Cold-adaptation of a methacrylamide gelatin towards the expansion of the biomaterial toolbox for specialized functionalities in tissue engineering.

Tissue regeneration is witnessing a significant surge in advanced medicine. It requires the interaction of scaffolds with different cell types for efficient tissue formation post-implantation. The presence of tissue subtypes in more complex organs demands the co-existence of different biomaterials showing different hydrolysis rate for specialized cell-dependent remodeling. To expand the available toolbox of biomaterials with sufficient mechanical strength and variable rate of enzymatic degradation, a cold-adapted methacrylamide gelatin was developed from salmon skin. Compared with mammalian methacrylamide gelatin (GelMA), hydrogels derived from salmon GelMA displayed similar mechanical properties than the former. Nevertheless, salmon gelatin and salmon GelMA-derived hydrogels presented characteristics common of cold-adaptation, such as reduced activation energy for collagenase, increased enzymatic hydrolysis turnover of hydrogels, increased interconnected polypeptides molecular mobility and lower physical gelation capability. These properties resulted in increased cell-remodeling rate in vitro and in vivo, proving the potential and biological tolerance of this mechanically adequate cold-adapted biomaterial as alternative scaffold subtypes with improved cell invasion and tissue fusion capacity.

Anti-inflammatory and antimicrobial activity of bioactive hydroxyapatite/silver nanocomposites.

Biomaterials are often used in orthopedic surgery like cavity fillings. However, related complications often require long-term systemic antibiotics, device removal, and extended rehabilitation. Hydroxyapatite/silver (HA/Ag) composites have been proposed as implantation biomaterials owing to the osteogenic properties of hydroxyapatite and to the antimicrobial efficiency of silver. Nevertheless, higher silver concentrations induce cytotoxic effects. The aim of this study was to synthesize and characterize HA/Ag nanocomposites that will allow us to use lower concentrations of silver nanoparticles with better antimicrobial efficiency and anti-inflammatory properties. The characterization of HA/Ag was performed by scanning electron microscopy, energy dispersive spectroscopy, X-ray diffraction, Fourier-transform infrared spectra, X-ray photoelectron spectroscopy, and laser diffraction. Bioactivity was evaluated under a simulated body fluid. The viability of osteoblast like-cells (MG-63) was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and the antimicrobial activity was evaluated by the standard McFarland method. The detection of nitric oxide was measured by a colorimetric assay and the inflammatory cytokines by flow cytometry. We obtained particulate composites of calcium phosphates identified as hydroxyapatite and silver nanoparticles. The bioactivity of the HA/Ag nanocomposites on SFB was confirmed by apatite formations. The viability of MG-63 cells was not affected. We also found antimicrobial activity against Escherichia coli, Staphylococcus aureus, and Candida albicans owing to the presence of silver nanoparticles at non-cytotoxic concentrations. HA/Ag reduced the release of nitric oxide and decreased the secretion of IL-1 and TNF-α in cells stimulated with Lipopolysaccharide (LPS). In conclusion, the inflammatory and antimicrobial capacity of the HA/Ag nanocomposites, as well as its bioactivity and low cytotoxicity make it a candidate as an implantation biomaterial for bone tissues engineering and clinical practices in orthopedic, oral and maxillofacial surgery.

Chitosan/poly-octanoic acid 2-thiophen-3-yl-ethyl ester blends as a scaffold to maintain myoblasts regeneration potential in vitro.

Satellite cells are a small cell population that function as muscle-specific adult stem cells. When muscle damage occurs, these cells are able to activate, proliferate, and ultimately fuse with each other in order to form new myofibers or fuse with existing ones. For tissue engineering applications, obtaining a sufficient number of myoblasts prior transplantation that maintains their regenerative capacity is critical. This can be obtained by in vitro expansion of autologous satellite cells. However, once plated, the self-renewal and regenerative capacity of myoblasts is rapidly lost, obtaining low yields per biopsy. For this purpose, we evaluated in vitro culture of the murine myoblast cell line C2C12 and mouse primary myoblasts with chitosan and chitosan/poly-octanoic acid 2-thiophen-3-yl-ethyl ester blends (poly(OTE)). The films of chitosan/poly(OTE) blends were heterogeneous and slightly rougher than chitosan and poly(OTE) films. Poly(OTE) presence improved myoblast adhesion in both cell types and prevented complete differentiation, but maintaining their differentiation potential in vitro. We identified that the polymer blend chitosan/poly(OTE) could be a suitable substrate to culture satellite cells/myoblasts in vitro preventing differentiation prior transplantation. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 118-130, 2017.

Improving myoblast differentiation on electrospun poly(ε-caprolactone) scaffolds.

Polymer scaffolds are used as an alternative to support tissue regeneration when it does not occur on its own. Cell response on polymer scaffolds is determined by factors such as polymer composition, topology, and the presence of other molecules. We evaluated the cellular response of murine skeletal muscle myoblasts on aligned or unaligned fibers obtained by electrospinning poly(ε-caprolactone) (PCL), and blends with poly(lactic-co-glycolic acid) (PLGA) or decorin, a proteoglycan known to regulate myogenesis. The results showed that aligned PCL fibers with higher content of PLGA promote cell growth and improve the quality of differentiation with PLGA scaffolds having the highest confluence at over 68% of coverage per field of view for myoblasts and more than 7% of coverage for myotubes. At the same time, the addition of decorin greatly improves the quantity and quality of differentiated cells in terms of cell fusion, myotube length and thickness, being 71, 10, and 51% greater than without the protein, respectively. Interestingly, our results suggest that at certain concentrations, the effect of decorin on myoblast differentiation exceeds the topological effect of fiber alignment. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2241-2251, 2017.

Evaluation of polymer-based granular formulations of Bacillus thuringiensis israelensis against larval Aedes aegypti in the laboratory.

A strategy to increase residual activity of Bacillus thuringiensis serovar. israelensis (Bti) extract through slow-release formulations and protection from solar radiation was studied. The median lethal concentration (LC50) and 90% lethal concentration (LC90) levels of laboratory-reared early 4th-stage larval Aedes aegypti after exposure to Bti extract were determined. Formulations with 4 polymers and 1 solar protectant were prepared, and their shelf lives were evaluated for year-long storage at 20-35 degrees C and 50-80% relative humidity. Also, the effect of ultraviolet light on unformulated (extract) and formulated Bti larvicidal activity persistence was determined. Laboratory bioassays were conducted with larval Ae. aegypti introduced into treated and control containers at 1, 2, 7, 14, 21, 28, 35, and 42 days after treatment, and larval mortalities were checked 24 h after introduction. Probit analysis of Bti extract showed LC50 and LC90 values of 0.016 and 0.051 mg/liter, respectively. The polymer-based Bti formulations showed no significant loss of insecticidal activity after 8 months of storage. Ultraviolet irradiation reduced activity of unformulated Bti extract after different exposure times, up to 40-46%, whereas Bti formulated with gelatin or acacia gum showed lower variation in larvicidal activity than formulations with sodium alginate and paraffin for protecting the activity of Bti toxin. Residual activity against 4th-stage Ae. aegypti in the laboratory for the formulation containing acacia gum at 10% (w/w) was 80% mortality at 14 days after treatment, whereas the Bti formulation containing gelatin (10%, w/w) caused 65% mortality. In addition, Bti formulations made with paraffin at 5% (w/w) sustained up to 60% mortality for 21 days. Unformulated Bti showed only 2.6% mortality, and a commercial preparation maintained 37% mortality, both at 14 days after treatment.