Conductive hydrogels based on tragacanth and silk fibroin containing dopamine functionalized carboxyl-capped aniline pentamer: Merging hemostasis, antibacterial, and anti-oxidant properties into a multifunctional hydrogel for burn wound healing.

Hydrogels possessing both conductive characteristics and notable antibacterial and antioxidant properties hold considerable significance within the realm of wound healing and recovery. The object of current study is the development of conductive hydrogels with antibacterial and antioxidant properties, emphasizing their potential for effective wound healing, especially in treating third-degree burns. For this purpose, various conductive hydrogels are developed based on tragacanth and silk fibroin, with variable dopamine functionalized carboxyl-capped aniline pentamer (CAP@DA). The FTIR analysis confirms that the CAP powder was successfully synthesized and modified with DA. The results show that the incorporation of CAP@DA into hydrogels can increase the porosity and swellability of the hydrogels. Additionally, the mechanical and viscoelastic properties of the hydrogels are also improved. The release of vancomycin from the hydrogels is sustained over time, and the hydrogels are effective in inhibiting the growth of Methicillin-resistant Staphylococcus aureus (MRSA). In vitro cell studies of the hydrogels show that all hydrogels are biocompatible and support cell attachment. The hydrogels' tissue adhesiveness yielded a satisfactory hemostatic outcome in a rat-liver injury model. The third-degree burn was created on the dorsal back paravertebral region of the rats and then grafted with hydrogels. The burn was monitored for 3, 7, and 14 days to evaluate the efficacy of the hydrogel in promoting wound healing. The hydrogels revealed treatment effect, resulting in enhancements in wound closure, dermal collagen matrix production, new blood formation, and anti-inflammatory properties. Better results were obtained for hydrogel with increasing CAP@DA. In summary, the multifunctional conducive hydrogel, featuring potent antibacterial properties, markedly facilitated the wound regeneration process.

Alginate/gum arabic-based biomimetic hydrogel enriched with immobilized nerve growth factor and carnosine improves diabetic wound regeneration.

Diabetic foot ulcers (DFUs) often remain untreated because they are difficult to heal, caused by reduced skin sensitivity and impaired blood vessel formation. In this study, we propose a novel approach to manage DFUs using a multifunctional hydrogel made from a combination of alginate and gum arabic. To enhance the healing properties of the hydrogel, we immobilized nerve growth factor (NGF), within specially designed mesoporous silica nanoparticles (MSN). The MSNs were then incorporated into the hydrogel along with carnosine (Car), which further improves the hydrogel's therapeutic properties. The hydrogel containing the immobilized NGF (SiNGF) could control the sustain release of NGF for >21 days, indicating that the target hydrogel (AG-Car/SiNGF) can serve as a suitable reservoir managing diabetic wound regeneration. In addition, Car was able to effectively reduce inflammation and significantly increase angiogenesis compared to the control group. Based on the histological results obtained from diabetic rats, the target hydrogel (AG-Car/SiNGF) reduced inflammation and improved re-epithelialization, angiogenesis, and collagen deposition. Specific staining also confirmed that AG-Car/SiNGF exhibited improved tissue neovascularization, transforming growth factor-beta (TGFß) expression, and nerve neurofilament. Overall, our research suggests that this newly developed composite system holds promise as a potential treatment for non-healing diabetic wounds.

Injectable hydrogel based on silk fibroin/carboxymethyl cellulose/agarose containing polydopamine functionalized graphene oxide with conductivity, hemostasis, antibacterial, and anti-oxidant properties for full-thickness burn healing.

Overcoming bacterial infections and promoting wound healing are significant challenges in clinical practice and fundamental research. This study developed a series of enzymatic crosslinking injectable hydrogels based on silk fibroin (SF), carboxymethyl cellulose (CMC), and agarose, with the addition of polydopamine functionalized graphene oxide (GO@PDA) to endow the hydrogel with suitable conductivity and antimicrobial activity. The hydrogels exhibited suitable gelation time, stable mechanical and rheological properties, high water absorbency, and hemostatic properties. Biocompatibility was also confirmed through various assays. After loading the antibiotic vancomycin hydrochloride, the hydrogels showed sustained release and good antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). The fast gelation time and desirable tissue-covering ability of the hydrogels allowed for a good hemostatic effect in a rat liver trauma model. In a rat full-thickness burn wound model, the hydrogels exhibited an excellent treatment effect, leading to significantly enhanced wound closure, collagen deposition, and granulation tissue formation, as well as neovascularization and anti-inflammatory effects. In conclusion, the antibacterial electroactive injectable hydrogel dressing, with its multifunctional properties, significantly promoted the in vivo wound healing process, making it an excellent candidate for full-thickness skin wound healing.

Injectable multifunctional hydrogel based on carboxymethylcellulose/polyacrylamide/polydopamine containing vitamin C and curcumin promoted full-thickness burn regeneration.

Burn injuries are a major global problem, with a high risk of infection and mortality. This study aimed to develop an injectable hydrogel for wound dressings, composed of sodium carboxymethylcellulose/polyacrylamide/polydopamine containing vitamin C (CMC/PAAm/PDA VitC) for its antioxidant and antibacterial properties. Simultaneously, silk fibroin/alginate nanoparticles (SF/SANPs) loaded with curcumin (SF/SANPs CUR) were incorporated into the hydrogel to enhance wound regeneration and reduce bacterial infection. The hydrogels were fully characterized and tested in vitro and in preclinical rat models for biocompatibility, drug release, and wound healing efficacy. Results showed stable rheological properties, appropriate swelling and degradation ratios, gelation time, porosity, and free radical scavenging capacity. Biocompatibility was confirmed through MTT, lactate dehydrogenase, and apoptosis evaluations. Hydrogels containing curcumin demonstrated antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). In the preclinical study, hydrogels containing both drugs showed superior support for full-thickness burn regeneration, with improved wound closure, re-epithelialization, and collagen expression. The hydrogels also showed neovascularization and anti-inflammatory effects, as confirmed by CD31 and TNF-α markers. In conclusion, these dual drug-delivery hydrogels showed significant potential as wound dressings for full-thickness wounds.

Theranostic silk sericin/SPION nanoparticles for targeted delivery of ROR1 siRNA: Synthesis, characterization, diagnosis and anticancer effect on triple-negative breast cancer.

Triple-negative breast cancer (TNBC) has the worst prognosis among all breast cancer subtypes. The lack of proper treatments prompted scientists to find a practical targeted therapy to treat this type of tumor. Based on previous studies, tyrosine kinase-like orphan receptor (ROR1) is overexpressed in TNBC cells. Here, we designed a system consisting of superparamagnetic iron oxide nanoparticles (SPIONs) decorated with silk sericin (SS NPs) for the targeted delivery of ROR1 siRNA, a gene silencer to knockdown the expression of human ROR1 gene. NPs exhibited spherical shape of about 193 nm with acceptable properties both in vitro and in vivo. The apoptosis study showed significant death of MDA-MB-231 cells after 24 h treatment with the prepared NPs. The real-time PCR study also demonstrated an almost complete shutdown of ROR1 expression. Guided by magnetic field, enhanced accumulation of NPs was observed in breast tumors induced by 4T1 cells in BALB/c mice. Histological evaluation of the tumor exhibited necrosis 14 days post-treatment with the siRNA-loaded NPs; whereas, the untreated tumor was proliferating. Also, the tumor growth rate was significantly decreased after treatment with siRNA-loaded NPs in vivo. In conclusion, the prepared delivery system could be considered as a potential therapeutic strategy for treating TNBC.

Advanced Multifunctional Wound Dressing Hydrogels as Drug Carriers.

Skin injuries, especially chronic wounds, remain a significant healthcare system problem. The number of burns, diabetic patients, pressure ulcers, and other damages is also growing, particularly in elderly populations. Several investigations are pursued in designing more effective therapeutics for treating different wound injuries. These efforts have resulted in developing multifunctional wound dressings to improve wound repair. For this, preparing multifunctional dressings using various methods has provided a new attitude to support effective skin regeneration. This review focuses on the recent developments in designing multifunctional hydrogel dressings with hemostasis, adhesiveness, antibacterial, and antioxidant properties.

Dual drug delivery system of teicoplanin and phenamil based on pH-sensitive silk fibroin/sodium alginate hydrogel scaffold for treating chronic bone infection.

For effective treatment of infected bone, it is essential to use local drug delivery systems with the ability to deliver both antibiotics and osteoinductive factors. Herein, a pH-sensitive silk fibroin (SF)/sodium alginate (SA) hydrogel scaffolds containing teicoplanin (TEC) and phenamil (PM) loaded SF nanoparticles (PMSFNPS) are introduced for treating chronic osteomyelitis. The TEC and PM showed a sustained- and pH-sensitive release behavior from SF/SA hydrogel. The higher release rate was seen in an alkaline pH in comparison to neutral and acidic pH during 10 days. The eluted TEC maintained its antibacterial activity of >75 % during 35 days and in three different pH values (5.5, 7.4, and 8.5). The cellular study indicated that the scaffolds containing PMSFNPs could promote the cell viability, ALP activity, and matrix mineralization. Moreover, the in vivo effectiveness of hydrogel scaffolds were analyzed with radiography, histological and Immunohistochemistry evaluations. The lower infection and higher regeneration were observed in methicillin-resistant Staphylococcus aureus (MRSA) infected rat bone treated with hydrogel scaffold containing PMSFNPs and TEC compared to other groups. Consequently, this dual-drug delivery system could be a hopeful approach for effective treatment of chronic bone infection.

Thermosensitive chitosan/poly(N-isopropyl acrylamide) nanoparticles embedded in aniline pentamer/silk fibroin/polyacrylamide as an electroactive injectable hydrogel for healing critical-sized calvarial bone defect in aging rat model.

Thermosensitive nanoparticles with phase transition abilities have been considered as suitable materials in biomedical fields, especially drug delivery systems. Moreover, electroactive injectable hydrogels supporting bone regeneration of the elderly will highly be desired in bone tissue engineering applications. Herein, thermosensitive nanoparticles were fabricated using chitosan/poly(N-isopropyl acrylamide) for simvastatin acid delivery. The nanoparticles were incorporated into electroactive injectable hydrogels based on aniline pentamer/silk fibroin/polyacrylamide containing vitamin C. The nanoparticles had thermosensitive properties as simvastatin acid had higher release rates at 37 than 23 °C without significant burst release. The hydrogels also revealed an appropriate gelation time, stable mechanical and rheological characteristics, high water absorbency, and proper biodegradability. In vitro studies indicated that the hydrogel was biocompatible and nontoxic, especially those containing drugs. Implantation of the hydrogels containing both simvastatin acid and vitamin C into the critical calvarial bone defect of the aged rat also demonstrated significant enhancement of bone healing after 4 and 8 weeks post-implantation. We found that the electroactive injectable hydrogels containing thermosensitive nanoparticles exhibited great potential for treating bone defects in the elderly rats.

Fabrication of Silk Scaffold Containing Simvastatin-Loaded Silk Fibroin Nanoparticles for Regenerating Bone Defects

Background: In the present study, a tissue engineered silk fibroin (SF) scaffold containing simvastatin-loaded silk fibroin nanoparticles (SFNPs) were used to stimulate the regeneration of the defected bone. Methods: At first, the porous SF scaffold was prepared using freeze-drying. Then simvastatin-loaded SFNPs were made by dissolvation method and embedded in the SF scaffold. Afterwards, the scaffold and the NPs were characterized in terms of physicochemical properties and the ability to release the simvastatin small molecule. Results: The results exhibited that the SF scaffold had a porous structure suitable for releasing the small molecule and inducing the proliferation and attachment of osteoblast cells. SFNPs containing simvastatin had spherical morphology and were 174 ± 4 nm in size with -24.5 zeta potential. Simvastatin was also successfully encapsulated within the SFNPs with 68% encapsulation efficiency. Moreover, the small molecule revealed a sustained release profile from the NPs during 35 days. The results obtained from the in vitro cell-based studies indicated that simvastatin-loaded SFNPs embedded in the scaffold had acceptable capacity to promote the proliferation and alkaline phosphatase production of osteoblast cells while inducing osteogenic matrix precipitation. Conclusion: The SF scaffold containing simvastatin-loaded SFNPs could have a good potential to be used as a bone tissue-engineered construct.

Combination Therapy of Breast Cancer by Codelivery of Doxorubicin and Survivin siRNA Using Polyethylenimine Modified Silk Fibroin Nanoparticles.

Here, polyethylenimine (PEI) modified silk fibroin nanoparticles (SFNPs) were prepared for codelivery of doxorubicin (DOX) and survivin siRNA. The prepared NPs were characterized in terms of stability and structural, functional, and physicochemical properties. Moreover, the ability of the conjugate to escape from the endosome and cellular uptake were assessed. Afterward, the in vivo therapeutic efficacy was analyzed in the mice model. The siRNA loaded PEI-SFNPs showed acceptable size, zeta potential, and stability in serum. It also effectively induced apoptosis in the 4T1 mouse mammary tumor cell line. Cellular uptake and endosomal escape analyses confirmed that PEI-SFNPs containing siRNA could escape from the endosome and accumulate in the cytoplasm of 4T1 cells. Real time-PCR indicated the significant decrease in the expression of survivin mRNA in the 4T1 cell line 48 h postincubation with siRNA loaded PEI-SFNPs. In vivo biodistribution of PEI-SFNPs confirmed higher accumulation of SFNPs in the tumor site compared with other organs. The codelivery systems remarkably reduced the growth rate of breast tumor in the mice model without any obvious weight lost. Histopathological and tunnel staining exhibited more apoptotic tumor cells in the group containing both DOX and survivin siRNA. Tumorigenic breast tissue resected from the animals after treatment with siRNA also exhibited significant suppression of survivin gene. In conclusion, the prepared drug delivery system had an acceptable potential in tumor removal, apoptosis induction in cancer cells, and therapeutic efficacy. Thus, it would be a good candidate for breast cancer therapy.

Conductive Biomaterials as Substrates for Neural Stem Cells Differentiation towards Neuronal Lineage Cells.

The injuries and defects in the central nervous system are the causes of disability and death of an affected person. As of now, there are no clinically available methods to enhance neural structural regeneration and functional recovery of nerve injuries. Recently, some experimental studies claimed that the injuries in brain can be repaired by progenitor or neural stem cells located in the neurogenic sites of adult mammalian brain. Various attempts have been made to construct biomimetic physiological microenvironment for neural stem cells to control their ultimate fate. Conductive materials have been considered as one the best choices for nerve regeneration due to the capacity to mimic the microenvironment of stem cells and regulate the alignment, growth, and differentiation of neural stem cells. The review highlights the use of conductive biomaterials, e.g., polypyrrole, polyaniline, poly(3,4-ethylenedioxythiophene), multi-walled carbon nanotubes, single-wall carbon nanotubes, graphene, and graphite oxide, for controlling the neural stem cells activities in terms of proliferation and neuronal differentiation. The effects of conductive biomaterials in axon elongation and synapse formation for optimal repair of central nervous system injuries are also discussed.

Dual drug delivery system based on pH-sensitive silk fibroin/alginate nanoparticles entrapped in PNIPAM hydrogel for treating severe infected burn wound.

Herein, the pH-sensitive vancomycin (VANCO) loaded silk fibroin-sodium alginate nanoparticles (NPs) embedded in poly(N-isopropylacrylamide) (PNIPAM) hydrogel containing epidermal growth factor (EGF) are introduced for treating chronic burn wound infections. The hybrid system was developed to control the release rates of an antibiotic and growth factor for optimal treatment of burn infections. VANCO had a pH responsive release behavior from the nanoparticle (NP) and showed higher release rate in an alkaline pH compared to the neutral pH during 10 d. About 30% of EGF was also released from the hydrogel within 20 d. The released VANCO and EGF preserved their bioactivity more than ∼ 80%. The suitable physico-chemical properties and cellular behaviors of PNIPAM hydrogel supported the proliferation and growth of the fibroblast cells. Furthermore, the higher re-epithelialization with good wound contraction rate, neovascular formation, and expression of transforming growth factor-beta were observed in S. aureus infected rat burn wound by using the hydrogel containing VANCO and EGF compared with untreated wounds and hydrogel alone. The wound infection was also significantly reduced in the groups treated with the hydrogels containing VANCO. Overall, in vitro and in vivo results suggested that developed hybrid system would be a promising construct to treat severe wound infection.

Bilayer Cylindrical Conduit Consisting of Electrospun Polycaprolactone Nanofibers and DSC Cross-Linked Sodium Alginate Hydrogel to Bridge Peripheral Nerve Gaps.

Herein, a bilayer cylindrical conduit (P-CA) is presented consisting of electrospun polycaprolactone (PCL) nanofibers and sodium alginate hydrogel covalently cross-linked with N,N'-disuccinimidyl carbonate (DSC). The bilayer P-CA conduit is developed by combining the electrospinning and outer-inner layer methods. Using DSC, as a covalent crosslinker, increases the degradation time of the sodium alginate hydrogel up to 2 months. The swelling ratio of the hydrogel is also 503% during the first 8 h. The DSC cross-linked sodium alginate in the inner layer of the conduit promotes the adhesion and proliferation of nerve cells, while the electrospun PCL nanofibers in the outer layer provide maximum tensile strength of the conduit during surgery. P-CA conduit promotes the migration of Schwann cells along the axon in a rat model based on functional and histological evidences. In conclusion, P-CA conduit will be a promising construct for repairing sciatic nerves in a rat model.

Agarose-based biomaterials for advanced drug delivery.

Agarose is a prominent marine polysaccharide representing reversible thermogelling behavior, outstanding mechanical properties, high bioactivity, and switchable chemical reactivity for functionalization. As a result, agarose has received particular attention in the fabrication of advanced delivery systems as sophisticated carriers for therapeutic agents. The ever-growing use of agarose-based biomaterials for drug delivery systems resulted in rapid growth in the number of related publications, however still, a long way should be paved to achieve FDA approval for most of the proposed products. This review aims at a classification of agarose-based biomaterials and their derivatives applicable for controlled/targeted drug delivery purposes. Moreover, it attempts to deal with opportunities and challenges associated with the future developments ahead of agarose-based biomaterials in the realm of advanced drug delivery. Undoubtedly, this class of biomaterials needs further advancement, and a lot of critical questions have yet to be answered.

Functionalized silk fibroin nanofibers as drug carriers: Advantages and challenges.

Electrospinning is well thought of as the most potent nanofiber producing technique, which is applicable in biomedical fields. The generation of electrospun nanofibers as drug carriers has widely been shown much interest over the past years. Electrospun nanofibers meet various advantages as drug delivery platforms including high surface area, acceptable mechanical properties based on the choice of the polymer, high processability, and the possibilities for surface modifications. Silk fibroin protein has gained a great attention as a drug delivery carrier due to ease of purification, sterilization, processability without using chemical crosslinkers, good biocompatibility, tailorable biodegradability, low immunogenicity, and high capacity to stabilize the loaded drugs. These characteristics along with advantageous benefits of electrospinning provide opportunities for producing suitable nanofibers based on silk fibroin for drug delivery purposes. It is also possible to incorporate various functional moieties to the electrospun silk fibroin nanofibers to enhance its biological activities. This review covers the progress in electrospinning of silk fibroin as a drug carrier in recent years.

Design and fabrication of dual-targeted delivery system based on gemcitabine-conjugated human serum albumin nanoparticles.

Dual-targeted drug delivery system has established their reputation as potent vehicles for cancer chemotherapies. Herein, gemcitabine (Gem) was conjugated to human serum albumin (HSA) via dithiodipropionic anhydride to fabricate Gem-HSA nanoparticles. It was hypothesized that this system can enhance the low stability of Gem and can improve its intracellular delivery. Furthermore, folate was applied as targeting agent on HSA nanoparticles for increasing the tumor selectivity of Gem. To evaluate the structural properties of synthesized products, 1 H NMR and FT-IR were performed. Moreover, HPLC was implemented for confirming the conjugation between HSA and Gem. Nanoparticles have shown spherical shape with negative charge. The release rate of Gem was dependent to the concentration of glutathione and pH. Folate-targeted HSA nanoparticles have shown higher cytotoxicity, cellular uptake, and apoptosis induction on folate receptor overexpressing MDA-MB-231 cells in comparison to non-targeted nanoparticles. Finally, it is considered that the developed dual-targeted nanoparticles would be potent in improving the stability and efficacy of intracellular delivery of Gem and its selective delivery to cancer cells.

Functionalized theranostic nanocarriers with bio-inspired polydopamine for tumor imaging and chemo-photothermal therapy.

Nanocarriers sensitive to near infrared light (NIR) are useful templates for chemo-photothermal therapy (PTT) and imaging of tumors due to the ability to change the absorbed NIR energy to heat. The conventional photo-absorbing reagents lack the efficient loading and release of drug before reaching the target site leading to insufficient therapeutic outcomes. To overcome these limitations, the surface of nanocarriers can be modified with different polymers with wide functionalities to provide systems with diagnostic, therapeutic, and theranostic capabilities. Among various polymers, polydopamine (PDA) has been more interested due to complex structure with various chemical moieties, and the capacity to be used through different coating mechanism. In this review, we describe the complex structure, chemical properties, and coating mechanisms of PDA. Moreover, the advantage and surface modification of some relevant nanosystems based on carbon materials, gold, iron oxide, manganese, and upconverting nanomaterials by using PDA will be discussed, in detail.

Carboxymethyl dextran-trimethyl chitosan coated superparamagnetic iron oxide nanoparticles: An effective siRNA delivery system for HIV-1 Nef.

Gene therapy, including small interfering RNA (siRNA) technology, is one of the leading strategies that help to improve the outcomes of the current therapeutic systems against HIV-1 infection. The successful therapeutic application of siRNAs requires their safe and efficient delivery to specific cells. Here, we introduce a superparamagnetic iron oxide nanoparticle (SPION) for delivering siRNA against HIV-1 nef (anti-nef siRNA) into two cell lines, HEK293 and macrophage RAW 264.7. SPIONs were coated with trimethyl chitosan (TMC), and thereafter, different concentrations of SPION-TMC were coated with different ratios of a carboxymethyl dextran (CMD) to modify the physicochemical properties and improve the biological properties of the nanocarriers. The nanoparticles exhibited a spherical shape with an average size of 112 nm. The obtained results showed that the designed delivery route enhanced the uptake of siRNA into both HEK293 and RAW 264.7 cells compared with control groups. Moreover, CMD-TMC-SPIONs containing anti-nef siRNA significantly reduced the expression of HIV-1 nef in HEK293 stable cells. The modified siRNA-loaded SPIONs also displayed no toxicity or apoptosis-inducing effects on the cells. The CMD-TMC-SPIONs are suggested as potential nanocarriers for siRNA delivery in gene therapy of HIV-1 infection.

Inhibiting hepatic gluconeogenesis by chitosan lactate nanoparticles containing CRTC2 siRNA targeted by poly(ethylene glycol)-glycyrrhetinic acid.

Diabetes mellitus is a chronic metabolic disorder characterized by insulin deficiency and impaired glucose metabolism. Overexpression of cAMP response element binding protein (CREB)-regulated transcriptional coactivator 2 (CRTC2) plays an important role in high gluconeogenesis in patients with diabetes type II. Using RNA interference technology for silencing CRTC2 gene expression could be helpful in controlling the level of blood glucose and gluconeogenesis. In this study, we designed a siRNA delivery platform comprising blended chitosan lactate (CT) and polyethylene glycol (PEG) conjugated with glycyrrhetinic acid (GA) for controlling gluconeogenesis. The nanoparticles showed spherical and smooth surface with ~ 189-nm size and + 5.1 zeta potential. Targeted nanoparticles were efficiently stable in serum and different levels of heparin media over 48 h. The gene knockdown efficiency of nanoparticles was comparable to Lipofectamine®, while they had no significant in vitro and in vivo toxicity. The in vivo therapeutic efficacy of targeted nanoparticles was also confirmed by reduced amount of fasting blood sugar in diabetic rat models. Furthermore, the nanoparticles were mostly accumulated in the liver after 2 h indicating the significant targeting ability of the prepared nanoparticles. Therefore, CT/PEG-GA nanoparticles can be considered as a potential candidate for targeted delivery of siRNA into hepatocytes in order to regulate gluconeogenesis in diabetes.

Gold nanorods reinforced silk fibroin nanocomposite for peripheral nerve tissue engineering applications.

Nowadays, regenerating peripheral nerves injuries (PNIs) remain a major clinical challenge, which has gained a great attention between scientists. Here, we represent a nanocomposite based on silk fibroin reinforced gold nanorods (SF/GNRs) to evaluate the proliferation and attachment of PC12 cells. The morphological characterization of nanocomposites with transmission electron microscopy (TEM) and Scanning electron microscopy (SEM) showed that the fabricated scaffolds have porous structure with interconnected pores that is suitable for cell adhesion and growth. GNRs significantly improved the poor electrical conductivity of bulk silk fibroin scaffold. Evaluating the morphology of PC12 cells on the scaffold also confirmed the normal morphology of cells with good rate of adhesion. SF/GNRs nanocomposites showed better cellular attachment, growth and proliferation without any toxicity compared with bulk SF scaffold. Moreover, immunostaining studies represented the overexpression of neural specific proteins like nestin and neuron specific enolase (NSE) in the cells cultured on SF/GNRs nanocomposites in comparison to neat SF scaffolds.