Use Of Sodium Dodecyl Sulfate Research Articles

A Pseudo-Surfactant Chemical Permeation Enhancer to Treat Otitis Media via Sustained Transtympanic Delivery of Antibiotics.

Chemical permeation enhancers (CPEs) represent a prevalent and safe strategy to enable noninvasive drug delivery across skin-like biological barriers such as the tympanic membrane (TM). While most existing CPEs interact strongly with the lipid bilayers in the stratum corneum to create defects as diffusion paths, their interactions with the delivery system, such as polymers forming a hydrogel, can compromise gelation, formulation stability, and drug diffusion. To overcome thischallenge, differing interactions between CPEs and the hydrogel system are explored, especially those with sodium dodecyl sulfate (SDS), an ionic surfactant and a common CPE, and those with methyl laurate (ML), a nonionic counterpart with a similar length alkyl chain. Notably, the use of ML effectively decouples permeation enhancement from gelation, enabling sustained delivery across TMs to treat acute otitis media (AOM),which is not possible with the use of SDS. Ciprofloxacin and ML are shown to form a pseudo-surfactant that significantly boosts transtympanic permeation. The middle ear ciprofloxacin concentration is increased by 70-fold in vivo in a chinchilla AOM model, yielding superior efficacy and biocompatibility than the previous highest-performing formulation. Beyond improved efficacy and biocompatibility, this single-CPE formulation significantly accelerates its progression toward clinical deployment.

Supercritical cartilage decellularization: Cosolvent effect on mechanical properties

Decellularized animal tissue may serve as scaffolds in tissue engineering. Despite the well-disseminated use of sodium dodecyl sulfate (SDS) for decellularization, SDS-based methods are time-consuming and may cause extracellular matrix (ECM) denaturation. This study explores supercritical CO2 (scCO2) extraction as an alternative to decellularize chicken sternum cartilage. Various scCO2 decellularization methods were tested, including pure scCO2, scCO2 with water, ethanol, or their mixture. SDS outperformed in decellularization but caused the highest extracellular matrix damage. Pure scCO2 treatment for 2 h reduced mechanical resistance due to dehydration. scCO2 with 99 % ethanol led to extensive tissue dehydration, while the addition of 70 % ethanol increased cell removal without significant tissue damage. Further research is recommended to optimize scCO2 decellularization of this tissue with other cosolvents while maintaining adequate hydration.

Ultrasonication‐assisted alkali treatment of hemp fibers to improve the fiber/matrix interface of hemp/epoxy composites: The influence of sodium dodecyl sulfate surfactant

AbstractAlkali pretreatment of fibers is an efficient technique to enhance fiber/matrix bonding in natural fiber composites. This study investigates if the addition of sodium dodecyl sulfate (SDS) surfactant increases the efficacy of alkali treatment of hemp fibers. SDS concentrations ranging between 2 and 10% wt/wt were used with a constant (10% wt/wt) NaOH concentration. Fiber surfaces were observed with scanning electron microscopy (SEM). Fiber surfaces were cleaner when SDS was used compared with alkali treatment with no SDS added indicating the elimination of substances such as lignin, waxes, hemicelluloses, and pectin. Fourier‐transform infrared spectroscopy and TGA studies also confirmed the removal of these substituents. Mechanical performance of the composites was evaluated via static and dynamic tests. The tensile strength and modulus improved 24.9% and 13.6% respectively with the addition of 10 wt% SDS. The flexural strength and modulus also showed an improvement of 22.4% and 26.3% respectively when 10 wt% SDS was added to alkali solution. Dynamic mechanical tests showed that the storage and loss modulus of the composites increase with SDS/alkali treatment. Results indicated that the use of SDS in alkali treatment notably improves the composite performance which is attributable to more efficient elimination of noncellulosic constituents from fiber structure.Highlights Sodium dodecyl sulfate (SDS) improves the efficacy of NaOH treatment of hemp. Removal of noncellulosic substances from fibers was facilitated by SDS. Ultrasound/SDS assisted alkalization of fibers improved composite properties.

Anticorrosion Protection of New Composite Coating for Cobalt-Based Alloy in Hydrochloric Acid Solution Obtained by Electrodeposition Methods

In this work, electrochemical deposition techniques (galvanostatic and potentiostatic) were used to obtain coatings of a new composite polymer, 3-methylpyrrole—sodium dodecyl sulfate/poly 2-methythiophene (P3MPY-SDS/P2MT), on cobalt-based alloy samples for anti-corrosion safety. The use of sodium dodecyl sulfate as a dopant ion in electrosynthesis can have a relevant effect on the anticorrosive property of the composite polymer layer by blocking the entry of corrosive ions. The cobalt alloy specimen had an important impact on the electrochemical performance of the composite coating and this together with the presence of the polymeric layer was achieved by simultaneously constitution of a complex oxides film and polymeric layers. The polymeric coatings were analyzed using scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, and cyclic voltammetry (CV) methods. The corrosion protection of the P3MPY-SDS/P2MT-covered cobalt-based alloy was explored using electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization procedures in a 1 M HCl solution. The corrosion speed of the P3MPY-SDS/P2MT-covered cobalt-based alloy was observed to be ~10 times less than an uncovered specimen, and the effectiveness of the composite layers of this coating is greater than 91%. This superior efficaciousness was obtained by the electropolymerization of P3MPY-SDS/P2MT at current densities of 1 mA/cm2 and 0.5 mA/cm2, applied potentials of 0.9 V and 1.0 V, and a molar ratio of 5:1. Corrosion test results indicate that the P3MPY-SDS/P2MT coatings provide a good result: protection against the corrosion of a cobalt-based alloy in aggressive solutions.

Effects of eco-friendly cyclodextrin on the methane hydrate formation in the complex systems

Due to the increasing demand for natural gas, solidified natural gas (SNG) technology provides an attractive option for natural gas storage and transportation. However, the use of sodium dodecyl sulfate (SDS) to promote hydrate formation will have a negative impact on the environment. To overcome this difficulty, the compounding system of environmentally friendly additives with SDS was intended to find the synergistic promotion effects aiming to reduce the usage of SDS in this study. Three kinds of cyclodextrins including αCD, βCD, hydroxypropyl-βCD (HPCD) with SDS were compounded to study the effect of their interaction on hydrate formation kinetics. The result showed that when used with SDS, the formation of hydrates was promoted by αCD and βCD but inhibited by HPCD. It is emphasized that the promoting effect depended on the water channel (completely hydrophilic zones) formed by H-bonds between cyclodextrin and SDS self-assembled channel-type complexes. Compared with 500 ppm SDS used along, it was possible to reduce the dosage of SDS by 40%. Finally, by observing the morphology of hydrate formation, it is found that the addition of cyclodextrins would affect the driving force for hydrate growth in the horizontal direction during hydrate formation. This study provides theoretical support for further understanding of cyclodextrin and surfactant compounding and shows an effective way for the application of SNG industry.

RATIONAL CONDITIONS OF PRODUCING THE ACTIVATED CARBON WITH WELL-DEVELOPED NANOPOROUS STRUCTURE FOR THE TREATMENT OF NATURAL AND WASTE WATERS

Rational conditions for the activation of coal of a high stage of metamorphism (anthracite) have been determined, which allow the development of a nanoporous structure. Obtained activate carbon with nanoporous structure can be use for the treatment of natural and waste waters have been carried out. The structurally sorptive characteristics and granulometric size composition of obtained samples were determined. It was established that in order to obtain a sorbent with specified properties, it is necessary to introduce oxygen into the reacting mixture at the second stage of anthracite activation and control the activation time. The activation in the first stage was carried out with steam at a temperature of 750–800 °C, and in the second stage – with steam in the presence of air oxygen at 280–300 °C. The rational content of oxygen in the activating mixture is in the range of 2.7–4.4 %, while the specific surface area of activated carbon increases, which corresponds to nano-sized pores and, accordingly, the volume of the adsorption space of coal. The process led to the development of mesoporosity ~ 0.2–0.3 cm3/g, after which the microporous sample of activated carbon was impregnated with a surfactant solution. Thus, the rational conditions of activation are the duration of the process of 1.5 hours in the presence of 4.4 % oxygen in the activating mixture. It is shown that the largest values of the total specific surface area and the specific surface area of mesopores are achieved when using ionic surfactants with alkyl chain length n = 10–16 as modifiers. In particular, the use of sodium dodecyl sulfate СН3(СН2)11SO3Na as a modifier allows to obtain the largest value of the total surface Stot 1030 m2/g and the surface of mesopores Smе 334 m2/g. It was established that the activation of modified samples of activated carbon leads to a decrease in the size of granules.The content of fractions with a size of 1.0–2.0 mm and 0.5–1.0 mm is 80–85 %, which are rational for use for water purification from organic compounds.

Enhanced Strength and Flexibility of Ni-Co Nanocomposites through Electrodeposition with SiC Nanoparticles and SDS Additive

This study focuses on the electrodeposition of Ni-Co nanocomposites using a modified Wattsbath. The bath is supplemented with silicon carbide (SiC) nanoparticles and an organic additive, Sodiumdodecyl sulfate (SDS). The objective is to investigate the influence of these additions on the structure,mechanical properties, and flexibility of the resulting coatings. Hardness and weight loss tests areconducted, and the coatings are examined through microscopy. In conclusion, this study successfullydemonstrates the electrodeposition of Ni-Co nanocomposites with improved strength and flexibility. Theincorporation of SiC nanoparticles and the use of SDS as a surfactant show promising results. Thesefindings have practical implications for enhancing electroplating techniques in various industries.

Enhanced adsorption of chlorogenic acid polyphenol by layered double hydroxide modified with sodium dodecyl sulfate

Polyphenols are a class of organic contaminants highly present in wastewater from industrial and agro-industrial processes. Adsorptive processes are employed to remove contaminants from effluents without the generation of by-products. Among the challenges in removal is the search for adsorbents that have affinity and favor the interaction with the contaminant. In this sense, this paper aimed to evaluate a layered double hydroxide modified with sodium dodecyl sulfate as an adsorbent material of the chlorogenic acid polyphenol. Carbonated magnesium-aluminum hydrotalcite was the layered double hydroxide employed for modification at different sodium dodecyl sulfate contents. The results showed that the modification with 50% w w-1 sodium dodecyl sulfate increased the adsorptive capacity of the modified layered double hydroxide from 0.030 mg g-1 to 2.3 mg g-1 after 24 hours of contact with the polyphenol. Thus, the use of sodium dodecyl sulfate for surface modification of layered double hydroxide indicated positive results that favor the application of the modified material in the removal of polyphenols.

Two green spectrofluorimetric methods for the assay of atomoxetine hydrochloride in pure form and commercial capsules with application to content uniformity testing.

Atomoxetine hydrochloride (ATX) is a potent and non-stimulant drug which was approved for the treatment of attention-deficit hyperactivity disorder. Owing to its importance, two green, simple and validated spectrofluorimetric methods were developed for its sensitive assay in pure and capsule forms. The first method (Method I) relied on measuring the enhanced fluorescence of ATX by the use of sodium dodecyl sulfate in alkaline medium at λ ex 227 nm/λ em 298 nm. The second method (Method II) involved complex formation of ATX with Erythrosine B (EB) in aqueous acidic solution resulting in quantitative quenching of the EB naive fluorescence. This complex was formed in the presence of Britton-Robinson buffer (pH 4.0). The difference in fluorescence intensity was measured at λ ex 527/λ em550 nm. The calibration curves were linear through the ranges of 0.2-2.0 µg ml-1 for Method I, 0.2-4.0 µg ml-1 for Method II with good correlation coefficient (r = 0.9998) for both methods. The suggested methods were perfectly applied for determination of ATX in its commercial capsules and content uniformity test. The greenness of the proposed methods was confirmed by three different assessment tools and it was found that both methods were green, eco-friendly and environmentally safe.

Copper metal elimination from polluted soil by electro-kinetic technique.

The electro kinetic method is one of the common methods in the process of pollutants removal. In this paper, the process of removing copper from contaminated soil has been studied. Some improved conditions were used in this process; the pH of the solution was changed for each experiment during the first three experiments. SDS (sodium dodecyl sulfate) has been used as an activator to improve the removal process by washing the soil with it. Date palm fibers (DPF) were also used as an adsorbent material to counteract the reverse flow that occurred during the removal process, thus increasing the removal value. Various experiments were performed, where it was observed that by decreasing the pH, the removal capacity was increased. The removal capacity in the three different experiments were 70% at pH 4, 57% at pH 7, and 45% at pH 10. The use of SDS as a solution in the process increased the dissolution and absorption of copper from the soil surface and then increased the removal capacity (74%). The use of DPF to counter the osmosis flow has been successful in adsorbing the pollutants (copper) returning from this flow, and thus this material can be considered good from an economic and environmental aspect when compared to other commercial adsorbents.

Electropositive far-ultraviolet carbon nanoparticles-based photoluminescent photoinduced-electron transfer sensor for sodium dodecyl sulfate and sodium lauryl sulfonate detection

Unrestrained use of sodium dodecyl sulfate (SDS) and sodium lauryl sulfonate (SLS) in industry, agriculture and household are causing its subsequent accumulation in waterways, which can cause the adverse effect on various living organisms, especially aquatic organisms. Therefore, an efficient and rapid technique to detect SDS and SLS in the environment is urgently demanded. Herein, a novel far-ultraviolet emission carbon nanoparticles (FCNPs, quantum yield of ∼15.47%) with positive charges was synthesized by one-pot simple hydrothermal route using 5-hydroxytryptamine as the precursor, and applied to the detection of SDS and SLS. When the exposure to SDS and SLS, the FCNPs displayed anionic detergent-sensitive fluorescent quenching. The mechanism was proved to be that enrichment-positive charge of –NH3+ groups can strongly combine with negatively charged SDS/SLS molecules through electrostatic adsorption, resulting in luminescent quenching of FCNPs via photoinduced-electron transfer mechanism. By using this sensing platform, there are two good linear relationships between the quenching degree of the FCNPs and the concentrations of SDS and SLS in the range of 5.0 nM–50 μM and 2.5 nM to 50 μM with almost the same LOD of ∼2.5 nM, respectively. Moreover, this sensing platform has been successfully applied to detect the SDS and SLS in real domestic wastewater samples. Therefore, the electropositive luminescent FCNPs might possess enormously potential utilization in the detection of anionic detergent for environmental protection.

Comparative study of the most commonly used methods for total protein determination in milk of different species and their ultrafiltration products.

Milk ultrafiltration is a widely used membrane filtration process that allows the recuperation of whey proteins in a concentrate high in total solids, which can later be transformed in multiple healthy dairy products with great prospects for the food industry. Protein content is a decisive factor for the technological performance of milk concentrates and currently, the ISO standard method for its determination is Kjeldahl, which is time-consuming and requires specific instrumentation. For this reason, the use of rapid methods to quantify protein would greatly facilitate the monitoring of the milk ultrafiltration process. In this study, the bicinchoninic acid assay (BCA), the detergent compatible Bradford assay and the Dumas method were compared to Kjeldahl protein determination to select a quick and accurate methodology suitable for milk of different species and its ultrafiltration products (retentates and permeates). The protein content obtained from Bradford assay and Dumas method in origin milk and retentate samples was consistent with Kjeldahl values. In contrast, BCA protein levels were significantly different when compared to Kjeldahl and no method was proved to be suitable for protein determination in permeate samples. The use of sodium dodecyl sulfate was also examined to improve protein measurements without success. In comparison with the official method, Bradford assay quantitatively provided the best results, and it would be recommended for a quick, economic and easy determination of total protein content in milk and retentate samples.

Application of green first derivative synchronous spectrofluorometric method for quantitative analysis of fexofenadine hydrochloride and pseudoephedrine hydrochloride in pharmaceutical preparation and spiked human plasma

Fexofenadine hydrochloride and pseudoephedrine hydrochloride are prescribed in a combined dosage form for the treatment of allergic rhinitis. In the present work, a sensitive synchronous fluorescence spectroscopic method was applied in conjunction with first derivative for quantitative estimation of fexofenadine hydrochloride and pseudoephedrine hydrochloride in pure form, pharmaceutical tablets and spiked human plasma. Fexofenadine hydrochloride showed its conventional emission spectrum at 294 nm when excited at 267 nm. On the other hand, pseudoephedrine hydrochloride showed its conventional emission spectra at 286 nm when excited at 261 nm. The fluorescence intensities were greatly enhanced by the use of sodium dodecyl sulphate as a micellar surfactant. Application of the synchronous mode to measure the fluorescence spectra of the above drugs provided sharp narrowing bands, but the overlap was not completely resolved. Derivatization of the synchronous spectra to the first order completely resolved the overlap of the fluorescence spectra and allowed simultaneous quantitative determination of the drugs under study. Fexofenadine hydrochloride and pseudoephedrine hydrochloride could be determined from their first-order synchronous spectra at 286 and 294 nm, respectively, without interfering with each other. The method showed linearity with an excellent correlation coefficient in the concentration range of 100–1500 ng/mL for Fexofenadine hydrochloride and 50–1000 ng/mL for pseudoephedrine hydrochloride. The method was successfully applied for the simultaneous determination of the studied drugs in pharmaceutical formulation, with mean percent recoveries for Fexofenadine hydrochloride and pseudoephedrine hydrochloride of 99.49 ± 0.931 and 98.67 ± 0.634, respectively, and in spiked human plasma, with mean percent recoveries for Fexofenadine hydrochloride and pseudoephedrine hydrochloride of 95.21 ± 1.938 and 94.89 ± 1.763, respectively. Furthermore, the greenness of the described method was assessed using four different tools namely, the national environmental method index, the analytical eco-scale, the green analytical procedure index and the AGREE evaluation method. The proposed method seemed to be superior to the reported HPLC method with respect to the metrics of the greenness characters.

Magnetically Recyclable -SO3--Coated Nanoparticles Promote Gas Storage via Forming Hydrates.

Efficient gas enrichment approaches are of great importance for the storage and transportation of clean energy and the sequestration of carbon dioxide. Of special interest is the regulated gas hydrate-based method; however, its operation requires adequate additives to overcome the low-storage capacity issue. Thus, this method is not economically feasible or environmentally friendly. In this work, a novel recyclable hydrate promoter of copolystyrene-sodium styrenesulfonate@Fe3O4 (PNS) nanoparticles with an integrated core-shell structure was synthesized through emulsion polymerization. This was found to effectively reduce the induction time of methane hydrate formation by one-third compared with the widely used sodium dodecyl sulfate (SDS); the corresponding gas storage capacity was also comparable, up to 155 v/v. In addition, the PNS nanoparticles showed a good performance in foam inhibition upon hydrate decomposition, which frequently occurred with the use of SDS and other surfactant-based promoters. In particular, the new promoters contributed to a more than 30% increase in CO2 storage capacity, coacting with the fine sediments that mimic a marine environment. This provided further possibilities of sequestering CO2 in the form a gas hydrate under the seafloor. The underlying mechanism was proposed to involve anchored surfactants on the surface and tiny channels between the nanoparticles that lead to rapid hydrate nucleation and controlled growth. The results showed that the integrated magnetically recovering nanoparticles developed in this study could improve the efficiency of gas storage by forming gas hydrates; the excellent recycling performance paved the way for solving the economic and environmental problems encountered in additive usage.

Comparison of protein characterization using In solution and S-Trap digestion methods for proteomics

Protein extraction and digestion are important analytical steps in the study of proteomics. The use of sodium dodecyl sulfate (SDS) buffer makes it possible to effectively analyze various proteins. Its use was evaluated using the S-Trap digestion method and compared to the traditional In solution digestion method. Differences in protein composition were examined for each protein preparation method. S-Trap digestion followed by SDS buffer extraction clearly increased the number of identified proteins, including more mitochondrial and membrane-related proteins. The S-Trap digestion method with 5% SDS buffer was applied to the pellet remaining from the removal of RIPA buffer-soluble proteins, which identified more extracellular space proteins than the conventional S-Trap digestion method. S-Trap digestion of the pellet was particularly advantageous for identifying proteins located inside multilayer membranes.

Exploring the effects of selected essential amino acids on the self-association of sodium dodecyl sulphate at different temperatures

Sodium dodecyl sulphate (SDS) has well established biological applications in the determination of molecular weights and structure determination of proteins, peptides etc. Due to these reasons, the study of SDS interactions with essential amino acids (AAs) can be valuable as they can diversify the use of SDS in applications of importance in biology and drug delivery designs etc. The aggregation process of SDS and its detailed physiochemical properties in aqueous system, in the presence and absence of three essential amino acids L-Threonine (THR), DL- Methionine (MET) and L-Phenylalanine (PHE), is studied in a temperature range of 293 K-323 K using conductivities, surface tension (γ), particle size and zetapotential measurements using dynamic light scattering (DLS) technique. From conductivity measurements, critical micelle concentration (CMC), counterion binding (β) and thermodynamic parameters such as standard free energy change of micellization (ΔGmo), standard enthalpy change of micellization (ΔHmo), standard entropy change of micellization (ΔSmo) etc., are calculated. The surface properties and interfacial parameters of SDS systems are studied using surface tensiometery. Results established that the presence of amino acids (AAs) reduced the CMC of SDS. The electrostatic interaction of AAs with charged micellar surface were also observed. The micellization process of SDS was found thermodynamically feasible in the aqueous amino acid (AAA) system. From the surface tension data, interfacial parameters such as Гmax, Amin,pC20 , and surface effectiveness of SDS in aqueous and AAA system have been calculated and compared. It was revealed that surface adsorption and rearrangement of SDS molecular ion at their micellar concentration at air/liquid interface are significantly affected by AA addition. Hydrodynamic diameter (Dh) and zetapotential (ζ) of aqueous SDS solution helped to check the effect of amino acid additives on the colloidal size and stability of SDS aggregates. The results from different techniques were consistent to establish the feasibility of SDS micellization process in aqueous as well as aqueous AA solution.

Surfactant-Assisted Synthesis of Pt Nanocubes Using Poly(N-isopropylacrylamide) Nanogels.

Poly(N-isopropylacrylamide) (PNIPAM) nanogels were prepared by emulsion polymerization using sodium dodecyl sulfate (SDS) and employed as a capping agent in platinum nanoparticle (Pt NP) synthesis by liquid-phase reduction with hydrogen gas. When the PNIPAM nanogels were used without removing SDS, that is, a slight amount of SDS was included in the reaction solution, Pt nanocubes (NCs) were predominantly produced (>80%). The proportion of the resultant Pt NCs was much higher than that obtained using the PNIPAM linear polymer (∼60%). To clarify the effects of the three-dimensional polymer network and SDS, we synthesized Pt NPs using the PNIPAM nanogel without SDS (SDS-free PNIPAM nanogel) and found that Pt NCs are rarely formed, and most NPs obtained have an irregular shape. When only SDS was used as a capping agent, NCs were hardly obtained, but other polyhedral NPs were formed. Furthermore, the use of SDS together with the PNIPAM polymer led to the decrease in the proportion of the Pt NCs compared with that obtained using only the linear polymer. These results indicate that the enhancement of the Pt NC proportion using the PNIPAM nanogel with SDS is attributable to not only the three-dimensional polymer network of the PNIPAM nanogel but also the assist of SDS as a capping agent.

An Integrated Approach for the Efficient Extraction and Solubilization of Rice Microsomal Membrane Proteins for High-Throughput Proteomics.

The preparation of microsomal membrane proteins (MPs) is critically important to microsomal proteomics. To date most research studies have utilized an ultracentrifugation-based approach for the isolation and solubilization of plant MPs. However, these approaches are labor-intensive, time-consuming, and unaffordable in certain cases. Furthermore, the use of sodium dodecyl sulfate (SDS) and its removal prior to a mass spectrometry (MS) analysis through multiple washing steps result in the loss of proteins. To address these limitations, this study introduced a simple micro-centrifugation-based MP extraction (MME) method from rice leaves, with the efficacy of this approach being compared with a commercially available plasma membrane extraction kit (PME). Moreover, this study assessed the subsequent solubilization of isolated MPs in an MS-compatible surfactant, namely, 4-hexylphenylazosulfonate (Azo) and SDS using a label-free proteomic approach. The results validated the effectiveness of the MME method, specifically in the enrichment of plasma membrane proteins as compared with the PME method. Furthermore, the findings showed that Azo demonstrated several advantages over SDS in solubilizing the MPs, which was reflected through a label-free quantitative proteome analysis. Altogether, this study provided a relatively simple and rapid workflow for the efficient extraction of MPs with an Azo-integrated MME approach for bottom-up proteomics.

POS-476 OPTIMIZATION OF SDS-BASED KIDNEY DECELLULARIZATION RESULTS IN ECM SCAFFOLDS APPLICABLE FOR 3D RECELLULARIZATION

Kidney decellularization is used to obtain extracellular matrix (ECM) scaffolds as a platform for 3D culturing and kidney tissue regeneration. ECM contains biological molecules such as growth factors necessary for cell survival, proliferation, and maturation during organogenesis. While decellularization allows for the complete removal of cells, it also causes the removal of specific growth factors from the ECM, which could have deleterious effects on cell survival, proliferation, and differentiation during recellularization. We hypothesize that using a mild decellularization protocol which employs the use of sodium dodecyl sulphate (SDS) below its critical micelle concentration (CMC) of 0.2% in water at room temperature will allow for the greater retention of bioactive molecules which will improve recellularization efficiencies. Porcine kidney cortex was cut into 1mm x 6mm slices and decellularized using 0.05%, 0.075%, or 0.1% SDS in a soak and agitation protocol for 24 hours at room temperature. Decellularization was determined through histological and immunofluorescent staining, and DNA quantification. The retention of ECM proteins and biomolecules was determined by histological and immunofluorescent staining, mass spectrometry, and protein array analysis. Four different cell lines were used for recellularization; human-induced pluripotent stem cells (hiPSC), mesoderm derived from hiPSC, intermediate mesoderm derived from hiPSC, and primary human renal epithelial cells. ECM scaffolds were dried on Transwell inserts and cells were pipetted on the surface of the scaffold. ECM-cell constructs were grown for two weeks in culture at air-liquid interface. Recellularization efficiency, cell organization, and differentiation were determined using whole mount staining. Complete decellularization was achieved when SDS was used in concentrations below its CMC. Decellularizing with 0.05% SDS resulted in the greatest retention of glycosaminoglycan chains (GAGs), revealing that GAGs are still significantly removed from the ECM when using SDS below its CMC. Upon protein quantification, scaffolds decellularized with higher SDS concentrations were more enriched with matrisome proteins compared to scaffolds decellularized with lower SDS concentrations, which correlated to a decrease in growth factors and cytokines present on the scaffolds as determined by protein array. Following 14-day recellularization, hiPSCs and kidney progenitor cells proliferated on the ECM and expressed markers such as LIM1, WT1, PBX1 and CD31. Primary renal epithelial cells maintained a mature phenotype, expressing LTL, cytokeratin, and KSP. Herein, we describe a novel protocol for the decellularization of kidney cortex scaffolds that allows for greater retention of biomolecules. This retention of growth factors and cytokines aids in the proliferation and directed differentiation of hiPS and kidney progenitor cells toward mature renal lineages. Thus, we determined that our ECM scaffolds are able to support the growth of both hiPS and organ-matched cells. This can be applied to other organ decellularization/ recellularization protocols, as well as for the generation of kidney tissue for further in vitro studies.

Dense Bone-Derived 3d Bioscaffold: Preparation, Characterization, and Assessment of Its Potential for Bone Marrow Mesenchymal Stem Cells (BMSCS) Growth and Differentiation

Background: Since bone defects can result in different disabilities, many efforts have been made to bone tissue engineering. In this case, scaffolds play an important role as a key element of tissue engineering in providing three-dimensional structure for cell growth in vitro Objective: The aim of the present study was to provide the three-dimensional biological bioscaffold from the bovine femur dense bone and investigate the possibility of its potential for application in tissue engineering as biological 3D ECM bioscaffold via mesenchymal stem cells seeding and differentiation toward bone tissue. Methods: For the preparation of bioscaffolds, after cutting bovine femur bone into small pieces, demineralization and decellularization were done. Bioscaffolds biocompatibility was evaluated using an MTT assay. The morphological and cell adhesion characteristics of Bone marrow mesenchymal stem cells (BMSCs) on the bioscaffolds were evaluated using Scanning Electron Microscopy (SEM) technique. Finally, the cells were treated with an osteogenic differentiation medium and then evaluated for differentiation. Results: Histological studies showed that the use of sodium dodecyl sulfate (2.5%) for 8 h eliminated the cells. Radiography and calcium oxalate test confirmed demineralization. MTT assay and SEM studies showed that the obtained bioscaffolds are biocompatible and could provide an optimum three-dimensional environment for cell adhesion and movement. Moreover, the Alizarin red staining showed a higher differentiation rate for BMSCs. Conclusion: In the present study, bone-derived 3D bioscaffold showed an important role in the growth and differentiation of BMSCs, due to the natural characteristics, cell adhesion properties, and potential to enhance differentiation toward bone tissue. It may have the potential for use as bioscaffold as supporting metrics for maintenance, growth in bone tissue engineering.