Bombyx Mori Cocoons Research Articles

Silk sericin-based functional food for osteoporosis prevention and therapy

The development of new functional foods that can potentially reduce the risk of diseases, such as osteoporosis, has received increasing attention in recent years. Here we demonstrated that silk sericin-based functional food (SSBF) exerted preventive and therapeutic effects against bone loss. Sericin isolated at 70 °C for 60 min from the cocoon of Bombyx mori stimulated osteoblast differentiation by increasing osteoblastic gene expression while inhibiting receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation in vitro. Micro-computed tomography analysis revealed that pre- and post-oral administration of SSBF increased bone mass in ovariectomized mouse model in vivo. Furthermore, dynamic bone histomorphometry analysis revealed that SSBF enhanced osteoblast activity and suppressed bone resorption by reducing the number of osteoclasts. These findings indicated the potential role of SSBF in preventing and being a therapeutic candidate for osteoporosis.

The Potential of Natural Carotenoids-Containing Sericin of the Domestic Silkworm Bombyx mori.

Sericin derived from the white cocoon of Bombyx mori has been attracting more attention for its utilization in food, cosmetics, and biomedicine. The potential health benefits of natural carotenoids for humans have also been well-established. Some rare strains of Bombyx mori (B. mori) produce yellow-red cocoons, which endow a potential of natural carotenoid-containing sericin. We hypothesized that natural carotenoid-containing sericin from yellow-red cocoons would exhibit better properties compared with white cocoon sericin. To investigate the physicochemical attributes of natural carotenoid-containing sericin, we bred two silkworm strains from one common ancestor, namely XS7 and XS8, which exhibited different cocoon colors as a result of the inconsistent distribution of lutein and β-carotene. Compared with white cocoon sericin, the interaction between carotenoids and sericin molecules in carotenoid-containing sericin resulted in a unique fluorescence emission at 530, 564 nm. The incorporation of carotenoids enhanced the antibacterial effect, anti-cancer ability, cytocompatibility, and antioxidant of sericin, suggesting potential wide-ranging applications of natural carotenoid-containing sericin as a biomass material. We also found differences in fluorescence characteristics, antimicrobial effects, anti-cancer ability, and antioxidants between XS7 and XS8 sericin. Our work for the first time suggested a better application potential of natural carotenoid-containing sericin as a biomass material than frequently used white cocoon sericin.

Fibroin Nanoparticles: Use in Drug Delivery

The silkworm Bombyx mori.L. is infected by various diseases viz; grasserie, flacherie, muscardine, and pebrine. Among all these diseases the grasserie causes major economic loss to the industry and is one of the main reasons for low silk productivity. It is caused by a virus known as Bombyx mori nucleopolyhedro virus (BmNPV). The impact of grasserie disease on silkworms is significant as it leads to reduced silk production and can result in economic losses for sericulture farmers. In India greater than 50% of silk cocoon crop loss is due to BmNPV [1] and in Kashmir valley, the loss is about 28-32% [2]. Silk obtained from the cocoons of silkworm Bombyx mori L. is a natural fibrous protein well known for being lightweight, having high mechanical strength, good flexibility, and luster making it ideal for the textile industry. In addition, fibroin extracted from the cocoons of domesticated silkworm Bombyx mori L. has gained growingly interest due to its excellent mechanical properties and high biocompatibility, biodegradability, inexpensiveness, and preparation flexibility [3]. These properties of silk fibroin lead to the formulation of fibroin nanoparticles (FNP’-s) which can be used to encapsulate different types of therapeutic compounds like proteins, vaccines, enzymes, etc. Fibroin has been approved as a biomaterial by the Food and Drug Administration (FDA) and has been popularly used in numerous medical applications such as sutures, tissue regeneration, coating devices, and drug delivery systems [4]. It has been studied that SF-derived curcumin nanoparticles show higher efficacy against breast cancer cells and have the potential to treat in vivo breast tumors by local, sustained, and long-term therapeutic delivery as a biodegradable system [5]. Therefore, fibroin-coated nanoparticles can be used effectively for disease management.

Impact of crystalline domains on long-term stability and mechanical performance of anisotropic silk fibroin sponges.

Sponge-like materials made from regenerated silk fibroin biopolymers are a tunable and advantageous platform for in vitro engineered tissue culture and in vivo tissue regeneration. Anisotropic, three-dimensional (3D) silk fibroin sponge-like scaffolds can mimic the architecture of contractile muscle. Herein, we use silk fibroin solution isolated from the cocoons of Bombyx mori silkworms to form aligned sponges via directional ice templating in a custom mold with a slurry of dry ice and ethanol. Hydrated tensile mechanical properties of these aligned sponges were evaluated as a function of silk polymer concentration (3% or 5%), freezing time (50% or 100% ethanol), and post-lyophilization method for inducing crystallinity (autoclaving, water annealing). Hydrated static tensile tests were used to determine Young's modulus and ultimate tensile strength across sponge formulations at two strain rates to evaluate rate dependence in the calculated parameters. Results aligned with previous reports in the literature for isotropic silk fibroin sponge-like scaffolds, where the method by which beta-sheets were formed and level of beta-sheet content (crystallinity) had the greatest impact on static parameters, while polymer concentration and freezing rate did not significantly impact static mechanical properties. We estimated the crystalline organization using molecular dynamics simulations to show that larger crystalline regions may be responsible for strength at low strain amplitudes and brittleness at high strain amplitudes in the autoclaved sponges. Within the parameters evaluated, extensional Young's modulus is tunable in the range of 600-2800 kPa. Dynamic tensile testing revealed the linear viscoelastic region to be between 0% and 10% strain amplitude and 0.2-2 Hz frequencies. Long-term stability was evaluated by hysteresis and fatigue tests. Fatigue tests showed minimal change in the storage and loss modulus of 5% silk fibroin sponges for more than 6000 min of continuous mechanical stimulation in the linear regime at 10% strain amplitude and 1 Hz frequency. Furthermore, we confirmed that these mechanical properties hold when decellularized extracellular matrix is added to the sponges and when the mechanical property assessments were performed in cell culture media. We also used nano-computed tomography (nano-CT) and simulations to explore pore interconnectivity and tortuosity. Overall, these results highlight the potential of anisotropic, sponge-like silk fibroin scaffolds for long-term (>6 weeks) contractile muscle culture with an in vitro bioreactor system that provides routine mechanical stimulation.

Microstructure of the silk fibroin-based hydrogel scaffolds derived from the orb-web spider Trichonephila clavata

Due to the unique properties of the silk fibroin (SF) made from silkworm, SF-based hydrogels have recently received significant attention for various biomedical applications. However, research on the SF-based hydrogels isolated from spider silks has been rtricted due to the limited collection and preparation of naïve silk materials. Therefore, this study focused on the microstructural characteristics of hydrogel scaffolds derived from two types of woven silk glands: the major ampullate gland (MAG) and the tubuliform gland (TG), in the orb-web spider Trichonephila clavate. We compared these spider glands with those of the silk fibroin (SF) hydrogel scaffold extracted from the cocoon of the insect silkworm Bombyx mori. Our FESEM analysis revealed that the SF hydrogel has high porosity, translucency, and a loose upper structure, with attached SF fibers providing stability. The MAG hydrogel displayed even higher porosity, as well as elongated fibrous structures, and improved mechanical properties: while the TG hydrogel showed increased porosity, ridge-like or wall-like structures, and stable biocapacity formed by physical crosslinking. Due to their powerful and versatile microstructural characteristics, the MAG and TG hydrogels can become tailored substrates, very effective for tissue engineering and regenerative medicine applications.

Exploring silk fibroin aerogels via different coagulation approaches

Aerogels are highly porous nanostructured materials with a high specific surface area, a very low density and a low thermal conductivity. Bio-aerogels, prepared from biopolymers, have gained interest for biomedical applications. While most bio-aerogels are polysaccharide-based, protein-based aerogels using silk fibroin (SF) from Bombyx mori cocoons remain underexplored. Our study delved into the impact of coagulation methods on SF aerogel properties, specifically, comparing ethanol treatment (Method 1) and sodium dihydrogen phosphate (NaH2PO4) (Method 2). Aerogels obtained through Method 1 exhibited a volume shrinkage of approximately 12–22 % and a density of about 0.06–0.07 g cm−3, while those obtained via Method 2 demonstrated a more substantial volume shrinkage of approximately 60–70 % and a higher density of around 0.2 g cm−3. These differences significantly influenced the internal structure of the aerogels, manifesting distinct morphological features of the materials. Mechanical tests revealed that SF aerogels derived from Method 2 displayed superior stress resistance at 80 % strain and higher elastic recovery when compared to samples of Method 1. In conclusion, the choice in coagulation methods broadens the mechanical property and density window for this type of aerogel which offers opportunities for a wide range of biomedical applications.

Conductive electrospun nanofiber based on silk fibroin/cellulose nanocrystals/reduced graphene oxide as a wound healing material

Composite electrospun nanofibers based on silk fibroin (SF), aminated reduced graphene oxide (NH2rGO), and oxidized-cellulose nanocrystals (AD-CNCs) were designed to promote the healing process by electrical conductivity. In this respect, CNCs were produced from cotton cellulose and oxidized in the presence of sodium periodate, and reduced GO was synthesized and functionalized with amine with p-anthranilic acid via an esterification reaction. Then the crosslink was prepared by the Schiff-base reaction between oxidized CNCs and NH2rGO. Electrospun nanofibers were prepared from silk fibroin, previously prepared from Bombyx Mori cocoons, AD-CNCs, and different ratios of NH2rGO. The SF/AD-CNCs/NH2rGO nanofibrous scaffold presented multiple propitious characteristics for cell proliferation and wound dressing including low cytotoxicity, a hydrophilic surface, thermal stability, and degradability. The physicochemical evaluations showed that with increasing the NH2rGO content, hydrophilicity and electroconductivity of nanofibers increased. The SF/AD-CNCs/NH2rGO biodegradable conductive scaffold with 2.5% NH2rGO content showed favorable nanofibrous structure, supporting and stimulating fibroblast cell proliferation and attachment. The SF/AD-CNCs/NH2rGO nanofibrous scaffold demonstrates the application potential in wound healing management.

Extraction and characterization of sericin obtained from unreelable cocoons of Bombyx mori L

Extraction and characterization of sericin obtained from unreelable cocoons of Bombyx mori L

Effect of Herbal Plant Extracts on the Economic Parameters of Silkworm Bombyx mori L.

Background: In the unorganized sector, the silk industry has grown to become a well-liked cottage business that employs more than 10 million rural residents. The development of new technologies that are efficient, labor-saving, and environmentally benign is being pursued. Objectives: To evaluate the secondary metabolites present in the selected Cucurbita moschata and Ipomoea batatas herbal plant (Leaf) extracts with five solvents namely ethanol, chloroform, acetone, petroleum ether, and water. To determine the antimicrobial activity of plant extracts against selected bacterial strains and assess the effect of fortified mulberry leaves with plants on the growth of silkworms and their weight of cocoon, pupa, shell, and shell ratio of silkworm Bombyx mori. Method: In this study, the selected herbal plants (Cucurbita moschata and Ipomoea batatas) were collected by hand-picking method from Singaneri village, Nanguneri Taluk, Tirunelveli districts, Tamil Nadu, India. The qualitative phytochemical analyses of selected herbal plant extracts were performed using standard procedures. Results: Phytochemicals results revealed that the different extracts (petroleum ether, chloroform, acetone, ethanol, and water extracts) revealed that, phenolic compounds, steroids, saponins, and xanthoproteins were present in the acetone extract. In petroleum ether tannins, saponins, pholobatanins, cardiac glycosides, phenolic compounds, flavanoids, carbohydrates, and xanthoproteins were present. In the antibacterial activity, ethanol and acetone extracts showed significant inhibitory effects against the pathogens viz., Enterococcus, E.coli, Aceto bactor and Bacillus cereus extracts did not show any activity. Moderate activity was expressed by ethanol extracts of I. batatasa gainst E.coli and Acetobactor (8.1±0.3mm diameter) concerning petroleum ether extracts. C. moschata revealed high activity against E.coli and Acetobactor with diameter of inhibition greater than 13.0±0.2 mm while C. moschata exhibited only moderate activity against E.coli and Acetobactor. Conclusion: The selected herbal plant extracts could increase some biological characteristics of silkworm B. mori, but this enhancement could economically improve the sericulture goals. The growers might be offered this substance to help them produce more silk.

Study on Structure and Anti-UV Properties of Sericin Cocoons

Abstract Through biogenetic engineering, the posterior silk glands of Bombyx mori larvae are degenerated, and only the middle silk glands are retained to secrete sericin to produce sericin fibers, so as to obtain sericin cocoons with high sericin content (>98.5%). For this paper, sericin cocoons were studied, and the morphological structure characteristics and properties of sericin cocoons and common Bombyx mori cocoons were compared and analyzed through such modern test techniques as SEM, FTIR, and XRD. Results show that sericin cocoons have the basic morphological structure of cocoons, but because of the brittleness and hardness of sericin, the ssericin cocoons have poor integrity with a thermal decomposition rate that is higher than that of cocoons; the two cocoons are of a similar protein structure, with both α-helix and β-sheet structures, and dominantly the β-sheet structure; and the crystallinity of sericin cocoons (10.48%) is lower than that of the common Bombyx mori cocoons (27.10%). Sericin was extracted from both cocoons to obtain a mixed sericin solution respectively, which were coated on base fabrics of polyester taffeta (210T) for coating finish and subjected to qualitative research for their anti-UV properties. The base fabrics coated with mixed sericin solutions of sericin cocoons and Bombyx mori cocoons have an anti-UV performance index (UPF) of 23.9% and 22.5% respectively, which are higher than that of the base fabric (8.9%)..

Composite resin reinforced with silk nanoparticles from Bombyx mori cocoon for dental applications.

The objective of this work was to evaluate the mechanical performance of Z350 resin composite modified with Bombyx mori cocoons silk nanoparticles for dental applications. Four experimental groups were analyzed G0% = Filtek Z350 resin composite (control); G1% = Filtek Z350 with 1% of silk nanoparticles; G3% = Filtek Z350 with 3% of silk nanoparticles; G5% = Filtek Z350 with 5% of silk nanoparticles. It was employed scanning electron microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction, 3-point flexural strength test, Knoop hardness test, and surface roughness. From 3-point flexural strength tests the control group presented the best results G0% = 113.33 MPa (±23.73). The higher flexural modulus was shown by groups G3% = 29.150 GPa (±5.191) and G5% = 34.101 GPa (±7.940), which are statistically similar. The Knoop microhardness test has shown statistical difference only among the G3% group between the top 80.78 (± 3.00) and bottom 68.80 (±3.62) and no difference between the groups. The roughness test presented no statistical difference between the groups. The incorporation of silk nanoparticles reduced the flexural strength of Z350 resin composite. The surface roughness and microhardness tests showed no changes in any of the groups studied.

Mini-Review on Analytical Methods Applied for Analysis and Characterization of Sericin

Abstract: Sericin is a serine-rich polydispersed glycoprotein found in Bombyx mori's cocoons. Sericin is extracted from cocoons as a protein, composed of amino acids like aspartic acid, glycine, tyrosine, serine, and glutamic acid with carboxyl, hydroxyl, and an amino group. Sericin has been explored for various pharmacological activities, such as antioxidant, anti-inflammatory, antiapoptotic, antiproliferative, antibacterial, anti-hypercholesteremia, and wound healing activity. Moreover, sericin has also been explored as a biopolymer for the preparation of nanoparticles, scaffolds, hydrogels, films, etc. This mini-review illustrates the reported methods for the characterization of extracted sericin and quantification in pharmaceutical formulations. The review covers analytical methods like UV-Visible Spectroscopy, Fouriertransform infrared spectroscopy, amino acid analysis, mass spectroscopy, and high-performance liquid chromatography with a brief explanation of every analytical method.

Automated Prototype for Bombyx mori Cocoon Sorting Attempts to Improve Silk Quality and Production Efficiency through Multi-Step Approach and Machine Learning Algorithms.

Cocoon sorting is one of the most labor-demanding activities required both at the end of the agricultural production and before the industrial reeling process to obtain an excellent silk quality. In view of the possible relaunch of European sericulture, the automatization of this production step is mandatory both to reduce silk costs and to standardize fiber quality. The described research starts from this criticality in silk production (the manual labor required to divide cocoons into different quality classes) to identify amelioration solutions. To this aim, the automation of this activity was proposed, and a first prototype was designed and built. This machinery is based on the use of three cameras and imaging algorithms identifying the shape and size of the cocoons and outside stains, a custom-made light sensor and an AI model to discard dead cocoons. The current efficiency of the machine is about 80 cocoons per minute. In general, the amelioration obtained through this research involves both the application of traditional sensors/techniques to an unusual product and the design of a dedicated sensor for the identification of dead/alive pupae inside the silk cocoons. A general picture of the overall efficiency of the new cocoon-sorting prototype is also outlined.

Ingestible capsules carrying 3D printed springs: a possible future prospective for SBS treatment?

Background Short Bowel Syndrome (SBS) is a malabsorption syndrome characterised by a severe reduction of the absorbent surface of the intestinal mucosa. Treatment of this condition needs multi-professional teams and different therapies, which are not always enough to ensure enteral autonomy. New techniques are being explored, particularly distraction enterogenesis, which can allow the lengthening of the residual intestines of these patients. This study aims to demonstrate the possibility of using biodegradable materials to design ingestible capsules which carry 3D-printed springs capable of reaching the patient’s intestine. These devices could be used as a slightly invasive distraction enterogenesis technique, stimulating cell proliferation and intestinal elongation without surgery. Materials and methods Capsules were realised with gelatin from pigskin type A from Sigma-Aldrich mixed with regenerated silk fibroin (RS) obtained by reverse engineering on Bombyx Mori cocoons. Springs are composed of a structure of regenerated silk (RS) modified with graphene nanoplatelets (GNP) externally covered with a biodegradable polyhydroxybutyrate-valerate (PHBV) shell. Springs were realised with 3D printing, through which, with an extruder, PHBV and RS compounds are deposited simultaneously in a 3D structure. The springs’ capsules were then analysed with solvents simulating the gastric and intestinal environment to verify their resistance to degradation. PBS (Phosphate Buffered Saline), composed of calcium chloride and magnesium chloride (CaCl 2 + MgCl 2 ), with a pH value of 7.4, was used as a degradative agent; for the gastric tract, we chose the acetic acid, CH 3 COOH, at 12% with a pH value of 2.3. Results While the gelatin-only capsules showed poor resistance to degradation in PBS, the new compound based on gelatin and regenerated silk showed excellent resistance in gastric and intestinal environments, allowing the pills to reach the intestine without dissolving. In addition, the results show variability in the release times of the springs as a function of the pH values and the elastic constants of the springs used: the latter determined that in acetic acid, the release time is increased at an increase of the elastic constant. In contrast, in PBS, an opposite trend was observed. Conclusions Our results confirm the possibility of using gelatin, silk fibroin and PHBV to design devices capable of transporting implantable endoluminal 3D structures, drugs, or growth factors, laying the foundations for a new approach to distraction enterogenesis in SBS patients.

Electrospun Silk Fibroin-CNT Composite Fibers: Characterization and Application in Mediating Fibroblast Stimulation.

Electrospinning is a simple, low-cost, and highly efficient technique to generate desirable nano/microfibers from polymer solutions. Silk fibroin (SF), a biopolymer found in Bombyx mori cocoons, has attracted attention for various biomedical applications. In this study, functionalized CNT was incorporated in SF to generate biocomposite fibers by electrospinning. The electrospun (E-spun) fibers were well aligned with morphology mimicking the locally oriented ECM proteins in connective tissues. While as-spun fibers dissolved in water in just two minutes, ethanol vapor post-treatment promoted β-sheet formation leading to improved fiber stability in an aqueous environment (>14 days). The addition of a minute amount of CNT effectively improved the E-spun fiber alignment and mechanical strength while retained high biocompatibility and biodegradability. The fibers’ electrical conductivity increased by 13.7 folds and 21.8 folds, respectively, in the presence of 0.1 w% and 0.2 w% CNT in SF fibers. With aligned SF-CNT 0.1 % fibers as a cell culture matrix, we found electrical stimulation effectively activated fibroblasts from patients of pelvic organ prolapse (POP), a connective tissue disorder. The stimulation boosted the fibroblasts’ productivity of collagen III (COLIII) and collagen I (COLI) by 74 folds and 58 folds, respectively, and reduced the COLI to COLIII ratio favorable for tissue repair. The developed material and method offer a simple, direct, and effective way to remedy the dysfunctional fibroblasts of patients for personalized cell therapeutic treatment of diseases and health conditions associated with collagen disorder.

Insights into the structure and composition of mineralized hard cocoons constructed by the oriental moth, Monema (Cnidocampa) flavescens Walker

Animals widely use minerals and organic components to construct biomaterials with excellent properties, such as teeth, bones, molluscan shells and eggshells. The larvae of the oriental moth, Monema (Cnidocampa) flavescens Walker, secrete silk proteins that combine closely with calcareous minerals to construct a hard cocoon, which is completely different from the mineral-free Bombyx mori cocoon. The cocoons of oriental moths are likely to be the hardest among the cocoons constructed by insect species. The cocoons of oriental moths were found to be mainly composed of calcium oxalates and Asx/Ser/Gly-rich cocoon proteins, but the types of calcium oxalates and cocoon proteins remain to be elucidated. In this study, we provide an in-depth explanation of the inorganic and organic components in the oriental moth cocoon. Microscopy and imaging technologies revealed that the cocoon is composed of mineral crystals, silk fibers and other organic matter. X-ray diffraction and infrared spectral analyses showed that the mineral crystals in the oriental moth cocoon were mainly CaC2H2O4·H2O. ICP-OES analysis suggested that the mineral crystals in the cocoons were mainly CaC2H2O4·H2O. LC-MS/MS-based proteomics allowed us to identify 467 proteins from the oriental moth cocoon, including 252 uncharacterized proteins, 87 enzymes, 36 small molecule binding proteins, and 5 silk proteins. Among the uncharacterized proteins, 25 of which were Asn-rich proteins because they contained a high proportion of Asn residues (19.1%–41.4%). Among the top 20 cocoon proteins with the highest abundance, 9 of which were Asn-rich proteins. The qPCR was used to investigate the expression patterns of the major cocoon protein-coding genes. Three fibroins and three Asn-rich proteins were expressed only in the silk gland but not in other tissues. The expression of Asn-rich proteins in the silk gland gradually increased from the anterior silk gland to the posterior silk gland. These findings provide important references for understanding the formation mechanism and mechanical properties of mineralized hard cocoons constructed by oriental moths.

Toxicological evaluation of water-extract sericin from silkworm (Bombyx mori) in pregnant rats and their fetus during pregnancy

Sericin is a natural protein produced by the silkworm Bombyx mori, which has a wide range of biological activities and has a broad application prospect in multiple areas. However, systemic toxicity and safety assessment of sericin is still rare. This study was aimed to evaluate the toxic effects of water-extract sericin from cocoons of Bombyx mori in pregnant rats and their fetuses during pregnancy. Eighty pregnant rats were randomly divided into three treatment groups, one negative and one positive control group. The treatment groups were administered water-extract sericin solutions at doses of 1,000, 500, and 250 mg/kg, while the negative and positive control groups were administered pure water and 300 mg/kg aspirin, respectively. Rats were exposed daily by oral gavage from the seventh day of gestation for 10 consecutive days and sacrificed on the 20th day of gestation. The results showed that water-extract sericin did not induce any treatment-related changes on pregnant rats (clinical signs, body weights, food consumption, ovarian and uterine weights) and fetuses (body weights, body lengths, tail lengths, visceral, and skeletal development). The no-observed-adverse-effect-level (NOAEL) of sericin was determined to be 1,000 mg/kg body weight in rats. These results indicated that water-extract sericin is of low teratogenic potential under the experimental conditions of this study.

Preparation and Physicochemical Properties of a Hemosorbent Derived from Bombyx mori Cocoon Fibroin

Preparation and Physicochemical Properties of a Hemosorbent Derived from Bombyx mori Cocoon Fibroin

Crystallite Shapes and Functional Data Analysis of Silk forms using X-ray Diffraction: Microwave Irradiation Effects

The silk cocoon of Bombyx mori, fiber, and thin films were considered in our study. The pure silk thin films were prepared by extracting silk from Bombyx mori cocoons. These samples were treated with microwave irradiation at different intervals, and X-ray diffraction data of these samples were obtained. XRD data of these samples were used as an essential input to compute crystallite size along with different Bragg reflections. Employing different statistical tools, the shape of the ordered region for different samples was obtained, and these shapes were analyzed and concluded that the obtained shapes are not ellipsoidal as assumed by earlier researchers and these are multi-shaped and strongly depend on the direction of Bragg reflections. The functional data analysis was also carried out to determine the correlation between physical and microstructural parameters.

Natural based reusable materials for microfluidic substrates: The silk road towards sustainable portable analytical systems

Portable analytical systems are versatile tools for application in areas including biomedicine, biosecurity, food safety and environmental monitoring. This work contributes to the increasing demand for low-cost, environmentally friendly substrates for portable analytical systems by using natural Bombyx mori cocoons. Further, silk fibroin is also extracted from these cocoons and electrospun into oriented and randomly oriented fiber substrates. Oxygen plasma treatment is applied to improve their hydrophilicity. Fiber morphology, mechanical properties, porosity, thermal characteristics and surface contact angle are extensively characterized and the ability of the samples for passive capillary flows demonstrated. Plasma treated pressed cocoons show superhydrophilicity, capillary flow rates of 44.8 ± 3.75 mm.min-1, and high mechanical resistance with Young's modulus values up to 592.13 ± 19.83 MPa.The developed materials are used as substrates for the colorimetric quantification of three commonly scrutinized clinical analytes. Hydrophobic barriers are first wax-printed on all samples with a proper design and albumin assays are performed on all substrates. Further assays for uric acid and glucose quantification are successfully accomplished on the pressed cocoons after a simple in between washing step, with overall high coefficient of determination, proving the suitability of the developed materials as low-cost, sustainable and reusable microfluidic substrates.