The differentiation potential of mesenchymal stem cells (MSC) has been extensively tested on electrospun scaffolds. However, this potential is often assessed with lineage-specific medium, making it difficult to interpret the real contribution of the properties of the scaffold in the cell response. In this study, we analyzed the ability of different polycaprolactone/polylactic acid PCL/PLA electrospun scaffolds (pure or blended compositions, random or aligned fibers, various fiber diameters) to drive MSC towards bone or tendon lineages in the absence of specific differentiation medium. C3H10T1/2 cells (a mesenchymal stem cell model) were cultured on scaffolds for 96 h without differentiation factors. We performed a cross-analysis of the cell–scaffold interactions (spreading, organization, and specific gene expression) with mechanical (elasticity), morphological (porosity, fibers diameter and orientation) and surface (wettability) characterizations of the electrospun fibers. We concluded that (1) osteogenic differentiation can be initiated on pure PCL-based electrospun scaffolds without specific culture conditions; (2) fiber alignment modified cell organization in the short term and (3) PLA added to PCL with an increased fiber diameter encouraged the stem cells towards the tendon lineage without additional tenogenic factors. In summary, the differentiation potential of stem cells on adapted electrospun fibers could be achieved in factor-free medium, making possible future applications in clinically relevant situations.

For decades, the unique regenerative properties of the human amniotic membrane (hAM) have been successfully utilized in ophthalmology. As a directly applied biomaterial, the hAM should be available in a ready to use manner in clinical settings. However, an extended period of time is obligatory for performing quality and safety tests. Hence, the low temperature storage of the hAM is a virtually inevitable step in the chain from donor retrieval to patient application. At the same time, the impact of subzero temperatures carries an increased risk of irreversible alterations of the structure and composition of biological objects. In the present study, we performed a comprehensive analysis of the hAM as a medicinal product; this is intended for a novel strategy of application in ophthalmology requiring a GMP production protocol including double freezing–thawing cycles. We compared clinically relevant parameters, such as levels of growth factors and extracellular matrix proteins content, morphology, ultrastructure and mechanical properties, before and after one and two freezing cycles. It was found that epidermal growth factor (EGF), transforming growth factor beta 1 (TGF-β1), hepatocyte growth factor (HGF), basic fibroblast growth factor (bFGF), hyaluronic acid, and laminin could be detected in all studied conditions without significant differences. Additionally, histological and ultrastructure analysis, as well as transparency and mechanical tests, demonstrated that properties of the hAM required to support therapeutic efficacy in ophthalmology are not impaired by dual freezing.

Magnesium is a promising metal for resorbable cardiovascular implants due to its high biocompatibility, high corrosion tendency, and mechanical properties. However, adapting its corrosion rate to the physiological healing processes is required to ascertain a safe graft function. A protective polymeric layer is supposed to slow down the corrosion rate of magnesium. Additionally, coatings can improve the host’s tissue interaction with the implant by implementing the local delivery of antibiotic drugs and growth or cell adhesion factors. However, little is known about the interaction of polymer-based coatings, their degradation, and magnesium corrosion. This study examines the corrosion mechanism of magnesium protected by spin coatings and electrospun fiber coatings under physiological conditions. Pure magnesium specimens were coated with polycaprolactone (PCL). The corrosion of the coated magnesium was evaluated using an immersion test in simulated body fluid. Spin coatings provided efficient protection against corrosive attacks and a significantly lower corrosion rate by 75% compared to uncoated magnesium. In contrast, fiber coatings did not provide relevant corrosion protection. On the other hand, magnesium corrosion caused the accelerated degradation of the PCL layer. A reliable and safe implant function is vital, especially in cardiovascular applications. Magnesium coating, therefore, should be carried out with spin coatings.

Microstructural responses to the mechanical load of polymers used in tissue engineering is notably important for qualification at in vivo testing, although insufficiently studied, especially regarding promising polycaprolactone (PCL). For further investigations, electrospun PCL scaffolds with different degrees of fiber alignment were produced, using two discrete relative drum collector velocities. Development and preparation of an adjusted sample geometry enabled in situ tensile testing in scanning electron microscopy. By analyzing the microstructure and the use of selected tracking techniques, it was possible to visualize and quantify fiber/fiber area displacements as well as local fractures of single PCL fibers, considering quasi-static tensile load and fiber alignment. The possibility of displacement determination using in situ scanning electron microscopy techniques for testing fibrous PCL scaffolds was introduced and quantified.

Assigning the hemocompatibility of small vascular implants is one of the great challenges in biomedical engineering. Due to the fact, that there are no widely approved test setups1, we decided to developed a modified CHANDLER-Loop system for dynamic in vitro hemocompatibility tests. The setup allows simultaneous testing of about 30 tube rings with an inner diameter of 2,4 mm. Cardiovascular implants can be placed into these tube rings. After filling them with anticoagulated blood and closing them to loops, they were placed on a rotating disc. While the loop is rotating, the blood remains in the lower part of the loop. The relative movement between the foreign surface and the blood sample induces a blood flow without the need of mechanical pumping. A first test series with the new test setup had been conducted to achieve a highly hemocompatible tube material. The results were compared to hemocompatibility tests done by LEMM, mentioned in the ISO 10993-42,3. We could prove, that Tygon S50HL® and Polyurethane achieve similar hemocompatibility values. In addition, we did in vitro tests of bare metal stents. We recorded a difference in the index of thrombocytes of about 17 % between the loops with three stents and the control. During further experiments the difference decreases to 8 %, which might be due to adhesion processes. The ability to measure these processes demonstrates the high sensitivity and very low background activation of the test setup. This can be assured by the recorded index of hemolysis, which had been less than 0,8 % in all our experiments. By developing and improving our modified CHANDLER-Loop system, we were able to establish a dynamic in vitro test setup for the hemocompatibility testing of small vascular implants. Because of its extremely low background activation and a high variability, the system serves as a benchmark for upcoming test setups.SFB-TR37 – Q2SFB-TR37 – B