Journal of Bioactive and Compatible Polymers

Temperature-responsive hydrogel culture system is considered as an ideal platform for cell sheet harvest, but its complex preparation methods and harsh reaction conditions limit its application. Inspired by the marine mussels, a biomimetic method presented here is to construct a novel temperature-responsive hydrogel culture system for stem cell sheet harvest. The tissue culture polystyrene is first modified with polydopamine coating, and then amine-terminated poly(N-isopropylacrylamide) is grafted onto the coating via the Schiff base or Michael addition reaction to construct the temperature-sensitive hydrogel culture system. Then, bone marrow stromal cells are cultured on the culture system to construct cell sheets. The prepared culture system shows significant temperature-sensitive property with the grafted concentrations of poly(N-isopropylacrylamide) ranging from 0.5 to 1 g/L. Meanwhile, the constructed culture system has low cytotoxicity and facilitates the stem cell adhesion, proliferation, and cell sheet formation at 37°C. When the culture system is placed in a 20°C environment, the cell sheet can be completely detached from the surface of tissue culture polystyrene without being treated with any enzymes. More importantly, the cell morphology, cell sheet thickness, and the fibril structure of the associated proteins are similar to the cells cultured on the tissue culture polystyrene without modification. The biomimetic, simple, inexpensive, and environmentally friendly preparation of the culture system enables it to be used for the harvest of cell sheet and even applied to tissue engineering for tissue regeneration.

Our study presents a novel collagen wound dressing prepared from freshwater fish skin (Cyprinus Carpio) collagen type I. Half of the sponges were cross-linked with carbodiimide. The cross-linked and non-cross-linked collagen sponges were subsequently impregnated with gentamicin and lyophilized thus allowing for the attainment of the appropriate gentamicin content without the removal thereof during the cross-linking stage. The structure was evaluated via micro-CT and infrared spectrometry and the structural stability and gentamicin release properties were evaluated in phosphate buffer solution. The sponges were further tested via a rat model of an infected wound with Pseudomonas aeruginosa inoculation and compared with a reference commercial product. The sponges thus prepared provided a degree of open porosity that was comparable to or higher than that of the reference commercial product. Spectrometry analysis revealed that the cross-linked collagen sponge and reference commercial product sponge preserved their secondary collagen structure after 168 h while early accelerated degradation was observed with respect to the non-cross-linked collagen sponge. Gentamicin was released rapidly from all the sponges. Compared to those animals with gentamicin-containing sponges or gentamicin administered intramuscularly, the animals with the cross-linked collagen sponge without gentamicin exhibited marked clinical and laboratory infection signs. Both the administration routes (intramuscular and via gentamicin-containing sponges) provided similar gentamicin plasma levels. The resulting highly homogeneous product which was characterized by excellent structural and clinical properties proved effective in terms of the treatment of a surgical wound infection in a rat model. We demonstrated that all the gentamicin was released from the sponge and was absorbed in the systemic circulation. This is the first time that Cyprinus Carpio collagen has been used in the preparation of wound dressings. Thus, gentamicin-containing sponges provide a promising tool for the treatment and prevention of surgical site infections.

The main aim of this study is to fabricate an electrospun citric acid modified polyvinyl alcohol polyester that is biodegradable with non-toxic by-products and can be used for the culture of vascular smooth muscle cells. In this study, we have optimized the conditions for the electrospinning process of this polyester. The fibre morphology was studied by scanning electron microscopy which indicated that the fibre diameter was optimum at a range of 200 to 700 µm at 5% concentration and flow rate of 0.3 mL/h. The membranes were characterized for the change in structural aspects at the molecular level. The results showed development of more crystalline domains on electrospinning. The surface characteristics were also explored. Cell culture studies confirmed that the electrospun scaffold supported the attachment and proliferation of smooth muscle cells, which was evident from the cell proliferation assay. Hence, the electrospun polyester scaffolds are non-toxic and biocompatible with vascular smooth muscle cells, and find promising potential as scaffolds for vascular tissue engineering.

In this study, a combined experimental and computational study of long-term human bladder epithelial cell line HBLAK adhesion and proliferation on five different polymeric surfaces, namely hyaluronic acid, amylose, collagen, polyhydroxybutyrate, and polylactic acid, was performed with the goal to understand the nature of the attraction between various surface materials and a simplified model of the cell surface (transmembrane protein integrin). These biodegradable polymers are frequently used as scaffolds for tissue engineering. During formation of the new tissue, the scaffold polymers are gradually replaced by the natural extracellular matrix of the proliferating cells. Cell adhesion and proliferation experiments were carried out employing thin polymer films prepared by solvent casting while for the computational approach three-dimensional molecular models of layers of ordered polymeric fibers were used as quasi-planar nano-sized models of polymeric surface patches. Experimental results indicated a good capability of amylose, polyhydroxybutyrate, and hyaluronic acid polymer films to foster cell adhesion. Proliferation experiment, carried out by incubating cells with the investigated polymer films for 72 h, showed that all the investigated polymers are able to sustain a good proliferation of HBLAK cells almost comparable to plain glass. Computational estimate of molecular mechanic interaction energies between three-dimensional models of polymeric films and the collagen-binding α₂ I domain of the cell adhesion receptor integrin α₂β₁ confirmed elevated affinity to amylose and polyhydroxybutyrate that is related to higher polarity of function groups on the film surface as documented by the maps of molecular electrostatic potential. This combined experimental and modeling approach can contribute to rational design and surface modifications of polymeric material suitable for forming the scaffolds of human urethra that can be effectively colonized by stem cells.

Melatonin, a hormone produced in the pineal gland, has been investigated for bone repair, remodeling, osteoporosis, as well as osseointegration of the implants. In this study, different concentrations of melatonin (0–2000-µM) were embedded into silk films annealed by methanol or water. Then, their capacity to differentiate human mesenchymal stem cells into osteoblasts was investigated for bone tissue regeneration. While methanol-annealed silk films have ~55% crystallinity, room-temperature water-annealed silk films have ~30% crystallinity by depending upon their different β-sheet contents. Melatonin-loaded silk films exhibited an initial burst release followed by a continuous release for up to 5 days, and the β-sheet content of silk films did not affect the release behavior of melatonin, an amphiphilic molecule. Moreover, human mesenchymal stem cells exhibited an increase in osteogenic markers such as alkaline phosphatase activity, osteocalcin, and runt-related transcription factor 2 expressions on the melatonin-loaded methanol-annealed silk films in both proliferation and osteogenic media. The bioactivity of the melatonin-modified silk films was further confirmed by the enhanced mineralization compared to silk films alone. This study demonstrated the feasibility of developing melatonin-loaded silk materials and the positive effect of releasing melatonin at micromolar concentrations on osteogenic differentiation of human mesenchymal stem cells cultured especially in osteogenic medium.