In Situ Gelling Scaffolds Loaded with Platelet Growth Factors to Improve Cardiomyocyte Survival after Ischemia.

Myocardial infarction is caused by prolonged ischemia and it is one of the main cause that leads to heart failures. The aim of the present work was the development of in situ gelling systems, based on poloxamer 407 (P407) or sodium alginate (Alg), loaded with platelet lysate (PL) to enhance cardiomyocyte survival after ischemia. Chondroitin sulfate (CS), a negatively charged glycosaminoglycan able to interact with different positively charged bioactive molecules, such as growth factors, was also investigated with both the systems. The gelation properties of both systems (viscosity, viscoelasticity, consistency by means of penetrometry, and injectability) were characterized in a physiological environment. In vitro evaluation of biocompatibility using fetal cardiac cells (cardiomyocytes and cardiac fibroblasts) demonstrated that the PL loaded alginate/chondroitin sulfate system retained the highest number of viable cells with equal distribution of the populations of cardiomyocytes and fibroblasts. Furthermore, the ability of the systems to improve cardiomyocyte survival after ischemia was also assessed. PL allowed for the highest degree of survival of cardiomyocytes after oxidative damage (simulating ischemic conditions due to MI) and both the Alg + CS PL and, to a greater extent, the PL alone demonstrated a considerable increase in survival of cardiomyocytes. In conclusion, an in situ gelling alginate-chondroitin sulfate system, loaded with platelet lysate, was able to improve the survival of cardiomyocytes after oxidative damage resulting in a promising system to improve cardiac cell viability after ischemia.

Freeze dried chitosan acetate dressings with glycosaminoglycans and traxenamic acid.

Bleeding control plays an important role to increase survival in the early phase after a traumatic event. The aim of present work was the development of hemostatic sponge-like dressings based on chitosan, in association with glycosaminoglycans (GAG) (chondroitin sulfate or hyaluronic acid) and the improvement of their hemostatic performance by loading tranexamic acid (TA). The dressings were prepared by lyophilizazion and were characterized for mechanical, hydration, bioadhesion properties and morphology. Moreover, FTIR analysis was performed to understand the interactions between the different polyelectrolytes present in the dressings. Clotting was investigated in vitro by using rat whole blood. Moreover, in vitro biocompatibility and proliferation were evaluated towards fibroblasts. Ex vivo proliferation properties were assessed by using human skin. All the dressings were characterised by mechanical, hydration and bioadhesion properties suitable to be applied on bleeding wounds and to absorb bleeding or wound exudate, avoiding tissue dehydration. TA release was fast; TA and chitosan showed a synergic effect to speed up clotting. The dressings were biocompatible and able to sustain cell proliferation in vitro and ex vivo in human skin. In conclusion, sponge-like dressings based on chitosan and GAG and loaded with TA are an effective tool to enhance hemostasis and healing in bleeding wounds.

Electrospun Gelatin⁻Chondroitin Sulfate Scaffolds Loaded with Platelet Lysate Promote Immature Cardiomyocyte Proliferation.

The aim of the present work was the development of heart patches based on gelatin (G) and chondroitin sulfate (CS) to be used as implants to improve heart recovery after corrective surgery for critical congenital heart defects (CHD). Patches were prepared by means of electrospinning to obtain nanofibrous scaffolds and they were loaded with platelet lysate (PL) as a source of growth factors to further enhance the repair process. Scaffolds were characterized for morphology and mechanical properties and for the capability to support in vitro adhesion and proliferation of dermal fibroblasts in order to assess the system's general biocompatibility. Adhesion and proliferation of endothelial cells and cardiac cells (cardiomyocytes and cardiac fibroblasts from rat fetuses) onto PL-loaded patches was evaluated. Patches presented good elasticity and high stiffness suitable for in vivo adaptation to heart contraction. CS improved adhesion and proliferation of dermal fibroblasts, as proof of their biocompatibility. Moreover, they enhanced the adhesion and proliferation of endothelial cells, a crucial mediator of cardiac repair. Cell adhesion and proliferation could be related to elastic properties, which could favor cell motility. The presence of platelet lysate and CS was crucial for the adhesion and proliferation of cardiac cells and, in particular, of cardiomyocytes: G/CS scaffold embedded with PL appeared to selectively promote proliferation in cardiomyocytes but not cardiac fibroblasts. In conclusion, G/CS scaffold seems to be a promising system to assist myocardial-repair processes in young patient, preserving cardiomyocyte viability and preventing cardiac fibroblast proliferation, likely reducing subsequent uncontrolled collagen deposition by fibroblasts following repair.

Essential oil-loaded lipid nanoparticles for wound healing.

Chronic wounds and severe burns are diseases responsible for severe morbidity and even death. Wound repair is a crucial process and tissue regeneration enhancement and infection prevention are key factors to minimize pain, discomfort, and scar formation. The aim of this work was the development of lipid nanoparticles (solid lipid nanoparticles and nanostructured lipid carriers [NLC]), to be loaded with eucalyptus or rosemary essential oils and to be used, as medical devices, to enhance healing of skin wounds. Lipid nanoparticles were based on natural lipids: cocoa butter, as solid lipid, and olive oil or sesame oil, as liquid lipids. Lecithin was chosen as surfactant to stabilize nanoparticles and to prevent their aggregation. The systems were prepared by high shear homogenization followed by ultrasound application. Nanoparticles were characterized for physical-chemical properties, bioadhesion, cytocompatibility, in vitro proliferation enhancement, and wound healing properties toward normal human dermal fibroblasts. Antimicrobial activity of nanoparticles was evaluated against two reference microbial strains, one of Staphylococcus aureus, the other of Streptococcus pyogenes. Finally, the capability of nanoparticles to promote wound healing in vivo was evaluated on a rat burn model. NLC based on olive oil and loaded with eucalyptus oil showed appropriate physical-chemical properties, good bioadhesion, cytocompatibility, in vitro proliferation enhancement, and wound healing properties toward fibroblasts, associated to antimicrobial properties. Moreover, the in vivo results evidenced the capability of these NLC to enhance the healing process. Olive oil, which is characterized by a high content of oleic acid, proved to exert a synergic effect with eucalyptus oil with respect to antimicrobial activity and wound repair promotion.

Innovative Methods and Applications in Mucoadhesion Research.

The present review is aimed at elucidating relatively new aspects of mucoadhesion/mucus interaction and related phenomena that emerged from a Mucoadhesion workshop held in Munster on 2-3 September 2015 as a satellite event of the ICCC 13th-EUCHIS 12th. After a brief outline of the new issues, the focus is on mucus description, purification, and mucus/mucin characterization, all steps that are pivotal to the understanding of mucus related phenomena and the choice of the correct mucosal model for in vitro and ex vivo experiments, alternative bio/mucomimetic materials are also presented. Then a selection of preparative techniques and testing methods are described (at molecular as well as micro and macroscale) that may support the pharmaceutical development of mucus interactive systems and assist formulators in the scale-up and industrialization steps. Recent applications of mucoadhesive systems (including medical devices) intended for different routes of administration (oral, gastrointestinal, vaginal, nasal, ocular, and intravesical) and for the treatment of difficult to treat pathologies or the alleviation of symptoms are described.

Platelet lysate embedded scaffolds for skin regeneration.

INTRODUCTION: The work presents the development of acellular scaffolds extemporaneously embedded with platelet lysate (PL), as an innovative approach in the field of tissue regeneration/reparation. PL embedded scaffolds should have a tridimensional architecture to support cell migration and growth, in order to restore skin integrity. For this reason, chondroitin sulfate (CS) was associated with sodium alginate (SA) to prepare highly porous systems. METHODS: The developed scaffolds were characterized for chemical stability to γ-radiation, morphology, hydration and mechanical properties. Moreover, the capability of fibroblasts and endothelial cells to populate the scaffold was evaluated by means of proliferation test 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and confocal laser scanning microscopy study. The scaffolds, not altered by sterilization, were characterized by limited swelling and high flexibility, by foam-like structure with bubbles that formed a high surface area and irregular texture suitable for cell adhesion. RESULTS: Cell growth and scaffold population were evident on the bubble surface, where the cells appeared anchored to the scaffold structure. CONCLUSION: Scaffold network based on CS and SA demonstrated to be an effective support to enhance and to allow fibroblasts and endothelial cells (human umbilical vein endothelial cells, HUVEC) adhesion and proliferation. In particular, it could be hypothesized that cell adhesion was facilitated by the synergic effect of PL and CS. Although further in vivo evaluation is needed, on the basis of in vitro results, PL embedded scaffolds seem promising systems for skin wound healing.