The main goal of our research was to fabricate electrospun scaffolds from three different silk proteins—silk fibroin from Bombyx mori silkworm cocoons and two recombinant spidroins, rS2/12 and rS2/12-RGDS—and to perform a comparative analysis of the structure, biological properties, and regenerative potential of the scaffolds in a full-thickness rat skin wound model. The surface and internal structures were investigated using scanning electron microscopy and scanning probe nanotomography. The structures of the scaffolds were similar. The average fiber diameter of the scaffolds was 315 ± 26 nm, the volume porosity was 94.5 ± 1.4%, the surface-to-volume ratio of the scaffolds was 25.4 ± 4.2 μm−1 and the fiber surface roughness was 3.8 ± 0.6 nm. The scaffolds were characterized by a non-cytotoxicity effect and a high level of cytocompatibility with cells. The scaffolds also had high regenerative potential—the healing of the skin wound was accelerated by 19 days compared with the control. A histological analysis did not reveal any fragments of the experimental constructions or areas of inflammation. Thus, novel data on the structure and biological properties of the silk fibroin/spidroin electrospun scaffolds were obtained.
Fluorescent imaging is a widely used technique for detecting and monitoring the distribution, interaction, and transformation processes at molecular, cellular, and tissue level in modern diagnostic and other biomedical applications. Unique photophysical properties of fluorescent semiconductor nanocrystals “quantum dots” (QDs) make them advanced fluorophores for fluorescent labeling of biomolecules or optical encoding of microparticles to be used as bioimaging and theranostic agents in targeted delivery, visualization, diagnostics, and imaging. This paper reports on the results of development of an improved approach to the optical encoding of polyelectrolyte microcapsules with stable, covered with the multifunctional polyethyleneglycol derivatives water-soluble QDs, as well as characterization of the optical properties, morphological and structural properties of the encoded microcapsules. The embedding of QDs into the polymer microcapsule membrane through layer-by-layer deposition on a preliminarily formed polymeric polyelectrolyte shell makes it possible to obtain bright fluorescent particles with an adapted charge and size distribution that are distinctly discernible by flow cytometry as individual homogeneous populations. The fluorescent microcapsules developed can be used in further designing bioimaging and theranostic agents sensitive to various external stimuli along with photoexcitation.
A comparative analysis of the structure and biological properties of silk fibroin constructions was performed. Three groups of constructions were obtained: films obtained by casting an aqueous solution of silk fibroin and electrospun microfibrous scaffolds based on silk fibroin, with the addition of 30% gelatin per total protein weight. The internal structures of the films and single fibers of the microfibrous scaffolds consisted of densely packed globule structures; the surface area to volume ratios and volume porosities of the microfibrous scaffolds were calculated. All constructions were non-toxic for cells and provide high levels of adhesion and proliferation. The high regenerative potential of the constructions was demonstrated in a rat full-thickness skin wound healing model. The constructions accelerated healing by an average of 15 days and can be considered to be promising constructions for various tasks of tissue engineering and regenerative medicine.
Silk fibroin has recently gained considerable attention as a promising biomaterial for use in medical and bioengineering technologies due to its biocompatibility and favorable mechanical properties. In this study, composite gel based on silk fibroin microparticles and carboxymethyl cellulose was developed, characterized by a viscous, homogeneous white mass containing uniformly distributed fibroin microparticles ranging from 1 to 20 μm in size. The gel exhibited a kinematic viscosity of 36.5 × 10−6 St, allowing for convenient application to wounds using a syringe or spatula while preventing uncontrolled spreading. The cytocompatibility of the gel was confirmed using the methylthiazol tetrazolium (MTT) assay, which showed no cytotoxic effects on 3T3 fibroblast cells. Furthermore, the gel remained stable for over one year when stored at 10 °C, in contrast to conventional fibroin solutions, which typically lose stability within a month under similar conditions. In a full-thickness skin wound model in rats, the application of the gel significantly accelerated skin regeneration, with complete wound closure observed by day 15, compared with 30 days in the control group. Histological analysis confirmed the restoration of all skin layers. These findings demonstrate the high potential of the gel for applications in regenerative medicine and tissue engineering.
The development of advanced biomaterials and constructs for accelerated recovery of damaged tissues is a key direction in regenerative medicine. Biocompatible scaffolds based on natural biopolymers are widely used for these tasks. Organ decellularization enables obtaining a cell-free extracellular matrix (ECM) with preserved composition and biological activity. The objectives of the present work were combining these two approaches for the development of a composite scaffold based on silk fibroin and ECM microparticles and assessing its structure, biological properties, and regenerative potential. ECM microparticles were obtained by grinding the decellularized matrix of Wistar rat liver in liquid nitrogen. Scaffolds in the form of films were prepared by the casting method. The sinuous and rough topography of the scaffold surface was assessed by the scanning probe nanotomography (SPNT) technique. The inclusion of ECM microparticles in the composition did not affect the elasticity and tensile strength of the scaffolds. The obtained scaffold was non-toxic to cells, maintained high levels of adhesion and proliferation of mouse 3T3 fibroblast and Hep-G2 cells, and showed high regenerative potential, which was studied in the experimental model of full-thickness rat skin wound healing. The wound healing was accelerated by 1.74 times in comparison with the control.