Search

Effective hemorrhage control is indispensable for life-threatening emergencies in defense fields and civilian trauma. During major injuries, hemostatic agents are applied externally to mimic and accelerate the natural hemostasis process. Commercially available topical hemostatic agents are associated with several limitations

The utility of plant tissues as scaffolding materials has been gaining significant interest in recent years owing to their unique material characteristics that are ideal for tissue regeneration. In this study, the degradation and biocompatibility of natural cellulosic scaffolds derived from

A hostile myocardial microenvironment post ischemic injury (myocardial infarction) plays a decisive role in determining the fate of tissue-engineered approaches. Therefore, engineering hybrid 3D printed platforms that can modulate the MI microenvironment for improving implant acceptance has surfaced as a critical requirement for reconstructing an infarcted heart. Here, we have employed a non-mulberry silk-based conductive bioink comprising carbon nanotubes (CNTs) to bioprint functional 3D vascularized anisotropic cardiac constructs. Immunofluorescence staining, polymerase chain reaction-based gene expression studies, and electrophysiological studies showed that the inclusion of CNTs in the bioink played a significant role in upregulating matured cardiac biomarkers, sarcomere formation, and beating rate while promoting cardiomyocyte viability. These constructs were then microinjected with calcium peroxide and IL-10-loaded gelatin methacryloyl microspheres. Measurements of oxygen concentration revealed that these microspheres upheld the oxygen availability for maintaining cellular viability for at least 5 days in a hypoxic environment. Also, the ability of microinjected IL-10 microspheres to modulate the macrophages to anti-inflammatory M2 phenotype in vitro was uncovered using immunofluorescent staining and gene expression studies. Furthermore, in vivo subcutaneous implantation of microsphere-injected 3D constructs provided insights toward the extended time frame that was achieved for dealing with the hostile microenvironment for promoting host neovascularization and implant acceptance.

Silk fibroin and silk microfibers, both derived from silk cocoon, have been widely used for prospective biomedical, energy, and environmental applications. However, various complex and catalyst-based approaches have been adopted for chemical modification and integration of different functionalities in silk fibroin-based materials. Here, both tailored water wettability and mechanical property have been associated with silk microfiber reinforced silk fibroin sponges (SMFRSFSs) through the strategic introduction of β-sheets and a facile and catalyst-free chemical reaction at ambient conditions. While the controlled tailoring of β-sheets in the silk fibroin skeletal framework of the sponges allowed us to modulate the compressive modulus, the 1,4-conjugate addition reaction between amine residues of silk (fiber and fibroin) and acrylate groups of a multifunctional cross-linker provided residual chemical reactivity. Further, the chemically "reactive" sponge was postmodified with the selected alkylamines to introduce a wide range of water wettability (from 36 to 161°) without affecting the mechanical property. Thereafter, the silk cocoon-derived and extremely water-repellent sponge was used for environment-friendly cleaning of oil spillages through selective absorption-based and filtration-based oil/water separation at different and severe aqueous conditions. This silk cocoon-derived mechanically tailorable and chemically reactive sponge could also be useful for various biomedical and energy-related applications.

The synthesis and characterization of chiral pincer-ruthenium complexes of the type (R2NNN)RuCl2 (PPh3) (R = 3-methylbutyl and 3,3-dimethylbutyl) is reported here. The cytotoxicity studies of these complexes were studied and compared with the corresponding activity of achiral complexes. The cytotoxic effect of pincer-ruthenium complexes on human dermal fibroblasts and human tongue carcinoma cells assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay displayed an inhibition of normal and cancer cell growth in a dose-dependent manner. Intracellular reactive oxygen species (ROS) level measurement, lactate dehydrogenase assay, DNA fragmentation, and necrosis studies revealed that treatment with pincer-ruthenium complexes induced a redox imbalance in SAS cells by upregulating ROS generation and caused necrotic cell death by disrupting the cellular membrane integrity.

DNA-based nanostructures have emerged as a versatile component for nanoscale construction of soft materials. Multiple structural, functional properties and versatility in conjugation with other biomolecules made DNA the material of choice to use in various biomedical applications. DNA-based hydrogels significantly attracted attention in recent years owing to their properties and applications in biosensing, bioimaging, and therapeutics. Here, we summarize the recent advances in the area of DNA hydrogels where these are used either as structural material or as functional entities to make hybrid constructs with various biomedical applications. Multiple synthetic routes for constructing DNA hydrogels are summarized first, where the structural motifs and spatial arrangements are considered for the classification of DNA materials. We then present the characterization and properties of DNA hydrogels using multiple imaging and biophysical techniques. Further, different biomedical applications of DNA hydrogels are presented such as biosensing, bioimaging, and targeted drug delivery and as scaffolds to program cellular systems. Last, we discuss the vision and potential of DNA based hydrogels as an emerging class of therapeutically important devices for theragnostic and other biological applications.

Localized cancer chemotherapy through injectable hydrogels is a next-generation advanced substitute for the currently operational systemic route of drug administration. Recently, several hydrogels have been developed for prospective drug delivery applications; however, no

Vascular tissue engineering is aimed at developing regenerative vascular grafts to restore tissue function by bypassing or replacing defective arterial segments with tubular biodegradable scaffolds. Scaffolds are often combined with stem or progenitor cells to prevent acute thrombosis and initiate scaffold remodeling. However, there are limitations to cell-based technologies regarding safety and clinical translation. Extracellular vesicles (EVs) are nanosized particles released by most cell types, including stem and progenitor cells, that serve to transmit protein and RNA cargo to target cells throughout the body. EVs have been shown to replicate the therapeutic effect of their parent cells; therefore, EVs derived from stem or progenitor cells may serve as a more translatable, cell-free, therapeutic base for vascular scaffolds. Our study aims to determine if EV incorporation provides a positive effect on graft patency and remodeling

Myocardial microenvironment plays a decisive role in guiding the function and fate of cardiomyocytes, and engineering this extracellular niche holds great promise for cardiac tissue regeneration. Platforms utilizing hybrid hydrogels containing various types of conductive nanoparticles have been a critical tool for constructing engineered cardiac tissues with outstanding mechanical integrity and improved electrophysiological properties. However, there has been no attempt to directly compare the efficacy of these hybrid hydrogels and decipher the mechanisms behind how these platforms differentially regulate cardiomyocyte behavior. Here, we employed gelatin methacryloyl (GelMA) hydrogels containing three different types of carbon-based nanoparticles: carbon nanotubes (CNTs), graphene oxide (GO), and reduced GO (rGO), to investigate the influence of these hybrid scaffolds on the structural organization and functionality of cardiomyocytes. Using immunofluorescent staining for assessing cellular organization and proliferation, we showed that electrically conductive scaffolds (CNT- and rGO-GelMA compared to relatively nonconductive GO-GelMA) played a significant role in promoting desirable morphology of cardiomyocytes and elevated the expression of functional cardiac markers, while maintaining their viability. Electrophysiological analysis revealed that these engineered cardiac tissues showed distinct cardiomyocyte phenotypes and different levels of maturity based on the substrate (CNT-GelMA: ventricular-like, GO-GelMA: atrial-like, and rGO-GelMA: ventricular/atrial mixed phenotypes). Through analysis of gene-expression patterns, we uncovered that the engineered cardiac tissues matured on CNT-GelMA and native cardiac tissues showed comparable expression levels of maturation markers. Furthermore, we demonstrated that engineered cardiac tissues matured on CNT-GelMA have increased functionality through integrin-mediated mechanotransduction (via YAP/TAZ) in contrast to cardiomyocytes cultured on rGO-GelMA.

Urokinase plasminogen activator (uPA) has been extensively used as a thrombolytic drug in cases of myocardial infarction, thromboembolism, and ischemic brain stroke. Media optimization and high-density perfusion culture are the decisive factors that facilitate enhanced urokinase production in a conditioned medium. In this study, we have aimed for a high-density perfusion culture of HT1080, a human fibrosarcoma cell line, by formulating optimal media for enhanced urokinase productivity. Four scaffold variants were fabricated from silk fibroin and microfibers of Bombyx mori (BM) and Antheraea assamensis (AA) and physico-chemically characterized. Field emission scanning electron microscopy studies revealed a heterogeneous distribution of pores with interconnected networks supporting cell infiltration, attachment, and long-term viability. AA-based fiber-reinforced scaffolds (ASAF) demonstrated superior mechanical strength, integral stability, and increased cell proliferation as compared to pure silk scaffolds. Media formulation was accomplished by limiting serum concentration (2% FBS) and supplementing with 20 μg/mL arginine and 20 ng/mL TGF-β1 to retain the stationary phase of cells and augment the urokinase production. A perfusion bioreactor culture of HT1080-laden scaffolds in the presence of formulated media was performed for improving the production of urokinase, with a maximum activity of 432 U/L. Also, gene expression analysis revealed that the individual silk scaffolds have different effects on regulating the expression of plasminogen activator urokinase and plasminogen activator urokinase receptor. In brief, our results suggest that a perfusion bioreactor culture of HT1080-laden ASAF scaffolds in formulated media promotes an increased urokinase production, such that it can be further used as a novel 3D matrix platform for industrial production of the lifesaving uPA drug.

In this study, the effect of cellular cross-talk on modulation of chondrogenesis and hypertrophy while minimizing the usage of articular chondrocytes (ACs) has been investigated. Herein, co-culture of ACs with adipose-derived human mesenchymal stem cells (ADhMSCs) was employed for cross-talk within silk fibroin (SF)-based three-dimensional (3D) scaffolds. The co-culture model was developed by co-culturing four different ratios of ADhMSCs to ACs: 1:0, 3:1, 1:1, and 0:1 on porous 3D SF scaffolds for 21 days. The co-culture groups were cultured in defined media without adding any exogenous growth factors except the monoculture group, ADhMSC-only controls. The co-cultured constructs indicated significantly higher cellular viability and proliferation than the control monoculture groups. The supernatants of co-culture groups indicated significantly higher levels of TGF-β1 and IL-10, which confirmed the production of the morphogens/signaling molecules by chondrocytes for induction of ADhMSCs differentiation toward the chondrogenic phenotype. Biochemical assays indicated enhanced accumulation of sulfated glycosaminoglycans, collagen, and high DNA content along with high cellularity in co-culture groups than chondrocyte-only controls. Co-culture groups revealed synergistic interactions between cells as indicated by the interaction index value ranging from 2-3. Furthermore, upregulation of putative chondrogenic markers-aggrecan, sox-9, and collagen II, and significantly reduced expression of hypertrophic genes-collagen type X and MMP-13 was revealed in co-culture constructs. Histological and immunohistochemical staining also demonstrated even distribution and deposition of ECM in co-cultured constructs. Taken together, this work presents the potential of the developed 3D co-culture model toward modulation of chondrogenesis and hypertrophy via 3D microenvironment induced by physicochemical and biological properties of SF scaffolds, synergistic interactions between cells, and paracrine signaling in the co-culture system.

Full-thickness cutaneous wounds, such as deep burns, are complex wounds that often require surgical interventions. Herein, we show the efficacy of acellular grafts that can be made available off-the-shelf at an affordable cost using silk biomaterials. Silkworm silk fibroin (SF), being a cost-effective and natural biopolymer, provides essential features required for the fabrication of three-dimensional constructs for wound-healing applications. We report the treatment of third-degree burn wounds using a freeze-dried microporous scaffold of Antheraea assama SF (AaSF) functionalized with a recombinant spider silk fusion protein FN-4RepCT (FN-4RC) that holds the fibronectin cell binding motif. In order to examine the healing efficiency of functionalized silk scaffolds, an in vivo burn rat model was used, and the scaffolds were implanted by a one-step grafting procedure. The aim of our work is to investigate the efficacy of the developed acellular silk grafts for treating full-thickness wounds as well as to examine the effect of recombinant spider silk coatings on the healing outcomes. Following 14-day treatment, AaSF scaffolds coated with FN-4RC demonstrated accelerated wound healing when compared to the uncoated counterpart, commercially used DuoDERM dressing patch, and untreated wounds. Histological assessments of wounds over time further confirmed that functionalized silk scaffolds promoted wound healing, showing vascularization and re-epithelialization in the initial phase. In addition, higher extent of tissue remodeling was affirmed by the gene expression study of collagen type I and type III, indicating advanced stage of healing by the silk treatments. Thus, the present study validates the potential of scaffolds of combined silkworm silk and FN-4RC for skin regeneration.

Complex cutaneous wounds like diabetic foot ulcers represent a critical clinical challenge and demand a large-scale and low-cost strategy for effective treatment. Herein, we use a rabbit animal model to investigate efficacy of bioactive wound dressings made up of silk biomaterials. Nanofibrous mats of Antheraea assama silkworm silk fibroin (AaSF) are coated with various recombinant spider silk fusion proteins through silk-silk interactions to fabricate multifunctional wound dressings. Two different types of spider silk coatings are used to compare their healing efficiency: FN-4RepCT (contains a cell binding motif derived from fibronectin) and Lac-4RepCT (contains a cationic antimicrobial peptide from lactoferricin). AaSF mats coated with spider silk show accelerated wound healing properties in comparison to the uncoated mats. Among the spider silk coated variants, dual coating of FN-4RepCT and Lac-4RepCT on top of AaSF mat demonstrated better wound healing efficiency, followed by FN-4RepCT and Lac-4RepCT single coated counterparts. The in vivo study also reveals excellent skin regeneration by the functionalized silk dressings in comparison to commercially used Duoderm dressing and untreated wounds. The spider silk coatings demonstrate early granulation tissue development, re-epithelialization, and efficient matrix remodelling of wounds. The results thus validate potential of bioactive silk matrices in faster repair of diabetic wounds.

Repair and regeneration of nasal and auricular cartilage thrust significant challenges in reconstructive surgery. The burgeoning clinical requirement is yet to endorse a satisfactory cartilage replacement matrix. In this regard, we have bioengineered cross-linked decellularized caprine conchal cartilage (DC) as biocompatible, durable, and nontoxic matrices. The DC matrices exhibited reduced DNA and sulfated glycosaminoglycan (sGAG) with a minimal effect on the collagen content. Further, histology and scanning electron micrographs revealed a significant loss of cellular bodies and the presence of a compact matrix consisting of intricate collagen fibers, when compared to unprocessed matrices. An

The major limitations of traditional methods of anticancer drug delivery include systemic distribution and frequent administration intravenously. To address these issues, in our present approach, we have fabricated a nano hybrid silk hydrogel system for localized, targeted, and on-demand delivery of anticancer drugs. The hybrid system contains a blend of two varieties of silk protein and doxorubicin (DOX)-loaded folic acid functionalized single-walled carbon nanotubes (SWCNT-FA/DOX). Owing to the single-walled carbon nanotube (SWCNT) incorporation, the mechanical strength of the hybrid silk hydrogel composite enhanced significantly. A slow and sustained DOX release was recorded over a 14 day study. The amount of DOX released was determined by concentration of the SWCNT-FA/DOX payload, rate of silk degradation, pH of the released medium, and incubation temperature. The intermittent exposure of near-infrared light to the hybrid gel system stimulated on-demand DOX release. The

Catalyst-free and readily chemically reactive functional coatings that have immense prospects in various relevant applications are unprecedentedly synthesized directly using naturally existing bovi...

Prodigious progress in the past decade has pronounced 3D printing as one of the most promising technique for assembling biological materials in a complex layout that mimics native human tissues. Wi...

Cell instructive scaffolding platforms displaying synergistic effects by virtue of their chemical and physical cues have tremendous scope in modulating cell phenotype and thus improving the success of any graft. In this regard, we report here the development of Si- and Zn-doped brushite cement composited with silk scaffolding that hierarchically emulated the cancellous bone. The composite scaffolds fabricated exhibited an open porous network capable of enhanced osteoblast survival as attested by increased alkaline phosphatase activity and also sustaining osteoclast activity affirmed by tartrate resistant acid phosphatase staining. Moreover, the chemical cues presented by dissolutions products from the composite scaffold enabled the osteoblasts to secrete proangiogenic factors which favored better endothelial cell survival, confirmed through in vitro experiments. Moreover, the efficacy of these composite biomimetic scaffolds was validated in vivo in volumetric femur defects in rabbits, which revealed that these matrices influenced vascular cell infiltration and favored the formation of matured bony plate. Fluorochrome labeling studies and microtomography analysis revealed that at the end of three months, the implanted composite scaffolds had completely resorbed, leaving behind neo-osseous tissue and vouching for clinical translation of these composite matrices as viable and affordable bone-graft substitutes.

The syntheses, characterizations, and biological activities of three organoplatinum compounds, each containing a triazole ring, are discussed. These organometallic compounds demonstrate superior cytotoxicity against osteosarcoma and human breast cancer cells relative to cisplatin, a well-known chemotherapeutic agent used for chemotherapy.

Functional impairment of vascular cells is associated with cardiovascular pathologies. Recent literature clearly presents evidence relating cell microenvironment and their function. It is crucial to understand the cell-material interaction while designing a functional tissue engineered vascular graft. Natural silk biopolymer has shown potential for various tissue-engineering applications. In the present work, we aimed to explore the combinatorial effect of variable innate physicochemical properties and topographies of silk films on functional behavior of vascular cells. Silk proteins from different varieties (mulberry Bombyx mori, BM; and non-mulberry Antheraea assama, AA) possess unique inherent amino acid composition that leads to variable surface properties (roughness, wettability, chemistry, and mechanical stiffness). In addition, we engineered the silk film surfaces and printed a microgrooved pattern to induce unidirectional cell orientation mimicking their native form. Patterned silk films induced unidirectional alignment of porcine vascular cells. Regardless of alignment, endothelial cells (ECs) proliferated favorably on AA films; however, it suppressed production of nitric oxide (NO), an endogenous vasodilator. Unidirectional alignment of smooth muscle cells (SMCs) encouraged contractile phenotype as indicated by minimal cell proliferation, increment of quiescent (G0) phase cells, and upregulation of contractile genes. Moderately hydrophilic flat BM films induced cell aggregation and augmented the expression of contractile genes (for SMCs) and endothelial nitric oxide synthase, eNOS (for ECs). Functional studies further confirmed SMCs' alignment improving collagen production, remodeling ability (matrix metalloproteinase, MMP-2 and MMP-9 production) and physical contraction. Altogether, this study confirms vascular cells' functional behavior is crucially regulated by synergistic effect of their alignment and cell-substrate interfacial properties.

Silk is considered to be a potential biomaterial for a wide number of biomedical applications. Silk fibroin (SF) can be retrieved in sufficient quantities from the cocoons produced by silkworms. While it is easy to formulate into scaffolds with favorable mechanical properties, the natural SF does not contain bioactive functions. Spider silk proteins, on the contrary, can be produced in fusion with bioactive protein domains, but the recombinant procedures are expensive, and large-scale production is challenging. We combine the two types of silk to fabricate affordable, functional tissue-engineered constructs for wound-healing applications. Nanofibrous mats and microporous scaffolds made of natural silkworm SF are used as a bulk material that are top-coated with the recombinant spider silk protein (4RepCT) in fusion with a cell-binding motif, antimicrobial peptides, and a growth factor. For this, the inherent silk properties are utilized to form interactions between the two silk types by self-assembly. The intended function, that is, improved cell adhesion, antimicrobial activity, and growth factor stimulation, could be demonstrated for the obtained functionalized silk mats. As a skin prototype, SF scaffolds coated with functionalized silk are cocultured with multiple cell types to demonstrate formation of a bilayered tissue construct with a keratinized epidermal layer under in vitro conditions. The encouraging results support this strategy of fabrication of an affordable bioactive SF-spider silk-based biomaterial for wound dressings and skin substitutes.

Artificial bioinspired superhydrophobicity, which is generally developed through appropriate optimization of chemistry and hierarchical topography, is being recognized for its immense prospective applications related to environment and healthcare. Nevertheless, the weak interfacial interactions that are associated with the fabrication of such special interfaces often provide delicate biomimicked wettability, and the embedded antifouling property collapses on exposure to harsh and complex aqueous phases and also after regular physical deformations, including bending, creasing, etc. Eventually, such materials with potential antifouling property became less relevant for practical applications. Here, a facile, catalyst-free, and robust 1,4-conjugate addition reaction has been strategically exploited for appropriate covalent integration of modified graphene oxide to developing polymeric materials with (1) tunable mechanical properties and (2) durable antifouling property, which are capable of performing both in air and under oil. Furthermore, this approach provided a facile basis for (3) engineering a superhydrophobic monolith into arbitrary free-standing shapes and (4) decorating various flexible (metal, synthetic plastic, etc.) and rigid (glass, wood, etc.) substrates with thick and durable three-dimensional superhydrophobic coatings. The synthesized superhydrophobic monoliths and polymeric coatings with controlled mechanical properties are appropriate to withstand different physical insults, including twisting, creasing, and even physical erosion of the material, without compromising the embedded antiwetting property. The materials are also equally resistant to various harsh chemical environments, and the embedded antifouling property remained unperturbed even after continuous exposure to extremes of pH (pH 1 and pH 11), artificial sea water for a minimum of 30 days. These flexible and formable free-standing monoliths and stable polymeric coatings that are extremely water-repellent both in air and under oil, are of utmost importance owing to their suitability in practical circumstances and robust nature.

Natural silk is easily accessible from silkworms and can be processed into different formats suitable as biomaterials and cell culture matrixes. Recombinant DNA technology enables chemical-free functionalization of partial silk proteins through fusion with peptide motifs and protein domains, but this constitutes a less cost-effective production process. Herein, we show that natural silk fibroin (SF) can be used as a bulk material that can be top-coated with a thin layer of the recombinant spider silk protein 4RepCT in fusion with various bioactive motifs and domains. The coating process is based on a silk assembly to achieve stable interactions between the silk types under mild buffer conditions. The assembly process was studied in real time by quartz crystal microbalance with dissipation. Coatings, electrospun mats, and microporous scaffolds were constructed from Antheraea assama and Bombyx mori SFs. The morphology of the fibroin materials before and after coating with recombinant silk proteins was analyzed by scanning electron microscopy and atomic force microscopy. SF materials coated with various bioactive 4RepCT fusion proteins resulted in directed antibody capture, enzymatic activity, and improved cell attachment and spreading, respectively, compared to pristine SF materials. The herein-described procedure allows a fast and easy route for the construction of bioactive materials.

Tunable repeated drug administration is often inevitable in a number of pathological cases. Reloadable 3D matrices for sustained drug delivery are predicted as a prospective avenue to realize this objective. This study was directed toward sonication-induced fabrication of novel reloadable Bombyx mori silk fibroin (SF) (4, 6, and 8 wt %) hydrogel, injected within 3D porous (8 wt %) scaffolds. The focus was to develop a dual-barrier reloadable depot system for sustained molecular cargo release. Both the varying SF concentration (4, 6, and 8 wt %) and the sonication time (30, 45, and 60 s) dictated the extent of cross-linking, β-sheet content, and porosity (1-10 μm) influencing the release behavior of model molecules. Release studies of model molecules (trypan blue, TB, 961 Da and bovine serum albumin, BSA, 66 kDa) for 28 days attested that the variations in their molecular weight, the matrix cross-linking density, and the scaffold-hydrogel interactions dictated the release behavior. The Ritger and Peppas equation was further fitted into the release behavior of model molecules from various SF matrices. The hybrid constructs exhibited high compressive strength along with in vitro compatibility using primary porcine chondrocytes and tunable enzymatic degradation as assessed for 28 days. The aptness of the constructs was evinced as a reloadable model molecule (BSA and fluorescein isothiocyanate-inulin, 3.9 kDa) depot system through UV-visible and fluorescence spectroscopic analyses. The novel affordable platform developed using silk scaffold-hydrogel hybrid constructs could serve as a sustained and reloadable drug depot system for administration of multiple and repeated drugs.

An osteoarthritis pandemic has accelerated exploration of various biomaterials for cartilage reconstruction with a special emphasis on silk fibroin from mulberry (Bombyx mori) and non-mulberry (Antheraea assamensis) silk worms. Retention of positive attributes of the agarose standard and nullification of its negatives are central to the current agarose/silk fibroin hydrogel design. In this study, hydrogels of mulberry and non-mulberry silk fibroin blended with agarose were fabricated and evaluated in vitro for two weeks for cartilaginous tissue formation. The fabricated hydrogels were physicochemically characterized and analyzed for cell viability, proliferation, and extra cellular matrix deposition. The amalgamation of silk fibroin with agarose impacted the pore size, as illustrated by field emission scanning electron microscopy studies, swelling behavior, and in vitro degradation of the hydrogels. Fourier transform infrared spectroscopy results indicated the blend formation and confirmed the presence of both components in the fabricated hydrogels. Rheological studies demonstrated enhanced elasticity of blended hydrogels with G' > G″. Biochemical analysis revealed significantly higher levels of sulfated glycosaminoglycans (sGAGs) and collagen (p ≤ 0.01) in blended hydrogels. More specifically, the non-mulberry silk fibroin blend showed sGAG and collagen content (∼1.5-fold) higher than that of the mulberry blend (p ≤ 0.05). Histological and immunohistochemical analyses further validated the enhanced deposition of sGAG and collagen, indicating maintenance of chondrogenic phenotype within constructs after two weeks of culture. Real-time PCR analysis further confirmed up-regulation of cartilage-specific aggrecan, sox-9 (∼1.5-fold) and collagen type II (∼2-fold) marker genes (p ≤ 0.01) in blended hydrogels. The hydrogels demonstrated immunocompatibility, which was evidenced by minimal in vitro secretion of tumor necrosis factor-α (TNF-α) by murine macrophages. Taken together, the results suggest promising attributes of blended hydrogels and particularly the non-mulberry silk fibroin/agarose blends as alternative biomaterial for cartilage tissue engineering.