS) as silica precursor, in different mass ratios up to a 30 wt% of silica. The nanocomposites are characterized as to their mechanical and thermal properties, water sorption, bioactivity and biocompatibility, reflecting the effect on the organic matrix provided by the silica network formation. The nanocomposites nucleate the growth of hydroxyapatite (HAp) on their surfaces when immersed in the simulated body fluid of the composition used in this work. Proliferation of the MC3T3 osteoblast-like cells on the materials was assessed with the MTS assay showing their biocompatibility. Immunocytochemistry reveals osteocalcin and type I collagen production, indicating that osteoblast differentiation was promoted by the materials, and calcium deposition was confirmed by von Kossa staining. The results indicate that these poly(CLMA-co-HEA)/silica nanocomposites could be a promising biomaterial for bone tissue engineering.

Cytotoxic effect of 4-hydroxytamoxifen conjugate material on human Schwann cells: Synthesis and characterization

In this study, the toxicity of 4-hydroxytamoxifen (4-OHT) on human Schwann cells (HSCs) was evaluated. Substantial alterations in the cell morphology and viability were observed at 4-OHT concentrations higher than 3 µg/mL. Therefore, we designed and synthesized a drug–polymer conjugate, based on N-(2-hydroxypropyl)methacrylamide (HPMA) and ethyl acrylate (EA) for delivering 4-OHT to the target tissue without the detrimental consequences of the systemic therapy currently used. The macromer carrier of 4-OHT (MATX), with a functionalization degree of 80%, was synthesized in two steps and verified by 1H-NMR and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectroscopy. MATX was conjugated to the poly(HPMA-co-EA) copolymer network via radical polymerization. The influence of MATX on the physical, chemical, and mechanical properties of poly(HPMA-co-EA-co-MATX) with a ratio of 69/29/2 wt% was compared to those of poly(HPMA-co-EA) networks with a similar feed mixture. The in vitro release of 4-OHT within 1 month was 6 wt% of the total amount of drug linked to the copolymer backbone.

One-Dimensional Migration of Olfactory Ensheathing Cells on Synthetic Materials: Experimental and Numerical Characterization

Olfactory ensheathing cells (OECs) are of great interest for regenerative purposes since they are believed to aid axonal growth. With the view set on the strategies to achieve reconnection between neuronal structures, it is of great importance to characterize the behaviour of these cells on long thread-like structures that may efficiently guide cell spread in a targeted way. Here, rat OECs were studied on polycaprolactone (PCL) long monofilaments, on long bars and on discs. PCL turns out to be an excellent substrate for OECs. The cells cover long distances along the monofilaments and colonize completely these structures. With the help of a one-dimensional (1D) analytical model, a migration coefficient, a net proliferation rate constant and the fraction of all cells which undergo migration were obtained. The separate effect of the three phenomena summarized by these parameters on the colonization patterns of the 1D path was qualitatively discussed. Other features of interest were also determined, such as the speed of the advance front of colonization and the order of the kinetics of net cell proliferation. Characterizing migration by means of these quantities may be useful for comparing and predicting features of the colonization process (such as times, patterns, advance fronts and proportion of motile cells) of different cell–substrate combinations.

Polymer chains incorporating caprolactone and arginine–glycine–aspartic acid functionalities: Synthesis, characterization and biological response in vitro of the Schwann cell

This study describes a strategy for the covalent immobilization of active adhesion peptide moieties onto polymers through the intermediacy of itaconic acid. The arginine–glycine–aspartic acid peptide was grafted to a novel poly(caprolactone 2-(methacryloyloxy) ethyl ester)-co-itaconic acid bulk biomaterial, in order to improve the cell adhesion of the polymer. First, the arginine–glycine–aspartic acid sequence was grafted onto itaconic acid via an amidation reaction using N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide as activation complex. The itaconic acid–arginine–glycine–aspartic acid macromer was characterized by Fourier transform infrared spectroscopy and 1H-NMR, yielding a functionalization degree of 85%. In a second step, poly(caprolactone 2-(methacryloyloxy) ethyl ester-co-itaconic acid–arginine–glycine–aspartic acid) (with a feed mixture of 90 wt% of caprolactone 2-(methacryloyloxy) ethyl ester and 10 wt% of itaconic acid–arginine–glycine–aspartic acid macromer) and a series of copolymers of caprolactone 2-(methacryloyloxy) ethyl ester and itaconic acid with different compositions (weight fractions of itaconic acid up to 20 wt%) were synthesized by radical copolymerization. The microstructure and network architecture of the new polymer systems were investigated. Mechanical moduli of poly(caprolactone 2-(methacryloyloxy) ethyl ester-co-itaconic acid), evaluated by dynamic–mechanical analysis, increase with the itaconic acid content. In poly(caprolactone 2-(methacryloyloxy) ethyl ester-co-itaconic acid–arginine–glycine–aspartic acid), the glass transition temperature and the mechanical moduli of the system are smaller than in the nonfunctionalized poly(caprolactone 2-(methacryloyloxy) ethyl ester-co-itaconic acid) copolymers, and the polymer is less hydrophilic. The results indicate that arginine–glycine–aspartic acid grafting of poly(caprolactone 2-(methacryloyloxy) ethyl ester-co-itaconic acid) copolymer networks can be useful for tissue engineering applications, because regenerative processes in the nervous system can be promoted and accelerated, thus, opening a possibility to generate materials with a high potential for clinical applicability.