Genipin-Cross-Linked Silk Fibroin/Alginate Dialdehyde Hydrogel with Tunable Gelation Kinetics, Degradability, and Mechanical Properties: A Potential Candidate for Tissue Regeneration.

Genipin-cross-linked silk fibroin (SF) hydrogel is considered to be biocompatible and mechanically robust. However, its use remains a challenge for in situ forming applications due to its prolonged gelation process. In our attempt to facilitate the in situ fabrication of a genipin-mediated SF hydrogel, alginate dialdehyde (ADA) was utilized as a reinforcement template. Here, SF/ADA-based hydrogels with different compositions were synthesized covalently and ionically. Incorporating ADA into the SF hydrogel increased pore size (44.66-174.66 µm), porosity (61.59-80.40%), and the equilibrium swelling degree (7.60-30.17). Moreover, a wide range of storage modulus and compressive modulus were obtained by adjusting the proportions of SF and ADA networks within the hydrogel. The in vitro cell analysis using preosteoblast cells (MC3T3-E1) demonstrated the cytocompatibility of all hydrogels. Overall, the covalently and ionically cross-linked SF/ADA hydrogel represents a promising solution for in situ forming hydrogels for applications in tissue regeneration.

Dual-crosslinked in-situ forming alginate/silk fibroin hydrogel with potential for bone tissue engineering.

This study aimed to improve the mechanical and biological properties of alginate-based hydrogels. For this purpose, in-situ forming hydrogels were prepared by dual crosslinking of Alginate (Alg)/Oxidized Alginate (OAlg)/Silk Fibroin (SF) through simultaneous ionic gelation using CaCO3-GDL and Schiff-base reaction. The resulting hydrogels were characterized by FTIR, SEM, compressive modulus, and rheological tests. Compared to the physically-crosslinked alginate hydrogel, the compressive modulus of dual-crosslinked Alg/OAlg/SF hydrogel increased from 28 to 67 kPa, due to the covalent imine bond formation. Then, MTT and DAPI staining assays were performed to demonstrate the biocompatibility of hydrogel. Furthermore, the differentiation potential of bone marrow mesenchymal stem cells encapsulated in hydrogel scaffolds to bone tissue was tested by ALP activity, Alizarin Red staining, and real-time PCR. The overall results showed the potential of Alginate/Oxidized Alginate/Silk Fibroin hydrogel scaffold for bone tissue engineering applications.

MLATE: Machine learning for predicting cell behavior on cardiac tissue engineering scaffolds.

Cardiovascular disease is one of the leading causes of mortality worldwide and is responsible for millions of deaths annually. One of the most promising approaches to deal with this problem, which has spread recently, is cardiac tissue engineering (CTE). Many researchers have tried developing scaffolds with different materials, cell lines, and fabrication methods to help regenerate heart tissue. Machine learning (ML) is one of the hottest topics in science and technology, revolutionizing many fields and changing our perspective on solving problems. As a result of using ML, some scientific issues have been resolved, including protein-folding, a challenging problem in biology that remained unsolved for 50 years. However, it is not well addressed in tissue engineering. An AI-based software was developed by our group called MLATE (Machine Learning Applications in Tissue Engineering) to tackle tissue engineering challenges, which highly depend on conducting costly and time-consuming experiments. For the first time, to the best of our knowledge, a CTE scaffold dataset was created by collecting specifications from the literature, including different materials, cell lines, and fabrication methods commonly used in CTE scaffold development. These specifications were used as variables in the study. Then, the CTE scaffolds were rated based on cell behaviors such as cell viability, growth, proliferation, and differentiation on the scaffold on a scale of 0-3. These ratings were considered a function of the variables in the gathered dataset. It should be stated that this study was merely based on information available in the literature. Then, twenty-eight ML algorithms were applied to determine the most effective one for predicting cell behavior on CTE scaffolds fabricated by different materials, compositions, and methods. The results indicated the high performance of XGBoost with an accuracy of 87%. Also, by implementing ensemble learning algorithms and using five algorithms with the best performance, an accuracy of 93% with the AdaBoost Classifier and Voting Classifier was achieved. Finally, the open-source software developed in this study was made available for everyone by publishing the best model along with a step-by-step guide to using it online at: https://github.com/saeedrafieyan/MLATE.

Preparation of gum tragacanth/poly (vinyl alcohol)/halloysite hydrogel using electron beam irradiation with potential for bone tissue engineering.

Nanocomposite hydrogels based on tyramine conjugated gum tragacanth, poly (vinyl alcohol) (PVA), and halloysite nanotubes (HNTs) were prepared by electron beam irradiation and characterized. The FTIR, 1H NMR, and TGA results confirmed the chemical incorporation of HNTs into gum tragacanth. Gel content and swelling of hydrogels decreased with HNTs loading up to 20 % wt. The mechanical strength of hydrogels increased by increasing HNTs content up to 10 % with 371 kPa fracture stress at 0.95 fracture strain, compared to 312 kPa stress at 0.79 strain for gum tragacanth/PVA hydrogel. Hydrogel's biocompatibility and osteogenic activity were tested by seeding rabbit bone marrow mesenchymal stem cells. The cell viability was >85 % after 7 days of culture. In vitro secretion of ALP and calcium deposition on hydrogels in alizarin red assay after 21 days of culture indicated hydrogel potential for bone tissue engineering.

Drug in adhesive transdermal patch containing antibiotic-loaded solid lipid nanoparticles.

The structure of the skin only allows those hydrophobic elements to penetrate through the depth of the skin with low molecular weight (less than 500 Da) and low daily dose (less than 100 mg/day). Skin penetration of many drugs such as antibiotics at a high daily dose remains an unresolved challenge. In this study a transdermal patch using cephalexin as an antibiotic drug model was developed. Cephalexin was loaded into α-tocopherol succinate-based solid lipid nanoparticles (SLNs). Cephalexin-loaded SLNs with a drug/lipid ratio of 20%, diameter of 180 ± 7 nm, and drug loading 7.9% led to the greatest inhibition zone of Staphylococcus aureus and showed the highest skin permeation capabilities. Cephalexin-loaded SLNs were distributed into poly-iso-butylene adhesive solution and final patches prepared using solvent casting. The physico-chemical characteristics, in vitro drug release, antimicrobial efficacy, and skin cell proliferation properties of patches were evaluated. Results indicated that the optimal transdermal patch formulation containing 90% adhesive solution, 7% cephalexin, and 3% cephalexin-loaded SLNs (with antibiotic content approximately 28% less) inhibited growth of S.aureus better than the formulation containing 90% adhesive solution and 10% cephalexin. In vitro evaluation of the growth of human fibroblast skin cells in media with the optimal patch exhibited greater proliferation (about 25.5%) than those in media without the patch.

Highly efficient porous magnetic polydopamine/copper phosphate with three-dimensional hierarchical nanoflower morphology as a selective platform for recombinant proteins separation.

The separation and purification of recombinant pharmaceutical proteins is a fundamental and challenging step in the biotechnology industry. Hierarchical nanostructures with unique features and high stability can be used as efficient adsorbents. In this study, hierarchical magnetic polydopamine-copper phosphate nanoflowers (Cu-PDA MNFs) were synthesized as high-performance magnetic adsorbents in a simple and low-cost method based on green chemistry. The prepared hybrid Cu-PDA MNFs revealed great performance for separating pure recombinant human growth hormone (rhGH) and the rhGH acquired from recombinant Pichia pastoris yeast fermentation. The analysis confirmed that Cu-PDA MNFs exhibited a high adsorption capacity of 257.4 mg rhGH g-1 Cu-PDA MNFs and a fast adsorption rate for approaching the adsorption equilibrium within less than 30 min with a recovery efficiency of 74% of rhGH from the real sample. In addition, recycling tests demonstrated the stability and recyclability of Cu-PDA MNFs for at least six cycles with almost constant adsorption capacity and no toxicity. Based on these results, Cu-PDA MNFs could be considered as a promising candidate for separation and purification of rhGH. This work presents a new approach to using organic-inorganic nanoflowers as the hierarchical nanostructure for purification of pharmaceutical proteins with high performance.

Injectable chitosan hydrogel embedding modified halloysite nanotubes for bone tissue engineering.

Low mechanical strength and untargeted osteoinduction of chitosan hydrogel limit its application for bone regeneration. This study aimed to develop an injectable chitosan hydrogel with enhanced mechanical strength and improved osteoinductivity for bone tissue engineering. For this purpose, chitosan-modified halloysite nanotubes (mHNTs) were synthesized first. Then, icariin as a bone inducer was loaded into mHNTs (IC@mHNTs), resulting in a sustained drug release system. Further, nanocomposite chitosan/mHNTs hydrogels were prepared by the sol-gel transition, leading to decreased gelation time and temperature and enhanced mechanical strength of the resulting scaffolds. The mesenchymal stem cells were encapsulated into the hydrogels, and in vitro viability assays showed scaffold biocompatibility. Moreover, embedded mHNTs or IC@mHNTs in the scaffold resulted in enhanced proliferation and bone differentiation of encapsulated cells. It was collectively demonstrated that the injectable in situ forming nanocomposite chitosan hydrogel loaded with IC@mHNTs is a promising candidate for bone regeneration.

Preparation, optimization, and evaluation of midazolam nanosuspension: enhanced bioavailability for buccal administration.

Midazolam is considered as one of the best first-line drugs in managing status epilepticus in children who require emergency drug treatment. Due to poor water solubility, oral bioavailability of midazolam is relatively low. To improve its dissolution and absorption, midazolam nano-suspensions were formulated with different stabilizers using the ultrasonic technique. A combination of Tween 80 and Poloxamer (TP) was considered as one stabilizer and 3-methyl chitosan (TMC) as another stabilizer. The ratio of the stabilizers was selected as an independent variable, and their effects on the particle size and the zeta potential were evaluated by the simplex lattice mixture method. The freeze-dried optimized midazolam nano-suspension powder was characterized by particle-size analysis, SEM, the stability test, and the dissolution test. The optimized midazolam nano-suspension (containing 76% TMC and 24% TP) had a mean particle size of 197 ± 7 nm and a zeta potential of 31 ± 4 (mV). The stability test showed that the midazolam nano-suspension is stable for 12 months. In the in vitro dissolution test, the midazolam nano-suspension showed a marked increase in the drug dissolution percentage versus coarse midazolam. In the in vivo evaluation, the midazolam nano-suspension exhibited a significant increase in the Cmax and the AUC0-5, and a major decrease in Tmax. The overall results indicate the nano-suspension of midazolam is a promising candidate for managing status epilepticus in children in emergency situation.

Physicomechanical, rheological and in vitro cytocompatibility properties of the electron beam irradiated blend hydrogels of tyramine conjugated gum tragacanth and poly (vinyl alcohol).

In the present study, preparation of blend hydrogels of tyramine conjugated gum tragacanth and poly (vinyl alcohol) was carried out by electron beam irradiation, and modification of hydrogel properties with poly (vinyl alcohol) was demonstrated. Gel content, swelling behavior, pore size and mechanical and rheological properties of hydrogels prepared at 14, 28 and 56 kilogray (kGy) with different ratios of polymers were investigated. Gel content increased from 67 ± 2% for pure tyramine conjugated gum tragacanth hydrogel to >92% for blend hydrogels. However, the corresponding equilibrium swelling degree decreased from 35.21 ± 1.51 to 9.14 ± 1.66 due to the higher crosslink density of blend hydrogel. The mechanical strength of the hydrogels with interconnected pores increased significantly in the presence of poly (vinyl alcohol) and increasing irradiation dose up to 28 kGy with a twenty-fold enhancement of stress fracture and excellent elastic recovery in cyclic compression analysis. The equilibrium swelling degree of blend hydrogel containing 3% w/v tyramine conjugated gum tragacanth and 2% w/v poly (vinyl alcohol) prepared at 28 kGy was 16.59 ± 0.81. The biocompatibility of hydrogels was tested in the presence of rabbit bone marrow mesenchymal stem cells. The viability of cells exposed to hydrogel extract was >92% after 7 days of culture and indicated hydrogel biocompatibility with potential biomedical applications.

Thermoresponsive Nanogels Based on Different Polymeric Moieties for Biomedical Applications.

Nanogels, or nanostructured hydrogels, are one of the most interesting materials in biomedical engineering. Nanogels are widely used in medical applications, such as in cancer therapy, targeted delivery of proteins, genes and DNAs, and scaffolds in tissue regeneration. One salient feature of nanogels is their tunable responsiveness to external stimuli. In this review, thermosensitive nanogels are discussed, with a focus on moieties in their chemical structure which are responsible for thermosensitivity. These thermosensitive moieties can be classified into four groups, namely, polymers bearing amide groups, ether groups, vinyl ether groups and hydrophilic polymers bearing hydrophobic groups. These novel thermoresponsive nanogels provide effective drug delivery systems and tissue regeneration constructs for treating patients in many clinical applications, such as targeted, sustained and controlled release.

Quantum dots-ßcyclodextrin-histidine labeled human adipose stem cells-laden chitosan hydrogel for bone tissue engineering.

Mesenchymal stem cells with differentiation ability to diverse cells play a crucial role in tissue engineering. Tracking the fate of these cells during the regeneration of tissue helps to obtain more information about their function. In this study, histidine conjugated ß-cyclodextrin as a cell-penetrating carrier with drug loading ability was attached to QDs nanoparticle (QD-ßCD-His) for stem cell labeling. Traceability of QD-ßCD-His labeled human adipose stem cells (hASCs) was monitored in 2D cell culture and 3D temperature-sensitive chitosan hydrogel scaffold. Dexamethasone (Dex) as an osteoinductive drug was loaded into QD-ßCD-His nano-carrier (QD-ßCD-His@Dex) to induce bone differentiation of labeled cells. Overall results indicated that QD-ßCD-His@Dex is a promising dual-purpose nano-carrier for stem cell labeling with osteoinductive potential in cell therapy as well as tissue engineering scaffolds.

Dual responsive chondroitin sulfate based nanogel for antimicrobial peptide delivery.

Poly (l-lactide)-graft-chondroitin sulfate (PLLA-g-CS) copolymers were synthesized with different l-lactide contents via ring-opening polymerization. Chemical structure of the synthesized copolymers was confirmed by FTIR and HNMR analyses. The degree of polymerization and substitution of PLLA was found to be 0.56 and 2.98, respectively. Nisin was loaded in PLLA-g-CS nanogels at 37 and 42 °C. The hydrodynamic radius of the nanogels was 181 and 399 nm, respectively. The release profile was studied at two different temperatures and pHs over 7 days. The results indicated a variation of the cumulative release of nisin from 25 to 98% depending on the pH and temperature of release media. Cytotoxicity test of nisin loaded nanogels on human dermis fibroblast cells, confirmed no toxic effect. Finally, Antimicrobial activity of the nanogel was evaluated against Staphylococcus aureus and Escherichia coli bacteria. Overall, this study indicated that the dual responsive nanocarrier could potentially be used for infection therapeutic applications.

Enhanced Cellular Uptake Of Phenamil Through Inclusion Complex With Histidine Functionalized ß-Cyclodextrin As Penetrative Osteoinductive Agent.

BACKGROUND: Phenamil (PH) is a small molecule that induces bone formation through upregulation of the TRB3 gene in the bone-regeneration process. ß-Cyclodextrins (ßCDs) with hydrophilic surfaces and a relatively hydrophobic cavity can form inclusion complexes with primarily hydrophobic small molecules such as PH, and increase their apparent solubility and dissolution rate. The hydrophilic surface of ßCDs prevents their interaction with the hydrophobic lipids of the cell membrane for penetration. Therefore, binding of penetrative groups, such as lysine, arginine, and histidine (His), to ßCDs for cell penetration is required. AIM: The aim of this study was to investigate the effect of His-conjugated ßCD on cellular uptake of PH for bone differentiation. METHODS: In this study, His-ßCDs were synthesized and used to prepare an inclusion complex of His-ßCD-PH. A hydroxypropyl-ßCD-PH (HP-ßCD-PH) inclusion complex for increasing PH solubility without a penetrative group was prepared for comparison. 3-D geometry of ßCD derivatives and PH-inclusion complexes was investigated by Fourier-transform infrared spectroscopy and molecular docking. Alizarin red staining and real-time PCR were performed to compare bone differentiation of His-ßCD-PH and HP-ßCD-PH. RESULTS: The results suggested that the benzene ring of PH was inserted into the wide side of both His-ßCD and HP-ßCD. Alizarin red staining at 14 days postculture in the presence of His-ßCD-PH at total concentration of 50 µM for PH showed that bone-matrix mineralization increased significantly compared with free PH and HP-ßCD-PH. Real-time PCR confirmed this result, and showed gene expression increased significantly (OPN 1.84-fold, OCN 1.69-fold) when stem cells were cultured with His-ßCD-PH. CONCLUSION: The overall results indicated that His-ßCD-PH is a promising carrier for osteoinductive PH with possible penetration ability and sustained release that reduces BMP2 consumption for differentiation of mesenchymal stem cells to bone tissue.

Osteogenic induction of human mesenchymal stem cells in multilayered electrospun scaffolds at different flow rates and configurations in a perfusion bioreactor.

Electrospun scaffolds are potentially interesting in bone tissue engineering due to a strong structural similarity to the natural bone matrix. To investigate the osteogenic behavior of cells on the scaffolds, dynamic culture of cells is essential to simulate the biological environment. In the present study, human mesenchymal stem cells (hMSCs) were cultured on multilayer nanohydroxyapatite-polycaprolactone electrospun scaffolds at different configurations (horizontal with or without pressure and parallel with the medium flow) and flow rates in a perfusion bioreactor. Alkaline phosphatase (ALP) activity, cell viability, Ca deposition and RUNX2 expression were determined in three different dynamic states, and compared with static culture after 1, 3, 7, and 14 days. Among dynamic groups, RUNX2 gene expression upregulated more in a horizontal state at a low flow rate without mechanical pressure (LF) and parallel flow (PF), than static group on day 7. At a high flow rate with mechanical pressure, Ca deposition and ALP activity increased 2.34 and 1.7 folds more than in static culture over 7 days, respectively. Furthermore, ALP activity, Ca deposition and RUNX2 gene expression increased in PF samples. PF provided longer culture time with higher cell differentiation. Therefore, high flow rate with mechanical pressure and PF are suggested for producing differentiated cell structure for bone tissue engineering.

Reduction of marginal mass required for successful islet transplantation in a diabetic rat model using adipose tissue-derived mesenchymal stromal cells.

BACKGROUND AIMS: Adipose tissue-derived mesenchymal stromal cells (AT-MSCs), widely known as multipotent progenitors, release several cytokines that support cell survival and repair. There are in vitro and in vivo studies reporting the regenerative role of AT-MSCs possibly mediated by their protective effects on functional islet cells as well as their capacity to differentiate into insulin-producing cells (IPCs). METHODS: On such a basis, our goal in the present study was to use three different models including direct and indirect co-cultures and islet-derived conditioned medium (CM) to differentiate AT-MSCs into IPCs and to illuminate the molecular mechanisms of the beneficial impact of AT-MSCs on pancreatic islet functionality. Furthermore, we combined in vitro co-culture of islets and AT-MSCs with in vivo assessment of islet graft function to assess whether co-transplantation of islets with AT-MSCs can reduce marginal mass required for successful islet transplantation and prolong graft function in a diabetic rat model. RESULTS: Our findings demonstrated that AT-MSCs are suitable for creating a microenvironment favorable for the repair and longevity of the pancreas ß cells through the improvement of islet survival and maintenance of cell morphology and insulin secretion due to their potent properties in differentiation. Most importantly, hybrid transplantation of islets with AT-MSCs significantly promoted survival, engraftment and insulin-producing function of the graft and reduced the islet mass required for reversal of diabetes. CONCLUSIONS: This strategy might be of therapeutic potential solving the problem of donor islet material loss that currently limits the application of allogeneic islet transplantation as a more widespread therapy for type 1 diabetes.

Development and Evaluation of an Anti-Epileptic Oral Fast-Dissolving Film with Enhanced Dissolution and In vivo Permeation.

OBJECTIVES: The main objective of this novel study was to develop midazolam hydrochloride fast dissolving oral films (FDFs) using solvent casting method and to evaluate the characteristics of the optimum formulation through in vitro and in vivo analysis. The FDFs are new favorable solid dosage forms that deliver drugs rapidly to the blood circulation system and have great advantages in the emergent control of severe neuropathic attacks in children. METHODS: Midazolam nanosuspensions were prepared using the ultrasonic method and then incorporated in the hydroxypropyl methyl cellulose (HPMC)/pullulan polymeric matrix with other excipients like glycerol and cellulose nanofiber as a softener and a compatibilizer, respectively. The prepared films were evaluated for mechanical properties, morphology study, disintegration time, and dissolution time. RESULTS: SEM images of FDFs showed the uniform distribution of spherical nanoparticles in the polymeric matrix. A film with 36% HPMC, 64% pullulan, and 21% glycerin was selected as the optimum formulation by the Design Expert 7 software. The optimum film was stable for three months. CONCLUSION: The pharmacokinetic parameters of midazolam oral film in comparison to coarse midazolam suspension exhibited significant increase in AUC, Cmax, and a good decrease in Tmax. The overall results showed the enhanced in vivo orotransmucosal absorption of poorly water-soluble drugs via the insertion of drugs nanosuspension in buccal films.

Self-assembled and pH-sensitive mixed micelles as an intracellular doxorubicin delivery system.

Nanocarrier-based drug delivery systems have been explored extensively in cancer therapy. Among the vast number of different nanocarrier systems applied to deliver chemotherapeutics to cancer tumor, intelligent systems which deliver drug to various sites in the body have attracted considerable attentions. Finding a specific stimulant that triggers the carrier to release its payload in the target tissue is a key parameter for efficacy of delivery systems. Acidic pH of cancer tumor helps a pH-sensitive carrier to release drug at the tumor site. In this study, a pH-sensitive mixed micellar system was developed using Dextran-Stearic Acid (Dex-SA) and Dextran-Histidine (Dex-His) conjugated polymers to deliver doxorubicin (DOX) to cancer cells. Drug release from this micellar system showed higher release rate at acidic pH than that of in neutral environment, where the release was 56 and 76% at pH 7.4 and acidic pH, respectively. Finally, the in vitro cytotoxicity and cell uptake of DOX-loaded micelles and free DOX on U87 MG cell line showed that micellar systems had more anti-proliferation effect and uptake compared to free drug.

Transferrin-conjugated magnetic dextran-spermine nanoparticles for targeted drug transport across blood-brain barrier.

Application of many vital hydrophilic medicines have been restricted by blood-brain barrier (BBB) for treatment of brain diseases. In this study, a targeted drug delivery system based on dextran-spermine biopolymer was developed for drug transport across BBB. Drug loaded magnetic dextran-spermine nanoparticles (DS-NPs) were prepared via ionic gelation followed by transferrin (Tf) conjugation as targeting moiety. The characteristics of Tf conjugated nanoparticles (TDS-NPs) were analyzed by different methods and their cytotoxicity effects on U87MG cells were tested. The superparamagnetic characteristic of TDS-NPs was verified by vibration simple magnetometer. Capecitabine loaded TDS-NPs exhibited pH-sensitive release behavior with enhanced cytotoxicity against U87MG cells, compared to DS-NPs and free capecitabine. Prussian-blue staining and TEM-imaging showed the significant cellular uptake of TDS-NPs. Furthermore, a remarkable increase of Fe concentrations in brain was observed following their biodistribution and histological studies in vivo, after 1 and 7 days of post-injection. Enhanced drug transport across BBB and pH-triggered cellular uptake of TDS-NPs indicated that these theranostic nanocarriers are promising candidate for the brain malignance treatment. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2851-2864, 2017.

Synthesis and application of magnetite dextran-spermine nanoparticles in breast cancer hyperthermia.

Cancer treatment has been very challenging in recent decades. One of the most promising cancer treatment methods is hyperthermia, which increases the tumor temperature (41-45 °C). Magnetic nanoparticles have been widely used for selective targeting of cancer cells. In the present study, magnetic dextran-spermine nanoparticles, conjugated with Anti-HER2 antibody to target breast cancer cells were developed. The magnetic dextran-spermine nanoparticles (DMNPs) were prepared by ionic gelation, followed by conjugation of antibody to them using EDC-NHS method. Then the Prussian blue method was used to estimate the targeting ability and cellular uptake. Cytotoxicity assay by MTT showed that antibody-conjugated MNPs (ADMNPs) have no toxic effect on SKBR3 and human fibroblast cells. Finally, the hyperthermia was applied to show that synthesized ADMNPs, could increase the cancer cells temperature up to 45 °C and kill most of them without affecting normal cells. These observations proved that Anti-HER2 conjugated magnetic dextran-spermine nanoparticles can target and destroy cancer cells and are potentially suitable for cancer treatment.

Fluorescein isothiocyanate-dyed mesoporous silica nanoparticles for tracking antioxidant delivery.

This study investigated the cellular uptake of fluorescein isothiocyanate-labelled mesoporous silica nanoparticles (FITC-MSNs), amine-functionalised FITC-MSNs (AP-FITC-MSNs) and their gallic acid (GA)-loaded counterparts. Mesoporous silica nanoparticles were labelled with fluorescein isothiocyanate, functionalised by 3-aminopropyltriethoxysilane (APTES) (AP-FITC-MSNs) and then loaded by GA. All nanoparticles were characterised by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy, and X-ray diffraction. The cytotoxicity of different concentrations of dyed nanoparticles was investigated using (3-(4,5-trihydroxybenzoic acid, dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and flow cytometry. TEM images showed that the average particle sizes of FITC-MSNs and AP-FITC-MSNs were about 100 and 110 nm, respectively. These nanoparticles were internalised by Caco-2 cells, accumulated and dispersed into the cytoplasm, nucleus, and subcellular organelles. Nanoparticles containing GA clearly decreased the viability of cells. FITC-MSNs showed no toxicity on Caco-2 cells at concentrations of ≤50 µg/ml. Functionalisation of FITC-MSNs using APTES decreased toxicity effects on the cells. It was found that FITC-MSNs can be applied at low concentrations as a marker in the cells. In addition, AP-FITC-MSNs showed better biocompatibility with Caco-2 cells than FITC-MSNs, because of their positive surface charges.