Gellan Gum Hydrogels Research Articles

Engineering retinal pigment epithelial cells regeneration for transplantation in regenerative medicine using PEG/Gellan gum hydrogels.

Retinal pigment epithelium (RPE) plays an important role in maintaining normal function and visual function of the retina, and the degeneration of RPE causes various retinal degenerative diseases. Currently, there is a lack of effective treatment for this, and it is being studied to produce a suitable scaffold for cell transplantation. In this experiment, Polyethylene glycol (PEG)/Gellan Gum (GG) hydrogel was prepared by adding harmless PEG and gellan gum, which is a biocompatible, degradable and widely used in modern tissue engineering. PEG/GG hydrogel was prepared with 0, 1, 3, 5 wt% PEG/GG according to the concentration of PEG, and ARPE-19 cells were used to confirm the cell attachment environment. As a result, it showed superior biocompatibility (>90%), cell adhesion and improved cell growth compared to gellan gum hydrogel. In addition, RT-PCR was used to confirm RPE-specific gene expression, and the result showed that it was positively influenced. As a result, it was observed that PEG/GG hydrogel promotes retinal regeneration compared to pure gellan gum. 3 wt% PEG/GG could be used as an alternative for retinal regeneration.

How Cross-Linking Mechanisms of Methacrylated Gellan Gum Hydrogels Alter Macrophage Phenotype.

In tissue engineering scaffolds, macrophages play a critical role in determining the host response to implanted biomaterials. Macrophage phenotype is dynamic throughout the host response, and a balance of phenotypes is essential for timely progression from injury to proper wound healing. Therefore, it is important to predict how materials will modulate the response of macrophages. In this study, we investigated the effect of methacrylated gellan gum (GG) hydrogels on macrophage phenotype and proliferation with the ultimate goal of improving rational design of biomedical implants. Naïve, along with classically and alternatively activated RAW 264.7 macrophages were seeded on methacrylated gellan gum hydrogels that were fabricated with different thiol-ene ratios and cross-linking mechanisms. Live/dead assays showed that all hydrogels supported cell attachment and proliferation. Stiffer substrates enhanced anti-inflammatory production of nitrites from both naïve and classically activated macrophages compared to the softer substrates. Moreover, arginine and CD206 expression-markers for alternatively activated macrophages-were inhibited by higher thiol content. Introducing ionic cross-links using calcium did not influence the proliferation or polarization for any of the three macrophage phenotypes. Our results suggest that the macrophage phenotype shift from M1 to M2 is controlled by the different cross-linking mechanisms, physical properties, and the chemistry of methacrylated gellan gum hydrogels.

Bone Marrow-Derived Mesenchymal Stem Cells Encapsulated in Functionalized Gellan Gum/Collagen Hydrogel for Effective Vascularization.

Gellan gum hydrogel holds great potential in tissue engineering, but the high phase transition temperature greatly inhibits the applications in biomedical field. In this study, gellan gum was modified with methacrylic anhydride, and then the phase transition temperature was reduced. The functionalized gellan gum together with type I collagen was gelled by ion/photo dual-cross-linking for fabricating bone marrow-derived mesenchymal stem cells (BMSCs)-encapsulating hydrogel for vascularization. After the ratio between gellan gum and collagen was optimized, the hydrogel with proper pore size and mechanical properties was prepared. The WST assay demonstrated that the hydrogel could offer excellent microenvironment for cell survival and proliferation. Finally, real-time quantitative polymerase chain reaction suggests that the hydrogel could promote BMSCs to differentiate into endothelial cells. Together, this work provides a general strategy for fabricating BMSCs-encapsulating hydrogel in one step, which has the potential for 3D-printing live cell scaffold for study of vasculogenic differentiation.

Preparation and Characterisation of Gellan Gum Hydrogel containing Curcumin and Limonene

Success in wound treatment requires careful assessment on the wound and suitable wound dressing. In this project a novel incorporation of both curcumin and limonene into gellan gum hydrogel have been done. This system is believed to have a better permeation of curcumin to the target area. The samples prepared are denoted as GGTF for gellan gum thin film, Cur-GGTF for curcumin gellan gum thin film. Samples of Cur-GGTF incorporated with limonene with three different concentrations of 0.0005 M, 0.0006 M and 0.0007 M are known as CurLim-GGTF05, CurLim-GGTF06 and CurLim-GGTF07, respectively. The hydrogels were crosslinked by calcium ion. From the FTIR analysis, all of the expected functional groups from the gellan gum, curcumin and limonene were found present in the hydrogel samples. The TGA-DTG analysis has proven that the incorporation of limonene into hydrogels has decreased the thermal stability of the hydrogels. The addition of limonene has caused a hike in the gel fractions of the hydrogels. The water vapour transmission rate (WVTR) and swelling test showed that the introductions of high concentrations of limonene into the hydrogels have hampered the absorption of water molecules thus causing the swelling degree to decrease.

Chemically Modified Gellan Gum Hydrogels with Tunable Properties for Use as Tissue Engineering Scaffolds.

Gellan gum is a naturally occurring polymer that can cross-link in the presence of divalent cations to form biocompatible hydrogels. However, physically cross-linked gellan gum hydrogels lose their stability under physiological conditions, thus restricting the applications of these hydrogels in vivo. To improve the mechanical strength of the gels, we incorporated methacrylate into the gellan gum and chemically cross-linked the hydrogel through three polymerization methods: step growth through thiol–ene photoclick chemistry, chain-growth via photopolymerization, and mixed model in which both mechanisms were employed. Methacrylation was confirmed and quantified by proton nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy. The mechanical properties and chemistry of the cross-linked gels were systematically altered by varying the reaction conditions. The compression moduli of the resulting hydrogels ranged between 6.4 and 17.2 kPa. The swelling ratios of the hydrogels were correlated with the compression moduli and affected by the addition of calcium. In vitro enzymatic degradation rate was found to depend on the degree of methacrylation. NIH/3T3 fibroblast cell proliferation and morphology were related to substrate stiffness, with a high stiffness leading generally to higher proliferation. The proliferation is further affected by the thiol–ene ratio. These results suggest that a hydrogel platform based on the gellan gum can offer versatile chemical modifications and tunable mechanical properties. The influence of these substrates on cell behavior suggests that the gellan gum hydrogels have the flexibility to be engineered for a variety of biomaterials applications.

Mineralization of gellan gum hydrogels with calcium and magnesium carbonates by alternate soaking in solutions of calcium/magnesium and carbonate ion solutions.

Mineralization of hydrogels is desirable prior to applications in bone regeneration. CaCO3 is a widely used bone regeneration material, and Mg, when used as a component of calcium phosphate biomaterials, has promoted bone-forming cell adhesion and proliferation and bone regeneration. In this study, gellan gum hydrogels were mineralized with carbonates containing different amounts of calcium (Ca) and magnesium (Mg) by alternate soaking in, firstly, a calcium and/or magnesium ion solution and, secondly, a carbonate ion solution. This alternate soaking cycle was repeated five times. Five different calcium and/or magnesium ion solutions, containing different molar ratios of Ca to Mg ranging from Mg free to Ca free were compared. Carbonate mineral formed in all sample groups subjected to the alternate soaking cycle. Ca : Mg elemental ratio in the mineral formed was higher than in the respective mineralizing solution. Mineral formed in the absence of Mg was predominantly CaCO3 in the form of a mixture of calcite and vaterite. Increasing the Mg content in the mineral formed led to the formation of magnesian calcite and decreased the total amount of the mineral formed and its crystallinity. Hydrogel mineralization and increasing Mg content in mineral formed did not obviously improve proliferation of MC3T3-E1 osteoblast-like cells or differentiation after 7days.

Generation of Gellan Gum-Based Adipose-Like Microtissues.

Adipose tissue is involved in many physiological processes. Therefore, the need for adipose tissue-like analogues either for soft tissue reconstruction or as in vitro testing platforms is undeniable. In this work, we explored the natural features of gellan gum (GG) to recreate injectable stable adipose-like microtissues. GG hydrogel particles with different percentages of polymer (0.5%, 0.75%, 1.25%) were developed and the effect of obtained mechanical properties over the ability of hASCs to differentiate towards the adipogenic lineage was evaluated based on the expression of the early (PPARγ) and late (FABP4) adipogenic markers, and on lipids formation and accumulation. Constructs were cultured in adipogenic induction medium up to 21 days or for six days in induction plus nine days in maintenance media. Overall, no significant differences were observed in terms of hASCs adipogenic differentiation within the range of Young’s moduli between 2.7 and 12.9 kPa. The long-term (up to six weeks) stability of the developed constructs supported its application in soft tissue reconstruction. Moreover, their ability to function as adipose-like microtissue models for drug screening was demonstrated by confirming its sensitivity to TNFα and ROCK inhibitor, respectively involved in the repression and induction of the adipogenic differentiation.

Evaluation of Saponin Loaded Gellan Gum Hydrogel Scaffold for Cartilage Regeneration

Several concentrations of saponin (Sa), which is water-soluble, were fabricated well with gellan gum (GG) solution. Sa, found in many plants like clematis and ginseng, has long been used in traditional chinese medicine to treat joint diseases including osteoarthritis. Also it has its ability of anti-inflammatory, antioxidant and anti-cancer. GG hydrogel is suitable as a cell encapsulating agent in the field of cartilage regeneration because of its easy processing and biocompatibility. In this study, GG hydrogel scaffolds with different concentration of Sa were conducted for analysis by SEM, FT-IR, compressive strength, water uptake, degradation rate, MTT assay, mRNA expression. It was observed that 0.025 wt% Sa/GG hydrogel scaffold shows good morphology, cell proliferation and mRNA expression results. The composite material supports cell growth covered with extracellular matrix (ECM) with maintaining its function. As a result, incorporation of Sa loaded with GG hydrogel scaffolds had positive result up to 0.025 wt% in cartilage regeneration.

Novel injectable gellan gum hydrogel composites incorporating Zn- and Sr-enriched bioactive glass microparticles: High-resolution X-ray microcomputed tomography, antibacterial and in vitro testing.

Mineralization of hydrogel biomaterials is desirable to improve their suitability as materials for bone regeneration. In this study, gellan gum (GG) hydrogels were formed by simple mixing of GG solution with bioactive glass microparticles of 45S5 composition, leading to hydrogel formation by ion release from the amorphous bioactive glass microparticles. This resulted in novel injectable, self-gelling composites of GG hydrogels containing 20% bioactive glass. Gelation occurred within 20min. Composites containing the standard 45S5 bioactive glass preparation were markedly less stiff. X-ray microcomputed tomography proved to be a highly sensitive technique capable of detecting microparticles of diameter approximately 8μm, that is, individual microparticles, and accurately visualizing the size distribution of bioactive glass microparticles and their aggregates, and their distribution in GG hydrogels. The widely used melt-derived 45S5 preparation served as a standard and was compared with a calcium-rich, sol-gel derived preparation (A2), as well as A2 enriched with zinc (A2Zn5) and strontium (A2Sr5). A2, A2Zn, and A2Sr bioactive glass particles were more homogeneously dispersed in GG hydrogels than 45S5. Composites containing all four bioactive glass preparations exhibited antibacterial activity against methicillin-resistant Staphylococcus aureus. Composites containing A2Zn5 and A2Sr5 bioactive glasses supported the adhesion and growth of osteoblast-like cells and were considerably more cytocompatible than 45S5. All composites underwent mineralization with calcium-deficient hydroxyapatite upon incubation in simulated body fluid. The extent of mineralization appeared to be greatest for composites containing A2Zn5 and 45S5. The results underline the importance of the choice of bioactive glass when preparing injectable, self-gelling composites.

Differentiation of osteoclast precursors on gellan gum-based spongy-like hydrogels for bone tissue engineering

Bone tissue engineering with cell-scaffold constructs has been attracting a lot of attention, in particular as a tool for the efficient guiding of new tissue formation. However, the majority of the current strategies used to evaluate novel biomaterials focus on osteoblasts and bone formation, while osteoclasts are often overlooked. Consequently, there is limited knowledge on the interaction between osteoclasts and biomaterials. In this study, the ability of spongy-like gellan gum and hydroxyapatite-reinforced gellan gum hydrogels to support osteoclastogenesis was investigated in vitro. First, the spongy-like gellan gum and hydroxyapatite-reinforced gellan gum hydrogels were characterized in terms of microstructure, water uptake and mechanical properties. Then, bone marrow cells isolated from the long bones of mice and cultured in spongy-like hydrogels were treated with 1,25-dihydroxyvitamin D3 to promote osteoclastogenesis. It was shown that the addition of HAp to spongy-like gellan gum hydrogels enables the formation of larger pores and thicker walls, promoting an increase in stiffness. Hydroxyapatite-reinforced spongy-like gellan gum hydrogels support the formation of the aggregates of tartrate-resistant acid phosphatase-stained cells and the expression of genes encoding DC-STAMP and Cathepsin K, suggesting the differentiation of bone marrow cells into pre-osteoclasts. The hydroxyapatite-reinforced spongy-like gellan gum hydrogels developed in this work show promise for future use in bone tissue scaffolding applications.

Rheological investigation of high-acyl gellan gum hydrogel and its mixtures with simulated body fluids.

Purpose Most of the studies concerning gellan have been focused on its application as a food ingredient, however, gellan is often considered as a candidate for the development of novel pharmaceutical formulations. Taking into account that gellan is ion-sensitive, it can be assumed that its initial mechanical properties can change upon contact with body secretions. Therefore, the aim of the work was to investigate the rheological properties of pure high-acyl gellan gum hydrogel (0.4%) and its mixtures with selected simulated body fluids. Methods The rheological investigations were performed on rotational rheometer and included oscillatory temperature, amplitude, and frequency sweeping. The results enabled estimation of the linear viscoelastic regime, calculation of the cross-over points, and percentage of structure recovery. Results In the case of pure hydrogel no evidence of thermosensitivity was observed in the range of 20-40°C. In pH = 1.2 (NaCl/HCl) the hydrogel structure was almost entirely destroyed. Mixing with phosphate buffer (pH = 4.5) resulted in higher gel strength than after dilution with deionized water. The opposite effect was observed in the case of pH = 7.4. The studies performed for the mixture of GG hydrogel and mucin indicated interaction between the components. The hydrogel elasticity increased in the presence of simulated tear, but decreased in simulated saliva and vaginal fluid. Conclusions In this study, it was shown that the stability of a three-dimensional gellan structure may be affected by pH and the presence of mucin which most probably competed with gellan gum in divalent cations binding. The observations presented in this study may be important in terms of potential application of gellan gum as a potential carrier in drug delivery systems.

Gellan Gum Hydrogels with Enzyme-Sensitive Biodegradation and Endothelial Cell Biorecognition Sites.

The survival of a biomaterial or tissue engineered construct is mainly hampered by the deficient microcirculation in its core, and limited nutrients and oxygen availability to the implanted or colonizing host cells. Aiming to address these issues, we herein propose bioresponsive gellan gum (GG) hydrogels that are biodegradable by metalloproteinase 1 (MMP-1) and enable endothelial cells adhesion and proliferation. GG is chemically functionalized with divinyl sulfone (DVS) and then biofunctionalized with thiol cell-adhesive peptides (T1 or C16) to confer GG endothelial cell biorecognition cues. Biodegradable hydrogels are then formed by Michael type addition of GGDVS or/and peptide-functionalized GGDVS with a dithiol peptide crosslinker sensitive to MMP-1. The mechanical properties (6 to 5580 Pa), swelling (17 to 11), MMP-1-driven degradation (up to 70%), and molecules diffusion coefficients of hydrogels are tuned by increasing the polymer amount and crosslinking density. Human umbilical cord vein endothelial cells depict a polarized elongated morphology when encapsulated within T1-containing hydrogels, in contrast to the round morphology observed in C16-containing hydrogels. Cell organization is favored as early as 1 d of cell culture within the T1-modified hydrogels with higher concentration of peptide, while cell proliferation is higher in T1-modified hydrogels with higher modulus. In conclusion, biodegradable and bioresponsive GGDVS hydrogels are promising endothelial cell responsive materials that can be used for vascularization strategies.

Phytase-mediated enzymatic mineralization of chitosan-enriched hydrogels

Hydrogels mineralized with calcium phosphate (CaP) are increasingly popular bone regeneration biomaterials. Mineralization can be achieved by phosphatase enzyme incorporation and incubation in calcium glycerophosphate (CaGP). Gellan gum (GG) hydrogels containing the enzyme phytase and chitosan oligomer were mineralized in CaGP solution and characterized with human osteoblast-like MG63 cells and adipose tissue-derived stem cells (ADSC). Phytase induced CaP formation. Chitosan concentration determined mineralization extent and hydrogel mechanical reinforcement. Phytase-induced mineralization promoted MG63 adhesion and proliferation, especially in the presence of chitosan, and was non-toxic to MG63 cells (with and without chitosan). ADSC adhesion and proliferation were poor without mineralization. Chitosan did not affect ADSC osteogenic differentiation.

Nanocellulose reinforced gellan-gum hydrogels as potential biological substitutes for annulus fibrosus tissue regeneration

Intervertebral disc (IVD) degeneration is associated with both structural damage and aging related degeneration. Annulus fibrosus (AF) defects such as annular tears, herniation and discectomy require novel tissue engineering strategies to functionally repair AF tissue. An ideal construct will repair the AF by providing physical and biological support, facilitating regeneration. The presented strategy herein proposes a gellan gum-based construct reinforced with cellulose nanocrystals (nCell) as a biological self-gelling AF substitute. Nanocomposite hydrogels were fabricated and characterized with respect to hydrogel swelling capacity, degradation rate in vitro and mechanical properties. Rheological evaluation on the nanocomposites demonstrated the GGMA reinforcement with nCell promoted matrix entanglement with higher scaffold stiffness observed upon ionic crosslinking. Compressive mechanical tests demonstrated compressive modulus values close to those of the human AF tissue. Furthermore, cell culture studies with encapsulated bovine AF cells indicated that nanocomposite constructs promoted cell viability and a physiologically relevant cell morphology for up to fourteen days in vitro.

Gellan gum–O,O′‐bis(2‐aminopropyl)‐polyethylene glycol hydrogel for controlled fertilizer release

ABSTRACTA gellan gum–Jeffamine superabsorbent hydrogel was obtained with different crosslink densities using different amounts of (1‐ethyl‐3‐(3‐dimethylaminopropyl)carbodiimide hydrochloride) and N‐hydroxysuccinimide. Infrared spectroscopy and thermal analysis confirm the crosslinking. A morphology analysis indicates denser structures for samples with higher crosslinking points. The swelling degree in high‐acyl gellan gum hydrogels was equivalent to 145 times their dry weight, and 77 times when low‐acyl gellan gum was used. Hydrogels also showed a 450 min water retention, as opposed to 280 min for pure water, evidencing good humidity control, suitable for use in arid climates. They also demonstrated a maximum release of commercial fertilizer of about 400 mg per gram for KH2PO4 and about 300 mg per gram for NPK 20‐5‐20. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018, 135, 45636.

Preparation and Characterisation of Carvacrol Encapsulated in Gellan Gum Hydrogel

Studies on plant materials as natural compound such as carvacrol (Carv) have gained much attention. Carv exhibits numerous potential as antimicrobial agent, food additives, antioxidant and etc. However, this free standing bioactive compound is unstable in the harsh environment conditions. Hence, the encapsulation technology provides protection to enhance the effectiveness in release manner. In this study, the preparation of Carv encapsulated in gellan gum hydrogel forming thin film (GG-Carv TF) was achieved by using 1.0 g of gellan gum at different concentrations of Carv (0.01-0.04 M). The FTIR spectra of GG-Carv TF revealed the combination of both functional groups from GG and Carv. The Carbon, Hydrogen and Nitrogen, CHN analysis further confirmed the encapsulation with the changes in the element percentage. Both swelling and degradation percentage increased with time and showed decreasing patterns in the range of 680.79-666.78 % and 26.83-19.15 % which can be observed as the concentration of Carv increased, respectively.

Enzymatic, urease-mediated mineralization of gellan gum hydrogel with calcium carbonate, magnesium-enriched calcium carbonate and magnesium carbonate for bone regeneration applications

Mineralization of hydrogel biomaterials is considered desirable to improve their suitability as materials for bone regeneration. Calcium carbonate (CaCO3 ) has been successfully applied as a bone regeneration material, but hydrogel-CaCO3 composites have received less attention. Magnesium (Mg) has been used as a component of calcium phosphate biomaterials to stimulate bone-forming cell adhesion and proliferation and bone regeneration in vivo, but its effect as a component of carbonate-based biomaterials remains uninvestigated. In the present study, gellan gum (GG) hydrogels were mineralized enzymatically with CaCO3 , Mg-enriched CaCO3 and magnesium carbonate to generate composite biomaterials for bone regeneration. Hydrogels loaded with the enzyme urease were mineralized by incubation in mineralization media containing urea and different ratios of calcium and magnesium ions. Increasing the magnesium concentration decreased mineral crystallinity. At low magnesium concentrations calcite was formed, while at higher concentrations magnesian calcite was formed. Hydromagnesite (Mg5 (CO3 )4 (OH)2 .4H2 O) formed at high magnesium concentration in the absence of calcium. The amount of mineral formed and compressive strength decreased with increasing magnesium concentration in the mineralization medium. The calcium:magnesium elemental ratio in the mineral formed was higher than in the respective mineralization media. Mineralization of hydrogels with calcite or magnesian calcite promoted adhesion and growth of osteoblast-like cells. Hydrogels mineralized with hydromagnesite displayed higher cytotoxicity. In conclusion, enzymatic mineralization of GG hydrogels with CaCO3 in the form of calcite successfully reinforced hydrogels and promoted osteoblast-like cell adhesion and growth, but magnesium enrichment had no definitive positive effect. Copyright © 2017 John Wiley & Sons, Ltd.

Design of nonionic micelle-laden polysaccharide hydrogels for controlled delivery of hydrophobic drugs

The incorporation of micelle self-assembly in hydrogels has been used to produce self-healing materials, materials with tunable mechanical properties, and hydrophilic or hydrophobic drug delivery systems. However, little is known about the connection among formulation – structure – properties (particularly transport) in these systems. This connection is explored in alkyl ethoxylate micelle-laden gellan gum hydrogels used as delivery system for the hydrophobic drug dexamethasone. Phase behavior maps and rheological characterization of the micelle-laden hydrogels indicate that their properties are largely dominated by the concentration and cloud point (CP) of the alkyl ethoxylate nonionic surfactant. The sol-gel temperature of the hybrid hydrogels was found to be close to the CP of the surfactant, and their storage modulus (G’) was found to increase with increasing surfactant concentration. A detailed analysis of the phase behavior maps, and evaluations of the mesh size of the hybrid hydrogels suggest a novel mechanism whereby micelles associate with gellan gum strands during high temperature hydration, hindering the double-helix assembly of the strands upon cooling. This increases the number of single strands and the density of physical cross-links, reducing the mesh size of the hydrogel. This mesh size reduction led to a decrease in the effective diffusion coefficient of micelles in the gel, and extended the release time of dexamethasone from 2h in surfactant-free hydrogel to more than 2days in the micelle-laden hydrogel.

Oxygen-Producing Gellan Gum Hydrogels for Dual Delivery of Either Oxygen or Peroxide with Doxorubicin.

Hypoxic environments in the core of tumors can give rise to resistance against anticancer therapeutics. Oxygen-producing biomaterials may be able to improve chemotherapeutic efficiency by locally disrupting the hypoxic environment. We hypothesized that gellan gum hydrogels could be loaded with both a solid peroxide and the chemotherapeutic drug doxorubicin, to release both oxygen and doxorubicin simultaneously. We show that calcium peroxide physically cross-links gellan gum into a hydrogel, which when loaded with catalase raises the dissolved oxygen content of media for up to 64 h. Additionally, doxorubicin could be loaded into the hydrogel in situ, allowing release in well-defined quantities.

* Synthesis and Characterization of Electroactive Gellan Gum Spongy-Like Hydrogels for Skeletal Muscle Tissue Engineering Applications.

Advances on materials' research for tissue engineering (TE) applications have shown that animal cells respond directly to the material physical, chemical, mechanical, and electrical stimuli altering a variety of cell signaling cascades, which consequently result in phenotypic and genotypic alterations. Gellan gum (GG) spongy-like hydrogels (SLH) with open microstructure, mechanical properties, and cell performance have shown promising results for soft TE applications. Taking advantage of intrinsic properties of GG-SLH and polypyrrole (PPy) electroactivity, we developed electroactive PPy-GG-SLH envisaging their potential use for skeletal muscle TE. Three different methods of in situ chemical oxidative polymerization were developed based on the availability of pyrrole: freely dissolved in solution (method I and III) or immobilized into GG hydrogels (method II). PPy was homogeneously distributed within (method I and III) and on the surface (method II) of GG-SLH, as also confirmed by Fourier Transform infrared spectra. PPy-GG-SLH showed higher conductivity than GG-SLH (p < 0.05) whereas PPy-GG-SLH (method I and II) showed the best conductivity among the 3 methods (∼1 to 2 × 10-4 S/cm). The microarchitecture of PPy-GG-SLH (method I) was similar to GG-SLH but PPy-GG-SLH (method II and III) presented smaller pore sizes and lower porosity. PPy-GG-SLH (method I and II) compressive modulus (∼450-500 KPa) and recovering capacity (∼75-90%) was higher than GG-SLH, nevertheless the mechanical properties of PPy-GG-SLH (method III) were lower. The water uptake of PPy-GG-SLH was rapidly up to 2500% and were stable along 60 days of degradation being the maximum weight loss 20%. Mechanically stable and electroactive PPy-GG-SLH (method I and II) were analyzed regarding cellular performance. PPy-GG-SLH were not cytotoxic for L929 cells. In addition, L929 and C2C12 myoblast cells were able to adhere and spread within PPy-GG-SLH, showing improved spreading in comparison to GG-SLH performance. Overall, PPy-GG-SLH show promising features as an alternative electroactive platform to analyze the influence of electrical stimulation on skeletal muscle cells.