The Arteriovenous Loop: Engineering of Axially Vascularized Tissue

Background: Most of the current treatment options for large-scale tissue defects represent a serious burden for the patients, are often not satisfying, and can be associated with significant side effects. Although major achievements have already been made in the field of tissue engineering, the clinical translation in case of extensive tissue defects is only in its early stages. The main challenge and reason for the failure of most tissue engineering approaches is the missing vascularization within large-scale transplants. Summary: The arteriovenous (AV) loop model is an in vivo tissue engineering strategy for generating axially vascularized tissues using the own body as a bioreactor. A superficial artery and vein are anastomosed to create an AV loop. This AV loop is placed into an implantation chamber for prevascularization of the chamber inside, e.g., a scaffold, cells, and growth factors. Subsequently, the generated tissue can be transplanted with its vascular axis into the defect site and anastomosed to the local vasculature. Since the blood supply of the growing tissue is based on the AV loop, it will be immediately perfused with blood in the recipient site leading to optimal healing conditions even in the case of poorly vascularized defects. Using this tissue engineering approach, a multitude of different axially vascularized tissues could be generated, such as bone, skeletal or heart muscle, or lymphatic tissues. Upscaling from the small animal AV loop model into a preclinical large animal model could pave the way for the first successful attempt in clinical application. Key Messages: The AV loop model is a powerful tool for the generation of different axially vascularized replacement tissues. Due to minimal donor site morbidity and the possibility to generate patient-specific tissues variable in type and size, this in vivo tissue engineering approach can be considered as a promising alternative therapy to current treatment options of large-scale defects

Tumor Cells Develop Defined Cellular Phenotypes After 3D-Bioprinting in Different Bioinks

Malignant melanoma is often used as a model tumor for the establishment of novel therapies. It is known that two-dimensional (2D) culture methods are not sufficient to elucidate the various processes during cancer development and progression. Therefore, it is of major interest to establish defined biofabricated three-dimensional (3D) models, which help to decipher complex cellular interactions. To get an impression of their printability and subsequent behavior, we printed fluorescently labeled melanoma cell lines with Matrigel and two different types of commercially available bioinks, without or with modification (RGD (Arginine-Glycine-Aspartate)-sequence/laminin-mixture) for increased cell-matrix communication. In general, we demonstrated the printability of melanoma cells in all tested biomaterials and survival of the printed cells throughout 14 days of cultivation. Melanoma cell lines revealed specific differential behavior in the respective inks. Whereas in Matrigel, the cells were able to spread, proliferate and form dense networks throughout the construct, the cells showed no proliferation at all in alginate-based bioink. In gelatin methacrylate-based bioink, the cells proliferated in clusters. Surprisingly, the modifications of the bioinks with RGD or the laminin blend did not affect the analyzed cellular behavior. Our results underline the importance of precisely adapting extracellular matrices to individual requirements of specific 3D bioprinting applications

Influence of the autotaxin-lysophosphatidic acid axis on cellular function and cytokine expression in different breast cancer cell lines

Previous studies provide high evidence that autotaxin (ATX)-lysophosphatidic acid (LPA) signaling through LPA receptors (LPAR) plays an important role in breast cancer initiation, progression, and invasion. However, its specific role in different breast cancer cell lines remains to be fully elucidated to offer improvements in targeted therapies. Within this study, we analyzed in vitro the effect of LPA 18:1 and the LPAR1, LPAR3 (and LPAR2) inhibitor Ki16425 on cellular functions of different human breast cancer cell lines (MDA-MB-231, MDA-MB-468, MCF-7, BT-474, SKBR-3) and the human breast epithelial cell line MCF-10A, as well as Interleukin 8 (IL-8), Interleukin 6 (IL-6) and tumor necrosis factor (TNF)-alpha cytokine secretion after LPA-incubation. ATX-LPA signaling showed a dose-dependent stimulatory effect especially on cellular functions of triple-negative and luminal A breast cancer cell lines. Ki16425 inhibited the LPA-induced stimulation of triple-negative breast cancer and luminal A cell lines in variable intensity depending on the functional assay, indicating the interplay of different LPAR in those assays. IL-8, IL-6 and TNF-alpha secretion was induced by LPA in MDA-MB-468 cells. This study provides further evidence about the role of the ATX-LPA axis in different breast cancer cell lines and might contribute to identify subtypes suitable for a future targeted therapy of the ATX-LPA axis

Tissue Viability of Free Flaps after Extracorporeal Perfusion Using a Modified Hydroxyethyl Starch Solution

Background: In free flap surgery, tissue is stored under hypothermic ischemia. Extracorporeal perfusion (EP) has the potential to extend storage time and the tissue's perspective of survival. In the present study, the aim is to improve a recently established, simplified extracorporeal perfusion system. Methods: Porcine musculus rectus abdominis were stored under different conditions. One group was perfused continuously with a simplified one-way perfusion system for six hours, while the other received only a single flush but no further treatment. A modified hydroxyethyl starch solution was used as a perfusion and flushing solution. Vitality, functionality, and metabolic activity of both groups were analyzed. Results: Perfused muscles, in contrast to the ischemically stored ones, showed no loss of vitality and significantly less functionality loss, confirming the superiority of storage under continuous perfusion over ischemic storage. Furthermore, in comparison to a previous study, the results were improved even further by using a modified hydroxyethyl starch solution. Conclusion: The use of EP has major benefits compared to the clinical standard static storage at room temperature. Continuous perfusion not only maintains the oxygen and nutrient supply but also removes toxic metabolites formed due to inadequate storage conditions

Complex wall modeling for hemodynamic simulations of intracranial aneurysms based on histologic images

Purpose For the evaluation and rupture risk assessment of intracranial aneurysms, clinical, morphological and hemodynamic parameters are analyzed. The reliability of intracranial hemodynamic simulations strongly depends on the underlying models. Due to the missing information about the intracranial vessel wall, the patient-specific wall thickness is often neglected as well as the specific physiological and pathological properties of the vessel wall. Methods In this work, we present a model for structural simulations with patient-specific wall thickness including different tissue types based on postmortem histologic image data. Images of histologic 2D slices from intracranial aneurysms were manually segmented in nine tissue classes. After virtual inflation, they were combined into 3D models. This approach yields multiple 3D models of the inner and outer wall and different tissue parts as a prerequisite for subsequent simulations. Result We presented a pipeline to generate 3D models of aneurysms with respect to the different tissue textures occurring in the wall. First experiments show that including the variance of the tissue in the structural simulation affect the simulation result. Especially at the interfaces between neighboring tissue classes, the larger influence of stiffer components on the stability equilibrium became obvious. Conclusion The presented approach enables the creation of a geometric model with differentiated wall tissue. This information can be used for different applications, like hemodynamic simulations, to increase the modeling accuracy.Peer reviewe

Prefabricated flaps and neoangiogenesis initiated via venous grafts in arteriovenous loops

New developments in regenerative medicine are bound to revolutionize the way we approach loss of function and form in human organisms. Especially in the field of reconstructive plastic surgery new biotechnologies find their way from bench to bed. Biofabrication is an evolving field that aims to combine natural biologic processes with bioartificial constructs with the scope of reconstituting tissue without having to rely on autotransplantation. In this brief review we present the concepts of intrinsic vs. extrinsic neovascularization and we discuss the use of neovascularization in three dimensional matrices. In a clinical context matrix flaps for application in reconstructive surgery can be fabricated this way

The challenge of musculoskeletal tissue engineering – from cell cultures to large animal models

Engineering functional skeletal muscle tissue still remains a major challenge. So far clinically relevant sizes of functional skeletal muscle tissue could not be engineered yet. One of the obstacles to overcome is the development of a suitable scaffold for muscle tissue engineering in vivo, another is the lack of differentiation in expanded adult muscle precursor cells. Materials and different architectures of scaffolds which are used for engineering functional skeletal muscle are presented here as well as approaches to the differentiation challenge. Finally the translation from cell culture over small to large animal models for engineering axially vascularized musculoskeletal tissues will be described