Stabilization of hyaluronan-based materials by peptide conjugation and its use as a cell-seeded scaffold in tissue engineering.

New materials based on molecules naturally occurred in body are assumed to be fully biocompatible and biodegradable. In our study, we used hyaluronic acid (HA) modified with peptides, which meet all this criterion and could be advantageously used in tissue engineering. Peptides with RGD, IKVAV or SIKVAV adhesive motif were attached to HA-based fiber or non-woven textile through ester bond in the term of solid phase peptide synthesis. A linker between HA and peptide containing three glycine or two 6-aminohexanoyl units was applied to make peptides more available for cell surface receptors. Dermal fibroblasts adhered readily on this material, preferentially to RGD peptide with 6-aminohexanoyl linker. Contrary, the absence of adhesive peptide did not allow the cell attachment but maintained the material stability.

Grafting of steroids to hyaluronan towards the design of delivery systems for antioxidants: The role of hydrophobic core.

In this work, amphiphilic hyaluronic acid (HA) was synthesized by the chemical bonding of steroids. Particularly, succinyl cholesterol (SCH), cholic acid (CA), deoxycholic acid (DOCA), and 18ß-glycyrrhetinic acid (GA) were activated by benzoyl chloride towards the esterification reaction of HA in water. The degree of substitution can be controlled by varying the feed ratio of mixed anhydride to HA and up to 25% (mol/mol) can be obtained. Due to mild reaction conditions, no degradation of the polysaccharide was observed. The prepared amphiphilic polymers were characterized by NMR, infrared spectroscopy (FT-IR) and SEC/MALLS, as well as turbidity and size of the aggregates. The developed system is proposed for the delivery of hydrophobic drugs; for this purpose, curcumin, vitamin E and coenzyme Q10 were used as hydrophobic models; these molecules were loaded into the conjugates with high efficiency. The loading capacity was a function of degree of substitution. Furthermore, the biocompatibility of the derivatives and the cellular uptake of the delivery system enabled us to demonstrate the potential of the prepared delivery systems.

Optimization of cell growth on palmitoyl-hyaluronan knitted scaffolds developed for tissue engineering applications.

Polysaccharides meet several criteria for a suitable biomaterial for tissue engineering, which include biocompatibility and ability to support the delivery and growth of cells. Nevertheless, most of these polysaccharides, for example dextran, alginate, and glycosaminoglycans, are highly soluble in aqueous solutions. Hyaluronic acid hydrophobized by palmitic acid and processed to the form of wet-spun fibers and the warp-knitted textile scaffold is water non-soluble, but biodegradable material, which could be used for the tissue engineering purpose. However, its surface quality does not allow cell attachment. To enhance the biocompatibility the surface of palmitoyl-hyaluronan was roughened by freeze drying and treated by different cell adhesive proteins (fibronectin, fibrinogen, laminin, methacrylated gelatin and collagen IV). Except for collagen IV, these proteins covered the fibers uniformly for an extended period of time and supported the adhesion and cultivation of dermal fibroblasts and mesenchymal stem cells. Interestingly, adipose stem cells cultivated on the fibronectin-modified scaffold secreted increasing amount of HGF, SDF-1, and VEGF, three key growth factors involved in cardiac regeneration. These results suggested that palmitoyl-hyaluronan scaffold may be a promising material for various applications in tissue regeneration, including cardiac tissue repair. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1488-1499, 2018.

Fabrication of biodegradable textile scaffold based on hydrophobized hyaluronic acid.

In this work, we report on the preparation of a novel biodegradable textile scaffold made of palmitoyl-hyaluronan (palHA). Monofilament fibres of palHA with a diameter of 120µm were prepared by wet spinning. The wet-spun fibres were subsequently processed into a warp-knitted textile. To find a compromise between swelling in water and degradability of the final textile scaffold, a series of palHA derivatives with different degrees of substitution of the palmitoyl chain was synthesized. Freeze-drying not only provided shape fixation, but also speeded up scaffold degradation in vitro. Fibronectin, fibrinogen, laminin and collagen IV were physically adsorbed on the textile surface to enhance cell adhesion on the material. The highest amount of adsorbed cell-adhesive proteins was achieved with fibronectin (89%), followed by fibrinogen (81%). Finally, textiles modified with fibronectin or fibrinogen both supported the adhesion and proliferation of normal human fibroblasts in vitro, proving to be a useful cellular scaffold for tissue engineering.

Influence of serum albumin on intracellular delivery of drug-loaded hyaluronan polymeric micelles.

Polymeric micelles are attractive drug delivery systems for intravenously administered nonpolar drugs. Although physical parameters like size, shape and loading capacity are considered as the most important for their efficiency, here we demonstrate that the effects of serum protein interaction and characteristics of loaded compound cannot be neglected during the micelle development and design of experimental set up. Polymeric micelles prepared from amphiphilic hyaluronic acid grafted with short (hexanoic) and long fatty acids (oleic) were tested after loading with two different hydrophobic models, Nile red and curcumin. The composition of micelles affected mainly the loading capacity. Both encapsulated compounds behaved differently in the in vitro cell uptake, which was also influenced by serum concentration, where serum albumin was found to be the primary destabilizing component. This destabilization was found to be influenced by polymeric micelle concentration. Thus, the chemical structure of micelle, the properties of non-covalently loaded substance and serum albumin/polymeric micelle ratio modulate the in vitro intracellular uptake of drugs loaded in nanocarriers.