Two-step Desolvation Method Research Articles

Functional Peptide-Loaded Gelatin Nanoparticles as Eyedrops for Cornea Neovascularization Treatment.

Corneal neovascularization (NV) is a process of abnormal vessel growth into the transparent cornea from the limbus and can disturb the light passing through the cornea, resulting in vision loss or even blindness. The use of nanomedicine as an effective therapeutic formulation in ophthalmology has led to higher drug bioavailability and a slow drug release rate. In this research, we designed and explored the feasibility of a new nanomedicine, gp91 ds-tat (gp91) peptide-encapsulated gelatin nanoparticles (GNP-gp91), for inhibiting corneal angiogenesis. GNP-gp91 were prepared by a two-step desolvation method. The characterization and cytocompatibility of GNP-gp91 were analyzed. The inhibition effect of GNP-gp91 on HUVEC cell migration and tube formation was observed by an inverted microscope. The drug retention test in mouse cornea was observed by in vivo imaging system, fluorescence microscope, and DAPI/TAMRA staining. Finally, the therapeutic efficacy and evaluation of neovascularization-related factors were conducted through the in vivo corneal NV mice model via topical delivery. The prepared GNP-gp91 had a nano-scale diameter (550.6 nm) with positive charge (21.7 mV) slow-release behavior (25%, 240hr). In vitro test revealed that GNP-gp91 enhanced the inhibition of cell migration and tube formation capacity via higher internalization of HUVEC. Topical administration (eyedrops) of the GNP-gp91 significantly prolongs the retention time (46%, 20 min) in the mouse cornea. In chemically burned corneal neovascularization models, corneal vessel area with a significant reduction in GNP-gp91 group (7.89%) was revealed when compared with PBS (33.99%) and gp91 (19.67%) treated groups via every two days dosing. Moreover, GNP-gp91 significantly reduced the concentration of Nox2, VEGF and MMP9 in NV's cornea. The nanomedicine, GNP-gp91, was successfully synthesized for ophthalmological application. These data suggest that GNP-gp91 contained eyedrops that not only have a longer retention time on the cornea but also can treat mice corneal NV effectively delivered in a low dosing frequency, GNP-gp91 eyedrops provides an alternative strategy for clinical ocular disease treatment in the culture.

Development of gelatin nanoparticles for the treatment of pathologies caused by Paraccocideoides brasiliensis

Introduction: Paraccocideoides brasiliensis (Pb) is a fungal pathology that causes serious lesions in the mouth in the form of ulcerations that destroy the gingiva, palate, reflecting on the immune system. For the treatment of Paracoccidioidomycosis (PMC) the choice of drug is based on the degree of the disease, with Amphotericin B (AB) being the most used due to its mechanism of action and interaction with the fungal membrane, causing its destruction. However, with all the history of AB toxicity, there is a need to reduce the serum levels of the drug in the body. The use of nanoparticles as controlled release systems (DDS) is an alternative for this purpose. The objective of this article was to develop gelatin nanoparticles for the treatment of pathologies caused by Pb. Method: In this work, gelatin nanoparticles were processed by the two-step desolvation method and loaded with AB. The nanoparticulate system was characterized by scanning electron microscopy (SEM) and dynamic light scattering (DLS). Finally, the effectiveness of DDS was evaluated against Pb yeasts by Disk Diffusion Test. Results: the synthesized particles presented suitable sizes and shapes for application in DDS with an average size of 169.4 and 172.0 nm, for the free and encapsulated system, respectively. Biological model tests of Pb showed positive results indicating that lower concentrations of AB can be efficiently administered if nanostructured DDS is used. Conclusion: This study is promising for future applications of AB-loaded gelatin nanoparticles for the treatment of pathologies caused by Paraccocideoides brasiliensis.

Dual-functional intelligent gelatin based packaging film for maintaining and monitoring the shrimp freshness

Dual-functional intelligent food packaging film for maintaining and monitoring food freshness has gained a growing number of attentions in food field. Herein, we introduced a smart gelatin-based packaging film integrating real-time monitoring and maintaining food freshness. In detail, through a two-step de-solvation method, gelatin particles were fabricated and further used for preparing oil-in-water (O/W) lavender essential oil Pickering emulsions (LEOPs). LEOPs and Alizarin (ALI) were integrated into the gelatin matrix of the three-dimensional (3D) network via intermolecular hydrogen bonding and dehydration condensation, respectively, to develop intelligent active packaging films (GL-ALI-LEOPs). GL-ALI-LEOPs exhibited excellent antibacterial and antioxidant properties and a sensitive, reversible, and rapid response to pH/NH3. Benefiting from good mechanical properties, light barrier properties and hydrophobicity of modified gelatin-based films, GL-ALI-LEOPs are readily processed into food packaging bags and smart tags. GL-ALI-LEOPs were successfully employed as an eco-friendly, low-cost, bacteriostatic and pH/NH3 quick-responsive smart trademark for real-timely monitoring and maintaining the freshness of shrimp.

Modified Desolvation Method Enables Simple One-Step Synthesis of Gelatin Nanoparticles from Different Gelatin Types with Any Bloom Values.

Gelatin nanoparticles found numerous applications in drug delivery, bioimaging, immunotherapy, and vaccine development as well as in biotechnology and food science. Synthesis of gelatin nanoparticles is usually made by a two-step desolvation method, which, despite providing stable and homogeneous nanoparticles, has many limitations, namely complex procedure, low yields, and poor reproducibility of the first desolvation step. Herein, we present a modified one-step desolvation method, which enables the quick, simple, and reproducible synthesis of gelatin nanoparticles. Using the proposed method one can prepare gelatin nanoparticles from any type of gelatin with any bloom number, even with the lowest ones, which remains unattainable for the traditional two-step technique. The method relies on quick one-time addition of poor solvent (preferably isopropyl alcohol) to gelatin solution in the absence of stirring. We applied the modified desolvation method to synthesize nanoparticles from porcine, bovine, and fish gelatin with bloom values from 62 to 225 on the hundreds-of-milligram scale. Synthesized nanoparticles had average diameters between 130 and 190 nm and narrow size distribution. Yields of synthesis were 62–82% and can be further increased. Gelatin nanoparticles have good colloidal stability and withstand autoclaving. Moreover, they were non-toxic to human immune cells.

Construction and performance evaluation of a sustained release implant material polyetheretherketone with antibacterial properties

ObjectiveThis study aimed to construct a tightly binding antibiotic sustained release system on the polyetheretherketone (PEEK) surface and investigate the cellular activity and antibacterial properties of the new oral implant materials. MethodsLow-temperature argon plasma under certain parameters was used to prepare P-PEEK with nano-topology, and chemical deposition technology was adopted to form a polydopamine (PDA) coating on the PEEK surface to build a biological binding platform, PDA/P-PEEK. Subsequently, vancomycin gelatin nanoparticles (Van-GNPs) were prepared by two-step desolvation method. Finally, Van-GNPs were combined with PEEK implant material surface to form a new composite material, Van-GNPs/PEEK. scanning electron microscope (SEM), atomic force microscope (AFM), energy dispersive spectrometer (EDS), and contact angle tester were used to comprehensively characterize the materials. The in vitro release test of Van was performed by dynamic dialysis with ultraviolet spectrophotometer. The cell cytotoxicity and adhesion tests were studied by mouse embryonic osteoblasts. The antibacterial properties were evaluated by bacterial adhesion test, plate colony counting, and antimicrobial ring test with Staphylococcus aureus and Streptococcus mutans. ResultsPEEK was treated with low-temperature argon plasma and attached to PDA to form a biological binding platform. The synthesized Van-GNPs were smooth, round, with uniform particle size distribution, and bound to PEEK to form a new composite material, which can release Van constantly. Cell experiments showed that Van-GNPs/PEEK had no cytotoxicity and had good interaction with osteoblasts. Bacterial experiments showed that surface conjugation with Van-GNPs could significantly improve the antibacterial performance of PEEK against S. aureus and S. mutans. SignificanceThis study demonstrated that Van-GNPs/PEEK have good cellular compatibility and autonomous antibacterial properties, which provide a theoretical basis for the wide application of PEEK in the field of stomatology.

Food-Grade Gelatin Nanoparticles: Preparation, Characterization, and Preliminary Application for Stabilizing Pickering Emulsions.

In this paper, the food-grade gelatin nanoparticles (GNPs) were prepared by a two-step desolvation method and using genipin as a cross-linker. The GNPs with narrow size distribution and good dispersion could be obtained only at pH 12. The effect of the genipin dosage (8–12 wt%) on the GNPs was systematically investigated. The results showed that the cross-linking degree of the GNPs increased with the increasing dosage of genipin, thus leading to a more obvious cross-linking morphology observed from scanning electron microscope (SEM). The obtained GNPs showed a good dispersibility with a size range of 386–438 nm. However, the GNPs cross-linked by 8 wt% genipin dosage revealed a relatively higher size because of the aggregation induced by hydrogen bond. The 10 wt% group had good thermal stability and storage stability. The optical microscopy results showed that the Pickering emulsions (30–50 vol% internal phase) stabilized by the GNPs had good uniformity and stability, even after 30 days of storage time, suggesting that the stable GNPs had great potential in food-grade Pickering emulsions.

Effect of preparation factors and storage temperature on fish oil-loaded crosslinked gelatin nanoparticle pickering emulsions in liquid forms

Fish oils are rich in omega-3 unsaturated fatty acids and have many important physiological functions and potential disease prevention abilities. It is necessary to develop novel dosage form and formulation such as food emulsions to encapsulate fish oils for protection, increase water solubility, and isolate the fishy taste. In this work, we mainly explore the effects of preparation factors and storage temperature on fish oil-loaded crosslinked nanoparticle Pickering emulsions in liquid form. Firstly, we prepared the crosslinked gelatin nanoparticles by a two-step desolvation method. Then we prepared fish oil-loaded gelatin nanoparticle Pickering emulsions by a homogenization processes. Subsequently, we explored the effects of preparation and storage factors (gelatin nanoparticle solution pH, homogenizing time, homogenizing speed, and storage temperature). Our results confirmed the successful preparation of crosslinked gelatin nanoparticles and fish oils-loaded gelatin nanoparticle Pickering emulsions. The Pickering emulsions mainly consist of three types of droplets with different sizes. The droplet sizes linearly decreased with the increase of gelatin nanoparticle solution pH and homogenizing time, whereas exponentially decreased with the increase of homogenizing speed. Storage temperatures had obviously different effects on the storage of the fish oil-loaded crosslinked gelatin nanoparticle Pickering emulsions. This work provides a way to encapsulate fish oils in emulsions and provides potential promising application prospectives in food beverages. It would be also beneficial to basic understanding of the formation of gelatin nanoparticle-based Pickering emulsions.

Synthesis of fish gelatin nanoparticles and their application for the drug delivery based on response surface methodology

Fish gelatin nanoparticle is produced using Tilapia fish skin for the first time by the two-step desolvation method. Fish gelatin is chosen for producing gelatin nanoparticles because no experiment have been done in using fish gelatin and to counter the problem associated with the use of mammalian gelatin, such as bovine spongiform encephalopathy disease. The effects of several factors on the particle size such as pH, acetone concentration, glutaraldehyde volume, stirring speed and stirring time are evaluated. Optimum conditions for the formation of gelatin nanoparticles are obtained using response surface method. Fish gelatin nanoparticles with optimum size of can be produced using pH of 2.45, acetone percentage of 16% (vol%), glutaraldehyde , stirring speed of 600 rpm, and stirring time for 6 h. The thermogram and molecular interaction of fish gelatin and fish gelatin nanoparticles are characterized using DSC and FTIR. In vitro drug release kinetic is examined using 5-fluorouracil as the model drug. The entrapment efficiency of 5-fluouracil as model drug is determined to be 40%. Fish gelatin could be used as a good alternative drug carrier for mammalian gelatin.

Electrophoretic deposition of dexamethasone-loaded gelatin nanospheres/chitosan coating and its dual function in anti-inflammation and osteogenesis.

Surface modification of metallic implants with bioactive and biodegradable coatings could be a promising approach for bone regeneration. The objective of this study was to prepare chitosan/gelatin nanospheres (GNs) composite coating for the delivery of dexamethasone (DEX). GNs with narrow size distribution and negative surface charge were firstly prepared by a two-step desolvation method. Homogeneous and stable gelatin nanospheres/chitosan (GNs/CTS) composite coatings were formed by electrophoretic deposition (EPD). Drug loading, encapsulation efficiency and in vitro release of DEX were estimated using high performance liquid chromatography (HPLC). The anti-inflammatory effect of DEX-loaded coatings on macrophage RAW 264.7 cells was assessed by the secretion of tumour necrosis factor (TNF) and inducible nitric oxide synthase (iNOS). Osteogenic differentiation of MC3T3-E1 osteoblasts on DEX-loaded coatings was investigated by osteogenic gene expression and mineralization. The DEX in GNs/CTS composite coating showed a two-stage release pattern could not only suppress inflammation during the burst release period, but also promote osteogenic differentiation in the sustained release period. This study might offer a feasible method for modifying the surface of metallic implants in bone regeneration.

Hierarchical Porous Protein Scaffold Templated from High Internal Phase Emulsion Costabilized by Gelatin and Gelatin Nanoparticles

Recently, three-dimensional (3D) scaffolds produced using poly-Pickering high internal phase emulsions (polyHIPEs) technology are particularly attractive in biomedical application. However, until now the most investigated polyHIPEs are hydrophobic composites originating from synthetic polymers. Here we present an investigation of a hierarchical porous protein scaffold templated from oil-in-water (O/W) HIPEs costabilized by fully natural materials, gelatin, and gelatin nanoparticles. Fairly monodispersed gelatin nanoparticles were first synthesized through a two-step desolvation method, and then they were used as emulsifiers together with gelatin to fabricate stable HIPEs with adjustable droplet size distribution and rheology. Monolithic scaffolds were formed by cross-linking the HIPEs with polymers as low as 2.5 wt % in the continuous phase, which appropriately presented a general high porosity and had an interconnected porous morphology with smooth pore walls and textured structures. Furthermore, the scaffolds were degradable and showed reasonably good biocompatibility; L929 cells could adhere to the surface of the materials and exhibited intensive growth and well-spread morphology. This hierarchical porous protein scaffold could, therefore, have important application as a 3D scaffold that offers enhanced cell adhesion and functionality.

Preparation and characterization of cardamom extract-loaded gelatin nanoparticles as effective targeted drug delivery system to treat glioblastoma

The purpose of this research study was to prepare and characterize cardamom extract-loaded gelatin nanoparticles (CE-loaded GNPs) with a diameter ≤200nm as a potent drug delivery system (DDS) for treatment of glioblastoma, which is the most common and aggressive type of brain tumor. The BBB poses physical and biological limitations to drug diffusion to reach target tissues. Polymeric nanoparticles, like gelatin, are suitable vehicles for drug delivery into the central nervous system (CNS). They are able to cross the BBB. Hence, we prepared CE-loaded GNPs by a two-step desolvation method. Seizures are detrimental secondary effects of brain tumors. Therefore, we used cardamom extract, which is an herbal anticancer and antiepileptic drug without any side effects, instead of synthetic drugs to load in gelatin nanoparticles during the particles preparation. We also prepared gelatin Type A and Type B nanoparticles for size comparison. Encapsulation efficiency, mean particle size, zeta potential and in vitro release profile were performed, and particle size analysis, dynamic light scattering (DLS), UV–Vis spectrophotometry, differential scanning calorimetry (DSC), X-Ray diffraction (XRD), scanning electron microscopy (SEM) and field emission scanning electron microscopy (FE-SEM) were employed to evaluate structural and physicochemical properties of the samples. CE-loaded GNPs were obtained with diameters of 40–200nm, zeta potential of −40.1mV and entrapment efficiency (EE) of 70%. The ratio of extract to polymer, 1:20, was revealed to be more suitable in obtaining smaller nanoparticles without any precipitate or aggregation. We also examined cytotoxic effects of CE and CE-loaded GNPs on human glioblastoma cancer U87MG cells.

Improved Stability and Enhanced Oral Bioavailability of Atorvastatin Loaded Stearic Acid Modified Gelatin Nanoparticles.

The present study evaluates the effects of stearic acid conjugation with gelatin and, its pharmaceutical potential to formulate novel atorvastatin (AT) loaded nanoparticles. AT loaded stearic acid modified gelatin nanoparticles (AT-MG NPs) were prepared via two-step desolvation method with extensive optimization of different process variables. Further, the developed nanoparticles where evaluated against in vitro Caco-2 cell model and in vivo bioavailability. Extensive optimization of nanoformulation resulted into the formation of AT-MG NPs with particle size 247.7±10.9nm, PDI 0.219±0.07, and entrapment efficiency 58.7±5.3%. Freeze dried nanoparticles were found to have spherical shape as determined by SEM and demonstrated excellent stability in simulated gastrointestinal conditions and during storage. Developed nanoparticles exhibited sustained release up to 24h and remarkably higher Caco-2 cell uptake. Mechanistic studies further revealed the clathrin and caveolae mediated endocytosis as principle mechanism. In line with Caco-2 cell uptake observations, AT-MG NPs showed ∼4.84-fold increase in the AUC0-∞ values of AT in comparison with free AT following oral administration. Overall, the stearic acid conjugated gelatin NPs demonstrates a promising potential in improving the drug payload of BCS class II drugs and enhancing oral bioavailability.

Controllable synthesis and characterisation of palladium (II) anticancer complex-loaded colloidal gelatin nanoparticles as a novel sustained-release delivery system in cancer therapy.

Over the past few years, there have been several attempts to deliver anticancer drugs into the body. It has been shown that compared to other available carriers, colloidal gelatin nanoparticles (CGNPs) have distinct properties due to their exceptional physico-chemical and biological characteristics. In this study, a novel water-soluble palladium (II) anticancer complex was first synthesised, and then loaded into CGNPs. The CGNPs were synthesised through a two-step desolvation method with an average particle size of 378 nm. After confirming the stability of the drug in the nanoparticles, the drug-loaded CGNPs were tested for in vitro cytotoxicity against human breast cancer cells. The results showed that the average drug encapsulating efficiency and drug loading of CGNPs were 64 and 10 ± 2.1% (w/w), respectively. There was a slight shift to higher values of cumulative release, when the samples were tested in lower pH values. In addition, the in vitro cytotoxicity test indicated that the number of growing cells significantly decreased after 48 h in the presence of different concentrations of drug. The results also demonstrated that the released drug could bind to DNA by a static mechanism at low concentrations (0.57 µM) on the basis of hydrophobic and hydrogen binding interactions.

Autofluorescent gelatin nanoparticles as imaging probes to monitor matrix metalloproteinase metabolism of cancer cells

In this paper, autofluorescent gelatin nanoparticles were synthesized as matrix metalloproteinase (MMP) responsive probes for cancer cell imaging. A modified two-step desolvation method was employed to generate these nanoparticles whose size was controllable and had stable autofluorescence. As glutaraldehyde was introduced as the crosslinking agent, the generation of Schiff base (CN) and double carbon bond (CC) between glutaraldehyde and gelatin endowed these gelatin nanoparticles distinct autofluorescence. Considering MMPs were usually overexpressed on the surface of cancer cells and they had degradation ability toward gelatin, we utilized these nanoparticles as imaging probes to responsively monitor the MMP metabolism of cancer cells according to the luminance change. As fluorescent probes, these nanoparticles had facile synthesis procedure and good biocompatibility, and provided a smart strategy to monitor cancer cell behaviors. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2854-2860, 2016.

A new method for the preparation of gelatin nanolayer: a possible approach to the in situ consolidation of damaged gelatin photographic emulsions

A major objective of all conservation treatments is to increase the physical and chemical stability of the objects being treated. Given that photographic gelatin emulsions are highly susceptible to different forms of damage such as flaking, cracking, and powdering, consolidation treatments often form an important part of the stabilization process. The application of nanomaterials is a new approach in photograph conservation, which will hopefully enable the control of the properties of traditional consolidants, thus improving their performance. In this study, gelatin nanoparticles and nanolayers were prepared and characterized by scanning electron microscopy, transmission electron microscopy, and dynamic light scattering for testing on damaged gelatin emulsion. The nanolayers were prepared by a two-step desolvation method, in order to obtain gelatin nanoparticles, followed by a third step involving rapid solvent evaporation. The prepared particles were found to have a spherical shape with sizes varying from 33 to 216 nm and a mean size of 42 nm depending on the conditions of synthesis. This research showed that both pH and temperature are very effective on particle size. Further studies are being planned to evaluate the benefits and potential problems of in situ consolidation of damaged gelatin emulsion using gelatin nanolayer technique.

Electrospun nanofibrous silk membranes embedding gelatin nanospheres for controlled drug delivery

Event Abstract Back to Event Electrospun nanofibrous silk membranes embedding gelatin nanospheres for controlled drug delivery Jiankang Song1, Alexey Klymov1, Wei Ji2, John A. Jansen1, Sander C. Leeuwenburgh1 and Fang Yang1 1 Radboud University Medical Center, Department of Biomaterials, Netherlands 2 KU Leuven, Department of Biomaterials, Belgium Introduction: Spatiotemporally controlled delivery of biomolecules such as growth factors and antibiotics has been proposed as an alternative to systemic delivery[1]. Fibrous silk membranes have been used to obtain such controlled and sustained delivery of biomolecules[2]. However, release of drugs from silk is hard to control due to lack of interactions between most biomolecules and silk. Gelatin nanospheres (GNs), on the other hand, are suitable vehicles for the controlled delivery of a variety of biomolecules due to their charged nature[3],[4]. Therefore, we have focused on the synthesis of silk-based drug release system containing oppositely charged GNs to improve control over the delivery of multiple bioactive molecules. To this end, we developed a simple core-shell electrospinning technique to fabricate silk nanofibers containing homogeneously distributed and oppositely charged GNs. Materials and Methods: Positively charged GNs (GANs, made of Gelatin Type A) and negatively charged GNs (GBNs, made of Gelatin Type B) were prepared by a two-step desolvation method[3] and were fluorescently labeled in PBS with DyLight™ 405 and 650 NHS esters, respectively. Regenerated silk fibroin was extracted from silk cocoons according to a previously developed protocol[5]. Nanofibrous membranes containing GANs and GBNs were prepared by core-shell electrospinning of silk/poly (ethylene oxide) (PEO) (3:1, wt/wt) suspensions containing GANs or GBNs at different feeding rates between core and shell suspensions, while nanofibers containing only GANs or GBNs were prepared by single nozzle electrospinning. The presence and distribution of GNs inside the nanofibers were characterized by confocal laser scanning microscopy. Moxifloxacin and vancomycin were added into GANs and GBNs suspensions, respectively, prior to preparation of drug-loaded nanofibrous membranes using electrospinning. The release kinetics of moxifloxacin and vancomycin from nanofibrous membranes were monitored by high performance liquid chromatography. Results and Discussion: Suspensions containing approximately 33.3 wt% of GNs could be electrospun into nanofibers without any structural inconsistencies (Figure 1e). GANs or GBNs were homogeneously distributed inside the nanofibers (Figure 1a-d). Core-shell electrospun nanofibers (Figure 2f) exhibited more co-localization of GANs and GBNs than single nozzle electrospun nanofibers (Figure 2e), while single nozzle electrospinning obtained more dispersed single GANs or GBNs. Moreover, the ratio and distribution of GANs and GBNs inside the nanofibers could be fine tuned by adjusting the feeding rates of core and shell suspensions, which offers the opportunity for independent control over the delivery of multiple biomolecules. Conclusion: Gelatin nanospheres were successfully incorporated into silk/PEO nanofibers to form fibrous “nano-in-nano” composites. The simple core-shell electrospinning technique can be utilized to fabricate nanofibers containing both GANs and GBNs, which enabled the fine tuning of the ratio and distribution of GANs and GBNs inside the nanofibers. The nanofibrous membranes can be used to delivery multiple biomolecules with adjustable release profile. China Scholarrship Council (CSC), project number: 201206150058; NutsOhra Foundation, project number: 1303-024

Evaluation of synovium-derived mesenchymal stem cells and 3D printed nanocomposite scaffolds for tissue engineering

Stem cells and scaffolds play a very important role in tissue engineering. Here, we isolated synovium-derived mesenchymal stem cells (SMSCs) from synovial membrane tissue and characterized stem-cell properties. Gelatin nanoparticles (NP) were prepared using a two-step desolvation method and then pre-mixed into different host matrix (silk fibroin (SF), gelatin (Gel), or SF–Gel mixture) to generate various 3D printed nanocomposite scaffolds (NP/SF, NP/SF–Gel, NP/Gel-1, and NP/Gel-2). The microstructure was examined by scanning electron microscopy. Biocompatibility assessment was performed through CCK-8 assay by coculturing with SMSCs at 1, 3, 7 and 14 days. According to the results, SMSCs are similar to other MSCs in their surface epitope expression, which are negative for CD45 and positive for CD44, CD90, and CD105. After incubation in lineage-specific medium, SMSCs could differentiate into chondrocytes, osteocytes and adipocytes. 3D printed nanocomposite scaffolds exhibited a good biocompatibility in the process of coculturing with SMSCs and had no negative effect on cell behavior. The study provides a strategy to obtain SMSCs and fabricate 3D printed nanocomposite scaffolds, the combination of which could be used for practical applications in tissue engineering.

Development of gemcitabine-adsorbed magnetic gelatin nanoparticles for targeted drug delivery in lung cancer

Magnetic iron oxide nanoparticles (IONPs) were coated with gelatin type B by means of the two-step desolvation method. Drug loading by adsorption was studied under various conditions such as different temperature, contact time, pH, and initial gemcitabine concentration. Further, Langmuir isotherm curves were constracted and constants were calculated. According to the Langmuir isotherm, the Gibbs free energy of the adsorption process at 25°C was − 4.74 kJ/mol. On the other hand, this value at 37°C was − 7.86 kJ/mol. In vitro drug release was performed at pH levels of 5 and 7.4, with gemcitabine-loaded magnetic gelatin nanoparticles and free gemcitabine, and both the results were subsequently compared.

Improved cisplatin delivery in cervical cancer cells by utilizing folate-grafted non-aggregated gelatin nanoparticles

PurposeCisplatin is highly effective in the treatment of cervical cancer. However, in therapeutic doses, cisplatin induces several adverse effects due to undesirable tissue distribution. Therefore, it is worth targeting cisplatin in cervical cancer cells by implicating non-aggregated ligand-modified nanotherapeutics. Methods and resultsHere, we report the preparation of non-aggregated folic acid-conjugated gelatin nanoparticles of cisplatin (Cis-GNs-FA) by two-step desolvation method with mean particle size of 210.6±9.6nm and 140.5±10.9nm for Cis-GNs to improve the drug delivery in cervical cancer, HeLa cells. FTIR and DSC spectra confirmed the presence and stability of cisplatin in gelatin matrix. Furthermore, amorphization of cisplatin in nanoparticles was ascertained by PXRD. Drug release followed a first-order release kinetic at both pH ∼ 5.6 (cervical cancer pH) and pH ∼ 7.4. In addition, a significant (P<0.05) decrease in IC50 value (8.3μM) and enhanced apoptosis were observed in HeLa cells treated with Cis-GNs-FA as compared to Cis-GNs (15.1μM) and cisplatin solution (40.2μM). In contrast, A549 lung cancer cells did not discriminate between Cis-GNs-FA and Cis-GNs due to the absence of folate receptors-α (FR-α). Consistently, higher cellular uptake, 80.54±7.60% was promoted by Cis-GNs-FA significantly (two-way ANOVA, P<0.05) greater than 51.68±9.78%, by Cis-GNs. This was also illustrated by CLSM images, which indicated that Cis-GNs-FA preferably accumulated in the cytoplasm of HeLa cells nearby nucleus by following receptor-mediated endocytosis pathway as compared to Cis-GNs. ConclusionTherefore, Cis-GNs-FA warrants further in-depth in vitro and in vivo investigations to scale up the technology for clinical translation.

Gelatin particle-stabilized high internal phase emulsions as nutraceutical containers.

In this paper, we report for the first time the use of a well-dispersed gelatin particle as a representative of natural and biocompatible materials to be an effective particle stabilizer for high internal phase emulsion (HIPE) formulation. Fairly monodispersed gelatin particles (∼200 nm) were synthesized through a two-step desolvation method and characterized by dynamic light scattering, ζ-potential measurements, scanning electron microscopy, and atomic force microscopy. Those protein latexes were then used as sole emulsifiers to fabricate stable oil-in-water Pickering HIPEs at different concentrations, pH conditions, and homogenization times. Most of the gelatin particles were irreversibly adsorbed at the oil-water interface to hinder droplet coalescence, such that Pickering HIPEs can be formed by a small amount of gelatin particles (as low as 0.5 wt % in the water phase) at pH far away from the isoelectric point of the gelatin particles. In addition, increasing homogenization time led to narrow size distribution of droplets, and high particle concentration resulted in more solidlike Pickering HIPEs. In vitro controlled-release experiments revealed that the release of the encapsulated β-carotene can be tuned by manipulating the concentration of gelatin particles in the formulation, suggesting that the stable and narrow-size-distributed gelatin-stabilized HIPEs had potential in functional food and pharmaceutical applications.