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Chondroitin sulfate (CS), a glycosaminoglycan, supports health through various physiological functions, including tissue protection, bone growth, and skin aging prevention. It also contributes to anticoagulant or anti-inflammatory processes, with its primary clinical use being osteoarthritis treatment. This study presents the results of the valorization of lipids and CS, both extracted from salmon co-products through enzymatic processes. The polar lipids, naturally rich in long-chain fatty acids (docosahexaenoic acid DHA C22:6 n-3 and eicosapentaenoic acid EPA C20:5 n-3), and the CS, primarily located in the nasal cartilage, were separated and concentrated before being characterized using various techniques to determine functional and lipid composition. These compounds were then used to formulate liposomes of 63 to 95 nm in size composed of 19.38% of DHA and 7.44% of EPA and encapsulating CS extract with a Δdi-4S/Δdi-6S ratio of 0.53 at 2 weight masses (10–30 kDa and >30 kDa) or CS standard all at two different concentrations. Liposomes were tested on human chondrocytes in inflamed conditions. Thus, compatibility tests, the expression of various inflammation markers at transcriptional and molecular levels, nitrites, and the amount of collagenase produced were analyzed. The results showed that CS, in synergy with the liposomes, played a positive role in combating chondrocyte inflammation even at a low concentration. [ABSTRACT FROM AUTHOR]

To meet the expectations of European consumers, who prioritize agro-environmental factors and local resources, the substitution of fats (palm, coconut, shea) and achieving a balanced fatty acid profile in spreadable fats are gaining more attention. The crystallization at 4 °C of a lipid blend composed of rapeseed oil, anhydrous dairy fats, and emulsifiers was studied using a multi-scale approach (DSC and X-ray diffraction techniques) to understand the emergence of polymorphic structures. Although the addition of PUFA from rapeseed oil reduces the atherogenicity and thrombogenicity indices in the blend, controlling the cooling kinetics influences the shapes (needles and spherulites) and sizes of the crystalline structures (small crystals form at a cooling rate of 1 °C min−1, while larger crystals form at higher rates of 5 and 10 °C min−1). The crystallization behavior revealed differences in polymorphic forms at 4 °C in the blend, with a transition to different forms occurring more rapidly compared to dairy fat (stop-and-return method). The study shows crystalline coexistence (α, β′, and β) in a 2L lamellar structure, with the β′ form being predominant. This structure is ideal for formulating a spreadable product, offering good spreadability (SFC < 32% at 10 °C), mouthfeel, and nutritional benefits compared to butter. [ABSTRACT FROM AUTHOR]

Given the importance of the extracellular medium during tissue formation, it was wise to develop an artificial structure that mimics the extracellular matrix while having improved physico-chemical properties. That is why the choice was focused on gelatin methacryloyl (GelMA), an inexpensive biocompatible hydrogel. Physicochemical and mechanical properties were improved by the incorporation of nanoparticles developed from two innovative fabrication processes: High shear fluid and low frequencies/high frequencies ultrasounds. Both rapeseed nanoliposomes and nanodroplets were successfully incorporated in the GelMA networks during the photo polymerization process. The impact on polymer microstructure was investigated by Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and enzymatic degradation investigations. Mechanical stability and viscoelastic tests were conducted to demonstrate the beneficial effect of the functionalization on GelMA hydrogels. Adding nanoparticles to GelMA improved the surface properties (porosity), tuned swelling, and degradability properties. In addition, we observed that nanoemulsion didn't change significantly the mechanical properties to shear and compression solicitations, whereas nanoliposome addition decreased Young's modulus under compression solicitations. Thus, these ways of functionalization allow controlling the design of the material by choosing the type of nanoparticle (nanoliposome or nanoemulsion) in function of the application. [ABSTRACT FROM AUTHOR]

Although the mechanical properties of ligament and tendon are well documented in research literature, very few unified mechanical formulations can describe a wide range of different loadings. The aim of this study was to propose a new model, which can describe tendon responses to various solicitations such as cycles of loading, unloading, and reloading or successive relaxations at different strain levels. In this work, experiments with cycles of loading and reloading at increasing strain level and sequences of relaxation were performed on white New Zealand rabbit Achilles tendons. We presented a local formulation of thermodynamic evolution outside equilibrium at a representative element volume scale to describe the tendon's macroscopic behavior based on the notion of relaxed stress. It was shown that the model corresponds quite well to the experimental data. This work concludes with the complexity of tendons' mechanical properties due to various microphysical mechanisms of deformation involved in loading such as the recruitment of collagen fibers, the rearrangement of the microstructure (i.e., collagens type I and III, proteoglycans, and water), and the evolution of relaxed stress linked to these mechanisms. [ABSTRACT FROM AUTHOR]

Bioreactors are defined as devices in which biological and/or biochemical processes develop under closely monitored and tightly controlled environmental and operating conditions (e.g. pH, temperature, mechanical conditions, nutrient supply and waste removal). In functional tissue engineering of musculoskeletal tissues, a bioreactor capable of controlling dynamic loading plays a determinant role. It has been shown that mechanical stretching promotes the expression of type I and III collagens, fibronectin, tenascin-C in cultured ligament fibroblasts (J.C.-H. Goh et al., Tissue Eng. 9 (2003), S31) and that human bone marrow mesenchymal stem cells (hBMMSC) – even in the absence of biochemical regulators – could be induced to differentiate into ligament-like fibroblast by the application of physiologically relevant cyclic strains (G. Vunjak-Novakovic et al., Ann. Rev. Biomed. Eng. 6 (2004), 131; H.A. Awad et al., Tissue Eng. 5 (1999), 267; R.G. Young et al., J. Orthop. Res. 16 (1998), 406). Different bioreactors are commercially available but they are too generic to be used for a given tissue, each tissue showing specific mechanical loading properties. In the case of ligament tissue engineering, the design of a bioreactor is still an open question. Our group proposes a bioreactor allowing cyclic traction–torsion on a scaffold seeded with stem cells. [ABSTRACT FROM AUTHOR]

Nanoliposomes are nano-sized vesicles that can be used as drug delivery carriers with the ability to encapsulate both hydrophobic and hydrophilic compounds. Moreover, their lipid compositions facilitate their internalization by cells. However, the interaction between nanoliposomes and the membrane barrier of the human body is not well-known. If cellular tests and animal testing offer a solution, their lack of physiological relevance and ethical concerns make them unsuitable to properly mimic human body complexity. Microfluidics, which allows the environment of the human body to be imitated in a controlled way, can fulfil this role. However, existing models are missing the presence of something that would mimic a basal membrane, often consisting of a simple cell layer on a polymer membrane. In this study, we investigated the diffusion of nanoliposomes in a microfluidic system and found the optimal parameters to maximize their diffusion. Then, we incorporated a custom made GelMA with a controlled degree of substitution and studied the passage of fluorescently labeled nanoliposomes through this barrier. Our results show that highly substituted GelMA was more porous than lower substitution GelMA. Overall, our work lays the foundation for the incorporation of a hydrogel mimicking a basal membrane on a drug delivery microfluidic platform.

There is a growing interest for complex in vitro environments that closely mimic the extracellular matrix and allow cells to grow in microenvironments that are closer to the one in vivo. Protein-based matrices and especially hydrogels can answer this need, thanks to their similarity with the cell microenvironment and their ease of customization. In this study, an experimental design was conducted to study the influence of synthesis parameters on the physical properties of gelatin methacryloyl (GelMA). Temperature, ratio of methacrylic anhydride over gelatin, rate of addition, and stirring speed of the reaction were studied using a Doehlert matrix. Their impact on the following parameters was analyzed: degree of substitution, mass swelling ratio, storage modulus (log(G')), and compression modulus. This study highlights that the most impactful parameter was the ratio of methacrylic anhydride over gelatin. Although, temperature affected the degree of substitution, and methacrylic anhydride addition flow rate impacted the gel's physical properties, namely, its storage modulus and compression modulus. Moreover, this experimental design proposed a theoretical model that described the variation of GelMA's physical characteristics as a function of synthesis conditions.

Nanoencapsulation of saffron extract (SE) components into the rapeseed lecithin nanoliposomes were performed by sonication of their aqueous dispersions as a green process. Dynamic light scattering (DLS) results exhibited that empty and SE loaded nanoliposomes (SENL) had average sizes in range of 118-138 nm, negative zeta potentials (-32.0 to -46.8 mV) and polydispersity index (PDI) less than 0.3 during storage for 28 days at 4 °C. Encapsulation efficiency of crocin was approximately 30%. The 70% of crocin released from SENLs within 5 h in PBS solution. Pullulan-based films were fabricated by incorporation of empty and SE loaded nanoliposomes into pullulan solution through casting method. The mechanical resistance and thermal stability of the films reduced by addition of nanoliposomes. FTIR and thermal characterizations indicated that SE was successfully encapsulated in the nanoliposomes and film matrix with high thermal stability. Incorporation of nanoliposomes enhanced the oxygen barrier properties of the films, while it didn't significantly affect the water vapor permeability (WVP) of the films. The obtained edible films or coatings can provide additional benefits due to unique flavor and color of saffron. In addition, the utilization of SE, can provide benefits for health-allegation from SE antioxidant capacity., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Curcumin (diferuloylmethane) is a natural bioactive compound with many health-promoting benefits. However, its poor water solubility and bioavailability has limited curcumin&rsquo;s biomedical application. In the present study, we encapsulated curcumin into liposomes, formed from natural sources (salmon lecithin), and characterized its encapsulation efficiency and release profile. The proposed natural carriers increased the solubility and the bioavailability of curcumin. In addition, various physico-chemical properties of the developed soft nanocarriers with and without curcumin were studied. Nanoliposome-encapsulated curcumin increased the viability and network formation in the culture of primary cortical neurons and decreased the rate of apoptosis.

Novel bioactive films were developed from the incorporation of Lactococcus lactis into polysaccharide films. Two different biopolymers were tested: cellulose derivative (hydroxylpropylmethylcellulose (HPMC)) and corn starch. Lactic acid bacteria (LAB) free or previously encapsulated in alginate-pectin composite hydrogel microbeads were added directly to the film forming solution and films were obtained by casting. In order to study the impact of the incorporation of the protective culture into the biopolymer matrix, the water vapour permeability, oxygen permeability, optical and mechanical properties of the dry films were evaluated. Furthermore, the antimicrobial effect of bioactive films against Listeria monocytogenes was studied in synthetic medium. Results showed that the addition of LAB or alginate-pectin microbeads modified slightly films optical properties. In comparison with HPMC films, starch matrix proves to be more sensitive to the addition of bacterial cells or beads. Indeed, mechanical resistance of corn starch films was lower but barrier properties were improved, certainly related to the possible establishment of interactions between alginate-pectin beads and starch. HPMC and starch films containing encapsulated bioactive culture showed a complete inhibition of listerial growth during the first five days of storage at 5 °C and a reduction of 5 logs after 12 days., Competing Interests: The authors declare no conflicts of interest.

Multi-scale characterization of structures and mechanical behavior of biological tissues are of huge importance in order to evaluate the quality of a biological tissue and/or to provide bio-inspired scaffold for functional tissue engineering. Indeed, the more information on main biological tissue structures we get, the more relevant we will be to design new functional prostheses for regenerative medicine or to accurately evaluate tissues. From this perspective, we have investigated the structures and their mechanical properties from nanoscopic to macroscopic scale of fresh ex-vivo white New-Zealand rabbit Achilles tendon using second harmonic generation (SHG) microscopy, atomic force microscopy (AFM) and tensile tests to provide a "simple" model whose parameters are relevant of its micro or nano structure. Thus, collagen fiber's crimping was identified then measured from SHG images as a plane sine wave with 28.4 ± 5.8 μm of amplitude and 141 ± 41 μm of wavelength. Young's moduli of fibrils (3.0 GPa) and amorphous phases (223 MPa) were obtained using TH-AFM. From these investigations, a non-linear Zener model linking a statistical Weibull's distribution of taut fibers under traction to crimp fibers were developed. This model showed that for small strain (<0.1), the amorphous inter-fibrils phase in collagen fibers is more solicited than collagen fibrils themselves. The results open the way to modeled macroscopic mechanical behavior of aligned-crimped collagen soft tissues using multi-scale tendon observations under static or dynamic solicitations., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

This study aims to investigate the in vitro degradation of a poly(L-lactic-co-glycolic acid)-poly(L-lactic-co-ϵ-caprolactone) (PLGA-PLCL) composite scaffold's mechanical properties under static culture condition and 2 h period per day of traction-torsion cyclic culture conditions of simultaneous 10% uniaxial strain and 90° of torsion cycles at 0.33 Hz. Scaffolds were cultured in static conditions, during 28 days, with or without cell seeded or under dynamic conditions during 14 days in a bioreactor. Scaffolds' biocompatibility and proliferation were investigated with Alamar Blue tests and cell nuclei staining. Scaffolds' mechanical properties were tested during degradation by uniaxial traction test. The PLGA-PLCL composite scaffold showed a good cytocompatibility and a high degree of colonization in static conditions. Mechanical tests showed a competition between two process of degradation which have been associated to hydrolytic and enzymatic degradation for the reinforce yarn in poly(L-lactic-co-glycolic acid) (PLGA). The enzymatic degradation led to a decrease effect on mechanical properties of cell-seeded scaffolds during the 21st days, but the hydrolytic degradation was preponderant at day 28. In conclusion, the structure of this scaffold is adapted to culture in terms of biocompatibility and cell orientation (microfiber) but must be improved by delaying the degradation of it reinforce structure in PLGA.

Health benefits of unsaturated fatty acids have been demonstrated over the last decades. Nanotechnology provided new process to produce particles such as liposomes and nanoliposomes made of pure phospholipids. These techniques are already used in pharmaceutics to augment the bioavailability and the bioefficiency of drugs. The aim of this paper is to characterize and evaluate the potential of nanoliposomes made of three lecithins (soya, rapeseed and salmon) on cell culture in order to use them in the future as drug delivery systems for tissue engineering. We began to measure, with zetasizer, the radius size of liposomes particles which are 125.5, 136.7 and 130.3 nm respectively for rapeseed, soya and salmon lecithin. Simultaneously, solutions observed by TEM demonstrated the particles were made much of liposomes than droplet (emulsion). Finally, we found that the solutions of lecithins were enough stable over 5 days at 37 °C to be used in culture medium. We investigated the effect of soya, rapeseed and salmon lecithin liposome from 2mg/mL to 5.2 μg/mL on metabolic activity and cell proliferation on rat bone marrow stem cells (rBMSC) during 14 days. The results showed that the three lecithins (soya, rapeseed and salmon) improve cell proliferation at different concentration., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Bone marrow mesenchymal stem cells (BMSC) have the potential to differentiate into a variety of cell types like osteoblasts, chondroblasts, adipocytes, etc. It is well known that mechanical forces regulate the biological function of cells. The aim of this study was to investigate the effect of uniaxial stretching on the orientation and biological functions of BMSC. Rat BMSCs were harvested from femoral and tibial bone marrow by density gradient centrifugation. Cells from passages 1-6 were characterized by flow cytometry using monoclonal antibodies. The recovered cells were stably positive for the markers CD90 and CD44 and negative for CD34 and CD45. A cyclic 10% uniaxial stretching at 1Hz was applied on rat BMSC for different time-courses. The length, width, and orientation of the cells were subsequently determined. Expression of collagen types I and III and tenascin-C mRNAs was measured by real-time RT-PCR, and the synthesis of these receptors was determined by radioimmunoassay. Results showed that uniaxial stretching lengthened and rearranged the cells. Compared with control groups, expression of collagen types I and III mRNAs was up-regulated after 12-h of stretching, while significant increase in synthesis of the two collagen protein types was not observed until after 24-h stretching. The expression of tenascin-C mRNA was significantly increased after a 24-h stretching. These data suggest that cyclic stretching promotes the synthesis of collagen types I and III and tenascin-C by the rat BMSC.