Polymeric nanoparticles containing kojic acid induce structural alterations and apoptosis-like death in Leishmania (Leishmania) amazonensis.

Leishmaniasis encompasses a cluster of neglected tropical diseases triggered by kinetoplastid phatogens belonging to the genus Leishmania. Current therapeutic approaches are toxic, expensive, and require long-term treatment. Nanoparticles are emerging as a new alternative for the treatment of neglected tropical diseases. Silk Fibroin is a biocompatible and amphiphilic protein that can be used for formulating nanoemulsions, while kojic acid is a secondary metabolite with antileishmanial actions. Thus, this study evaluated the efficacy of a nanoemulsion, formulated with silk fibroin as the surfactant and containing kojic acid (NanoFKA), against promastigote and amastigote forms of Leishmania (Leishmania) amazonensis. The NanoFKA had an average particle size of 176 nm, Polydispersity Index (PDI) of 0.370, and a Zeta Potential of -32.3 mV. It presented inhibitory concentration (IC50) values of >56 µg/mL and >7 µg/mL for the promastigote and amastigote forms, respectively. Ultrastructural analysis, cell cycle distribution and phosphatidylserine exposure showed that NanoFKA treatment induces apoptosis-like cell death and cell cycle arrest in L. (L.) amazonensis. In addition, NanoFKA exhibited no cytotoxicity against macrophages. Given these results, NanoFKA present leishmanicidal activity against L. (L.) amazonensis.

Silk fibroin-based dressings with antibacterial and anti-inflammatory properties.

Silk fibroin is a fibrillar protein obtained from arthropods such as mulberry and non-mulberry silkworms. Silk fibroin has been used as a dressing in wound treatment for its physical, chemical, mechanical, and biological properties. This systematic review analyzed studies from PubMed, Web of Science, and Scopus databases to identify the molecules preferred for functionalizing silk fibroin-based dressings and to describe their mechanisms of exhibiting anti-inflammatory and antibacterial properties. The analysis of the selected articles allowed us to classify the dressings into different conformations, such as membranes, films, hydrogels, sponges, and bioadhesives. The incorporation of various molecules, including antibiotics, natural products, peptides, nanocomposites, nanoparticles, secondary metabolites, growth factors, and cytokines, has allowed the development of dressings that promote wound healing with antibacterial and immunomodulatory properties. In addition, silk fibroin-based dressings have been established to have the potential to regenerate wounds such as venous ulcers, arterial ulcers, diabetic foot, third-degree burns, and neoplastic ulcers. Evaluation of the efficacy of silk fibroin-based dressings in tissue engineering is an area of great activity that has shown significant advances in recent years.

Silk fibroin-derived electrospun materials for biomedical applications: A review.

Electrospinning is a versatile technique for fabricating polymeric fibers with diameters ranging from micro- to nanoscale, exhibiting multiple morphologies and arrangements. By combining silk fibroin (SF) with synthetic and/or natural polymers, electrospun materials with outstanding biological, chemical, electrical, physical, mechanical, and optical properties can be achieved, fulfilling the evolving biomedical demands. This review highlights the remarkable versatility of SF-derived electrospun materials, specifically focusing on their application in tissue regeneration (including cartilage, cornea, nerves, blood vessels, bones, and skin), disease treatment (such as cancer and diabetes), and the development of controlled drug delivery systems. Additionally, we explore the potential future trends in utilizing these nanofibrous materials for creating intelligent biomaterials, incorporating biosensors and wearable sensors for monitoring human health, and also discuss the bottlenecks for its widespread use. This comprehensive overview illuminates the significant impact and exciting prospects of SF-derived electrospun materials in advancing biomedical research and applications.

Poly(vinyl alcohol)/Silk Fibroin/Ag-NPs Composite Nanofibers as a Substrate for MG-63 Cells' Growth.

Nanofiber scaffolds of polyvinyl alcohol, silk fibroin from Bombyx mori cocoons, and silver nanoparticles were developed as a substrate for MG-63 growth. The fiber morphology, mechanical properties, thermal degradation, chemical composition, and water contact angle were investigated. In vitro tests were performed by the cell viability MTS test of MG-63 cells on electrospun PVA scaffolds, mineralization was analyzed by alizarin red, and the alkaline phosphatase (ALP) assay was evaluated. At higher PVA concentrations, Young's modulus (E) increased. The addition of fibroin and silver nanoparticles improved the thermal stability of PVA scaffolds. FTIR spectra indicated characteristic absorption peaks related to the chemical structures of PVA, fibroin, and Ag-NPs, demonstrating good interactions between them. The contact angle of the PVA scaffolds decreased with the incorporation of fibroin and showed hydrophilic characteristics. In all concentrations, MG-63 cells on PVA/fibroin/Ag-NPs scaffolds had higher cell viability than PVA pristine. On day ten of culture, PVA18/SF/Ag-NPs showed the highest mineralization, observed by the alizarin red test. PVA10/SF/Ag-NPs presented the highest alkaline phosphatase activity after an incubation time of 37 h. The achievements indicate the potential of the nanofibers of PVA18/SF/Ag-NPs as a possible substitute for bone tissue engineering (BTE).

Pro-angiogenic photo-crosslinked silk fibroin hydrogel: a potential candidate for repairing alveolar bone defects

Abstract Objective: This study aimed to develop a pro-angiogenic hydrogel with in situ gelation ability for alveolar bone defects repair. Methodology: Silk fibroin was chemically modified by Glycidyl Methacrylate (GMA), which was evaluated by proton nuclear magnetic resonance (1H-NMR). Then, the photo-crosslinking ability of the modified silk fibroin was assessed. Scratch and transwell-based migration assays were conducted to investigate the effect of the photo-crosslinked silk fibroin hydrogel on the migration of human umbilical vein endothelial cells (HUVECs). In vitro angiogenesis was conducted to examine whether the photo-crosslinked silk fibroin hydrogel would affect the tube formation ability of HUVECs. Finally, subcutaneous implantation experiments were conducted to further examine the pro-angiogenic ability of the photo-crosslinked silk fibroin hydrogel, in which the CD31 and α-smooth muscle actin (α-SMA) were stained to assess neovascularization. The tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were also stained to evaluate inflammatory responses after implantation. Results: GMA successfully modified the silk fibroin, which we verified by our 1H-NMR and in vitro photo-crosslinking experiment. Scratch and transwell-based migration assays proved that the photo-crosslinked silk fibroin hydrogel promoted HUVEC migration. The hydrogel also enhanced the tube formation of HUVECs in similar rates to Matrigel®. After subcutaneous implantation in rats for one week, the hydrogel enhanced neovascularization without triggering inflammatory responses. Conclusion: This study found that photo-crosslinked silk fibroin hydrogel showed pro-angiogenic and inflammation inhibitory abilities. Its photo-crosslinking ability makes it suitable for matching irregular alveolar bone defects. Thus, the photo-crosslinkable silk fibroin-derived hydrogel is a potential candidate for constructing scaffolds for alveolar bone regeneration.

Performance of Colombian Silk Fibroin Hydrogels for Hyaline Cartilage Tissue Engineering.

The development and evaluation of scaffolds play a crucial role in the engineering of hyaline cartilage tissue. This work aims to evaluate the performance of silk fibroin hydrogels fabricated from the cocoons of the Colombian hybrid in the in vitro regeneration of hyaline cartilage. The scaffolds were physicochemically characterized, and their performance was evaluated in a cellular model. The results showed that the scaffolds were rich in random coils and ß-sheets in their structure and susceptible to various serine proteases with different degradation profiles. Furthermore, they showed a significant increase in ACAN, COL10A1, and COL2A1 expression compared to pellet culture alone and allowed GAG deposition. The soluble portion of the scaffold did not affect chondrogenesis. Furthermore, they promoted the increase in COL1A2, showing a slight tendency to differentiate towards fibrous cartilage. The results also showed that Colombian silk could be used as a source of biomedical devices, paving the way for sericulture to become a more diverse economic activity in emerging countries.

Electrochemical and Electroconductive Behavior of Silk Fibroin Electrospun Membrane Coated with Gold or Silver Nanoparticles.

The surface modification of materials obtained from natural polymers, such as silk fibroin with metal nanoparticles that exhibit intrinsic electrical characteristics, allows the obtaining of biocomposite materials capable of favoring the propagation and conduction of electrical impulses, acting as communicating structures in electrically isolated areas. On that basis, this investigation determined the electrochemical and electroconductive behavior through electrochemical impedance spectroscopy of a silk fibroin electrospun membrane from silk fibrous waste functionalized with gold or silver nanoparticles synthetized by green chemical reduction methodologies. Based on the results obtained, we found that silk fibroin from silk fibrous waste (SFw) favored the formation of gold (AuNPs-SFw) and silver (AgNPs-SFw) nanoparticles, acting as a reducing agent and surfactant, forming a micellar structure around the individual nanoparticle. Moreover, different electrospinning conditions influenced the morphological properties of the fibers, in the presence or absence of beads and the amount of sample collected. Furthermore, treated SFw electrospun membranes, functionalized with AuNPs-SFw or AgNPS-SFw, allowed the conduction of electrical stimuli, acting as stimulators and modulators of electric current.

Silk Fibroin-g-Polyaniline Platform for the Design of Biocompatible-Electroactive Substrate.

The structural modification of biopolymers is a current strategy to develop materials with biomedical applications. Silk fibroin is a natural fiber derived from a protein produced by the silkworm (Bombyx mori) with biocompatible characteristics and excellent mechanical properties. This research reports the structural modification of silk fibroin by incorporating polyaniline chain grafts through a one-pot process (esterification reaction/oxidative polymerization). The structural characterization was achieved by 1H-NMR and FT-IR. The morphology was studied by scanning electron microscopy and complemented with thermogravimetric analysis to understand the effect of the thermal stability at each step of the modification. Different fibroin silk (Fib): polyaniline (PAni) mass ratios were evaluated. From this evaluation, it was found that a Fib to PAni ratio of at least 1 to 0.5 is required to produce electroactive polyaniline, as observed by UV-vis and CV. Notably, all the fibroin-g-PAni systems present low cytotoxicity, making them promising systems for developing biocompatible electrochemical sensors.

Silk Fibroin/Poly(vinyl Alcohol) Microneedles as Carriers for the Delivery of Singlet Oxygen Photosensitizers.

Photodynamic therapy (PDT) is a medical treatment in which a combination of a photosensitizing drug and visible light produces highly cytotoxic reactive oxygen species (ROS) that leads to cell death. One of the main drawbacks of PDT for topical treatments is the limited skin penetration of some photosensitizers commonly used in this therapy. In this study, we propose the use of polymeric microneedles (MNs) prepared from silk fibroin and poly(vinyl alcohol) (PVA) to increase the penetration efficiency of porphyrin as possible applications in photodynamic therapy. The microneedle arrays were fabricated from mixtures in different proportions (1:0, 7:3, 1:1, 3:7, and 0:1) of silk fibroin and PVA solutions (7%); the polymer solutions were cast in polydimethylsiloxane (PDMS) molds and dried overnight. Patches containing grids of 10 × 10 microneedles with a square-based pyramidal shape were successfully produced through this approach. The polymer microneedle arrays showed good mechanical strength under compression force and sufficient insertion depth in both Parafilm M and excised porcine skin at different application forces (5, 20, 30, and 40 N) using a commercial applicator. We observe an increase in the cumulative permeation of 5-[4-(2-carboxyethanoyl) aminophenyl]-10,15,20-tris-(4-sulphonatophenyl) porphyrin trisodium through porcine skin treated with the polymer microneedles after 24 h. MNs may be a promising carrier for the transdermal delivery of photosensitizers for PDT, improving the permeation of photosensitizer molecules through the skin, thus improving the efficiency of this therapy for topical applications.

Confecção, caracterização e avaliação do efeito de membranas à base de fibroína carregadas ou não com neurotensina em feridas abertas em palato: estudo clínico randomizado / Manufacturing, characterization, and evaluation of silk fibroin-based membranes with or without neurotensin on palatal wound healing: in vitro and randomized clinical trial

A utilização de enxertos autógenos provenientes do palato para reconstrução de tecidos gengivais, apesar de serem considerados o padrão-ouro, causam grande morbidade e dor pós-operatória. Assim, com o intuito de reduzir essas complicações, alguns biomateriais podem ser explorados, acelerando a cicatrização e trazendo maior conforto ao paciente. Dessa forma, os objetivos do presente estudo foram: confeccionar e caracterizar membranas à base de fibroína de seda sem carregamento (F) ou carregadas com neurotensina (FN) e avaliar clinicamente seus efeitos no reparo de feridas abertas no palato. Para a primeira parte, as membranas foram confeccionadas e caracterizadas quanto à espessura, propriedades mecânicas, textura superficial, permeabilidade ao vapor de água, degradação enzimática, citotoxicidade, genotoxicidade e intumescimento. Para a segunda parte, sessenta e um pacientes com necessidade de exodontia e preservação alveolar foram randomizados em três grupos: Controle: exodontia e selamento do alvéolo com EGL removido do palato; Grupo F: exodontia com EGL e inserção da membrana F na ferida palatina; Grupo FN: exodontia com EGL e membrana FN na ferida palatina. A reparação da área doadora palatina foi avaliada por parâmetros clínicos centrados nos pacientes. A partir dos resultados de caracterização, pode-se constatar a homogeneidade das membranas, alta resistência mecânica e maior intumecimento, permeabilidade e rugosidade superficial, características favoráveis para feridas intraorais. Em adição, não apresentaram citotoxicidade (HFF-1, HaCaT e fibroblastos gengivais humanos) e genotoxicidade (HaCaT) in vitro. Em relação aos resultados clínicos, em 14 dias FN mostrou maior diminuição de ARF (p<0,001) e melhor ET (p = 0,022) do que F e GC; ao analisar-se o ICPF, ambos os biomateriais F e FN apresentaram melhores resultados em 14 dias comparados ao GC (p= 0,009 e p = 0,001 respectivamente); não houve diferença entre os grupos em relação ao ED (p= 0,312) e ET (p = 0,120). OHIP em F e FN se mostrou superior a GC a partir do 5º dia, NA e DPO mantiveram-se mais baixas em F e FN. Conclui-se que o uso de membranas à base de fibroína de seda carregadas ou não com NT apresenta benefícios clínicos no reparo de feridas e na qualidade de vida dos pacientes (AU)

The use of autogenous grafts from the palate for gingival tissue reconstruction, despite being considered the gold standard, causes great morbidity and postoperative pain. Thus, in order to reduce these complications, biomaterials can be explored, accelerating healing and bringing greater comfort to the patient. The objectives of the present study were: to manufacture and characterize unloaded (F) or neurotensinloaded (FN) silk fibroin-based membranes and to clinically evaluate their effects on the repair of palatal open wounds. For the first part, the membranes were made and characterized in terms of thickness, mechanical properties, surface texture, water vapor permeability, enzymatic degradation, cytotoxicity, genotoxicity and swelling. For the second part, sixty-one patients in need of extraction and alveolar preservation were randomized into three groups: Control: extraction and alveolar socket sealing with FGG removed from the palate; Group F: extraction with FGG and F membrane incorporation in the palatal wound; Group FN: extraction with EGL and FN membrane in the palatal wound. Palatal donor site repair was assessed by patient-centered and clinical parameters. From the characterization results, membranes' high homogeneity, mechanical resistance, swelling, permeability and surface roughness, can be verified, being favorable characteristics for intraoral wounds. In addition, they showed no cytotoxicity (HFF-1, HaCaT and human gingival fibroblasts) and genotoxicity (HaCaT) in vitro. Regarding the clinical results, at 14 days FN showed a greater decrease in ARF (p<0.001) and better ET (p = 0.022) than F and GC; when analyzing the ICPF, both biomaterials F and FN showed better results at 14 days compared to GC (p=0.009 and p=0.001 respectively); there was no difference between the groups regarding ED (p=0.312) and ET (p=0.120). OHIP in F and FN was superior to GC from the 5th day, NA and DPO remained lower in F and FN. It is concluded that the use of silk fibroinbased membranes loaded or not with NT has clinical benefits in wound repair and in the quality of life of patients (AU)

Chitosan/ß-TCP composites scaffolds coated with silk fibroin: a bone tissue engineering approach.

Bone regeneration and natural repair are long-standing processes that can lead to uneven new tissue growth. By introducing scaffolds that can be autografts and/or allografts, tissue engineering provides new approaches to manage the major burdens involved in this process. Polymeric scaffolds allow the incorporation of bioactive agents that improve their biological and mechanical performance, making them suitable materials for bone regeneration solutions. The present work aimed to create chitosan/beta-tricalcium phosphate-based scaffolds coated with silk fibroin and evaluate their potential for bone tissue engineering. Results showed that the obtained scaffolds have porosities up to 86%, interconnectivity up to 96%, pore sizes in the range of 60-170 µm, and a stiffness ranging from 1 to 2 MPa. Furthermore, when cultured with MC3T3 cells, the scaffolds were able to form apatite crystals after 21 d; and they were able to support cell growth and proliferation up to 14 d of culture. Besides, cellular proliferation was higher on the scaffolds coated with silk. These outcomes further demonstrate that the developed structures are suitable candidates to enhance bone tissue engineering.

Recovery of motor function in rats with complete spinal cord injury following implantation of collagen/silk fibroin scaffold combined with human umbilical cord-mesenchymal stem cells

SUMMARY OBJECTIVE: This study aimed to assess the effect of the collagen/silk fibroin scaffolds seeded with human umbilical cord-mesenchymal stem cells on functional recovery after acute complete spinal cord injury. METHODS: The fibroin and collagen were mixed (mass ratio, 3:7), and the composite scaffolds were produced. Forty rats were randomly divided into the Sham group (without spinal cord injury), spinal cord injury group (spinal cord transection without any implantation), collagen/silk fibroin scaffolds group (spinal cord transection with implantation of the collagen/silk fibroin scaffolds), and collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group (spinal cord transection with the implantation of the collagen/silk fibroin scaffolds co-cultured with human umbilical cord-mesenchymal stem cells). Motor evoked potential, Basso-Beattie-Bresnahan scale, modified Bielschowsky's silver staining, and immunofluorescence staining were performed. RESULTS: The BBB scores in the collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group were significantly higher than those in the spinal cord injury and collagen/silk fibroin scaffolds groups (p<0.05 or p<0.01). The amplitude and latency were markedly improved in the collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group compared with the spinal cord injury and collagen/silk fibroin scaffolds groups (p<0.05 or p<0.01). Meanwhile, compared to the spinal cord injury and collagen/silk fibroin scaffolds groups, more neurofilament positive nerve fiber ensheathed by myelin basic protein positive structure at the injury site were observed in the collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group (p<0.01, p<0.05). The results of Bielschowsky's silver staining indicated more nerve fibers was observed at the lesion site in the collagen/silk fibroin scaffolds + human umbilical cord-mesenchymal stem cells group compared with the spinal cord injury and collagen/silk fibroin scaffolds groups (p<0.01, p< 0.05). CONCLUSION: The results demonstrated that the transplantation of human umbilical cord-mesenchymal stem cells on a collagen/silk fibroin scaffolds could promote nerve regeneration, and recovery of neurological function after acute spinal cord injury.

Silk fibroin nanocomposites as tissue engineering scaffolds - A systematic review.

Silk fibroin is a protein with intrinsic characteristics that make it a good candidate as a scaffold for tissue engineering. Recent works have enhanced its benefits by adding inorganic phases that interact with silk fibroin in different ways. A systematic review was performed in four databases to study the physicochemical and biological performance of silk fibroin nanocomposites. In the last decade, only 51 articles contained either in vitro cell culture models or in vivo tests. The analysis of such works resulted in their classification into the following scaffold types: particles, mats and textiles, films, hydrogels, sponge-like structures, and mixed conformations. From the physicochemical perspective, the inorganic phase imbued in silk fibroin nanocomposites resulted in better stability and mechanical performance. This review revealed that the inorganic phase may be associated with specific biological responses, such as neovascularisation, cell differentiation, cell proliferation, and antimicrobial and immunomodulatory activity. The study of nanocomposites as tissue engineering scaffolds is a highly active area mostly focused on bone and cartilage regeneration with promising results. Nonetheless, there are still many challenges related to their application in other tissues, a better understanding of the interaction between the inorganic and organic phases, and the associated biological response.

Dissolution of Silk Fibroin in Mixtures of Ionic Liquids and Dimethyl Sulfoxide: On the Relative Importance of Temperature and Binary Solvent Composition.

We studied the dependence of dissolution of silk fibroin (SF) in mixtures of DMSO with ionic liquids (ILs) on the temperature (T = 40 to 80 °C) and DMSO mole fraction (χDMSO = 0.5 to 0.9). The ILs included BuMeImAcO, C3OMeImAcO, AlBzMe2NAcO, and Bu4NAcO; see the names and structures below. We used design of experiments (DOE) to determine the dependence of mass fraction of dissolved SF (SF-m%) on T and χDMSO. We successfully employed a second-order polynomial to fit the biopolymer dissolution data. The resulting regression coefficients showed that the dissolution of SF in BuMeImAcO-DMSO and C3OMeImAcO-DMSO is more sensitive to variation of T than of χDMSO; the inverse is observed for the quaternary ammonium ILs. Using BuMeImAcO, AlBzMe2NAcO, and molecular dynamics simulations, we attribute the difference in IL efficiency to stronger SF-IL hydrogen bonding with the former IL, which is coupled with the difference in the molecular volumes and the rigidity of the phenyl ring of the latter IL. The order of SF dissolution is BuMeImAcO-DMSO > C3OMeImAcO-DMSO; this was attributed to the formation of intramolecular H-bonding between the ether oxygen in the side chain of the latter IL and the relatively acidic hydrogens of the imidazolium cation. Using DOE, we were able to predict values of SF-m%; this is satisfactory and important because it results in economy of labor, time, and material.

Engineering of sustainable biomaterial composites from cellulose and silk fibroin: Fundamentals and applications.

This review addresses composites prepared from cellulose (Cel) and silk fibroin (SF) to generate multifunctional, biocompatible, biodegradable materials such as fibers, films and scaffolds for tissue engineering. First, we discuss briefly the molecular structures of Cel and SF. Their structural features explain why certain solvents, e.g., ionic liquids, inorganic electrolyte solutions dissolve both biopolymers. We discuss the mechanisms of Cel dissolution because in many cases they also apply to (much less studied) SF dissolution. Subsequently, we discuss the fabrication and characterization of Cel/SF composite biomaterials. We show how the composition of these materials beneficially affects their mechanical properties, compared to those of the precursor biopolymers. We also show that Cel/SF materials are excellent and versatile candidates for biomedical applications because of the inherent biocompatibility of their components.

Effects of Chemical Post-treatments on Structural and Physicochemical Properties of Silk Fibroin Films Obtained From Silk Fibrous Waste.

Silk fibroin (SF) is a protein polymer claimed to have outstanding potential for medical applications. However, because of the manufacturing process, materials from regenerated SF exhibit a higher percentage of amorphous structures. The amorphous structures cause the material to be water soluble and can significantly limit its applications in wet biological environments. In order to increase the amount of crystalline structures and decrease the water solubility of SF materials, post-treatment with alcohols is usually employed. SF can be obtained from silk fibrous wastes (SFW), usually discarded in silk textile processes. This represents an opportunity to produce materials with high added value from low-cost natural sources. In this study, SF was obtained from SFW, and films were made thereof followed by a post-treatment by immersion or in a saturated atmosphere of methanol (MeOH) or ethanol (EtOH), using different exposure times. The resulting films were analyzed according to crystallinity, the percentage of crystalline and amorphous structures, and thermal stability. Also, water absorption and weight loss in aqueous media were determined. The results showed a significant increase in crystalline structures in all treated samples, varying according to the type and time of exposure to post-treatment conducted. The highest increase was shown in the case of the post-treatment by immersion in MeOH for 1 h, with a 23% increase over the untreated sample. This increase in crystallinity was reflected in an increase in the degradation temperature and a degradation rate of 5.3% on day 7. The possibility of tuning the degree of crystallinity, as well as thermal stability and aqueous integrity of thin films of SFW, can be applied to adjust these materials to the requirements of specific biomedical applications.

Direct Femtosecond Laser Printing of Silk Fibroin Microstructures.

Fabrication of functional silk fibroin microstructures has extensive applications in biotechnology and photonics. Considerable progress has been made based on lithographic methods and self-assembly approaches. However, most methods require chemical modification of silk fibroin, which restricts the functionalities of the designed materials. At the same time, femtosecond laser-induced forward transfer (fs-LIFT) has been explored as a simple and attractive processing tool for microprinting of high-resolution structures. In this paper, we propose the use of LIFT with fs-pulses for creating high-resolution structures of regenerated silk fibroin (SF). Furthermore, upon adding Eu3+/Tb3+ complexes to SF, we have been able to demonstrate the printing by LIFT of luminescent SF structures with a resolution on the order of 2 µm and without material degradation. This approach provides a facile method for printing well-defined two-dimensional (2D) micropatterns of pure and functionalized SF, which can be used in a wide range of optical and biomedical applications.

Phase Diagram and Estimation of Flory-Huggins Parameter of Interaction of Silk Fibroin/Sodium Alginate Blends.

Silk fibroin (SF) and sodium alginate (SA) are natural polymers used to produce biomaterials. One of the strategies to improve the properties of these products is to prepare blends with them, which are partially miscible. Phase separation is observed, therefore, the thermodynamic analysis of this system is important to predict the final state and composition of this blends. This study explored blends with a different initial composition of SF, SA, and water (WA) at 25°C and neutral pH. After phase separation, two phases were identified, one rich in SF and other rich in SA. The Flory-Huggins parameters of interaction of polymer-solvent and polymer-polymer were estimated using the extended equation and data of phase equilibrium, their values indicates the partial miscibility of the polymers.

A Cellularized Biphasic Implant Based on a Bioactive Silk Fibroin Promotes Integration and Tissue Organization during Osteochondral Defect Repair in a Porcine Model.

In cartilage tissue engineering, biphasic scaffolds (BSs) have been designed not only to influence the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone, promoting the implant's integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a BS based on the assembly of a cartilage phase constituted by fibroin biofunctionalyzed with a bovine cartilage matrix, cellularized with differentiated autologous pre-chondrocytes and well attached to a bone phase (decellularized bovine bone) to promote cartilage regeneration in a model of joint damage in pigs. BSs were assembled by fibroin crystallization with methanol, and the mechanical features and histological architectures were evaluated. The scaffolds were cellularized and matured for 12 days, then implanted into an osteochondral defect in a porcine model (n = 4). Three treatments were applied per knee: Group I, monophasic cellular scaffold (single chondral phase); group II (BS), cellularized only in the chondral phase; and in order to study the influence of the cellularization of the bone phase, Group III was cellularized in chondral phases and a bone phase, with autologous osteoblasts being included. After 8 weeks of surgery, the integration and regeneration tissues were analyzed via a histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular BSs reached a Young's modulus of 805.01 kPa, similar to native cartilage. In vitro biological studies revealed the chondroinductive ability of the BSs, evidenced by an increase in sulfated glycosaminoglycans and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, in Group I, the defects were not reconstructed. In Groups II and III, a good integration of the implant with the surrounding tissue was observed. Defects in group II were fulfilled via hyaline cartilage and normal bone. Group III defects showed fibrous repair tissue. In conclusion, our findings demonstrated the efficacy of a biphasic and bioactive scaffold based on silk fibroin and cellularized only in the chondral phase, which entwined chondroinductive features and a biomechanical capability with an appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Effect of Silk Fibroin on Cell Viability in Electrospun Scaffolds of Polyethylene Oxide.

In this study, a coating from electrospun silk fibroin was performed with the aim to modify the surface of breast implants. We evaluated the effect of fibroin on polymeric matrices of poly (ethylene oxide) (PEO) to enhance cell viability, adhesion, and proliferation of HaCaT human keratinocytes to enhance the healing process on breast prosthesis implantation. We electrospun six blends of fibroin and PEO at different concentrations. These scaffolds were characterized by scanning electron microscopy, contact angle measurements, ATR-FTIR spectroscopy, and X-ray diffraction. We obtained diverse network conformations at different combinations to examine the regulation of cell adhesion and proliferation by modifying the microstructure of the matrix to be applied as a potential scaffold for coating breast implants. The key contribution of this work is the solution it provides to enhance the healing process on prosthesis implantation considering that the use of these PEO⁻fibroin scaffolds reduced (p < 0.05) the amount of pyknotic nuclei. Therefore, viability of HaCaT human keratinocytes on PEO⁻fibroin matrices was significantly improved (p < 0.001). These findings provide a rational strategy to coat breast implants improving biocompatibility.