Influence of PCL and PHBV on PLLA Thermal and Mechanical Properties in Binary and Ternary Polymer Blends.

PLLA, PCL and PHBV are aliphatic polyesters which have been researched and used in a wide range of medical devices, and all three have advantages and disadvantages for specific applications. Blending of these materials is an attractive way to make a material which overcomes the limitations of the individual polymers. Both PCL and PHBV have been evaluated in polymer blends with PLLA in order to provide enhanced properties for specific applications. This paper explores the use of PCL and PHBV together with PLLA in ternary blends with assessment of the thermal, mechanical and processing properties of the resultant polymer blends, with the aim of producing new biomaterials for orthopaedic applications. DSC characterisation is used to demonstrate that the materials can be effectively blended. Blending PCL and PHBV in concentrations of 5-10% with PLLA produces materials with average modulus improved by up to 25%, average strength improved by up to 50% and average elongation at break improved by 4000%, depending on the concentrations of each polymer used. PHBV impacts most on the modulus and strength of the blends, whilst PCL has a greater impact on creep behaviour and viscosity. Blending PCL and PHBV with PLLA offers an effective approach to the development of new polyester-based biomaterials with combinations of mechanical properties which cannot be provided by any of the materials individually.

Strategies for Enhancing Polyester-Based Materials for Bone Fixation Applications.

Polyester-based materials are established options, regarding the manufacturing of bone fixation devices and devices in routine clinical use. This paper reviews the approaches researchers have taken to develop these materials to improve their mechanical and biological performances. Polymer blending, copolymerisation, and the use of particulates and fibre bioceramic materials to make composite materials and surface modifications have all been studied. Polymer blending, copolymerisation, and particulate composite approaches have been adopted commercially, with the primary focus on influencing the in vivo degradation rate. There are emerging opportunities in novel polymer blends and nanoscale particulate systems, to tune bulk properties, and, in terms of surface functionalisation, to optimise the initial interaction of devices with the implanted environment, offering the potential to improve the clinical performances of fracture fixation devices.

A Comparison of Osteoblast and Osteoclast In Vitro Co-Culture Models and Their Translation for Preclinical Drug Testing Applications.

As the population of western societies on average ages, the number of people affected by bone remodeling-associated diseases such as osteoporosis continues to increase. The development of new therapeutics is hampered by the high failure rates of drug candidates during clinical testing, which is in part due to the poor predictive character of animal models during preclinical drug testing. Co-culture models of osteoblasts and osteoclasts offer an alternative to animal testing and are considered to have the potential to improve drug development processes in the future. However, a robust, scalable, and reproducible 3D model combining osteoblasts and osteoclasts for preclinical drug testing purposes has not been developed to date. Here we review various types of osteoblast-osteoclast co-culture models and outline the remaining obstacles that must be overcome for their successful translation.

Multilayer nanoscale functionalization to treat disorders and enhance regeneration of bone tissue.

The coatings application onto medical devices has experienced a continuous growth in the last few years. Medical device coating market is expected to grow at a CAGR of 5.16% to reach USD 10 million by 2023 due to the increasing geriatric population and the growing demand for continuous innovation. Layer-by-Layer (LbL) assembly represents a versatile method to modify the surface properties, in order to control cell interaction and thus enhance biological functions. Furthermore, LbL is environmentally friendly, able to coat all types of surfaces with the creation of homogenous film and to include and control the release of biomolecules/drugs. This feature review provides a critical overview on recent progresses in functionalizing materials by LbL assembly for bone regeneration and disorder treatment. An overview of emerging and visionary opportunities on LbL technologies and further combination with other existing methods used in biomedical field, is also discussed to evidence the new challenges and potential developments in bone regenerative medicine.

Recent Approaches to the Manufacturing of Biomimetic Multi-Phasic Scaffolds for Osteochondral Regeneration.

Cartilage lesions of the knee are common disorders affecting people of all ages; as the lesion progresses, it extends to the underlying subchondral bone and an osteochondral defect appears. Osteochondral (OC) tissue compromises soft cartilage over hard subchondral bone with a calcified cartilage interface between these two tissues. Osteochondral defects can be caused by numerous factors such as trauma and arthritis. Tissue engineering offers the possibility of a sustainable and effective treatment against osteochondral defects, where the damaged tissue is replaced with a long-lasting bio-manufactured replacement tissue. This review evaluates both bi-phasic and multi-phasic scaffold-based approaches of osteochondral tissue regeneration, highlighting the importance of having an interface layer between the bone and cartilage layer. The significance of a biomimetic approach is also evidenced and shown to be more effective than the more homogenous design approach to osteochondral scaffold design. Recent scaffold materials and manufacturing techniques are reviewed as well as the current clinical progress with osteochondral regeneration scaffolds.

Influence of Parathyroid Hormone-Loaded PLGA Nanoparticles in Porous Scaffolds for Bone Regeneration.

Biodegradable poly(lactide-co-glycolide) (PLGA) nanoparticles, containing human parathyroid hormone (PTH (1-34)), prepared by a modified double emulsion-solvent diffusion-evaporation method, were incorporated in porous freeze-dried chitosan-gelatin (CH-G) scaffolds. The PTH-loaded nanoparticles (NPTH) were characterised in terms of morphology, size, protein loading, release kinetics and in vitro assessment of biological activity of released PTH and cytocompatibility studies against clonal human osteoblast (hFOB) cells. Structural integrity of incorporated and released PTH from nanoparticles was found to be intact by using Tris-tricine SDS-PAGE. In vitro PTH release kinetics from PLGA nanoparticles were characterised by a burst release followed by a slow release phase for 3-4 weeks. The released PTH was biologically active as evidenced by the stimulated release of cyclic AMP from hFOB cells as well as increased mineralisation studies. in vitro and cell studies demonstrated that the PTH bioactivity was maintained during the fabrication of PLGA nanoparticles and upon release. Finally, a content of 33.3% w/w NPTHs was incorporated in CH-G scaffolds, showing an intermittent release during the first 10 days and, followed by a controlled release over 28 days of observation time. The increased expression of Alkaline Phosphatase levels on hFOB cells further confirmed the activity of intermittently released PTH from scaffolds.

An overview of poly(lactic-co-glycolic) acid (PLGA)-based biomaterials for bone tissue engineering.

Poly(lactic-co-glycolic) acid (PLGA) has attracted considerable interest as a base material for biomedical applications due to its: (i) biocompatibility; (ii) tailored biodegradation rate (depending on the molecular weight and copolymer ratio); (iii) approval for clinical use in humans by the U.S. Food and Drug Administration (FDA); (iv) potential to modify surface properties to provide better interaction with biological materials; and (v) suitability for export to countries and cultures where implantation of animal-derived products is unpopular. This paper critically reviews the scientific challenge of manufacturing PLGA-based materials with suitable properties and shapes for specific biomedical applications, with special emphasis on bone tissue engineering. The analysis of the state of the art in the field reveals the presence of current innovative techniques for scaffolds and material manufacturing that are currently opening the way to prepare biomimetic PLGA substrates able to modulate cell interaction for improved substitution, restoration, or enhancement of bone tissue function.

Layer-by-layer assembly of nanotheranostic particles for simultaneous delivery of docetaxel and doxorubicin to target osteosarcoma.

Osteosarcoma (OS) is a rare form of primary bone cancer, impacting approximately 3.4 × 106 individuals worldwide each year, primarily afflicting children. Given the limitations of existing cancer therapies, the emergence of nanotheranostic platforms has generated considerable research interest in recent decades. These platforms seamlessly integrate therapeutic potential of drug compounds with the diagnostic capabilities of imaging probes within a single construct. This innovation has opened avenues for enhanced drug delivery to targeted sites while concurrently enabling real-time monitoring of the vehicle's trajectory. In this study, we developed a nanotheranostic system employing the layer-by-layer (LbL) technique on a core containing doxorubicin (DOXO) and in-house synthesized carbon quantum dots. By utilizing chitosan and chondroitin sulfate as polyelectrolytes, we constructed a multilayered coating to encapsulate DOXO and docetaxel, achieving a coordinated co-delivery of both drugs. The LbL-functionalized nanoparticles exhibited an approximate size of 150 nm, manifesting a predominantly uniform and spherical morphology, with an encapsulation efficiency of 48% for both drugs. The presence of seven layers in these systems facilitated controlled drug release over time, as evidenced by in vitro release tests. Finally, the impact of the LbL-functionalized nanoparticles was evaluated on U2OS and Saos-2 osteosarcoma cells. The synergistic effect of the two drugs was found to be crucial in inducing cell death, particularly in Saos-2 cells treated with nanoparticles at concentrations higher than 10 µg/ml. Transmission electron microscopy analysis confirmed the internalization of the nanoparticles into both cell types through endocytic mechanisms, revealing an underlying mechanism of necrosis-induced cell death.

Enhancing analysis of neo-formed contaminants in two relevant food global commodities: Coffee and cocoa.

Neo-formed contaminants (NFCs) are common in many foods, especially those subjected to high-temperature processing. Among these contaminants, products arising from the Maillard reaction, sugar reduction, thermal degradation of polyphenols and lipid oxidation, including acrylamide, furan, furfuryl alcohol, and hydroxymethylfurfural, are consistently linked to potential neoplastic effects. NFCs are found in globally traded commodities like coffee and cocoa, posing a significant risk due to their frequent consumption by consumers. A direct correlation exists between consumption frequency, exposure levels, and health risks. Hence, it's crucial to establish reliable methods to determine levels in both matrices, aiming to mitigate their formation and minimise risks to consumers. This review offers a comprehensive examination, discussion, and identification of emerging trends and opportunities to enhance existing methodologies for extracting and quantifying NFCs in coffee and cocoa. By presenting an in-depth analysis of performance parameters, we aim to guide the selection of optimal extraction techniques for quantifying individual NFCs. Based on the reviewed data, headspace extraction is recommended for furan, while solid and dispersive solid phase extractions are preferred for acrylamide when quantified using gas and liquid chromatography, respectively. However, it is worth noting that the reported linearity tests for certain methods did not confirm the absence of matrix effects unless developed through standard addition, leading to uncertainties in the reported values. There is a need for further research to verify method parameters, especially for determining NFCs like furfuryl alcohol. Additionally, optimising extraction and separation methods is essential to ensure complete compound depletion from samples. Ideally, developed methods should offer comprehensive NFC determination, reduce analysis time and solvent use, and adhere to validation parameters. This review discusses current methods for extracting and quantifying NFCs in coffee and cocoa, highlighting emerging trends and emphasising the need to improve existing techniques, especially for compounds like furfuryl alcohol.

Cell seeding via bioprinted hydrogels supports cell migration into porous apatite-wollastonite bioceramic scaffolds for bone tissue engineering.

Cell seeding via cell-laden hydrogels offers a rapid way of depositing cells onto a substrate or scaffold. When appropriately formulated, hydrogels provide a dense network of fibres for cellular encapsulation and attachment, creating a protective environment that prevents cells to be washed away by media. However, when incorporating hydrogels into a cell seeding strategy the cellular capacity for migration from a hydrogel network and subsequent biofunctionality must be assessed. Here, we compare cell seeding via a bioprinted hydrogel with conventional manual cell seeding in media. To this end, we use a binder jet 3D printed bioceramic scaffold as a model system for bone tissue engineering and the reactive jet impingement (ReJI) bioprinting system to deliver high cell density cell-laden hydrogels onto the surface of the scaffolds. The bioceramic scaffolds were produced in apatite-wollastonite (AW) glass-ceramic, with a total porosity of ~50 %, with pore size predominantly around 50-200 µm. Bone marrow-derived mesenchymal stromal cells were seeded onto the porous AW substrate both in media and via ReJI bioprinting. Cell seeding in media confirmed the osteoinductive nature and the ability of the scaffold to support cell migration within the porous structure. Cell seeding via ReJI bioprinting demonstrated that the cell-laden hydrogel penetrated the porous AW structure upon hydrogel deposition. Furthermore, cells would then migrate out from the hydrogel network and interact with the bioceramic substrate. Overall, levels of cell migration and mineralisation were significant and comparable for both seeding approaches. However, cell seeding via bioprinted hydrogels may serve as an effective strategy for in situ cell seeding into implants, which is desired in clinical tissue engineering procedures, avoiding the time taken for cell attachment from media, and the requirement to maintain a specific orientation until attachment has occurred.

Layer-by-Layer Coatings of Collagen-Hyaluronic acid Loaded with an Antibacterial Manuka Honey Bioactive Compound to Fight Metallic Implant Infections.

Implant-associated severe infections can result in catastrophic implant failures; thus, advanced antibacterial coatings are needed to combat infections. This study focuses on harnessing nature-inspired self-assembly of extracellular matrix (ECM)-like coatings on Ti alloy with a combination of jellyfish-derived collagen (J-COLL) and hyaluronic acid (HA) using our customized automated hybrid layer-by-layer apparatus. To improve the anti-infection efficacy of coatings, we have incorporated a natural antibacterial agent methylglyoxal (MGO, a Manuka honey compound) in optimized multilayer coatings. The obtainment of MGO-loaded multilayer coatings was successfully assessed by profilometry, contact angle, attenuated total reflectance (ATR)-Fourier transform infrared spectroscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy. In vitro degradation confirmed the controlled release activity of MGO with a range of concentrations from 0.90 to 2.38 mM up to 21 days. A bacterial cell culture study using Escherichia coli (E. coli) and Staphylococcus epidermidis (S. epidermidis) confirmed that the MGO incorporated within layers 7 and 9 had a favorable effect on preventing bacterial growth and colonization on their surfaces. An in vitro cytocompatibility study confirmed that MGO agents included in the layers did not affect or reduce the cellular functionalities of L929 fibroblasts. In addition, MGO-loaded layers with Immortalized Mesenchymal Stem Cells (Y201 TERT-hMSCs) were found to favor the growth and differentiation of Y201 cells and promote calcium nodule formation. Overall, these surface coatings are promising candidates for delivering antimicrobial activity with bone-inducing functions for future bone tissue engineering applications.

Exploring the effect of utilising organic acid solutions in ultrasound-assisted extraction of pectin from apple pomace, and its potential for biomedical purposes.

Biomass resulting from food production represents valuable material to recover different biomolecules. In our study, we used apple pomace to obtain pectin, which is traditionally extracted using mineral acids. Our hypothesis consisted of carrying out extractions with organic acids, assisted by ultrasound, by varying processing parameters including time, temperature, and type of acid. The analytical determinations of galacturonic acid content, methoxylation and esterification degree, ζ-potential and extraction yield were used as pectin quality indicators. Pectins extracted using treatment conditions with better performance were assessed biologically in vitro for their potential to be used in biomedical applications. Overall, the extracted pectin presented a galacturonic acid content, methoxylation and esterification degree ranged from 19.7 to 67%, 26.8-41.4% and 58-65.2% respectively, and were negatively charged (-24.1 to -13.2 mV). It was found that factors of time and temperature greatly influenced the response variables excepting the esterification degree, while the acid type influenced the ζ-potential, methoxylation and esterification degrees. Additionally, it was seen that the longer extraction time (50 min) and higher temperature (50 °C) exhibited the better extraction yield (∼10.9%). Finally, the selected pectin showed high cytocompatibility up to 500 µg/mL of concentration when seeded with Neonatal Normal Human Dermal Fibroblasts.

An In Vitro Engineered Osteochondral Model as Tool to Study Osteoarthritis Environment.

Osteoarthritis (OA) is a joint degenerative pathology characterized by mechanical and inflammatory damages affecting synovium, articular cartilage (AC), and subchondral bone (SB). Several in vitro, in vivo, and ex vivo models are developed to study OA, but to date the identification of specific pharmacological targets seems to be hindered by the lack of models with predictive capabilities. This study reports the development of a biomimetic in vitro model of AC and SB interface. Gellan gum methacrylated and chondroitin sulfate/dopamine hydrogels are used for the AC portion, whereas polylactic acid functionalized with gelatin and nanohydroxyapatite for the SB. The physiological behavior of immortalized stem cells (Y201s) and Y201s differentiated in chondrocytes (Y201-Cs), respectively, for the SB and AC, is demonstrated over 21 days of culture in vitro in healthy and pathological conditions, whilst modeling the onset of cytokines-induced OA. The key metrics are: lower glycosaminoglycans production and increased calcification given by a higher Collagen X content, in the AC deep layer; higher expression of pro-angiogenic factor (vegf) and decreased expression of osteogenic markers (coll1, spp1, runx2) in the SB. This novel approach provides a new tool for studying the development and progression of OA.

A cytokine-induced spheroid-based in vitro model for studying osteoarthritis pathogenesis.

Given the lack of in vitro models faithfully reproducing the osteoarthritis (OA) disease on-set, this work aimed at manufacturing a reliable and predictive in vitro cytokine-based Articular Cartilage (AC) model to study OA progression. Cell spheroids of primary human fetal chondrocytes (FCs) and h-TERT mesenchymal stem cells differentiated chondrocytes (Y201-C) were analysed in terms of growth kinetics, cells proliferation and apoptosis over 10 days of culture, in healthy condition or in presence of cytokines (interleukin-1ß, -6 and TNF-α). Then, the spheroids were assembled into chondrospheres using a bottom-up strategy, to obtain an in vitro cytokines-induced OA model. The resulting chondrospheres were evaluated for gene expression and anabolic ECM proteins. Compared to the healthy environment, the simulated OA environment induced chondrocyte hyperproliferation and apoptotic pathway, decreased expression of anabolic ECM proteins, and diminished biosynthetic activity, resembling features of early-stage OA. These characteristics were observed for both Y201-C and HC at high and low concentrations of cytokines. Both HC and Y201-C demonstrated the suitability for the manufacturing of a scaffold-free in vitro OA model to facilitate studies into OA pathogenesis and therapeutic strategies. Our approach provides a faithful reproduction of early-stage osteoarthritis, demonstrating the ability of obtaining different disease severity by tuning the concentration of OA-related cytokines. Given the advantages in easy access and more reproducible performance, Y201-C may represent a more favourable source of chondrocytes for establishing more standardized protocols to obtain OA models.

Bioprinting of Cell-Laden Hydrogels onto Titanium Alloy Surfaces to Produce a Bioactive Interface.

The surface of metal implants serves as a powerful signaling cue for cells. Its properties play an essential role in stabilizing the bone-implant interface and facilitating the early osseointegration by encouraging bone deposition on the surface. However, effective strategies to deliver cells to the metal surfaces are yet to be explored. Here, a bioprinting process, called reactive jet impingement (ReJI), is used to deposit high concentrations (4 × 107  cells mL-1 ) of mesenchymal stromal cells (MSCs) within hydrogel matrices directly onto the titanium alloy surfaces that vary in surface roughness and morphology. In this proof-of-concept study, cell-hydrogel-metal systems are fabricated with the aim of enhancing bioactivity through delivering MSCs in hydrogels at the bone-implant interface. These results show that the high cell concentrations encourage quick cell-biomaterial interactions at the hydrogel-metal surface interface, and cell morphology is influenced by the surface type. Cells migrate from the hydrogels and deposit mineralized matrix rich in calcium and phosphorus on the titanium alloy surfaces. The authors demonstrate that ReJI bioprinting is a promising tool to deliver cells in a 3D environment before implantation that can be used when developing a new generation of medical devices for bone tissue engineering.

Adhesive Bioinspired Coating for Enhancing Glass-Ceramics Scaffolds Bioactivity.

Bioceramic scaffolds, composed of a biphasic composite containing bioactive glass and hydroxyapatite, were prepared in this work to overcome the intrinsic limits of the two components taken separately (in particular, their specific reactivities and dissolution rates, which should be tunable as a function of the given clinical requirements). To mimic the biological environment and tune the different stages of cellular response, a coating with gelatin and chondroitin sulphate via Layer-by-Layer (LbL) assembly was presented and discussed. The resulting functionalized scaffolds were affected by the coating in terms of microstructure and porosity. In addition, the LbL coating significantly enhanced the seeded cell behaviour, with high adhesion, proliferation and osteogenic activity, as revealed by the alkaline phosphatase activity and overexpression of osteopontin and osteocalcin.

A Chondrosphere-Based Scaffold Free Approach to Manufacture an In Vitro Articular Cartilage Model.

In vitro engineering of human articular cartilage (AC) is a regenerative medicine challenge. The main objective of this study was the development of a repeatable scaffold-free in vitro model of chondrocyte spheroid-based treatments of cartilage defects, to allow for systematic study and further optimization of this type of treatment. Human articular chondrocytes (HC) and immortalized mesenchymal cells differentiated in chondrocytes (Y201-Cs) were cultured in round-bottom 96-well plates to produce multicellular spheroids and their growth kinetics, and viability was evaluated over 7 days of culture. Then, the spheroids were assembled and cultured for 21 days on a gelatin-coated poly(lactic-co-glycolic acid) electrospun membrane (10 spheroids/cm2), following a protocol in line with the clinically approved Chondrosphere® (CO.DON AG) technique. Both HC and Y201-C cells formed compact and viable spheroids after 7 days of culture with a reduction of diameter over the 7 days from 1300 ± 150 µm to 600 ± 90 µm and from 1250 ± 60 µm to 800 ± 20 µm for HC and Y201-C, respectively. When the spheroids were transferred onto the support membrane, these adhered on the membrane itself and fused themselves, producing collagen type II (COL2A1) and aggrecan (ACAN), according to gene expression and glycosaminoglycans quantification analyses. We detected higher expression of COL2A1 in HC cells, while the Y201-C constructs were characterized by an increased ACAN expression. The approach we presented allows a standardizable production of spheroids with predictable geometry and the creation of a reproducible scaffold-free in vitro AC-like construct showing high expression of chondrogenic markers, using both HC and Y201-C. In addition, the bankable Y201-C cells provide an effective base model for experimentation with the spheroid approach to further enhance the process. Impact statement This is first work on the development of a repeatable scaffold-free in vitro model based on an optimized protocol in line with a recent clinically approved Chondrosphere® (CO.DON AG) technique. In addition, we demonstrated that a bankable cell type (Y201-C) could produce an engineered cartilage-like construct, giving a repeatable model as a key tool for experimentation of therapeutic treatment ahead of studies with heterogeneous cell populations.

A bioprintable gellan gum/lignin hydrogel: a smart and sustainable route for cartilage regeneration.

In this work a hydrogel, based on a blend of two gellan gums with different acyl content embedding lignin (up to 0.4%w/v) and crosslinked with magnesium ions, was developed for cartilage regeneration. The physico-chemical characterizations established that no chemical interaction between lignin and polysaccharides was detected. Lignin achieved up to 80 % of ascorbic acid's radical scavenging activity in vitro on DPPH and ABTS radicals. Viability of hMSC onto hydrogel containing lignin resulted comparable to the lignin-free one (>70 % viable cells, p > 0.05). The presence of lignin improved the hMSC 3D-constructs chondrogenesis, bringing to a significant (p < 0.05) up-regulation of the collagen type II, aggrecan and SOX 9 chondrogenic genes, and conferred bacteriostatic properties to the hydrogel, reducing the proliferation of S. aureus and S. epidermidis. Finally, cellularized 3D-constructs were manufactured via 3D-bioprinting confirming the processability of the formulation as a bioink and its unique biological features for creating a physiological milieu for cell growth.

Valuable effect of Manuka Honey in increasing the printability and chondrogenic potential of a naturally derived bioink.

Hydrogel-based bioinks are the main formulations used for Articular Cartilage (AC) regeneration due to their similarity to chondral tissue in terms of morphological and mechanical properties. However, the main challenge is to design and formulate bioinks able to allow reproducible additive manufacturing and fulfil the biological needs for the required tissue. In our work, we investigated an innovative Manuka honey (MH)-loaded photocurable gellan gum methacrylated (GGMA) bioink, encapsulating mesenchymal stem cells differentiated in chondrocytes (MSCs-C), to generate 3D bioprinted construct for AC studies. We demonstrated the beneficial effect of MH incorporation on the bioink printability, leading to the obtainment of a more homogenous filament extrusion and therefore a better printing resolution. Also, GGMA-MH formulation showed higher viscoelastic properties, presenting complex modulus G∗ values of ∼1042 â€‹Pa, compared to ∼730 â€‹Pa of GGMA. Finally, MH-enriched bioink induced a higher expression of chondrogenic markers col2a1 (14-fold), sox9 (3-fold) and acan (4-fold) and AC ECM main element production (proteoglycans and collagen).

Impact of heterogeneously crosslinked calcium alginate networks on the encapsulation of ß-carotene-loaded beads.

This study investigated the impact of heterogeneity of crosslinking on a range of physical and mechanical properties of calcium alginate networks formed via external gelation with 0.25-2% sodium alginate and 2.5 and 5% CaCl2. Crosslinking in films with 1-2% alginate was highly heterogeneous, as indicated by their lower calcium content (35-7 mg Ca·g alginate-1) and apparent solubility (5-6%). Overall, films with 1-2% alginate showed higher resistance (tensile strength = 51-147 MPa) but lower elasticity (Elastic Modulus = 2136-10,079 MPa) than other samples more homogeneous in nature (0.5% alginate, Elastic Modulus = 1918 MPa). Beads with 0.5% alginate prevented the degradation of ß-carotene 1.5 times more efficiently than 1% beads (5% CaCl2) at any of the storage temperatures studied. Therefore, it was postulated that calcium alginate networks crosslinked to a greater extent and in a more homogeneous manner showed better mechanical performance and barrier properties for encapsulation applications.