Alginate Dialdehyde Research Articles

Cell specificity of magnetic cell seeding approach to hydrogel colonization.

Tissue-engineered scaffolds require an effective colonization with cells. Superparamagnetic iron oxide nanoparticles (SPIONs) can enhance cell adhesion to matrices by magnetic cell seeding. We investigated the possibility of improving cell attachment and growth on different alginate-based hydrogels using fibroblasts and endothelial cells (ECs) loaded with SPIONs. Hydrogels containing pure alginate (Alg), alginate dialdehyde crosslinked with gelatin (ADA-G) and Alg blended with G or silk fibroin (SF) were prepared. Endothelial cells and fibroblasts loaded with SPIONs were seeded and grown on hydrogels for up to 7 days, in the presence of magnetic field during the first 24 h. Cell morphology (fluorescent staining) and metabolic activity (WST-8 assay) of magnetically-seeded versus conventionally seeded cells were compared. Magnetic seeding of ECs improved their initial attachment and further growth on Alg/G hydrogel surfaces. However, we did not achieve an efficient and stable colonization of ADA-G films with ECs even with magnetic cell seeding. Fibroblast showed good initial colonization and growth on ADA-G and on Alg/SF. This effect was further significantly enhanced by magnetic cell seeding. On pure Alg, initial attachment and spreading of magnetically-seeded cells was dramatically improved compared to conventionally-seeded cells, but the effect was transient and diminished gradually with the cessation of magnetic force. Our results demonstrate that magnetic seeding improves the strength and uniformity of initial cell attachment to hydrogel surface in cell-specific manner, which may play a decisive role for the outcome in tissue engineering applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2948-2956, 2017.

Insights into the rheological behaviors evolution of alginate dialdehyde crosslinked collagen solutions evaluated by numerical models

The elaboration of the rheological behaviors of alginate dialdehyde (ADA) crosslinked collagen solutions, along with the quantitative analysis via numerical models contribute to the controllable design of ADA crosslinked solution system's fluid mechanics performance during manufacturing process for collagen biomaterials. In the present work, steady shear flow, dynamical viscoelasticity, creep-recovery, thixotropy tests were performed to characterize the rheological behaviors of the collagen solutions incorporating of ADA from the different aspects and fitted with corresponding numerical models. It was found that pseudoplastic properties of all samples enhanced with increasing amounts of ADA, which was confirmed by the parameters calculated from the Ostwald-de Waele model, Carreau and Cross model, for instance, the non-Newtonian index (n) decreased from 0.786 to 0.201 and a great increase by 280 times in value of viscosity index (K) was obtained from Ostwald-de Waele model. The forth-mode Leonov model was selected to fit all dynamic modulus-frequency curves due to its higher fitting precision (R2>0.99). It could be found that the values of correlation shear viscosity (ηk) increased and the values of relaxation time (θk) decreased with increasing ADA at the fixed k value, suggesting that the incorporation of ADA accelerated the transformation of the collagen solutions from liquid-like to gel-like state due to more formation of CN linkages between aldehyde groups and lysine residues. Also, the curves of creep and recovery phase of the native and crosslinked collagen solutions were simulated well using Burger model and a semi-empirical model, respectively. The ability to resist to deformation and elasticity strengthened for the samples with higher amounts of ADA, accompanied with the important fact that compliance value (J50) decreased from 56.317Pa−1 to 2.135Pa−1 and the recovery percentage (Rcreep) increased from 2.651% to 28.217%. Finally, it was found that the area of thixotropic loop increased from 8.942Pa/s to 17.823Pa/s with increasing introduction of ADA, suggesting that stronger thixotropic behavior was associated with higher amount of ADA. Furthermore, Herschel–Bulkley model was employed to describe the up and down curves of all samples and it was confirmed that all collagen solutions belonged to pseudoplastic fluid (the flow index<1) without apparent yield stress and shear-thinning behaviors were more obvious with increasing additions of ADA according to the increasing consistency coefficient K values. Overall, this work contributed a new insight into the interactions between collagen and ADA based on quantitative rheological methods reflecting the different rheological properties and the results obtained should be of great utility in the extensive application of ADA crosslinked collagen solutions into diverse collagen-based materials.

Fabrication of Cell-Loaded Two-Phase 3D Constructs for Tissue Engineering.

Hydrogel optimisation for biofabrication considering shape stability/mechanical properties and cell response is challenging. One approach to tackle this issue is to combine different additive manufacturing techniques, e.g., hot-melt extruded thermoplastics together with bioplotted cell loaded hydrogels in a sequential plotting process. This method enables the fabrication of 3D constructs mechanically supported by the thermoplastic structure and biologically functionalised by the hydrogel phase. In this study, polycaprolactone (PCL) and polyethylene glycol (PEG) blend (PCL-PEG) together with alginate dialdehyde gelatine hydrogel (ADA-GEL) loaded with stromal cell line (ST2) were investigated. PCL-PEG blends were evaluated concerning plotting properties to fabricate 3D scaffolds, namely miscibility, wetting behaviour and in terms of cell response. Scaffolds were characterised considering pore size, porosity, strut width, degradation behaviour and mechanical stability. Blends showed improved hydrophilicity and cell response with PEG blending increasing the degradation and decreasing the mechanical properties of the scaffolds. Hybrid constructs with PCL-PEG blend and ADA-GEL were fabricated. Cell viability, distribution, morphology and interaction of cells with the support structure were analysed. Increased degradation of the thermoplastic support structure and proliferation of the cells not only in the hydrogel, but also on the thermoplastic phase, indicates the potential of this novel material combination for biofabricating 3D tissue engineering scaffolds.

Dynamic release of gentamicin sulfate (GS) from alginate dialdehyde (AD)-crosslinked casein (CAS) films for antimicrobial applications

In the present work, antibiotic drug gentamicin sulfate (GS) has been loaded into alginate dialdehyde-crosslinked casein (CAS) films for wound dressing applications. The films have been characterized by Fourier transform infrared spectroscopy, X-ray diffraction analysis and scanning electron microscopy. The dynamic release of model drug GS has been investigated in the physiological fluid at 37 °C. The drug release data has been interpreted in the terms of various kinetic models such as Power function model, first order model and Schott model. The release data was found to be well fitted by Schott model. The various diffusion coefficients are also evaluated. The adsorption of model therapeutic protein BSA on the film has been investigated. The maximum adsorption is found to be 5.7 mg/cm2.The films were tested for their antibacterial and anti-fungal action. Finally, the in vivo wound healing study was carried out on Albino wistar rats.

One-Pot Synthesis of Diphenylalanine-Based Hybrid Nanospheres for Controllable pH- and GSH-Responsive Delivery of Drugs

Nano drug delivery systems have attracted much research and clinical interest but they remain challenging with respect to developing controllable, nontoxic, biocompatible and biodegradable nanocarriers. Herein, we report the first pH- and glutathione (GSH)-responsive diphenylalanine (FF)-based hybrid spheres using natural alginate dialdehyde (ADA) as the cross-linker to induce self-assembly of FF and an in situ reducer of Au3+ ions into Au nanoparticles (Au NPs). Not only is the synthesis simple and high-yielding, but also these biocompatible hybrid spheres can encapsulate more than 95% of hydrophobic drug (camptothecin, CPT). Moreover, the CPT-loaded carriers are stable under normal physiological environments, and have excellent pH- and GSH-responsive release at pH 5.0 with 10 mM GSH, which is similar to the tumor microenvironment. Also, these nanocarriers can be taken up by cancer cells and have greater cytotoxicity than free drugs. These attributes make nanospheres very promising for drug loading and d...

Reactive electrospinning of composite nanofibers of carboxymethyl chitosan cross-linked by alginate dialdehyde with the aid of polyethylene oxide

We have prepared carboxymethyl chitosan-alginate dialdehyde (CMCS-ADA) nanofibers via a reactive electrospinning process with the aid of polyethylene oxide (PEO). The presence of PEO delayed the gelation of CMCS and ADA, thus providing ease of use to adjust the mixing of CMCS-PEO and ADA-PEO blended solution. The mixed solution can be adjusted to come out from the needle before the gel formation or when the gel was just about to form. Defect-free CMCS-ADA-PEO nanofibers with average diameters ranging from 100nm to 900nm were obtained using water as a solvent. The in situ cross-linked CMCS-ADA nanofibers were then obtained following the extraction of water-soluble PEO. After immersion in phosphate-buffered saline (PBS) at a pH of 7.4 for up to 15days, the as-spun CMCS-ADA-PEO composite nanofibers maintained structural integrity, confirming the success of the crosslinking. The PEO-extracted CMCS-ADA nanofibers promoted the adhesion, proliferation and alkaline phosphatase activity of bone marrow stromal cells.

Alginate dialdehyde (AD)-crosslinked casein films: synthesis, characterization and water absorption behavior

In this work, for the first time, alginate dialdehyde (AD) has been used as a cross linker to prepare casein films. The films, synthesized, have been characterized by Fourier transform infrared spectroscopy, X-ray diffraction analysis and scanning electron microscopy and thermogravimetric analysis. The films were investigated for their water absorption capacity in the physiological fluid of pH 7.4 at 37 °C. The film samples (AD-X-CAS)12, (AD-X-CAS)24 and (AD-X-CAS)36 demonstrated an equilibrium water absorption of 30.86, 26.15 and 11.93 g/g, respectively. Moreover, the swelling exponents for the films were in the range of 0.66–0.75, thus indicating non-Fickian water transport mechanism. The dynamic water uptake data were analysed by various kinetic models. The water vapour transmission rates (WVTR) for the three film samples were found to be 16587, 15900 and 14316 g/m2/day, respectively. The higher values of WVTR, obtained for all the three samples, indicated their suitability for high exudating wounds. The results of expansion study in 4% gelatin medium revealed that the three samples exhibited an almost 2.54, 2.09 and 1.90-fold increase in their diameter in 90, 75 and 60 min, respectively.

Evaluation of alginate dialdehyde as a suitable crosslinker on modifying porcine acellular dermal matrix: The aggregation of collagenous fibers

ABSTRACTPorcine acellular dermal matrix (PADM) has been investigated widely as a natural biomaterial. However, the success of PADM has been limited by insufficient stability and poor physicochemical properties. In our work, alginate dialdehyde (ADA) with various oxidation degrees (OD = 25%, 45%, and 65%) were explored to modify PADM, while glutaraldehyde (GA), dehydrathermal treatment (DHT), and carbodiimide (EDC) were used as the control. The efficacy of ADA on modifying PADM increased along with the rising of oxidation degree. The ADA (OD = 65% and 45%) groups showed better mechanical and thermal stability, crosslink density, and resistance to enzymatic degradation than ADA (OD = 25%) and DHT + EDC group. Meanwhile, the structure of PADM crosslinked by ADA (OD = 45% and 65%) were maintained largely. Further, ADA (OD = 45%) group revealed better cytocompatibility than DHT + EDC, ADA (OD = 65%) and GA group. Considering the balance of cytocompatibility and physicochemical behavior, ADA (OD = 45%) was more suitable as a natural derived crosslinker to modify PADM in tissue engineering. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 43550.

Effects of pH on the alginate dialdehyde (ADA)-crosslinking of natural biological tissues and in vitro study of the endothelial cell compatibility of ADA-crosslinked biological tissues

Investigating the optimal pH-environment for ADA fixation and the HUVECs compatibility of the ADA-fixed biological vascular scaffolds.

Modification of collagen–chitosan matrix by the natural crosslinker alginate dialdehyde

In the present study, collagen (Coll) was mixed with the natural crosslinker chitosan (CTS), and then, alginate dialdehyde (ADA) was added to crosslink the mixtures. The properties of these Coll matrix sponges were investigated afterwards. Fourier transform infrared spectroscopy (FTIR) analysis and in vitro fiber formation analysis showed the intact retention of the classical triple-helical structure after crosslinking. Scanning electron microscopy (SEM) showed that microfibril structural interactions between Coll structures became more compact. Significant improvement in the thermostability of the crosslinked mixtures was observed with the pure mixtures of Coll and CTS. Antibacterial activity measurements indicated no affect of ADA on Coll/CTS sponges. In conclusion, the modification of the Coll/CTS mixtures with ADA preserves the classical triple-helical structure, enhances stabilization, maintains good biocompatibility and may pave the way for new medical applications.

Interactions between Chitosan and Alginate Dialdehyde Biopolymers and Their Layer-by-Layer Assemblies.

Biopolymers are researched extensively for their applications in biomaterials science and drug delivery including structures and complexes of more than one polymer. Chemical characterization of complexes formed between chitosan (CHI) and alginate dialdehyde (ADA) biopolymers established that while electrostatic interactions dominate (as determined from X-ray photoelectron spectroscopy (XPS)) covalent cross-linking between these biopolymers also contribute to their stability (evidenced from immersion in salt solution). It was furthermore found that imine bond formation could not be directly detected by any of the techniques XPS, FTIR, (1)H NMR, or fluorescence. The layer-by-layer assemblies of the biopolymers formed on silica colloids, glass slides, and alginate hydrogel beads were evaluated using XPS, as well as zeta potential measurements for the silica colloids and changes to hydration properties for the hydrogels. It was found that the degree of oxidation of ADA affected the LbL assemblies in terms of a greater degree of CHI penetration observed when using the more conformationally flexible biopolymer ADA (higher degree of oxidation).

Evaluation of an alginate–gelatine crosslinked hydrogel for bioplotting

Using additive manufacturing to create hydrogel scaffolds which incorporate homogeneously distributed, immobilized cells in the context of biofabrication approaches represents an emerging and expanding field in tissue engineering. Applying hydrogels for additive manufacturing must consider the material processing properties as well as their influence on the immobilized cells. In this work alginate-dialdehyde (ADA), a partially oxidized alginate, was used as a basic material to improve the physico-chemical properties of the hydrogel for cell immobilization. At first, the processing ability of the gel using a bioplotter and the compatibility of the process with MG-63 osteoblast like cells were investigated. The metabolic and mitochondrial activities increased at the beginning of the incubation period and they balanced at a relatively high level after 14–28 days of incubation. During this incubation period the release of vascular endothelial growth factor-A also increased. After 28 days of incubation the cell morphology showed a spreading morphology and cells were seen to move out of the scaffold struts covering the whole scaffold structure. The reproducible processing capability of alginate–gelatine (ADA–GEL) and the compatibility with MG-63 cells were proven, thus the ADA–GEL material is highlighted as a promising matrix for applications in biofabrication.

Preparation of core–shell magnetic polydopamine/alginate biocomposite for Candida rugosa lipase immobilization

A flexible, biocompatible and bioadhesive enzyme immobilizing material, which was synthesized based on the covalent assembly of biomimetic polymer and oxidized polysaccharide on magnetic nanoparticles (NPs), has been developed in this feasibility study. In this work, the bio-inspired polymer, polydopamine (PDA), was used to modify the well-monodispersed Fe3O4 NPs (mPDA NPs) with a controllable thickness via a dip-coating process, then the alginate di-aldehyde (ADA) was covalently assembled on the mPDA NPs and employed as a naturally occurring linking agent for Candida rugosa lipase (CRL) immobilization. The resulting support material was characterized by means of the transmission electron microscope (TEM), Fourier transform infrared spectra (FT-IR), X-ray diffraction (XRD), thermogravimetry (TG) analyser, and vibrating sample magnetometer (VSM). It was verified that the prepared mPDA NPs possessed distinct core–shell structure with uniform size and high saturation magnetization. For further application, the mPDA NPs was utilized in CRL immobilizing procedures and demonstrated can facilitate improving the enzyme activities. The optimum amount of lipase was 200mgg−1 support, the optimal pH and temperature for the catalyse condition of the immobilized CRL was 7.0 and 40°C, respectively. Moreover, the immobilized CRL kept the high activity at 77% after 12 times of recycling for batch hydrolysis of olive oil emulsion. This magnetic bioadhesive composite with functionalized properties and adhesion strength presents a general strategy for the immobilization of macromolecules.

Evaluation of fibroblasts adhesion and proliferation on alginate-gelatin crosslinked hydrogel.

Due to the relatively poor cell-material interaction of alginate hydrogel, alginate-gelatin crosslinked (ADA-GEL) hydrogel was synthesized through covalent crosslinking of alginate di-aldehyde (ADA) with gelatin that supported cell attachment, spreading and proliferation. This study highlights the evaluation of the physico-chemical properties of synthesized ADA-GEL hydrogels of different compositions compared to alginate in the form of films. Moreover, in vitro cell-material interaction on ADA-GEL hydrogels of different compositions compared to alginate was investigated by using normal human dermal fibroblasts. Viability, attachment, spreading and proliferation of fibroblasts were significantly increased on ADA-GEL hydrogels compared to alginate. Moreover, in vitro cytocompatibility of ADA-GEL hydrogels was found to be increased with increasing gelatin content. These findings indicate that ADA-GEL hydrogel is a promising material for the biomedical applications in tissue-engineering and regeneration.

In vitro and in vivo Biocompatibility of Alginate Dialdehyde/Gelatin Hydrogels with and without Nanoscaled Bioactive Glass for Bone Tissue Engineering Applications.

In addition to good mechanical properties needed for three-dimensional tissue engineering, the combination of alginate dialdehyde, gelatin and nano-scaled bioactive glass (45S5) is supposed to combine excellent cellular adhesion, proliferation and differentiation properties, good biocompatibility and predictable degradation rates. The goal of this study was to evaluate thein vitro and in vivo biocompatibility as a first step on the way to its use as a scaffold in bone tissue engineering. In vitro evaluation showed good cell adherence and proliferation of bone marrow derived mesenchymal stem cells seeded on covalently crosslinked alginate dialdehyde-gelatin (ADA-GEL) hydrogel films with and without 0.1% nano-Bioglass®(nBG). Lactate dehydrogenase (LDH)- and mitochondrial activity significantly increased in both ADA-GEL and ADA-GEL-nBG groups compared to alginate. However, addition of 0.1% nBG seemed to have slight cytotoxic effect compared to ADA-GEL. In vivo implantation did not produce a significant inflammatory reaction, and ongoing degradation could be seen after four weeks. Ongoing vascularization was detected after four weeks. The good biocompatibility encourages future studies using ADA-GEL and nBG for bone tissue engineering application.

Fabrication of alginate-gelatin crosslinked hydrogel microcapsules and evaluation of the microstructure and physico-chemical properties.

Microencapsulation of cells by using biodegradable hydrogels offers numerous attractive features for a variety of biomedical applications including tissue engineering. This study highlights the fabrication of microcapsules from an alginate-gelatin crosslinked hydrogel (ADA-GEL) and presents the evaluation of the physico-chemical properties of the new microcapsules which are relevant for designing suitable microcapsules for tissue engineering. Alginate di-aldehyde (ADA) was synthesized by periodate oxidation of alginate which facilitates crosslinking with gelatin through Schiff's base formation between the free amino groups of gelatin and the available aldehyde groups of ADA. Formation of Schiff's base in ADA-GEL and aldehyde groups in ADA was confirmed by FTIR and NMR spectroscopy, respectively. Thermal degradation behavior of films and microcapsules fabricated from alginate, ADA and ADA-GEL was dependent on the hydrogel composition. The gelation time of ADA-GEL was found to decrease with increasing gelatin content. The swelling ratio of ADA-GEL microcapsules of all compositions was significantly decreased, whereas the degradability was found to increase with the increase of gelatin ratio. The surface morphology of the ADA-GEL microcapsules was totally different from that of alginate and ADA microcapsules, observed by SEM. Two different buffer solutions (with and without calcium salt) have an influence on the stability of microcapsules which had a significant effect on the gelatin release profile of ADA-GEL microcapsules in these two buffer solutions.

Modification of collagen with a natural derived cross-linker, alginate dialdehyde

The interaction between collagen and a natural derived cross-linker alginate dialdehyde (ADA) was investigated. Fourier transform infrared (FTIR) spectroscopy and the circular dichroism (CD) measurements indicate that the structure integrity of collagen is still maintained after the ADA treatment, while the differential scanning calorimetry (DSC) study suggests that ADA could promote collagen–ADA membrane's thermostability compared to pure collagen. And the atomic force microscopy (AFM) of cross-linked collagen reveals a denser network structure. Besides, the water contact angle test indicates that the hydrophilic property of collagen–ADA membrane is promoted, which is favorable for cell's attachment and proliferation. Meanwhile, the cytocompatibility results imply that not only no extra cytotoxicity is introduced into the collagen–ADA membrane after ADA treatment, but also collagen–ADA membrane facilitates cell's proliferation when the content of ADA is less than 20%. In conclusion, our study reveals that ADA stabilizes collagen as a cross-linker and preserves its triple helical structure.

Assembled Microcapsules by Doxorubicin and Polysaccharide as High Effective Anticancer Drug Carriers

Doxorubicin, together with the modified polysaccharide (alginate dialdehyde), was used as a wall material to fabricate microcapsules through self-cross-linking by a template method. The microcapsules as-prepared are pH-responsive. Relevant scanning electronic microscopy, atom force microscopy and confocal laser scanning microscopy confirm the morphology of the uniform microcapsules. The spectroscopic results show that the microcapsules are assembled through electrostatic interaction and Schiff's base covalent bonding. Doxorubicin can be released sustainably from the capsules in buffer solution at a lower pH value. The cellular uptake of the microcapsules and drug release induced by acidic microenvironment are time-dependent processes. The cell cytotoxicity experiments in vitro demonstrate that the doxorubicin-based microcapsules have high efficiency to kill the cancer cells.

Preparation of chitosan/silk fibroin blending membrane fixed with alginate dialdehyde for wound dressing

The objective of this work was to prepare chitosan/silk fibroin (CS/SF) blending membranes crosslinked with alginate dialdehyde (ADA) as wound dressings and to evaluate the physical properties and biocompatibility of the membranes. The morphology of membrane was observed by scanning electron microscopy (SEM) which showed that the well consistency of these two compositions. Further, the stability, water absorption and water vapor permeability of the ADA fixed CS/SF membranes could meet the needs of wound dressing. Furthermore, the biocompatibility of ADA fixed membranes was investigated by MTT assays and SEM in vitro, and the membranes were found to promote the cell attachment and proliferation. These results suggest that ADA fixed CS/SF blending membranes with a suitable ratio could be a promising candidate for wound healing applications.

In vitro enzymatic degradation of a biological tissue fixed by alginate dialdehyde

Biological tissues must be chemically fixed before they can be implanted in humans as tissue engineering scaffolds. To provide an ideal tissue engineering scaffold material, which is biodegradable and cytocompatible, a novel crosslinking agent, alginate dialdehyde (ADA), was employed to fix biological tissues by our group. The study mainly investigated the enzymatic degradation of ADA fixed biological tissues in vitro. Glutaraldehyde, the most commonly used crosslinking agent for biological tissue fixation, was employed as a control. The results suggested that, the ADA fixation could enhance the resistance against enzymatic degradation of biological tissues effectively. Meanwhile, compared to glutaraldehyde-fixed tissues, the ADA-fixed tissues could also degrade gradually over time. Moreover, the ADA crosslinking reagent itself had a stimulatory effect on cell proliferation when at an appropriate concentration. The results obtained in this study demonstrate that ADA fixation might provide a successful example of the biodegradable scaffold materials in tissue engineering.