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Melt electrowriting (MEW) is an aspiring 3D printing technology with an unprecedented resolution among fiber-based printing technologies. It offers the ability to direct-write predefined designs utilizing a jet of molten polymer to fabricate constructs composed of fibers with diameters of only a few micrometers. These dimensions enable unique construct properties. Poly(ɛ-caprolactone) (PCL), a semicrystalline polymer mainly used for biomedical and life science applications, is the most prominent material for MEW and exhibits excellent printing properties. Despite the wealth of melt electrowritten constructs that have been fabricated by MEW, a detailed investigation, especially regarding fiber analysis on a macro- and microlevel is still lacking. Hence, this study systematically examines the influence of process parameters such as spinneret diameter, feeding pressure, and collector velocity on the diameter and particularly the topography of PCL fibers and sheds light on how these parameters affect the mechanical properties and crystallinity. A correlation between the mechanical properties, crystallite size, and roughness of the deposited fiber, depending on the collector velocity and applied feeding pressure, is revealed. These findings are used to print constructs composed of fibers with different microtopography without affecting the fiber diameter and thus the macroscopic assembly of the printed constructs., Competing Interests: No competing financial interests exist., (Copyright 2021, Mary Ann Liebert, Inc., publishers.)

Ligaments and tendons are comprised of aligned, crimped collagen fibrils that provide tissue-specific mechanical properties with non-linear extension behaviour, exhibiting low stress at initial strain (toe region behaviour). To approximate this behaviour, we report fibrous scaffolds with sinusoidal patterns by melt electrowriting (MEW) below the critical translation speed (CTS) by exploitation of the natural flow behaviour of the polymer melt. More specifically, we synthesised photopolymerizable poly(L-lactide-co-ε-caprolactone-co-acryloyl carbonate) (p(LLA-co-ε-CL-co-AC)) and poly(ε-caprolactone-co-acryloyl carbonate) (p(ε-CL-co-AC)) by ring-opening polymerization (ROP). Single fibre (fØ = 26.8 ± 1.9 µm) tensile testing revealed a customisable toe region with Young's Moduli ranging from E = 29 ± 17 MPa for the most crimped structures to E = 314 ± 157 MPa for straight fibres. This toe region extended to scaffolds containing multiple fibres, while the sinusoidal pattern could be influenced by printing speed. The synthesized polymers were cytocompatible and exhibited a tensile strength of σ = 26 ± 7 MPa after 10 4 cycles of preloading at 10% strain while retaining the distinct toe region commonly observed in native ligaments and tendon tissue., Statement of Significance: Damaged tendons and ligaments are serious and frequently occurring injuries worldwide. Recent therapies, including autologous grafts, still have severe disadvantages leading to a demand for synthetic alternatives. Materials envisioned to induce tendon and ligament regeneration should be degradable, cytocompatible and mimic the ultrastructural and mechanical properties of the native tissue. Specifically, we utilised photo-cross-linkable polymers for additive manufacturing (AM) with MEW. In this way, we were able to direct-write cytocompatible fibres of a few micrometres thickness into crimp-structured elastomer scaffolds that mimic the non-linear biomechanical behaviour of tendon and ligament tissue., (Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Reinforcing hydrogels with micro-fibre scaffolds obtained by a Melt-Electrospinning Writing (MEW) process has demonstrated great promise for developing tissue engineered (TE) constructs with mechanical properties compatible to native tissues. However, the mechanical performance and reinforcement mechanism of the micro-fibre reinforced hydrogels is not yet fully understood. In this study, FE models, implementing material properties measured experimentally, were used to explore the reinforcement mechanism of fibre-hydrogel composites. First, a continuum FE model based on idealized scaffold geometry was used to capture reinforcement effects related to the suppression of lateral gel expansion by the scaffold, while a second micro-FE model based on micro-CT images of the real construct geometry during compaction captured the effects of load transfer through the scaffold interconnections. Results demonstrate that the reinforcement mechanism at higher scaffold volume fractions was dominated by the load carrying-ability of the fibre scaffold interconnections, which was much higher than expected based on testing scaffolds alone because the hydrogel provides resistance against buckling of the scaffold. We propose that the theoretical understanding presented in this work will assist the design of more effective composite constructs with potential applications in a wide range of TE conditions.

The viscoelastic properties of blood clot have been studied most commonly using thrombelastography (TEG) and thromboelastometry (ROTEM). ROTEM-based bleeding treatment algorithms recommend administering platelets to patients with low EXTEM clot strength (e.g., clot amplitude at 10 minutes [A10] <40 mm) once clot strength of the ROTEM® fibrin-based test (FIBTEM) is corrected. Algorithms based on TEG typically use a low value of maximum amplitude (e.g., <50 mm) as a trigger for administering platelets. However, this parameter reflects the contributions of various blood components to the clot, including platelets and fibrin/fibrinogen. The platelet component of clot strength may provide a more sensitive indication of platelet deficiency than clot amplitude from a whole blood TEG or ROTEM® assay. The platelet component of the formed clot is derived from the results of TEG/ROTEM® tests performed with and without platelet inhibition. In this article, we review the basis for why this calculation should be based on clot elasticity (e.g., the E parameter with TEG and the CE parameter with ROTEM®) as opposed to clot amplitude (e.g., the A parameter with TEG or ROTEM®). This is because clot elasticity, unlike clot amplitude, reflects the force with which the blood clot resists rotation within the device, and the relationship between clot amplitude (variable X) and clot elasticity (variable Y) is nonlinear. A specific increment of X (ΔX) will be associated with different increments of Y (ΔY), depending on the initial value of X. When calculated correctly, using clot elasticity data, the platelet component of the clot can provide a valuable insight into platelet deficiency in emergency bleeding.

Background: Viscoelastic tests such as functional fibrinogen polymerization assays (FFPAs) in thrombelastography (TEG®) or thromboelastometry (ROTEM®) measure clot elasticity under platelet inhibition. Incomplete platelet inhibition influences maximum clot firmness (MCF) of FFPAs. We compared the ability of existing and newly developed FFPAs to eliminate the platelet contribution to clot strength., Methods: MCF of whole blood (WB), platelet-rich plasma (PRP), and platelet-poor plasma samples was recorded using a ROTEM device with different FFPAs, including the TEG functional fibrinogen test (FFTEG) and different ROTEM-based assays: the standard fib-tem reagent (FIBTEM), a lyophilized single-portion reagent fib-tem S (FIBTEM-S), a newly developed reagent FIBTEM PLUS, as well as FIBTEM or the standard extrinsic activation reagent ex-tem® (EXTEM) combined with 10-μg abciximab (FIBTEM-ABC/EXTEM-ABC)., Results: In WB (platelet count [mean ± SD], 183 ± 37 × 10/μL; plasma fibrinogen concentration, 2.49 ± 0.58 g/L), FFTEG and EXTEM-ABC showed higher MCF (15.7 ± 2.8 mm) than FIBTEM or FIBTEM-S (11.4 ± 3.3 mm, P < 0.001), whereas FIBTEM-ABC and FIBTEM PLUS resulted in lower MCF (9.3 ± 2.8 mm, P < 0.001). In 2 different PRP samples, with platelet counts of 407 ± 80 × 10/μL and 609 ± 127 × 10/μL, FIBTEM-ABC and FIBTEM PLUS reduced platelet contribution to clot strength within 95% confidence interval limits of -1.4 to 0.1 mm and -1.2 to 0.4 mm, respectively. Using all FFPAs it was observed that the Pearson correlation coefficient between plasma fibrinogen concentration and WB MCF was high (range, 0.75-0.93) and significant, regardless of the underlying platelet inhibiting component. Evaluating differences in the interception of regression lines by using analysis of covariance, we compared platelet-poor plasma and both PRP samples within the same assays and found that in contrast to the FIBTEM-ABC and FIBTEM PLUS assays, the FFTEG, EXTEM-ABC, FIBTEM, and FIBTEM-S methods still detected residual platelet activity and grossly overestimated fibrin clot strength in samples with high platelet counts., Conclusions: FFPAs based solely on glycoprotein-IIb/IIIa inhibition, such as FFTEG or EXTEM-ABC, are less effective than cytochalasin D-based assays, such as FIBTEM or FIBTEM-S, at inhibiting the platelet component of clot strength. The FIBTEM PLUS assay, and the combination of FIBTEM and abciximab, sufficiently inhibits platelet contribution to clot elasticity. The combination of a glycoprotein-IIb/IIIa receptor blocker and cytochalasin D allows evaluation of functional fibrinogen polymerization without platelet "noise." In a clinical setting, the significance of potent platelet inhibition ensures a more accurate assessment of MCF and therefore the need for fibrinogen supplementation therapy. Further studies are necessary to investigate the application and impact of these tests in a clinical situation.

Background: Viscoelastic tests such as functional fibrinogen polymerization assays (FFPAs) in thrombelastography (TEG(®)) or thromboelastometry (ROTEM(®)) measure the elasticity of extrinsically activated clotting under conditions of platelet inhibition. There are no reports on whether components of the FFPAs have any effects on fibrin polymerization, aside from the effects of platelet inhibition., Methods: Using various platelet-free plasma (PFP) preparations, we compared the extrinsically activated EXTEM thromboelastometric assay with 3 FFPAs: FIBTEM, FIBTEM PLUS, and the Functional Fibrinogen Test(®) (FFTEG). These FFPAs activate coagulation extrinsically but additionally inhibit platelet function. We used calibration plasma (Instrumentation Laboratory and Siemens), pooled fresh-frozen plasma (Octaplas) and freshly prepared PFP from a healthy volunteer. EXTEM and all FFPAs were run in parallel on a ROTEM device., Results: Median (interquartile range) maximum clot firmness (MCF) values for all plasma preparations were: 20.5 mm (17.25-22.0 mm) in EXTEM, 23.0 mm (18.5-24.0 mm) in FIBTEM, 23.0 mm (18.25-24.75 mm) in FIBTEM PLUS, and 18.0 mm (16.0-19.0 mm) in FFTEG. Compared with EXTEM, FIBTEM and FIBTEM PLUS (P < 0.01) showed increased MCF values whereas FFTEG (P < 0.001) showed decreased MCF values. Further experiments in PFP showed that the platelet inhibitors used in the FFPAs (cytochalasin D or the glycoprotein-IIb/IIIa inhibitor abciximab) were not causing these alterations in MCF. However, reducing the activating tissue factor concentration (by diluting the extrinsic assay) decreased the MCF., Conclusions: We speculate that FIBTEM and FIBTEM PLUS may contain stabilizing agents that enhance fibrin polymerization whereas FFTEG might contain less tissue factor than the ROTEM assays.

Background: Fibrin-based clot firmness is measured as maximum amplitude (MA) in the functional fibrinogen (FF) thrombelastographic assay and maximum clot firmness (MCF) in the FIBTEM thromboelastometric assay. Differences between the assays/devices may be clinically significant. Our objective was to compare clot firmness parameters through standard (FF on a thrombelastography device [TEG®]; FIBTEM on a thromboelastometry device [ROTEM®]) and crossover (FF on ROTEM®; FIBTEM on TEG®) analyses., Methods: Whole-blood samples from healthy volunteers were subjected to thrombelastography and thromboelastometry analyses. Samples were investigated native and following stepwise dilution with sodium chloride solution (20%, 40%, and 60% dilution). Samples were also assessed after in vitro addition of medications (heparin, protamine, tranexamic acid) and 50% dilution with hydroxyethyl starch, gelatin, sodium chloride, and albumin., Results: FF produced higher values than FIBTEM, regardless of the device, and TEG® produced higher values than ROTEM®, regardless of the assay. With all added medications except heparin 400 U/kg bodyweight, FF MA remained significantly higher (P < 0.05) than FIBTEM MCF, which was largely unchanged. FF MA was significantly reduced (P = 0.04) by high-dose heparin and partially restored with protamine. Fifty percent dilution with hydroxyethyl starch, albumin, and gelatin decreased FIBTEM MCF and FF MA by >50%., Conclusions: These results demonstrate differences when measuring fibrin-based clotting via the FF and FIBTEM assays on the TEG® and ROTEM® devices. Point-of-care targeted correction of fibrin-based clotting may be influenced by the assay and device used. For the FF assay, data are lacking.

Introduction: Thromboelastometry (TEM)-guided haemostatic therapy with fibrinogen concentrate and prothrombin complex concentrate (PCC) in trauma patients may reduce the need for transfusion of red blood cells (RBC) or platelet concentrate, compared with fresh frozen plasma (FFP)-based haemostatic therapy., Methods: This retrospective analysis compared patients from the Salzburg Trauma Centre (Salzburg, Austria) treated with fibrinogen concentrate and/or PCC, but no FFP (fibrinogen-PCC group, n = 80), and patients from the TraumaRegister DGU receiving ≥ 2 units of FFP, but no fibrinogen concentrate/PCC (FFP group, n = 601). Inclusion criteria were: age 18-70 years, base deficit at admission ≥ 2 mmol/L, injury severity score (ISS) ≥ 16, abbreviated injury scale for thorax and/or abdomen and/or extremity ≥ 3, and for head/neck < 5., Results: For haemostatic therapy in the emergency room and during surgery, the FFP group (ISS 35.5 ± 10.5) received a median of 6 units of FFP (range: 2, 51), while the fibrinogen-PCC group (ISS 35.2 ± 12.5) received medians of 6 g of fibrinogen concentrate (range: 0, 15) and 1200 U of PCC (range: 0, 6600). RBC transfusion was avoided in 29% of patients in the fibrinogen-PCC group compared with only 3% in the FFP group (P< 0.001). Transfusion of platelet concentrate was avoided in 91% of patients in the fibrinogen-PCC group, compared with 56% in the FFP group (P< 0.001). Mortality was comparable between groups: 7.5% in the fibrinogen-PCC group and 10.0% in the FFP group (P = 0.69)., Conclusions: TEM-guided haemostatic therapy with fibrinogen concentrate and PCC reduced the exposure of trauma patients to allogeneic blood products.