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Spheroids based on chondrocytes are gaining interest in the treatment of injured cartilage tissue due to their regenerative potential. In this study, chondrocyte-based aggregates were formed by culturing cells on a polymeric coating of a lactose-modified chitosan (CTL) and their evolution was followed for 14 days of culture. Morphological analyses through scanning electron microscopy indicated that these cell aggregates formed by a process of cell aggregation rather than proliferation, resulting in structures with an irregular morphology that were up to 1 mm in size. Chondrocytes interact very closely with CTL. In the early days of cellular aggregation, CTL is distributed among the cells, while in later stages, it is localized in the inner part of the aggregates, forming an amorphous matrix. The initial outer position of the polymer is replaced over time by an increased matrix deposition whose main component is type II collagen. Transmission electron microscopy pointed out that the chondrocytes internalized the polymer during the first days of culture. The presence of CTL does not affect the ability of cells to migrate from chondro-aggregates when transferred to uncoated wells, mimicking the ability of cells to colonize a cartilage defect. Overall, these results suggest that CTL plays a role as a temporary matrix for chondrocytes aggregation during chondro-aggregates formation. This opens up innovative therapeutic approaches for cartilage regeneration without the need to use a scaffold to deliver cells to the damaged site.KEYWORDS: chondrocytes, spheroids, lactose-modified chitosan, cartilage regeneration, bioactive polysaccharide

Alginate-based hydrogels with tunable mechanical properties are developed by chemical methylation of the polysaccharide backbone, which was performed either in homogeneous phase (in solution) or in heterogeneous phase (on hydrogels). Nuclear Magnetic Resonance (NMR) and Size Exclusion Chromatography (SEC-MALS) analyses of methylated alginates allow to identify the presence and location of methyl groups on the polysaccharide, and to investigate the influence of methylation on the stiffness of the polymer chains. The methylated polysaccharides are employed for the manufacturing of calcium-reticulated hydrogels for cell growth in 3D. The rheological characterization shows that the shear modulus of hydrogels is dependent on the amount of cross-linker used. Methylated alginates represent a platform to explore the effect of mechanical properties on cell activity. As an example, the effect of compliance is investigated using hydrogels displaying similar shear modulus. An osteosarcoma cell line (MG-63) was encapsulated in the alginate hydrogels and the effect of material compliance on cell proliferation and localization of YAP/TAZ protein complex is investigated by flow cytometry and immunohistochemistry, respectively. The results point out that an increase of material compliance leads to an increase of the proliferative rate of cells and correlates with the translocation of YAP/TAZ inside the cell nucleus.

Strain hardening has recently emerged as a near-universal response of biological tissues to mechanical stimulation as well as a powerful regulator of cell fate. Understanding the mechanistic basis for this nonlinear elasticity is crucial for developing bioinspired materials that mimic extracellular matrix mechanics. Here, we show that covalent networks built from highly acetylated chitosans exhibit strain hardening at physiological pH and osmolarity. While varying the chitosan physical-chemical composition and network connectivity, we provide evidence that temporary nodes arising from the entangling of chains between stable cross-links are at the root of nonlinear elasticity. The contour length (Lc) of the said chains revealed that the larger the chain length between the cross-links, the greater is the entanglement over disentanglement upon network stretching. To this end, we calculated that the minimum number of Khun's segments in Lc that contributes to the onset of strain hardening is 15. Furthermore, we identified a relationship between critical strain marking nonlinear elasticity and the network connectivity, being similar to that found for the cytoskeletal collagen matrix, indicating the potential use of semiflexible (neutral pH-soluble) chitosans in assembling extracellular matrix mimics.

Memona Khan,1 Hui Liu,2 Pasquale Sacco,3,4 Eleonora Marsich,3,4 Xiaowu Li,2 Nadia Djaker,1 Jolanda Spadavecchia1 1CNRS, UMR 7244, NBD-CSPBAT, Laboratory of Chemistry, Structures and Properties of Biomaterials and Therapeutic Agents University Paris13, Sorbonne Paris Nord, Bobigny, France; 2Department of Hepatobiliary Surgery, Guangdong Provincial Key Laboratory of Regional Immunity and Diseases& Carson International Cancer Center, Shenzhen University General Hospital & Shenzhen University Clinical Medical Academy Center, Shenzhen University, Shenzhen, People’s Republic of China; 3Department of Life Sciences, University of Trieste, Trieste, I-34127, Italy; 4Department of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, I-34129, ItalyCorrespondence: Nadia Djaker; Jolanda Spadavecchia, CNRS, UMR 7244, NBD-CSPBAT, Laboratory of Chemistry, Structures and Properties of Biomaterials and Therapeutic Agents University Paris13, Sorbonne Paris Nord, Bobigny, France, Email nadia.djaker@univ-paris13.fr; jolanda.spadavecchia@univ-paris13.frIntroduction: The realization of MRI contrast agents through chemical protocols of functionalization is a strong domain of research. In this work, we developed and formulated a novel hybrid gold nanoparticle system in which a gold salt (HAuCl4) is combined with dotarem, an MRI contrast agent (DOTA) by chelation (Method IN) and stabilized by a lactose-modified chitosan polymer (CTL; Chitlac) to form DOTA IN-CTL AuNPs.Result and Discussion: The authors demonstrate the biological efficiency of these nanoparticles in the case of three cell lines: Mia PaCa-2 (human pancreatic cancer cell line), TIB-75 (murine liver cell line) and KKU-M213 (cholangiocarcinoma cell line). DOTA IN-CTL AuNPs are stable under physiological conditions, are nontoxic, and are very efficient as PTT agents. The highlights, such as high stability and preliminary MRI in vitro and in vivo models, may be suitable for diagnosis and therapy.Conclusion: We proved that DOTA IN-CTL AuNPs have several advantages: i) Biological efficacy on three cell lines: MIA PaCa-2 (human pancreatic cancer cell line), TIB-75 (murine liver cell line) and KKU-M213 (cholangiocarcinoma cell line); ii) high stability, and no-toxicity; iii) high efficiency as a PPT agent. The study conducted on MRI in vitro and in vivo models will be suitable for diagnosis and therapy.Keywords: nanotheranostic, MRI, hybrid nanoparticles, dotarem, biomedical applications

Natural tissues and extracellular matrices (ECMs) are not purely elastic materials but exhibit dissipative properties. Although it has recently emerged as a novel regulator of cellular responses, the contribution of material dissipation to guiding cell-fate decisions is still in its infancy. Here, a strategy for tuning the dissipation rate of viscoplastic substrates while precisely regulating linear elasticity is reported. Semi-interpenetrating substrates consisting of a rigid hydrogel network intertwined with a branched biopolymer are described. The release of these weak physical entanglements under loading dissipates the applied stress and leads to the extension of the linear elasticity. These results reveal a crucial link between this material property and cell response in 2D cultures, impacting cell migration mode and speed, vinculin-dependent focal adhesion geometry and size, F-actin organization, the transmission of forces, and Yes-associated protein nuclear translocation. It is shown that cells require joint actomyosin contractility and microtubule tension to probe the substrate and decide whether or not to adhere, revealing a clear correlation between force transmission, substrate dissipation rate, and amount of anchoring points. Overall, these findings introduce linear elasticity as a novel design parameter for assembling tunable dissipative materials to study cell mechanosensing in 2D and possibly also in 3D cultures.

Polysaccharide electrospun wound dressings should be an effective strategy in the field of wound care, as they combine an extracellular matrix-like structure with excellent biomimicry. However, their high hydrophilicity and large surface area cause a rapid dissolution in aqueous environments, compromising their clinical employment. In the present paper, electrospun membranes prepared using hyaluronic acid, a bioactive lactose-modified chitosan (CTL), and polyethylene oxide have been crosslinked using glutaraldehyde, genipin, EDC/NHS or thermal treatments, obtaining very poor results in terms of membrane stability. Therefore, carbonyldiimidazole (CDI) and methacrylic anhydride were investigated in an innovative way, where CDI proved to be the best compromise between nanofiber water resistance, architecture maintenance and degradability. Indeed, the swelling and degradation behavior as well as the water vapor permeability of these matrices were tested, revealing the effectiveness of the electrospun products in absorbing large amount of liquid while maintaining the balance between water retention and gas permeability.

Developing synthetic materials able to mimic micro- and macrorheological properties of natural networks opens up to novel applications and concepts in materials science. The present contribution describes an active network based on a semi-synthetic polymer, a lactitol-bearing chitosan derivative (Chitlac), and a transient inorganic cross-linker, boric acid. Due to the many and diverse anchoring points for boric acid on the flanking groups of Chitlac, the cross-links constantly break and reform in a highly dynamic fashion. The consequence is a network with unusual non-equilibrium and mechanical properties closely resembling the rheological behavior of natural three-dimensional arrangements and of cytoskeleton. Concepts like network nucleation, reorganization and disassembly are declined in terms of amount of the cross-linker, which acts as a putative motor for remodeling of the network upon application of energy. The out-of-equilibrium and non-linear behavior render the semi-synthetic system of great interest for tissue engineering and for developing in-vitro mimics of natural active matrices.

Correction for ‘CTL–doxorubicin (DOX)–gold complex nanoparticles (DOX–AuGCs): from synthesis to enhancement of therapeutic effect on liver cancer model’ by Qiqian Liu et al., Nanoscale Adv., 2020, 2, 5231–5241. https://doi.org/10.1039/D0NA00758G.

Lactose-modified chitosan (CTL) is sulfated using SO3·py or SO3·DMF as sulfating agents. The two products are characterized by elemental analysis, FT-IR, 1H,13C-DEPT-HSQC and 1H,13C-HSQC-TOCSY experiments which allow the extent and selectivity of chemical sulfation to be determined. Dynamic Light Scattering shows a pH-dependent association of the sulfated polysaccharides which are described as flexible by the Smidsrød's B parameter and the intrinsic viscosity at infinite ionic strength. Shear viscosity and intrinsic viscosity show that sulfation protocols lead to chain scission which is more pronounced when SO3·DMF is used. The sulfated samples are able to induce aggregation of human bone marrow mesenchymal stem cells, resulting in the formation of smaller nodules compared to the unmodified CTL sample. Over time, the sample with the higher degree of sulfation allows further aggregation between cell clusters while the sample with the lower degree of sulfation shows dissolution of the aggregates.

Bioactive and biodegradable porous scaffolds can hasten the healing of bone defects; moreover, patient stem cells seeded onto scaffolds can enhance the osteoinductive and osteoconductive properties of these biomaterials. In this work, porous alginate/hydroxyapatite scaffolds were functionalized with a bioactive coating of a lactose-modified chitosan (CTL). The highly interconnected porous structure of the scaffold was homogeneously coated with CTL. The scaffolds showed remarkable stability up to 60 days of aging. Human Dental Pulp Stem Cells (hDPSCs) cultured in the presence of CTL diluted in culture medium, showed a slight and negligible increase in terms of proliferation rate; on the contrary, an effect on osteogenic differentiation of the cells was observed as a significant increase in alkaline phosphatase activity. hDPSCs showed higher cell adhesion on CTL-coated scaffolds than on uncoated ones. CTL coating did not affect cell proliferation, but stimulated cell differentiation as shown by alkaline phosphatase activity analysis.

Nine antibacterial di-methacrylate monomers based on bis-quaternary ammonium salts (bis-QAMs) were synthesized and structurally characterized. The biological activity of the bis-QAMs was tested in terms of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) on different bacterial strains achieving promising results and, in most cases, a complete bactericidal effect using a bis-QAM concentration lower than 1 mg/mL. Two of the structures showed comparable and superior activity against S. mutans than the commercial monomer 12-methacryloyloxydodecyl pyridinium bromide (MDBP). All the bis-QAMs here described were able to inhibit S. mutans biofilm formation at a concentration equal to the MIC value. From the analysis of the obtained data, some correlation regarding the structure and the antibacterial activity of the bis-QAMs could be drawn: a flexible alkyl C12 spacer between the two quaternary ammonium moieties increased the monomer antibacterial effect in comparison to the aromatic ones; the equilibrium between hydrophobic and hydrophilic moieties was directly correlated to the bactericidal range of action; the increase of the steric hindrance of the ammonium side groups might be both advantageous or disadvantageous to the antibacterial efficacy depending on the whole monomer chemical structure. Even though the possible correlation between the monomer structures and their bacteriostatic or bactericidal effect is under investigation, the monomers exhibited low cytotoxicity on human dental pulp stem cells, confirming their promising potential in the dental materials' field. Statement of significance The use of dental resins with antibacterial monomers might prevent the formation of secondary caries at the restoration margins. For this purpose, a series of di-methacrylate bis-quaternary ammonium monomers (QAMs) was developed. Unlike antibacterial mono-methacrylate monomers already described in the literature, the synthesized di-methacrylate monomers have the potential of acting as cross-linkers stabilizing the polymeric network and bear two quaternary ammonium groups that increase their antibacterial ability. The QAMs exert bactericidal activity on both Gram(+) and Gram(−) bacterial strains maintaining at the same time good biocompatibility with the oral environment. Some structural elements of the monomers were clearly related to high antibacterial properties, and this can help design new active structures and better understand their mechanism of action.

Chronic wounds deeply invalidate patient's quality of life, involving very high costs for the medical system. Numerous wound dressings have been studied over the years, and active wound dressings replaced traditional passive dressings to promote tissue regeneration and wound closure. Aiming at an optimal reproduction of the physiological environment, electrospun wound dressings are emerging since they mimic the architecture of the extracellular matrix and provide a large contact surface area, enabling exudate absorption and permeability as well as good conformability to the wound site. The use of polysaccharides offers an excellent biomimicry, as they ensure good biocompatibility, biodegradability, and non-immunogenicity. Furthermore, they possess bioactive properties, such as antimicrobial, anti-inflammatory, and antioxidant properties, which can promote and enhance the healing process. The aim of this review is to present the morphological, physical, and chemical features of an ideal wound dressing together with the traditional and the current strategies, and the already commercialized wound dressings. Moreover, the review is focused on the preparation of polysaccharide-based electrospun nanofibrous devices and on the strategies for the modulation and improvement of membrane stability and bioactivity. Lastly, a comprehensive consideration on the process and requirements that lead to the commercialization of the wound dressings is reported.

The present manuscript deals with the elucidation of the mechanism of genipin binding by primary amines at neutral pH. UV-VIS and CD measurements both in the presence of oxygen and in oxygen-depleted conditions, combined with computational analyses, led to propose a novel mechanism for the formation of genipin derivatives. The indications collected with chiral and achiral primary amines allowed interpreting the genipin binding to a lactose-modified chitosan (CTL or Chitlac), which is soluble at all pH values. Two types of reaction and their kinetics were found in the presence of oxygen: (i) an interchain reticulation, which involves two genipin molecules and two polysaccharide chains, and (ii) a binding of one genipin molecule to the polymer chain without chain&ndash, chain reticulation. The latter evolves in additional interchain cross-links, leading to the formation of the well-known blue iridoid-derivatives.

A miscibility study between oppositely charged polyelectrolytes, namely hyaluronic acid and a lactose-modified chitosan, is here reported. Experimental variables such as polymers&rsquo, weight ratios, pH values, ionic strengths and hyaluronic acid molecular weights were considered. Transmittance analyses demonstrated the mutual solubility of the two biopolymers at a neutral pH. The onset of the liquid-liquid phase separation due to electrostatic interactions between the two polymers was detected at pH 4.5, and it was found to be affected by the overall ionic strength, the modality of mixing and the polymers&rsquo, weight ratio. Thorough Dynamic Light Scattering (DLS) measurements were performed to check the quality of the formed coacervates by investigating their dimensions, homogeneity and surface charge. The whole DLS results highlighted the influence of the hyaluronic acid molecular weight in affecting coacervates&rsquo, dispersity and size.

Chitosan derivatives, and more specifically, glycosylated derivatives, are nowadays attracting much attention within the scientific community due to the fact that this set of engineered polysaccharides finds application in different sectors, spanning from food to the biomedical field. Overcoming chitosan (physical) limitations or grafting biological relevant molecules, to mention a few, represent two cardinal strategies to modify parent biopolymer; thereby, synthetizing high added value polysaccharides. The present review is focused on the introduction of oligosaccharide side chains on the backbone of chitosan. The synthetic aspects and the effect on physical-chemical properties of such modifications are discussed. Finally, examples of potential applications in biomaterials design and drug delivery of these novel modified chitosans are disclosed.

The present paper explores the effect of boric acid on Chitlac, a lactose-modified chitosan which had previously shown interesting biological and physical-chemical features. The herewith-reported experimental evidences demonstrated that boric acid binds to Chitlac, producing conformational and association effects on the chitosan derivative. The thermodynamics of boric acid binding to Chitlac was explored by means of 11B NMR, circular dichroism (CD), and UV–vis spectroscopy, while macromolecular effects were investigated by means of viscometry and dynamic light scattering (DLS). The experimental results revealed a chain–chain association when limited amounts of boric acid were added to Chitlac. However, upon exceeding a critical boric acid limit dependent on the polysaccharide concentration, the soluble aggregates disentangle. The rheological behavior of Chitlac upon treatment with boric acid was explored showing a dilatant behavior in conditions of steady flow. An uncommonly high dependence in the scaling law between the zero-shear viscosity and the concentration of Chitlac was found, i.e., η0 ∝ CCTL5.8, pointing to interesting potential implications of the present system in biomaterials development.

Adhesive coatings for implantable biomaterials can be designed to prevent material displacement from the site of implant. In this paper, a strategy based on the use of melanin-like nanoparticles (MNPs) for the development of adhesive coatings for polysaccharidic membranes was devised. MNPs were synthesized in vitro and characterized in terms of dimensions and surface potential, as a function of pH and ionic strength. The in vitro biocompatibility of MNPs was investigated on fibroblast cells, while the antimicrobial properties of MNPs in suspension were evaluated on E. coli and S. aureus cultures. The manufacturing of the adhesive coatings was carried out by spreading MNPs over the surface of polysaccharidic membranes; the adhesive properties of the nano-engineered coating to the target tissue (intestinal serosa) were studied in simulated physiological conditions. Overall, this study opens for novel approaches in the design of naturally inspired nanostructured adhesive systems.

Introduction Despite extensive research, anastomotic leakage (AL) remains one of the most dreaded complications after colorectal surgery. Since butyrate enemas are known to enhance anastomotic healing, several administration routes have been explored in this study. Methods Three intraluminal approaches involving butyrate were investigated: (1) butyrin-elucidating patch, (2) a single injection of hyaluronan-butyrate (HA-But) prior to construction of the proximal anastomosis and (3) rectal hyaluronan-butyrate (HA-But) enemas designed for distal anastomoses. The main outcome was AL and secondary outcomes were bursting pressure, histological analysis of the anastomosis, zymography to detect MMP activity and qPCR for gene expression of MMP2, MMP9, MUC2 and TFF3. Results Neither the patches nor the injections led to a reduction of AL in experiments 1 and 2. In experiment 3, a significant reduction of AL was accomplished with the (HA-But) enema compared to the control group together with a higher bursting pressure. Histological analysis detected only an increased inflammation in experiment 2 in the hyaluronan injection group compared to the control group. No other differences were found regarding wound healing. Zymography identified a decreased proenzyme of MMP9 when HA-But was administered as a rectal enema. qPCR did not show any significant differences between groups in any experiment. Conclusion Butyrate enemas are effective in the enhancement of colonic anastomosis. Enhanced butyrate-based approaches designed to reduce AL in animal models for both proximal and distal anastomoses were not more effective than were butyrate enemas alone. Further research should focus on how exogenous butyrate can improve anastomotic healing after gastrointestinal surgery. Electronic supplementary material The online version of this article (doi:10.1007/s00384-016-2718-z) contains supplementary material, which is available to authorized users.

Osteoarthritis (OA) is a chronic disease affecting joint functionality and often managed with hyaluronic acid (HA) administration. In this study, a hydrogel based on a lactose-modified chitosan (CTL) reticulated with boric acid has been developed as a viscosupplement for OA treatment. The rheological characterization allowed to identify a composition whose properties were in line with those of commercial products (in the order of tens of Pascal). The selected CTL-hydrogel showed biocompatibility and antioxidant activity in vitro, and it did not influence cytokines release by macrophages. Degradation studies carried out over 24 h pointed out its higher resistance to chemical degradation with respect to HA samples. Overall, this study underlines the advantages of the CTL-hydrogel to address the treatment of OA and shed light on an innovative application of CTL polymer, which is one of the main component of the proposed hydrogel system and not used in mixture with other molecules.

The present contribution deals with the synthesis and characterization of N-isopropyl chitosan in which the introduction of hydrophobic groups leads to an increased flexibility of the polysaccharide backbone. The isopropyl groups extend the solubility of the modified-chitosan samples and render the modified chitosan a pH- and thermo-sensitive system for hydrogel formation. Indeed, upon varying the pH of the system and/or its temperature within a range compatible with biological applications, a non-reversible sol-gel transition occurs, as determined through extended rheological analyses. The modified chitosan samples show a very good biocompatibility as determined through preliminary viability and cell growth experiments.

The intravaginal route of administration can be exploited to treat local diseases and for systemic delivery. In this work, we developed an alginate/chitosan membrane sufficiently stable in a simulated vaginal fluid and able to dissolve over time at a very slow and linear rate. The membrane demonstrated good mechanical properties both in its swollen and dry form. As a study case, we evaluated the viability of this potential drug delivery system for the treatment of bacterial vaginosis, a common disease affecting women in their reproductive age. Metronidazole was effectively included in the alginate/chitosan membrane and its bactericide effect was demonstrated against Staphylococcus aureus and Gardnerella vaginalis, simultaneously showing good biocompatibility with a cervix epithelial cell line. Since this alginate/chitosan membrane is stable in a simulated vaginal environment, is easy to fabricate and can be used for the controlled release of a model drug, it represents a promising drug delivery system for local intravaginal applications.

In this work, we bring back a rapid way to conceive doxorubicin (DOX) hybrid gold nanoparticles, in which DOX and Au(III) ions were complexed with a hydrochloride-lactose-modified chitosan, named CTL and dicarboxylic acid-terminated polyethylene-glycol (PEG), leading to hybrid polymer-sugar-metal nanoparticles (DOX–AuGSs). All formulations were assessed by spectroscopic techniques (Raman and UV-Vis) and transmission electron microscopy (TEM). To estimate the therapeutic effect of DOX–AuGSs in liver cancer, murine HepG2 cells were used to induce a hepatic carcinoma model in nude mice. The survival time of the tumor-bearing mice, body weight and tumor volume were measured and recorded. The cytokines were used to detect the serum inflammatory factors, and the blood cell analyzer was used to determine the blood cell content of different groups of nude mice. The outcomes demonstrate that DOX–AuGCs significantly suppressed the tumor growth derived from human HepG2 injection and reduce the tumor index without affecting the body weight of mice. Moreover, DOX–AuGCs significantly reduced the serum levels of cytokines IL-6, TNF-α and IL-12 P70. Finally, a histological analysis of the heart tissue sections indicated that DOX–AuGCs significantly reduce the chronic myocardial toxicity of DOX during the period of treatment.

Mounting evidences have recognized that dual cross-link and double-network gels can promisingly recapitulate the complex living tissue architecture and overcome mechanical limitations of conventional scaffolds used hitherto in regenerative medicine. Here, dual cross-link gels formed of a bioactive lactose-modified chitosan reticulated via both temporary (boric acid-based) and permanent (genipin-based) cross-linkers are reported. While boric acid rapidly binds to lactitol flanking diols increasing the overall viscosity, a slow temperature-driven genipin binding process takes place allowing for network strengthening. Combination of frequency and stress sweep experiments in the linear stress-strain region shows that ultimate gel strength, toughness, and viscoelasticity depend on polymer-to-genipin molar ratio. Notably, herewith it is demonstrated that linear stretching correlates with strain energy dissipation through boric acid binding/unbinding dynamics. Strain-hardening effect in the nonlinear regime, along with good biocompatibility in vitro, points at an interesting role of present system as biological extracellular matrix substitute.

Galectins (Gal) are a family of dimeric lectins, composed by two galactoside-binding sites implicated in the regulation of cancer progression and immune responses. In this study, we report for the first time the synthesis and the physical-chemical characterization of galectin-1-complex-gold COOH-terminated polyethlenglicole (PEG)-coated NPs (Gal-1 IN PEG-AuNPs) and their ability to recognize glucose in an aqueous solution with a concentration varying from 10 mM to 100 pM. The chemical protocol consistsof three steps: (i) complexation between galectin-1Gal-1 and tetrachloroauric acid (HAuCl4) to form gold-protein grains; (ii) staking process of COOH-terminated polyethlenglicole molecules (PEG) onto Gal-1-Au complex and (iii) reduction of hybrid metal ions to obtain a colloidal stable solution. During the complexation, the spectral signatures related to the Gal-1 orientation on the gold surface have been found to change due to its protonation state. The effective glucose monitoring was detected by UV–vis, Raman spectroscopy and Transmission Electron Microscopy (TEM). Overall, we observed that the interaction is strongly dependent on the Gal-1 conformation at the surface of gold nanoparticles.

Biodegradable membranes for cartilage applications were manufactured starting from polymeric networks of a lactose-modified chitosan (CTL), previously proposed for chondrocytes stimulation. This implantable biomaterial was conceived as a reservoir of a bioactive polymer that could promote the activity of chondrocytes and the healing of cartilage defects. Freeze-drying of reticulated hydrogels enabled to obtain pliable membranes with a homogeneous polymeric texture, as pointed out by scanning electron microscopy analyses. Swelling tests and dimensional evaluations showed that the material is able to absorb physiological fluids and expand gradually upon rehydration. This feature was evaluated on a simulated cartilage defect on pig's humerus (ex vivo), which revealed the capability of the membranes to progressively fit the tissue voids on the damaged cartilage. The rheological properties of the rehydrated membranes pointed out their peculiar strain-stiffening behavior, which represents a promising feature for the regeneration of tissues subjected to variable mechanical loads and deformations. Biological in vitro studies demonstrated the biocompatibility of the membranes in contact with primary chondrocytes and osteoblasts. Taken together, these results represent a starting point for the development of a novel generation of implantable biomaterials for cartilage treatment based on CTL.

An emerging strategy to improve adhesiveness of biomaterials in wet conditions takes inspiration from the adhesive features of marine mussel, which reside in the chemical reactivity of catechols. In this work, a catechol-bearing molecule (dopamine) was chemically grafted onto alginate to develop a polysaccharide-based membrane with improved adhesive properties. The dopamine-modified alginates were characterized by NMR, UV spectroscopy and in vitro biocompatibility. Mechanical tests and in vitro adhesion studies pointed out the effects of the grafted dopamine within the membranes. The release of HA from these resorbable membranes was shown to stimulate fibroblasts activities (in vitro). Finally, a preliminary in vivo test was performed to evaluate the adhesiveness of the membrane on porcine intestine (serosa). Overall, this functionalized membrane was shown to be biocompatible and to possess considerable adhesive properties owing to the presence of dopamine residues grafted on the alginate backbone. Statement of Significance This article describes the development of a mussels-inspired strategy for the development of an adhesive polysaccharide-based membrane for wound healing applications. Bioadhesion was achieved by grafting dopamine moieties on the structural component on the membrane (alginate): this novel biomaterial showed improved adhesiveness to the intestinal tissue, which was demonstrated by both in vitro and in vivo studies. Overall, this study points out how this nature-inspired strategy may be successfully exploited for the development of novel engineered biomaterials with enhanced bioadhesion, thus opening for novel applications in the field of general surgery.

Alginate-based membranes containing hyaluronic acid (HA) were manufactured by freeze-drying calcium-reticulated hydrogels. The study of the distribution of the two macromolecules within the hydrogel enabled to highlight a polymer demixing mechanism that tends to segregate HA in the external parts of the constructs. Resistance and pliability of the membranes were tuned, while release and degradation studies enabled to quantify the diffusion of both polysaccharides in physiological solution and to measure the viable lifetime of the membranes. Biological studies in vitro proved that the liquid extracts from the HA-containing membranes stimulate wound healing and that fibroblasts are able to colonize the membranes. Overall, such novel alginate-HA membranes represent a promising solution for several medical needs, in particular for wound treatment, giving the possibility to provide an in situ administration of HA from a resorbable device.

Recent evidence has led to the hypothesis that dissipation of energy through the viscoelastic extracellular matrix (ECM) can play a cardinal role in directing cell-fate decisions, but whether and how it correlates with specific cell response is at present unclear. Here, viscoelastic and plastic 2D chitosan-based substrates endowed with different dissipative energies are developed and cell behavior studied in terms of adhesion and spreading. While keeping constant stress relaxation and systematically decoupling overall stiffness from linear elongation, an energy dissipation term (J mol−1) is introduced, that is the molar energy required to deviate from linear stress–strain regime and enter into plastic region. Strikingly, an inverse relationship is unveiled between substrate dissipation energy and cell response, with high adhesion/high spreading and low adhesion/no spreading detected for substrates at low and high dissipation energy, respectively. It is concluded that cells decide how to react depending on the effective energy they can earmark for their functions.

Implantable membranes based on alginate and hyaluronic acid (HA) were manufactured to obtain a rapidly resorbing pliable mesh for the in situ administration of HA to intestinal tissue. Morphological analyses of this interpenetrated matrix pointed out a homogeneous polymeric texture while degradation studies demonstrated that the material is able to dissolve in physiological solutions within few days. Biological studies in vitro showed that the membrane is biocompatible towards human dermal fibroblasts and that liquid extracts from the HA-containing membrane can stimulate wound healing. A preliminary in vivo biocompatibility study on rats showed that the membranes in direct contact with the intestine did not elicit any acute adverse reaction or immune response, while only a mild inflammatory reaction was noticed at the mesenteric or serosal region. Overall, these results appear to support the application of these polysaccharide-based materials for intestinal wound healing.[GRAPHICS].

The aim of the present contribution is twofold as it reports (i) on the role played by chitosan acetylation degree for the stability of nanoparticles (NPs) formed with hyaluronan and (ii) on the effect of the interaction of such NPs with immune cells. Chitosans with similar viscosity-average molecular weight, Mv, (i.e., 200 000) and different fractions of acetylated units (F A ) together with low-molecular-weight hyaluronan were chosen for developing a select library of formulations via electrostatic complex coacervation. The resulting NPs were analyzed in terms of size, polydispersity, surface charge, and stability in physiological-mimicked media by dynamic light scattering. Only medium acetylated chitosan (F A = 0.16) guaranteed the stability of NPs. To explore the effect of NPs interaction with immune cells, the release of proinflammatory cytokines and the reactive oxygen species production by human macrophages and neutrophils, respectively, were evaluated. Strikingly, a structure-function relationship emerged, showing that NPs made of chitosans with F A = 0.02, 0.25, 0.46, and 0.63 manifested a proinflammatory activity, linked to the instability of the system. Conversely, NPs made of chitosan with F A = 0.16 neither modified the functional response of macrophages nor that of neutrophils. Of note, such NPs were found to possess additional properties potentially advantageous in applications such as delivery of therapeutics to target inflamed sites: (i) they are devoid of cytotoxic effects, (ii) they avoid engulfment during the early stage of interaction with macrophages, and (iii) they are muco-adhesive, thereby providing for site-specificity and long-residence effects

L’invenzione concerne un procedimento per la preparazione di idrogeli di derivati oligosaccaridici del chitosano e di acido borico, che permette di formare idrogeli omogenei evitando la formazione di precipitati. Sotto un altro aspetto viene descritto un idrogelo ottenibile da tale procedimento. Vengono altresì descritti gli usi degli idrogeli in campo biomedico e farmaceutico, nelle applicazioni di viscosupplementazione, come sostituto della matrice extracellulare e come dispositivo di somministrazione di farmaci.

Galectins (Gal) are a family of glycan-binding proteins characterized by their affinity for β-galactosides. Galectin-1 (Gal-1), a dimeric lectin with two galactoside-binding sites, regulates cancer progression and immune responses. Coordination chemistry has been engaged to develop versatile multivalent neoglycoconjugates for binding Gal-1. In this study we report a fast and original method to synthesize hybrid gold nanoparticles in which a hydrochloride lactose-modified chitosan, named CTL, is mixed with dicarboxylic acid-terminated polyethylene glycol (PEG), leading to shell-like hybrid polymer-sugar-metal nanoparticles (CTL-PEG-AuNPs). The aim of this paper is to preliminarily study the interaction of the CTL-PEG-AuNPs with a target protein, namely, Gal-1, under specific conditions. The molecular interaction has been measured by Transmission Electron Microscopy (TEM), UV-vis, and Raman Spectroscopy on a large range of Gal-1 concentrations (from 0 to 10-12 M). We observed that the interaction was strongly dependent on the Gal-1 concentration at the surface of the gold nanoparticles.

Alginate/hydroxyapatite composites, with isotropic or anisotropic porosity, were developed using two different freezing methods. Morphological analysis highlighted the differences in their three-dimensional network. Swelling and in vitro degradation studies revealed that both structures are highly hydrophilic and stable. Static compression tests showed that isotropic scaffolds possess higher modulus and toughness than anisotropic ones. Cyclic compression tests revealed that wet scaffolds are able to withstand high deformations with a peculiar shape-recovery behaviour. In vitro biological tests showed that osteoblast-like cells proliferate on both types of structures in a comparable manner.

The main functions of biological adhesives and sealants are to repair injured tissues, reinforce surgical wounds, or even replace common suturing techniques. In general surgery, adhesives must match several requirements taking into account clinical needs, biological effects, and material features; these requirements can be fulfilled by specific polymers. Natural or synthetic polymeric materials can be employed to generate three-dimensional networks that physically or chemically bind to the target tissues and act as hemostats, sealants, or adhesives. Among them, fibrin, gelatin, dextran, chitosan, cyanoacrylates, polyethylene glycol, and polyurethanes are the most important components of these interfaces; various aspects regarding their adhesion mechanisms, mechanical performance, and resistance to body fluids should be taken into account to choose the most suitable formulation for the target application. This review aims to describe the main adhesives and sealant materials for general surgery applications developed in the past decades and to highlight the most important aspects for the development of future formulations.

Chronic non-healing wounds are a clinically important problem in terms of number of patients and costs. Wound dressings such as hydrogels, hydrocolloids, polyurethane films and foams are commonly used to manage these wounds since they tend to maintain a moist environment which is shown to accelerate re-epithelialization. The use of antibacterial compounds is important in the management of wound infections. A novel wound-dressing material based on a blended matrix of the polysaccharides alginate, hyaluronic acid and Chitlac-silver nanoparticles is here proposed and its application for wound healing is examined. The manufacturing approach to obtain membranes is based on gelling, foaming and freeze-casting of alginate, hyaluronic acid and Chitlac-silver nanoparticles mixtures using calcium ions as the cross-linking agent. Comprehensive evaluations of the morphology, swelling kinetics, permeability, mechanical characteristics, cytotoxicity, capability to inhibit metalloproteinases and of antibacterial property were conducted. Biological in vitro studies demonstrated that hyaluronic acid released by the membrane is able to stimulate the wound healing meanwhile the metal silver exploits an efficient antibacterial activity against both planktonic bacteria and biofilms. Overall, the experimental data evidence that the studied material could be used as antibacterial wound dressing for wound healing promotion.

The effect of transient cross-links has been explored on a lactose-modified chitosan, which previously had shown interesting biological features. The presence of galactose side chains and of the polyol spacer resulted particularly appealing for the reticulation by borate ions. The interaction between chitlac and borax was investigated by means of 11B NMR while rheology pointed to a marked non-linear behavior depending on the amount of borax added to the system. The presence of limited amount of cross-linking ion led to dilatant behavior when the steady flow curve was measured. In addition, strain stiffening was noticed on elastic response upon exceeding a critical stress, indicating a transient nature in the formation of the cross-links. The non-linear response of chitlac in the presence of borax compared surprisingly well with the one showed by proteins composing the natural ECM pointing at a possible role of mechanotransduction in the biological significance of the modified chitosan.

Objective The present study evaluated the influence of time, mass and surface area of demineralized dentin collagen matrices on telopeptides release. The hypotheses tested were that the rates of ICTP and CTX release by matrix bound endogenous proteases are 1) not time-dependent, 2) unrelated to specimen mass, 3) unrelated to specimen surface area. Methods Non-carious human molars (N = 24) were collected and randomly assigned to three groups. Dentin slabs with three different thicknesses: 0.37 mm, 0.75 mm, and 1.50 mm were completely demineralized and stored in artificial saliva for one week. Collagen degradation was evaluated by sampling storage media for ICTP and CTX telopeptidases. Activity of MMPs in the aging medium was evaluated using fluorometric activity assay kit. Results A statistically significant (p < 0.05) decrease in the release of both ICTP and CTX fragments over time was observed irrespective of the specimen thickness. When data were normalized by the specimen mass, no significant differences were observed. Releases of ICTP and CTX were significantly related to the aging time as a function of surface area for the first 12 h. Total MMP activity, mainly related to MMP-2 and -9, decreased with time (p < 0.05). Significance Because the release of collagen fragments was influenced by specimen storage time and surface area, it is likely that cleaved collagen fragments closer to the specimen surface diffuse into the incubation medium; those further away from the exposed surface are still entrapped within the demineralized dentin matrix. Bound MMPs can only degrade the substrate within the limited zone of their molecular mobility.