Michael Edelmayer

Continuous Monitoring of Cardiac Rhythms in Left Ventricular Assist Device Patients

Monitoring of cardiac rhythms is of major importance in the treatment of heart failure patients with left ventricular assist devices (LVADs) implanted. A continuous surveillance of these rhythms could improve out-of-hospital care in these patients. The aim of this study was to investigate cardiac rhythms using available pump data only. Datasets (n = 141) obtained in the normal ward, in the intensive care unit, and during bicycle ergometry were analyzed in 11 recipients of a continuous flow LVAD (59.1 ± 9.7 years; male 82%). Tachograms and arrhythmic patterns derived from the pump flow waveform, and a simultaneously recorded ECG were compared, as well as heart rate variability parameters such as: the average heart beat duration (RR interval), the standard deviation of the beat duration (SDNN), the root-mean-square of the difference of successive beat durations (RMSSD), and the number of pairs of adjacent beat duration differing by >50 ms divided by the number of all beats (pNN50). A very good agreement of cardiac rhythm parameters from the pump flow compared with ECG was found. Tachycardia, atrial fibrillation, and extrasystoles could be accurately identified from the tachograms derived from the pump flow. Also, Bland-Altman analysis comparing pump flow with ECG indicated a very small difference in average RR interval of 0.3 ± 1.0 ms, in SSDN of 0.5 ± 2.7 ms, in RMSSD of 1.0 ± 5.6 ms, and in pNN50 of 0.3 ± 1.0%. Continuous monitoring of cardiac rhythms from available pump data is possible. It has the potential to reduce the out-of-hospital diagnostic burden and to permit a more efficient adjustment of the level of mechanical support.

L-mimosine and hypoxia can increase angiogenin production in dental pulp-derived cells

BackgroundAngiogenin is a key molecule in the healing process which has been successfully applied in the field of regenerative medicine. The role of angiogenin in dental pulp regeneration is unclear. Here we aimed to reveal the impact of the hypoxia mimetic agent L-mimosine (L-MIM) and hypoxia on angiogenin in the dental pulp.MethodsHuman dental pulp-derived cells (DPC) were cultured in monolayer and spheroid cultures and treated with L-MIM or hypoxia. In addition, tooth slice organ cultures were applied to mimic the pulp-dentin complex. We measured angiogenin mRNA and protein levels using qPCR and ELISA, respectively. Inhibitor studies with echinomycin were performed to reveal the role of hypoxia-inducible factor (HIF)-1 signaling.ResultsBoth, L-MIM and hypoxia increased the production of angiogenin at the protein level in monolayer cultures of DPC, while the increase at the mRNA level did not reach the level of significance. The increase of angiogenin in response to treatment with L-MIM or hypoxia was reduced by echinomycin. In spheroid cultures, L-MIM increased angiogenin at protein levels while the effect of hypoxia was not significant. Angiogenin was also expressed and released in tooth slice organ cultures under normoxic and hypoxic conditions and in the presence of L-MIM.ConclusionsL-MIM and hypoxia modulate production of angiogenin via HIF-1 differentially and the response depends on the culture model. Given the role of angiogenin in regeneration the here presented results are of high relevance for pre-conditioning approaches for cell therapy and tissue engineering in the field of regenerative endodontics.

Collagen barrier membranes do not adsorb hypoxia mimetic activity‐Activity of gingival fibroblasts cultured directly on collagen barrier membranes loaded with hypoxia mimetic agents

Hypoxia-based strategies for applications in oral surgery and periodontology have been proposed where collagen barrier membranes (CBM) are loaded with hypoxia mimetic agents (HMA) to induce a pro-angiogenic response. While it was found that CBM release HMA, it remained unclear if CBM adsorb HMA activity. Here we evaluated the response of oral cells cultured on CBM, supplemented with the HMA dimethyloxalylglycine (DMOG), desferrioxamine (DFO), and l-mimosine (l-MIM). Gingival fibroblasts (GF) were cultured on unwashed CBM as well as on CBM that had been washed with serum-free medium for 48 hours. The pro-angiogenic response was measured based on vascular endothelial growth factor (VEGF) production. Viability and proliferation were assessed based on MTT and BrdU assays. We found that GF seeded onto CBM loaded with DFO and l-MIM, but not DMOG, showed an increase in VEGF to 6.1-fold and 7.7-fold compared to unloaded CBM, respectively. Cells remained vital, but a trend for decreased proliferation was observed on DMOG and DFO-loaded CBM which did not reach the level of significance. Evaluation of washed CBM revealed no difference between the unloaded CBM and CBM supplemented with DMOG, DFO, or l-MIM. In conclusion, our results suggest that CBM do not adsorb hypoxia mimetic activity but release HMA within the first hours.

Hypoxia-based strategies for regenerative dentistry—Views from the different dental fields

The understanding of the cell biological processes underlying development and regeneration of oral tissues leads to novel regenerative approaches. Over the past years, knowledge on key roles of the hypoxia-based response has become more profound. Based on these findings, novel regenerative approaches for dentistry are emerging, which target cellular oxygen sensors. These approaches include hypoxia pre-conditioning and pharmacologically simulated hypoxia. The increase in studies on hypoxia and hypoxia-based strategies in regenerative dentistry highlights the growing attention to hypoxias role in regeneration and its underlying biology, as well as its application in a therapeutic setting. In this narrative review, we present the current knowledge on the role of hypoxia in oral tissues and review the proposed hypoxia-based approaches in different fields of dentistry, including endodontics, orthodontics, periodontics, and oral surgery.

Effect of prolyl hydroxylase inhibitor-loaded collagen barrier membranes on osteoclastogenesis and osteoblastogenesis

Prolyl hydroxylase inhibitors induce a proangiogenic response and are therefore proposed to optimize regenerative approaches in periodontics and oral surgery. Here the effect of the prolyl hydroxylase inhibitors dimethyloxalylglycine and deferoxamine, released from collagen barrier membranes, on osteoclastogenesis and osteoblastogenesis was evaluated. Collagen barrier membranes were loaded with dimethyloxalylglycine and deferoxamine. Release studies were performed and supernatants were taken after 1, 3, 6, 24, and 48 h. The effect of these supernatants on osteoblast- and osteoclast-precursor cells was evaluated. Furthermore, dose response studies for dimethyloxalylglycine and deferoxamine were performed. Osteoclastogenesis was evaluated with RAW 264.7 cells based on the number of multinuclear tartrate-resistant acid phosphatase positive cells. Osteoblastogenesis was evaluated with MC3T3-E1 cells based on alkaline phosphatase. Metabolic activity and cell proliferation were assessed based on MTT and BrdU assays. Vascular endothelial growth factor production was evaluated using an immunoassay. We found that supernatants taken in the first hour from collagen barrier membranes loaded with dimethyloxalylglycine or deferoxamine reduced osteoclastogenesis. Osteoblastogenesis was not reduced significantly. Cell proliferation and metabolic activity of RAW 264.7 and MC3T3-E1 cells were inhibited by supernatants of collagen barrier membranes loaded with deferoxamine but not dimethyloxalylglycine. In RAW 264.7 cell culture, vascular endothelial growth factor production was increased only by supernatants of collagen barrier membranes loaded with dimethyloxalylglycine, but not deferoxamine. In MC3T3-E1 cell culture, supernatants of collagen barrier membranes loaded with dimethyloxalylglycine and deferoxamine both increased vascular endothelial growth factor production. Direct measurements showed that the majority of dimethyloxalylglycine and deferoxamine is released in the first hours. Dose-response studies supported the divergent effects of deferoxamine and dimethyloxalylglycine in RAW 264.7 and MC3T3-E1 cultures. Our findings show diverse effects of dimethyloxalylglycine- and deferoxamine-loaded collagen barrier membranes during osteoclastogenesis and osteoblastogenesis. Preclinical studies will reveal if the increase in vascular endothelial growth factor together with the inhibitory effect on osteoclasts can stimulate oral tissue regeneration.

Evaluation of Resins for Stereolithographic 3D-Printed Surgical Guides: The Response of L929 Cells and Human Gingival Fibroblasts

Additive manufacturing is becoming increasingly important in dentistry for the production of surgical guides. The development of cost-effective desktop stereolithography (SLA) printing systems and the corresponding resins makes this novel technique accessible to dental offices and dental laboratories. The aim of the study was to reveal the response of soft tissue cells to Clear and Dental SG resins used in desktop SLA printing systems at different stages of processing. Cell activity of L929 cells and gingival fibroblasts (GF) in response to the materials was examined in indirect and direct monolayer culture models and a direct spheroid culture model based on MTT, resazurin-based toxicity assays, and live-dead staining. Overall we found that the impact of Clear and Dental SG resins on L929 and GF depends on the processing stage of the materials. Liquid Clear resin induced a stronger reduction of cell activity compared to Dental SG resin. Printing and postcuring reduced the impact on cell activity and viability. As in-house 3D printing for surgical guides is getting integrated in the digital workflow, our data suggest that careful adherence to processing guidelines—especially postcuring—is of clinical relevance.

Synthetic Clay–based Hypoxia Mimetic Hydrogel for Pulp Regeneration: The Impact on Cell Activity and Release Kinetics Based on Dental Pulp–derived Cells In Vitro

Introduction: Thixotropic synthetic clays have been successfully used for tissue engineering in regenerative medicine. The impact of these clays on the dental pulp, in particular in combination with hypoxia‐based approaches using hypoxia mimetic agents (HMAs), is unknown. Our aim was to reveal the response of dental pulp–derived cells (DPCs) to a synthetic clay–based hydrogel and evaluate the release of HMAs. Methods: Using resazurin‐based toxicity assays, live‐dead staining, and 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide staining, the viability of human DPCs seeded onto a synthetic clay–based hydrogel of 5%–0.15% as well as onto the hydrogels loaded with the HMAs dimethyloxalylglycine (DMOG), desferrioxamine, L‐mimosine, and CoCl2 was evaluated. Furthermore, supernatant of the hydrogels loaded with HMAs were generated. Vascular endothelial growth factor (VEGF) production of DPCs in response to the supernatant was measured to reveal the cellular response to the HMAs. Results: We found that the synthetic clay–based hydrogel did not impair the viability of DPCs. Cell monolayer and cell cluster formations were observed on the hydrogel. No significant increase of VEGF levels was observed in the supernatant when DPCs were cultured on hydrogels loaded with HMAs. Supernatant of DMOG‐loaded hydrogels stimulated VEGF production in DPCs in the first hour, whereas the effect of desferrioxamine, L‐mimosine, and CoCl2 did not reach a level of significance. Conclusions: The synthetic clay–based hydrogel represents a promising biomaterial that does not induce prominent toxic effects in DPCs. It can be loaded with DMOG to induce hypoxia mimetic activity. Overall, we provided first insights into the impact of synthetic clays on DPCs for tissue engineering purposes in regenerative endodontics.

Difference in release kinetics of unwashed and washed platelet‐released supernatants from bone substitute materials: the impact of platelet preparation modalities

BACKGROUND AND OBJECTIVE In regenerative dentistry, platelet preparations are applied to stimulate bone healing and periodontal regeneration. Here, we pursue a strategy where bone substitutes are used as carriers for platelet-released supernatants. The mitogenic capacity and release kinetics of loaded bone substitutes were assessed. MATERIAL AND METHODS Platelet-released supernatants of washed platelets (washed PRS) and platelet-released supernatants of unwashed platelets (unwashed PRS) were lyophilized onto the bone substitutes deproteinized bovine bone mineral, hydroxyapatite and β-tricalcium phosphate. Scanning electron microscopy images were taken. Supernatants of bone substitutes were collected at hours 1, 3, 6, 24, and 48 and medium was replaced. We evaluated the protein content with the bicinchoninic acid assay and the effect on proliferation using bioassays with human periodontal fibroblasts. Release of growth factors from the loaded bone substitutes was measured based on the platelet-derived growth factor isoform (PDGF-BB) and thrombin immunoassays. Furthermore, we assessed DNA and RNA content of washed PRS and unwashed PRS. RESULTS Unwashed PRS showed higher total protein concentrations than washed PRS, while the concentration of PDGF-BB, thrombin, DNA, RNA and their mitogenic effect was not significantly different. The bone substitute materials adsorbed protein over time but no significant changes in overall appearance was found. Supernatants collected from unwashed PRS-loaded bone substitute after 1 h induced a potent mitogenic response in periodontal fibroblasts. This pro-mitogenic capacity of the supernatants decreased over the observation period. Supernatants of washed PRS-loaded bone substitutes did not induce a substantial mitogenic effect. Levels of PDGF-BB, thrombin and protein were higher in supernatants of unwashed PRS-loaded bone substitutes than of washed PRS-loaded bone substitutes. CONCLUSION Bone substitutes loaded with unwashed PRS, but not bone substitutes loaded with washed PRS show continuously declining release kinetics. These data suggest that plasma components in platelet preparations can modify the release kinetics profile.

Release kinetics and mitogenic capacity of collagen barrier membranes supplemented with secretome of activated platelets - the in vitro response of fibroblasts of the periodontal ligament and the gingiva

BackgroundPlatelet preparations can stimulate the healing process and have mitogenic properties. We hypothesized that collagen barrier membranes (CBM), clinically used in guided bone regeneration and guided tissue regeneration, can serve as carriers for platelet secretome.MethodsSecretome was generated from washed platelets and unwashed platelets (washed/unwashed PSEC) and lyophilized onto CBM. Overall appearance of CBM was evaluated by scanning electron microscopy. The impact of PSEC on cell attachment was measured based on fluorescence microscopy with DiI-labeled cells. To assess the release kinetics, supernatants of CBM were collected and medium was replaced at hour 1–48. The mitogenic effect was evaluated with periodontal fibroblasts. Furthermore, the release of total protein, platelet-derived growth factor (PDGF)-BB, and transforming growth factor (TGF) β1 was measured.ResultsCBM overall appearance and cell attachment was not modulated by PSEC. Supernatants taken after one hour induced a mitogenic response in fibroblasts and showed the highest levels of total protein, TGFβ1 and PDGF-BB. These effects decreased rapidly in subsequent supernatants. While supernatants of CBM loaded with unwashed PSEC induced a stronger mitogenic response than supernatants of CBM loaded with washed PSEC this difference between the PSEC preparations was not observed when cells were seeded on 48–hours-washed CBM.ConclusionsCBM release platelet-derived factors in continuously declining release kinetics.