Cow blood - A superior storage option in forensics?

Given the use of modified blood products (e.g. leucocyte depleted erythrocyte concentrates in SAG-mannitol, dehydrated blood powder, defibrinated blood), drawing blood from conscious animals while minimizing their stress is a good option to obtain blood for bloodstain pattern analysis. Nevertheless, the blood must be well described since individual differences in quality can occur, and storage will influence blood components qualitatively and quantitatively. Cow has been discussed as a suitable source of blood supply, but current data lack hematological and full rheological perspectives. This project includes the respective parameters in combination with passive drip pattern experiments during refrigerated storage in multiple study arms. Cow blood displayed a constant increase in viscosity (at high shear rate: 1000s-1), reflecting the expected reduction in red blood cell (RBC) flexibility. RBCs shrank but remained intact with very few irregular shapes, therefore there was no evidence of hemolysis. Influence of storage on stain size in passive drip pattern experiments with different substrates was minimal. However in cows, it is not hemolysis but an early change in suspension properties that indicates storage lesion. Viscosity (at low shear rate: 1s-1) of some blood samples increased three-fold (peaking at day 14), transitioning sharply to near-Newtonian (almost shear-independent) behavior thereafter. The higher this increase in viscosity, the greater the increase in the number of satellite spatter on glass. In order to ensure high quality simulations in the future, comprehensive rheological analyses to detect gradual changes in blood pseudoplasticity should be implemented in the forensic discipline of bloodstain pattern analysis.

Systematic Comparison of Commercial Hydrogels Revealed That a Synergy of Laminin and Strain-Stiffening Promotes Directed Migration of Neural Cells.

Hydrogels have shown potential in replacing damaged nerve tissue, but the ideal hydrogel is yet to be found. In this study, various commercially available hydrogels were compared. Schwann cells, fibroblasts, and dorsal root ganglia neurons were seeded on the hydrogels, and their morphology, viability, proliferation, and migration were examined. Additionally, detailed analyses of the gels' rheological properties and topography were conducted. Our results demonstrate vast differences on cell elongation and directed migration on the hydrogels. Laminin was identified as the driver behind cell elongation and in combination with a porous, fibrous, and strain-stiffening matrix structure responsible for oriented cell motility. This study improves our understanding of cell-matrix interactions and thereby facilitates tailored fabrication of hydrogels in the future.

A Bayesian approach to blood rheological uncertainties in aortic hemodynamics.

Computational hemodynamics has received increasing attention recently. Patient-specific simulations require questionable model assumptions, for example, for geometry, boundary conditions, and material parameters. Consequently, the credibility of these simulations is much doubted, and rightly so. Yet, the matter may be addressed by a rigorous uncertainty quantification. In this contribution, we investigated the impact of blood rheological models on wall shear stress uncertainties in aortic hemodynamics obtained in numerical simulations. Based on shear-rheometric experiments, we compare the non-Newtonian Carreau model to a simple Newtonian model and a Reynolds number-equivalent Newtonian model. Bayesian Probability Theory treats uncertainties consistently and allows to include elusive assumptions such as the comparability of flow regimes. We overcome the prohibitively high computational cost for the simulation with a surrogate model, and account for the uncertainties of the surrogate model itself, too. We have two main findings: (1) The Newtonian models mostly underestimate the uncertainties as compared to the non-Newtonian model. (2) The wall shear stresses of specific persons cannot be distinguished due to largely overlapping uncertainty bands, implying that a more precise determination of person-specific blood rheological properties is necessary for person-specific simulations. While we refrain from a general recommendation for one rheological model, we have quantified the error of the uncertainty quantification associated with these modeling choices.

Chorion-derived extracellular matrix hydrogel and fibronectin surface coatings show similar beneficial effects on endothelialization of expanded polytetrafluorethylene vascular grafts.

The endothelium plays an important regulatory role for cardiovascular homeostasis. Rapid endothelialization of small diameter vascular grafts (SDVGs) is crucial to ensure long-term patency. Here, we assessed a human placental chorionic extracellular matrix hydrogel (hpcECM-gel) as coating material and compared it to human fibronectin in-vitro. hpcECM-gels were produced from placental chorion by decellularization and enzymatic digestion. Human umbilical vein endothelial cells (HUVECs) were seeded to non-, fibronectin- or hpcECM-gel-coated expanded polytetrafluorethylene (ePTFE) SDVGs. Coating efficiency as well as endothelial cell proliferation, migration and adhesion studies on grafts were performed. hpcECM-gel depicted high collagen and glycosaminoglycan content and neglectable DNA amounts. Laminin and fibronectin were both retained in the hpcECM-gel after the decellularization process. HUVEC as well as endothelial progenitor cell attachment were both significantly enhanced on hpcECM-gel coated grafts. HUVECs seeded to hpcECM-gel depicted significantly higher platelet endothelial cell adhesion molecule-1 (PECAM-1) expression in the perinuclear region. Cell retention to flow was enhanced on fibronectin and hpcECM-gel coated grafts. Since hpcECM-gel induced a significantly higher endothelial cell adhesion to ePTFE than fibronectin, it represents a possible alternative for SDVG modification to improve endothelialization.

Capillary-size flow of human blood plasma: Revealing hidden elasticity and scale dependence.

The dynamical mechanical analysis of blood generally uses models inspired by conventional flows, assuming scale-independent homogeneous flows and without considering fluid-surface boundary interactions. The present experimental study highlights the relevance of using an approach in line with physiological reality providing a strong interaction between the fluid and the boundary interface. New dynamic properties of human blood plasma are found: a finite shear elastic response (solid-like property) is identified in nearly static conditions, which also depends on the scale (being reinforced at small scales). The elastic behavior is confirmed by the induction, without heat transfer, of local hot and cold thermodynamic states evidencing a thermo-mechanical coupling in blood plasma so far known only in elastic materials. This finding opens new routes for medical diagnosis and device fabrication.

Red Blood Cell Stiffness and Adhesion Are Species-Specific Properties Strongly Affected by Temperature and Medium Changes in Single Cell Force Spectroscopy.

Besides human red blood cells (RBC), a standard model used in AFM-single cell force spectroscopy (SCFS), little is known about apparent Young's modulus (Ea) or adhesion of animal RBCs displaying distinct cellular features. To close this knowledge gap, we probed chicken, horse, camel, and human fetal RBCs and compared data with human adults serving as a repository for future studies. Additionally, we assessed how measurements are affected under physiological conditions (species-specific temperature in autologous plasma vs. 25 °C in aqueous NaCl solution). In all RBC types, Ea decreased with increasing temperature irrespective of the suspension medium. In mammalian RBCs, adhesion increased with elevated temperatures and scaled with reported membrane sialic acid concentrations. In chicken only adhesion decreased with higher temperature, which we attribute to the lower AE-1 concentration allowing more membrane undulations. Ea decreased further in plasma at every test temperature, and adhesion was completely abolished, pointing to functional cell enlargement by adsorption of plasma components. This halo elevated RBC size by several hundreds of nanometers, blunted the thermal input, and will affect the coupling of RBCs with the flowing plasma. The study evidences the presence of a RBC surface layer and discusses the tremendous effects when RBCs are probed at physiological conditions.

Animal blood in translational research: How to adjust animal blood viscosity to the human standard.

Animal blood is used in mock circulations or in forensic bloodstain pattern analysis. Blood viscosity is important in these settings as it determines the driving pressure through biomedical devices and the shape of the bloodstain. However, animal blood can never exactly mimic human blood due to erythrocyte properties differing among species. This results in the species-specific shear thinning behavior of blood suspensions, and it is therefore not enough to adjust the hematocrit of an animal blood sample to mimic the behavior of human blood over the entire range of shear rates that are present in the body. In order to optimize experiments that require animal blood, we need models to adapt the blood samples. We here offer mathematical models derived for each species using a multi linear regression approach to describe the influence of shear rate, hematocrit, and temperature on blood viscosity. Results show that pig blood cannot be recommended for experiments at low flow conditions (<200 s-1 ) even though erythrocyte properties are similar in pigs and humans. However, pig blood mimics human blood excellently at high flow condition. Horse blood is unsuitable as experimental model in this regard. For several studied conditions, sheep blood was the closest match to human blood viscosity among the tested species.

Gingipain R1 and Lipopolysaccharide From Porphyromonas gingivalis Have Major Effects on Blood Clot Morphology and Mechanics.

Background:Porphyromonas gingivalis and its inflammagens are associated with a number of systemic diseases, such as cardiovascular disease and type 2 diabetes (T2DM). The proteases, gingipains, have also recently been identified in the brains of Alzheimer's disease patients and in the blood of Parkinson's disease patients. Bacterial inflammagens, including lipopolysaccharides (LPSs) and various proteases in circulation, may drive systemic inflammation. Methods: Here, we investigate the effects of the bacterial products LPS from Escherichia coli and Porphyromonas gingivalis, and also the P. gingivalis gingipain [recombinant P. gingivalis gingipain R1 (RgpA)], on clot architecture and clot formation in whole blood and plasma from healthy individuals, as well as in purified fibrinogen models. Structural analysis of clots was performed using confocal microscopy, scanning electron microscopy, and AFM-Raman imaging. We use thromboelastography® (TEG®) and rheometry to compare the static and dynamic mechanical properties of clots. Results: We found that these inflammagens may interact with fibrin(ogen) and this interaction causes anomalous blood clotting. Conclusions: These techniques, in combination, provide insight into the effects of these bacterial products on cardiovascular health, and particularly clot structure and mechanics.

Blood Clot Phenotyping by Rheometry: Platelets and Fibrinogen Chemistry Affect Stress-Softening and -Stiffening at Large Oscillation Amplitude.

(1) Background: Together with treatment protocols, viscoelastic tests are widely used for patient care. Measuring at broader ranges of deformation than currently done will add information on a clot's mechanical phenotype because fibrin networks follow different stretching regimes, and blood flow compels clots into a dynamic non-linear response. (2) Methods: To characterize the influence of platelets on the network level, a stress amplitude sweep test (LAOStress) was applied to clots from native plasma with five platelet concentrations. Five species were used to validate the protocol (human, cow, pig, rat, horse). By Lissajous plots the oscillation cycle for each stress level was analyzed. (3) Results: Cyclic stress loading generates a characteristic strain response that scales with the platelet quantity at low stress, and that is independent from the platelet count at high shear stress. This general behavior is valid in the animal models except cow. Here, the specific fibrinogen chemistry induces a stiffer network and a variant high stress response. (4) Conclusions: The protocol provides several thresholds to connect the softening and stiffening behavior of clots with the applied shear stress. This points to the reversible part of deformation, and thus opens a new route to describe a blood clot's phenotype.

Storability of porcine blood in forensics: How far should we go?

Previous studies on the storability of porcine blood for bloodstain pattern analysis (BPA) focused on abattoir blood only and did not include measurements of viscoelasticity. Although known to provoke echinocyte formation, EDTA is widely used for BPA issues. We compared ageing samples taken from live pigs with abattoir blood and detected considerable differences in hematocrit (HCT), total protein and shear viscosity that even worsened with time. Upon storage, high shear viscosity continuously increased, resulting in a partial loss of the typical shear thinning property of blood. Furthermore, we explored CPDA-1, the gold standard in preserving red blood cells (RBCs), for storage of forensic samples. We found it to be a superior choice for anticoagulation, as the rise of high shear viscosity was attenuated compared to EDTA. When performing oscillation measurements, we found a sudden change of viscoelasticity of blood after 22 days, providing a cut-off for storage time. To highlight the importance of hematological and hemorheological changes upon cold storage, we performed simple drip pattern experiments. These tests revealed a tendency to smaller stain diameters and higher numbers of satellite spatter. While this contradicts expectations from elevated viscosity values, we associate this trend to microscopic inhomogeneities due to storage. We recommend CPDA-1 for prolonged storage of BPA samples and suggest the use of comprehensive test protocols including viscoelasticity for determination of the maximum shelf life of pig blood.

Near-Newtonian Blood Behavior - Is It Good to Be a Camel?

From a certain level of exercise-intensity onward, hematocrit increases in horses, which brings more oxygen carriers into the bloodstream. Camels, however, when used in competitive racing could be even in need of iron supplementation and blood transfusions due to a severe reduction of their available hematocrit compared to their resting hematocrit. Since the extrinsic and intrinsic mechanical properties of camel erythrocytes (RBC) are so different compared to RBCs of other mammals, the question arises whether this observation might be a response to endurance exercise aiming at keeping the RBC count low. Rheometry indicated dromedary camel blood to behave almost Newtonian, which is unique amongst mammals. Shear thinning did increase with the hematocrit, but remained marginal compared to horses. As a result, camel whole blood viscosity (WBV) exceeded horse WBV at high shear rates, an effect, which was significantly augmented when the packed cell volume (PCV) was increased. Therefore, in camels any infusion of RBCs into the bloodstream can increase the cardiac work and the energy input into the endothelium more effectively, which should generate vascular remodeling in the long term. Yielding, however, was completely absent in camel blood, confirming low cohesion between its components at quasi-static flow. Camel blood remained a viscous liquid without a threshold even at unphysiologically high PCVs. This can help to washout lactate when camels start to dehydrate and might contribute to the sustained working ability of these animals. The subtle pseudoplastic behavior and the high viscosity contrast across the RBC membrane point to weak coupling between blood flow and red cell behavior. We predict that RBCs flow as separate entities and can show various types of motion, which can lead to friction instead of being collectively aligned to the flow direction. In comparison to horses, this behavior will become relevant at higher RBC counts in front of flow obstacles and possibly cause vascular remodeling if the PCV rises during strenuous exercise, a matter that should be avoided.

Altered membrane rigidity via enhanced endogenous cholesterol synthesis drives cancer cell resistance to destruxins.

Destruxins, secondary metabolites of entomopathogenic fungi, exert a wide variety of interesting characteristics ranging from antiviral to anticancer effects. Although their mode of action was evaluated previously, the molecular mechanisms of resistance development are unknown. Hence, we have established destruxin-resistant sublines of HCT116 colon carcinoma cells by selection with the most prevalent derivatives, destruxin (dtx)A, dtxB and dtxE. Various cell biological and molecular techniques were applied to elucidate the regulatory mechanisms underlying these acquired and highly stable destruxin resistance phenotypes. Interestingly, well-known chemoresistance-mediating ABC efflux transporters were not the major players. Instead, in dtxA- and dtxB-resistant cells a hyper-activated mevalonate pathway was uncovered resulting in increased de-novo cholesterol synthesis rates and elevated levels of lanosterol, cholesterol as well as several oxysterol metabolites. Accordingly, inhibition of the mevalonate pathway at two different steps, using either statins or zoledronic acid, significantly reduced acquired but also intrinsic destruxin resistance. Vice versa, cholesterol supplementation protected destruxin-sensitive cells against their cytotoxic activity. Additionally, an increased cell membrane adhesiveness of dtxA-resistant as compared to parental cells was detected by atomic force microscopy. This was paralleled by a dramatically reduced ionophoric capacity of dtxA in resistant cells when cultured in absence but not in presence of statins. Summarizing, our results suggest a reduced ionophoric activity of destruxins due to cholesterol-mediated plasma membrane re-organization as molecular mechanism underlying acquired destruxin resistance in human colon cancer cells. Whether this mechanism might be valid also in other cell types and organisms exposed to destruxins e.g. as bio-insecticides needs to be evaluated.

Viscoelasticity and structure of blood clots generated in-vitro by rheometry: A comparison between human, horse, rat, and camel.

BACKGROUND: Although the coagulation system is evolutionary well preserved, profound species differences exist in viscoelastic as well as in common laboratory tests of coagulation. OBJECTIVE: Evaluating differences in clot formation and material characterisation of clots of four mammalian species on macro-, micro- and nanoscales by the means of rheometry, scanning electron microscopy (SEM) and small angle x-ray scattering (SAXS). METHODS: Blood samples were collected from healthy human volunteers, laboratory rats (HL/LE inbred strain), warmblood horses and dromedary camels. Clot formation was observed by oscillating shear rheometry until plateau formation of the shear storage modulus G', at which point selected clots were prepared for scanning electron microscopy. SEM images were analysed for fibre diameter and fractal dimension. Additionally, scattering profiles for plasma and whole blood samples were obtained with SAXS. RESULTS: Viscoelasticity of clots showed great interspecies variation: clots of rats and horses exhibited shorter clotting times and higher G' plateau values, when compared to human clots. Camel clots showed unique clotting characteristics with no G' plateau formation in the timeframe observed. Less differentiating features were found with SEM and SAXS, although the rat fibre network appears to be more convoluted and dense, which resulted in a higher fractal dimension. CONCLUSION: Clotting kinetic differs between the species, which is not only of clinical interest, but could also be an important finding for animal models of blood coagulation.

Temperature dependency of whole blood viscosity and red cell properties in desert ungulates: Studies on scimitar-horned oryx and dromedary camel.

Background: The dromedary camel and the oryx antelope are exposed to excessive heat and solar radiation in their desert habitat. Desertification of areas with by now little rainfall may occur eventually. Well-adapted large animal species show us what is needed to survive in scorching regions. Methods: Four scimitar-horned oryx antelopes (Oryx dammah), 10 camels (Camelus dromedarius), nine South African Merino sheep, and 17 Nguni cows were tested for RBC aggregation, RBC elongation, and plasma viscosity. The temperature dependency of blood viscosity was tested in 10 camels and compared to human reference values. Results: Unlike sheep, Nguni cow, and dromedary camel, oryx RBCs aggregate in native plasma (M0:5.2 (3.3/6.7); M1:18.1 (16.7/27.9); Myrenne MA1). Elongation indices of oryx RBCs were intermediate to low (EImax: 22.6 (19.2/25.3); SS1/2 3.67 (2.52/4.95); Rheodyn SSD). Camel RBCs did not display the typical SS/EI curve by rotational ektacytometry. In-vitro blood viscosity (Physica MCR302) was lower in camels than in human blood at equal hematocrit. A decrease of temperature had only little effect on camel blood. At 10 s−1, blood viscosity in camel increased from 2.18mPa*s (2.01/2.37) at 42◦C to 4.39mPa*s (4.22/4.51) at 12◦C. In human blood, viscosity ranged from 8.21mPa*s (6.95/8.25) at 37◦C to 15.52mPa*s (14.25/16.03) at 12◦C. At 1000 s−1, blood viscosity in camel ranged from 2.00mPa*s (1.95/2.04) at 42◦C to 3.98mPa*s (3.88/4.08) at 12◦C. In human blood, viscosity ranged from 5.35mPa*s (4.96/5.87) at 37◦C to 11.24mPa*s (10.06/11.17) at 12◦C. Conclusions: Desert ungulates may need RBC membranes, which are fortified to withstand changes in osmolality during dehydration-rehydration cycles. This reduces RBC deformability. Dromedary camel blood does not undergo stark changes in viscosity with changes in temperature. Therefore, blood fluidity could be rather maintained during the day and night cycle. This should reduce the need of the vascularity to rhythmically adapt to changing shear forces when camels experience heterothermy.

Very large and giant microsurgical bifurcation aneurysms in rabbits: Proof of feasibility and comparability using computational fluid dynamics and biomechanical testing.

BACKGROUND: Giant aneurysms are challenging lesions with unacceptable high rates of aneurysm recanalization and rerupture following embolization. Reliable in vivo models are urgently needed to test the performance of new more efficient endovascular devices. MATERIALS AND METHODS: Aneurysms were created in 11 New Zealand white rabbits (4.5-5.5kg): A long venous pouch (length 25-30mm) mimicking the aneurysm sac was derived from the external jugular vein and sutured into a microsurgically created bifurcation between both common carotid arteries. After 4 weeks the rabbits underwent 3T Magnetic resonance angiography (3T-MRA). Exemplary computational fluid dynamics (CFD) simulations were performed to compare the flow conditions of giant rabbit and human aneurysms. We used species-related boundary conditions, in particular, we measured blood viscosity values. Biaxial mechanical tests were performed for the mechanical characterization and comparison. COMPARISON WITH EXISITING METHOD(S): None. RESULTS: No peri- or postoperative mortality was observed. 3T-MRA showed aneurysm patency in 10 out of 11 aneurysms (90.9%). Aneurysm lengths ranged from 21.5-25.6mm and aneurysm necks from 7.3-9.8mm. CFD showed complex flow profiles with multiple vortices in both, rabbit and human aneurysms. Lower blood viscosity values of the rabbit (3.92mPas vs. human 5.34mPas) resulted in considerable lower wall shear stress rates (rabbit 0.38Pa vs. human 1.66Pa). Mechanical tests showed lower stiffness of rabbit aneurysms compared to unruptured human aneurysms. CONCLUSIONS: The proposed model showed favorable aneurysm patency rates, low morbidity and good hemodynamic comparability with complex flow patterns. Biomechanical testing suggests that experimental aneurysms might be even more fragile compared to human aneurysms.

Evaluation of canine red blood cell quality after processing with an automated cell salvage device.

OBJECTIVE: To evaluate the properties of RBC concentrate harvested after processing fresh whole blood units from healthy dogs with an automated cell salvage device. DESIGN: Prospective, in vitro, experimental study. SETTING: University teaching hospital. ANIMALS: Sixteen healthy, privately owned dogs of various breeds. INTERVENTIONS: Fresh canine whole blood collected in bags with citrate phosphate dextrose adenine solution was processed with an automated cell salvage device and analyzed in vitro. Laboratory values determined before (baseline, from a catheter sample) and after processing RBCs (procRBCs) included a complete blood count, selected blood chemistry analytes, erythrocyte osmotic resistance, whole blood viscosity, RBC aggregation, and RBC deformability. MEASUREMENTS AND MAIN RESULTS: Total recovery of RBCs was 80% ± 12%. Hematocrit of the procRBCs yielded by the device was 77% ± 3.7% (mean ± standard deviation). Gross morphology of the RBCs remained unchanged. The mean corpuscular volume, erythrocyte osmotic resistance, RBC deformability, RBC aggregation, and the activity of lactate dehydrogenase showed minor but statistically significant changes from baseline. No differences in the concentrations of free hemoglobin were observed. Whole blood viscosity was less in the procRBCs. Seventy-seven percent (mean) of the platelets were washed out, while a mean of 57% of the leukocytes remained in the procRBCs. CONCLUSIONS: Although processing canine blood with this automated cell salvage device leads to slight changes in some properties of RBCs, most of these changes are comparable to changes seen in human blood after processing. Present data indicate that the use of this cell salvage device does not induce changes in canine RBC concentrate that would preclude its use for transfusion.

Hemorheology in experimental research: is it necessary to consider blood fluidity differences in the laboratory rat?

This study was designed to identify whether blood fluidity differs between commercially available laboratory rat strains. The hemorheological profiles of seven clinically healthy wild-type rat strains were analyzed to determine whether any diversity in blood fluidity might affect the outcome of cardiovascular studies. Study 1: 65 healthy adult rats (Lewis, Long-Evans, Hairless, Wistar and Fisher; mixed gender and comparable ages) were compared. In order to determine the greatest possible difference, the two strains with the greatest hematocrit (HCT) differences were selected for more detailed evaluation. Red blood cell (RBC) deformability (maximum elongation index, shear stress for half-maximal deformation of RBC; both P < 0.0001), and the effect of plasma protein concentration upon plasma viscosity (P < 0.0001) were different between Lewis and Long-Evans strains. Whole blood viscosity - although different at native HCT (P < 0.004) - was unaltered following HCT standardization of samples. Differences in RBC aggregation were statistically significant but these were small and may not be clinically relevant. Study 2: these 65 animals were compared with 21 animals (10-16 weeks old; both sexes) from mutant strains (Dahl SS/JrHsdMcwiCrl, n = 10; ZDF-Lepr(fa)/Crl, n = 11). In both mutant strains, plasma and whole blood viscosity were increased compared with commonly used strains at native and standardized HCT (P < 0.001). Unusually high RBC aggregation values were seen in the ZDF rat strain (P < 0.001). It was concluded that the variability in blood fluidity among clinically healthy adult laboratory rat strains was both statistically and clinically significant. A hemorheological profile should be added to a routine phenotyping process, since both variables can significantly influence study outcomes.

Anticancer activity of methyl-substituted oxaliplatin analogs.

Oxaliplatin is successfully used in systemic cancer therapy. However, resistance development and severe adverse effects are limiting factors for curative cancer treatment with oxaliplatin. The purpose of this study was to comparatively investigate in vitro and in vivo anticancer properties as well as the adverse effects of two methyl-substituted enantiomerically pure oxaliplatin analogs [[(1R,2R,4R)-4-methyl-1,2-cyclohexanediamine] oxalatoplatinum(II) (KP1537), and [(1R,2R,4S)-4-methyl-1,2-cyclohexanediamine]oxalatoplatinum(II) (KP1691)] and to evaluate the impact of stereoisomerism. Although the novel oxaliplatin analogs demonstrated in multiple aspects activities comparable with those of the parental compound, several key differences were discovered. The analogs were characterized by reduced vulnerability to resistance mechanisms such as p53 mutations, reduced dependence on immunogenic cell death induction, and distinctly attenuated adverse effects including weight loss and cold hyperalgesia. Stereoisomerism of the substituted methyl group had a complex and in some aspects even contradictory impact on drug accumulation and anticancer activity both in vitro and in vivo. To summarize, methyl-substituted oxaliplatin analogs harbor improved therapeutic characteristics including significantly reduced adverse effects. Hence, they might be promising metal-based anticancer drug candidates for further (pre)clinical evaluation.