Institute of Fundamental Technological Research

Extreme loss of skeletal muscle overwhelms the natural regenerative capability of the body, results in permanent disability and substantial economic burden. Current surgical techniques result in poor healing, secondary injury to the autograft donor site, and incomplete recuperation of muscle function. Most current tissue engineering and regenerative strategies fail to create an adequate mechanical and biological environment that enables cell infiltration, proliferation, and myogenic differentiation. In this study, we present a nanoengineered skeletal muscle scaffold based on functionalized gelatin methacrylate (GelMA) hydrogel, optimized for muscle progenitors’ proliferation and differentiation. The scaffold was capable of controlling the release of insulin-like growth factor 1 (IGF-1), an important myogenic growth factor, by utilizing the electrostatic interactions with LAPONITE® nanoclays (NCs). Physiologically relevant levels of IGF-1 were maintained during a controlled release over two weeks. The NC was able to retain 50% of the released IGF-1 within the hydrogel niche, significantly improving cellular proliferation and differentiation compared to control hydrogels. IGF-1 supplemented medium controls required 44% more IGF-1 than the comparable NC hydrogel composites. The nanofunctionalized scaffold is a viable option for the treatment of extreme muscle injuries and offers scalable benefits for translational interventions and the growing field of clean meat production.

Engineering tissue-like scaffolds that can mimic the microstructure, architecture, topology, and mechanical properties of native tissues while offering an excellent environment for cellular growth has remained an unmet need. To address these challenges, multi-compartment composite fibers are fabricated. These fibers can be assembled through textile processes to tailor tissue-level mechanical and electrical properties independent of cellular level components. Textile technologies also allow controlling the distribution of different cell types and microstructure of fabricated constructs and directing cellular growth within 3D microenvironment. Here, we engineered composite fibers from biocompatible cores and biologically relevant hydrogel sheaths. The fibers are mechanically robust to be assembled using textile processes and could support adhesion, proliferation and maturation of cell populations important for engineering of skeletal muscles. We also demonstrated that the changes in the electrical conductivity of the multi-compartment fibers could significantly enhance myogenesis in vitro.

reinforced fibers, biotextiles, tissue engineering, organ weaving, interpenetrating network hydrogels, skeletal muscles

Liquid-crystal-based biomaterials provide promising platforms for the development of dynamic and responsive interfaces for tissue engineering. Cholesteryl ester liquid crystals (CLCs) are particularly well suited for these applications, due to their roles in cellular homeostasis and their intrinsic ability to organize into supramolecular helicoidal structures on the mesoscale. Here, we developed a nonwoven CLC electrospun scaffold by dispersing three cholesteryl ester-based mesogens within polycaprolactone (PCL). We tuned the ratio of our mesogens so that the CLC would be in the mesophase at the cell culture incubator temperature of 37°C. In these scaffolds, the PCL polymer provided an elastic bulk matrix while the homogeneously dispersed CLC established a viscoelastic fluidlike interface. Atomic force microscopy revealed that the 50% (w/v) cholesteryl ester liquid crystal scaffold (CLC-S) exhibited a mesophase with topographic striations typical of liquid crystals. Additionally, the CLC-S favorable wettability and ultrasoft fiber mechanics enhanced cellular attachment and proliferation. Increasing the CLC concentration within the composites enhanced myoblast adhesion strength promoted myofibril formationin vitrowith mouse myoblast cell lines.

Tendon injuries are frequent and occur in the elderly, young, and athletic populations. The inadequate number of donors combined with many challenges associated with autografts, allografts, xenografts, and prosthetic devices have added to the value of engineering biological substitutes, which can be implanted to repair the damaged tendons. Electrospun scaffolds have the potential to mimic the native tissue structure along with desired mechanical properties and, thus, have attracted noticeable attention. In order to improve the biological responses of these fibrous structures, we designed and fabricated 3D multilayered composite scaffolds, where an electrospun nanofibrous substrate was coated with a thin layer of cell-laden hydrogel. The whole construct composition was optimized to achieve adequate mechanical and physical properties as well as cell viability and proliferation. Mesenchymal stem cells (MSCs) were differentiated by the addition of bone morphogenetic protein 12 (BMP-12). To mimic the natural function of tendons, the cell-laden scaffolds were mechanically stimulated using a custom-built bioreactor. The synergistic effect of mechanical and biochemical stimulation was observed in terms of enhanced cell viability, proliferation, alignment, and tenogenic differentiation. The results suggested that the proposed constructs can be used for engineering functional tendons.

tendon tissue engineering, composite scaffolds, nanofibrous materials, mechanical stimulation, stem cell differentiation

Chronic, non-healing wounds place a significant burden on patients and healthcare systems, resulting in impaired mobility, limb amputation, or even death. Chronic wounds result from a disruption in the highly orchestrated cascade of events involved in wound closure. Significant advances in our understanding of the pathophysiology of chronic wounds have resulted in the development of drugs designed to target different aspects of the impaired processes. However, the hostility of the wound environment rich in degradative enzymes and its elevated pH, combined with differences in the time scales of different physiological processes involved in tissue regeneration require the use of effective drug delivery systems. In this review, we will first discuss the pathophysiology of chronic wounds and then the materials used for engineering drug delivery systems. Different passive and active drug delivery systems used in wound care will be reviewed. In addition, the architecture of the delivery platform and its ability to modulate drug delivery are discussed. Emerging technologies and the opportunities for engineering more effective wound care devices are also highlighted.

Wound healing, Drug delivery, Transdermal delivery, Microtechnologies, Nanotechnologies

Tissue engineering holds great potential in the production of functional substitutes to restore, maintain or improve the functionality in defective or lost tissues. So far, a great variety of techniques and approaches for fabrication of scaffolds have been developed and evaluated, allowing researchers to tailor precisely the morphological, chemical and mechanical features of the final constructs. Electrospinning of biocompatible and biodegradable polymers is a popular method for producing homogeneous nanofibrous structures, which might reproduce the nanosized organization of the tendons. Moreover, composite scaffolds obtained by incorporating nanoparticles within electrospun fibers have been lately explored in order to enhance the properties and the functionalities of the pristine polymeric constructs. The present study is focused on the design and fabrication of biocompatible electrospun nanocomposite fibrous scaffolds for tendon regeneration. A mixture of poly(amide 6) and poly(caprolactone) is electrospun to generate constructs with mechanical properties comparable to that of native tendons. To improve the biological activity of the constructs and modify their topography, wettability, stiffness and degradation rate, we incorporated silica particles into the electrospun substrates. The use of nanosize silica particles enables us to form bead-on-fiber topography, allowing the better exposure of ceramic particles to better profit their beneficial characteristics. In vitro biocompatibility studies using L929 fibroblasts demonstrated that the presence of 20 wt% of silica nanoparticles in the engineered scaffolds enhanced cell spreading and proliferation as well as extracellular matrix deposition. The results reveal that the electrospun nanocomposite scaffold represents an interesting candidate for tendon tissue engineering.

Periodontitis is a prevalent chronic, destructive inflammatory disease affecting tooth‐supporting tissues in humans. Guided tissue regeneration strategies are widely utilized for periodontal tissue regeneration generally by using a periodontal membrane. The main role of these membranes is to establish a mechanical barrier that prevents the apical migration of the gingival epithelium and hence allowing the growth of periodontal ligament and bone tissue to selectively repopulate the root surface. Currently available membranes have limited bioactivity and regeneration potential. To address such challenges, an osteoconductive, antibacterial, and flexible poly(caprolactone) (PCL) composite membrane containing zinc oxide (ZnO) nanoparticles is developed. The membranes are fabricated through electrospinning of PCL and ZnO particles. The physical properties, mechanical characteristics, and in vitro degradation of the engineered membrane are studied in detail. Also, the osteoconductivity and antibacterial properties of the developed membrane are analyzed in vitro. Moreover, the functionality of the membrane is evaluated with a rat periodontal defect model. The results confirmed that the engineered membrane exerts both osteoconductive and antibacterial properties, demonstrating its great potential for periodontal tissue engineering.

electrospinning, guided tissue regeneration, osteoconductive, periodontal regeneration, zinc oxide