Lung protection in cardio-pulmonary bypass.

Since the invention of the heart-lung machine paediatric cardiac surgery developed rapidly. For correction of complex cardiac malformations the application of a cardio-pulmonary bypass (CPB) has become indispensable but possible negative effects of this technique should not be neglected. Especially, both bypassed organs i.e. heart and lung are not perfused during the procedure and therefore are threatened by ischemia and reperfusion injury. Additionally, CPB was developed with a non-pulsatile flow but there are clinical observations that pulsatile flow might be superior with improved patient outcomes. Thus, the aim of our study was to evaluate the effect of CPB on lung structure and to assess whether different flow modalities (pulsatile vs. non-pulsatile flow) or application of the antibiotic minocycline might be advantageous. Thirty five piglets of four weeks age were examined and divided into five experimental groups: control (no CPB) without or with minocycline, CPB (non-pulsatile flow) without or with minocycline and CPB with pulsatile flow. CPB was performed for 90 min followed by a 120 min reperfusion and recovery phase. Thereafter, adenosine triphosphate-content of lung biopsies and histology was carried out. We found that CPB was associated with a significant thickening of alveolar wall accompanied by an infiltration of neutrophil leucocytes. Moreover, markers for hypoxia, apoptosis, nitrosative stress, inflammation and DNA damage were significantly elevated after CPB. These cellular damages could be partially inhibited by minocycline or pulsatile flow. Both, minocycline and pulsatile flow attenuate lung damage after CPB.

In Vitro Expression of the Extracellular Matrix Components Aggrecan, Collagen Types I and II by Articular Cartilage-Derived Chondrocytes.

The extracellular matrix (ECM) of hyaline cartilage is perfectly suited to transmit articular pressure load to the subchondral bone. Pressure is transferred by a high amount of aggrecan-based proteoglycans and collagen type II fibres in particular. After any injury, the hyaline cartilage is replaced by fibrocartilage, which is low in proteoglycans and contains collagen type I predominantly. Until now, long-term results of therapeutic procedures including cell-based therapies like autologous chondrocyte transplantation (ACT) lead to a replacement tissue meeting the composition of fibrocartilage. Therefore, it is of particular interest to discover how and to what extent isolation and in vitro cultivation of chondrocytes affect the cells and their expression of ECM components. Hyaline cartilage-derived chondrocytes were cultivated in vitro and observed microscopically over a time period of 35 days. The expression of collagen type I, collagen type II and aggrecan was analysed using RT-qPCR and Western blot at several days of cultivation. Chondrocytes presented a longitudinal shape for the entire cultivation period. While expression of collagen type I prevailed within the first days, only prolonged cultivation led to an increase in collagen type II and aggrecan expression. The results indicate that chondrocyte isolation and in vitro cultivation lead to a dedifferentiation at least to the stage of chondroprogenitor cells.

Parapagus conjoined twin calf: a case study - focused on CT and cardiac abnormalities.

Congenital duplication anomalies occasionally occur in both humans and animals. Although various forms of classification of these conjoined twins exist, each case should be considered as an individual. In the case study presented a Holstein Frisian calf, born alive after 281 days of normal gravidity was investigated by computed tomography (CT) and subsequent dissection. The calf could be classified as a parapagus dicephalus tetrabrachius. It exhibited two heads each of them with a complete cervical spine leading to a complete thoracic and lumbar spine and separate tails. Looking at the point of fusion, the twin was conjoined in the ventrolateral thoracic part of the body. The calf had two thoraces with four forelimbs and two pelvic limbs on a single pelvis. As two heads and thoraces were present, beginning at the oral cavity, two intestinal systems were observable leading to a fusion point at the ascending part of the duodenum been continued caudally as a single system. Within each thorax, two lungs and a heart were present. However, only the heart in the left thorax was normal shape, exhibiting a strong myocardium and increased size. In contrast, the heart within the right thorax was considerably smaller, round shaped, and it appeared to be spongy and not fully developed during the foetal period. Commonly, classification of conjoined twins is only based on the appearance of the skeletal system. However, in the case presented, the point of fusion of the skeletal system did not allow conclusions regarding the intestinal or cardiovascular system.

Angiogenesis and collagen type IV expression in different endothelial cell culture systems.

In vitro angiogenesis assays constitute an important tool for studying the mechanisms of angiogenesis and for identification of pro- and anti-angiogenic substances. Therefore, endothelial cell and media systems used for in vitro angiogenesis assays are required to mimic the angiogenic process in vivo including endothelial capability to express collagen type IV as a component of the basement membrane. In this study, the expression of collagen type IV and its α chains (α1-6) was investigated in different endothelial cell culture systems in vitro qualitatively and quantitatively. These systems included four different batches of microvascular endothelial cells derived from the human skin, heart and lung, from which only two batches were found to be angiogenic and two batches were classified as non-angiogenic. Distribution of the transcripts of the α chains of collagen type IV was similar in all cell and media systems investigated. However, secretion and deposition of a stable extracellular network of collagen type IV could only be observed in the angiogenic cultures. In conclusion, the consecutive steps of the angiogenic cascade in vivo as well as in vitro depend on an increasing secretion and subsequent extracellular deposition of collagen type IV.

In vivo evaluation of the angiogenic effects of the multiblock copolymer PDC using the hen's egg chorioallantoic membrane test.

Multiblock copolymers with shape-memory capability attracted tremendous interest as promising candidate materials for smart, degradable implants. In the present study the hen's egg-chorioallantoic membrane test (HET-CAM test) was used to investigate the angiogenic properties of a thermoplastic, biodegradable multiblock copolymer PDC composed of poly(p-dioxanone) hard segments (PPDO) and crystallizable poly(ε-caprolactone) switching segments (PCL), whereby PPDO and PCL homopolymers were investigated as controls. According to our HET-CAM test data, only PDC induced significant microvessel attraction and formation in the contact area of the test specimen after 48 hours of incubation showing newly formed blood vessels along the outer edge of the material. In contrast, no newly formed blood vessels were observed around the PPDO or PCL specimen after the same incubation period. These in vivo results indicate that the multiblock copolymer PDC possibly possesses an angiogenic effect and it can induce blood vessel formation in its direct vicinity when it is implanted in vivo.

Active participation instead of passive behaviour opens up new vistas in education of veterinary anatomy and histology.

Teaching morphology, a fundamental part of medicine curricula is traditionally based on lectures and practical trainings. We introduced peer-assisted learning (PAL) and student expert teams to the courses to give the students the possibility to improve their free speech and self-confidence. We involved students in active preparation of online materials such as labelled e-slides and e-pics. We offered online digital microscopy (Zoomify) and dissection (CyberPrep) allowing repeating the learned material and studying veterinary morphology outside the dissection theatre. Over 60% of first and third semester students profited from being a peer or being taught by a peer and 50% said the expert teams were an excellent method to learn the topographic anatomy. Almost all students applied Zoomify and CyberPrep and 75% of them found the digital microscopy and dissection to be a helpful or very helpful learning tool. In face of reduced contact hours, these forms of education compensated in part the lost teaching time. We observed improvement of rhetoric and presentation skills and self-confidence. The approaches should therefore find their constant place in the veterinary medicine curricula.

New insights in vascular development: vasculogenesis and endothelial progenitor cells.

In the course of new blood vessel formation, two different processes--vasculogenesis and angiogenesis--have to be distinguished. The term vasculogenesis describes the de novo emergence of a vascular network by endothelial progenitors, whereas angiogenesis corresponds to the generation of vessels by sprouting from pre-existing capillaries. Until recently, it was thought that vasculogenesis is restricted to the prenatal period. During the last decade, one of the most fascinating innovations in the field of vascular biology was the discovery of endothelial progenitor cells and vasculogenesis in the adult. This review aims at introducing the concept of adult vasculogenesis and discusses the efforts to identify and characterize adult endothelial progenitors. The different sources of adult endothelial progenitors like haematopoietic stem cells, myeloid cells, multipotent progenitors of the bone marrow, side population cells and tissue-residing pluripotent stem cells are considered. Moreover, a survey of cellular and molecular control mechanisms of vasculogenesis is presented. Recent advances in research on endothelial progenitors exert a strong impact on many different disciplines and provide the knowledge for functional concepts in basic fields like anatomy, histology as well as embryology.