Mauro Vaccarezza

Teaching Anatomy in the XXI Century: New Aspects and Pitfalls

Anatomy has historically been a cornerstone in medical education regardless of nation, racial background, or medical school system. By learning gross anatomy, medical students get a first “impression” about the structure of the human body which is the basis for understanding pathologic and clinical problems. Although the importance of teaching anatomy to both undergraduate and postgraduate students remains undisputed, there is currently a relevant debate concerning methods of anatomy teaching. In the past century, dissection and lectures were its sole pedagogy worldwide. Recently, the time allocated for anatomy teaching was dramatically reduced to such an extent that some suggest that it has fallen below an adequate standard. Traditional anatomy education based on topographical structural anatomy taught in lectures and gross dissection classes has been replaced by a multiple range of study modules, including problem-based learning, plastic models or computer-assisted learning, and curricula integration. “Does the anatomical theatre still have a place in medical education?” And “what is the problem with anatomic specimens?” We endeavor to answer both of these questions and to contribute to the debate on the current situation in undergraduate and graduate anatomy education. Doctors without anatomy are like moles.They work in the dark and the work of their hands are mounds. Friedrich Tiedemann The foundation of the study of the art of operating must be laid in the dissecting room. Robert Liston

3D printing: a valuable resource in human anatomy education

Anatomy is essential to the health and medical professions: by learning anatomy, medical students learn about the structure of the human body, providing them with the basic tools needed for understanding pathology and clinical problems. In the past century, dissection and lectures formed the basis of anatomy education worldwide. More recently, traditional anatomy education based on topographical structural anatomy taught in lectures and in gross dissection classes, has been replaced by a multiple range of study modules, including problem-based learning, plastic models and/or computer-assisted learning and curricula integration (Louw et al. 2009). The anatomy field is strongly confident that donated bodies can still benefit new medical students significantly, and that dissection and pro-section procedures cannot be underestimated in a modern medical curriculum (Louw et al. 2009). Nevertheless, dissection and light microscopy are not problem-free. Storing human bodies is expensive, and other issues such as preservation and reduced suitability for dissection due to illness, age or obesity could be a problem; moreover, careful dissection is time-consuming and microscopy equipment can be expensive. Aside from biological and methodological matters, dissection and prosection have also issues concerning ethical convictions and legal restrictions or simply logistical problems due to lack of space, funds, recruitment, or proper furniture and equipment. Considerable variations in the legal and ethical frameworks concerning body bequests for anatomical examination exist worldwide based on cultural and religious variations as well as different legal and constitutional backgrounds. For instance, there are different views concerning the ‘‘ownership’’ of cadavers or the acceptability of using unclaimed bodies that have not given informed consent (McHanwell et al. 2008). In addition to known methods such as plastination and Thiel method embalming, a new three-dimensional printing system (3D printing) has been developed recently—an innovative approach that could become a valuable resource in anatomy education. 3D printing (also known as additive manufacturing or rapid prototyping) has existed since the late 1980s but has seen rapid advancements more recently because of decreased cost, computer engineering, and expanding applications. Rapid prototyping involves creating a physical 3D model from a computerised mould. The technology has been used in industrial processes to create forerunners of intended final products; models can be also analysed and modified before production is planned (Gibson et al. 2010). Basically, the principle of rapid prototyping is to use 3D computer models for the reconstruction of a 3D physical model by the addition of material layers (Gibson et al. 2010). With additive fabrication, the machine reads in data from a CAD drawing and lays down successive layers of liquid, powder, or other sheet material, and in this way builds up the model from a series of cross sections. These M. Vaccarezza (&) V. Papa Department of Human, Social and Health Sciences, University of Cassino and Southern Lazio, Campus Folcara, via S. Angelo in Theodice, 03043 Cassino, FR, Italy e-mail: m.vaccarezza@uq.edu.au; m.vaccarezza@unicas.it

Bone Marrow-Derived Mesenchymal Cell Differentiation toward Myogenic Lineages: Facts and Perspectives

Bone marrow-derived mesenchymal stem cells (BM-MSCs) are valuable platforms for new therapies based on regenerative medicine. BM-MSCs era is coming of age since the potential of these cells is increasingly demonstrated. In fact, these cells give origin to osteoblasts, chondroblasts, and adipocyte precursors in vitro, and they can also differentiate versus other mesodermal cell types like skeletal muscle precursors and cardiomyocytes. In our short review, we focus on the more recent manipulations of BM-MSCs toward skeletal and heart muscle differentiation, a growing field of obvious relevance considering the toll of muscle disease (i.e., muscular dystrophies), the heavier toll of heart disease in developed countries, and the still not completely understood mechanisms of muscle differentiation and repair.

PKCε as a novel promoter of skeletal muscle differentiation and regeneration

INTRODUCTIONnSatellite cells are muscle resident stem cells and are responsible for muscle regeneration. In this study we investigate the involvement of PKCε during muscle stem cell differentiation in vitro and in vivo. Here, we describe the identification of a previously unrecognized role for the PKCε-HMGA1 signaling axis in myoblast differentiation and regeneration processes.nnnMETHODSnPKCε expression was modulated in the C2C12 cell line and primary murine satellite cells in vitro, as well as in an in vivo model of muscle regeneration. Immunohistochemistry and immunofluorescence, RT-PCR and shRNA silencing techniques were used to determine the role of PKCε and HMGA1 in myogenic differentiation.nnnRESULTSnPKCε expression increases and subsequently re-localizes to the nucleus during skeletal muscle cell differentiation. In the nucleus, PKCε blocks Hmga1 expression to promote Myogenin and Mrf4 accumulation and myoblast formation. Following in vivo muscle injury, PKCε accumulates in regenerating, centrally-nucleated myofibers. Pharmacological inhibition of PKCε impairs the expression of two crucial markers of muscle differentiation, namely MyoD and Myogenin, during injury induced muscle regeneration.nnnCONCLUSIONnThis work identifies the PKCε-HMGA1 signaling axis as a positive regulator of skeletal muscle differentiation.

Physical exercise as chemosensitizer

The benefits of physical exercise for the maintenance of human health, for the prevention and cure of metabolic syndrome and diabetes as well as for the prevention of cardiovascular diseases are well known [1]. Recently, a provocative study demonstrates an enhanced tumour perfusion and diminished tumour hypoxia in a model of orthotopic prostate cancer in rats exposed to treadmill exercise [2]. Of note, the measurements were performed on the conscious animal, which avoids the effects of anaesthetics on central haemodynamics and tumour blood flow, and provide an accurate evaluation of the resting versus exercising condition [2]. The reported blood flow increase of 200 % and the 50 % reduction in tumour hypoxia during exercise are unique findings [2] that are of particular interest to effectively enhance compound delivery to tumours. Intriguingly, a proposed mechanism is the ‘‘normalization’’ of the endothelial layer that is defective in tumours, and it represents a formidable barrier to efficient drug delivery [3, 4]. It would be interesting to assess the relative contribution of several types of exercise to these changes in endothelial permeability as well potential molecular mechanisms, keeping in mind a potential translation to these findings to clinical oncology. Of note, this provocative data are of interest even for blood cancers, considering the established role of bone marrow angiogenesis in the outcome of solid cancers and haematologic malignancies [3–5]. More experimental proof is definitely needed in animal models and in humans to support these results. Nevertheless, this is a possible inexpensive and valuable way to enhance drug potency [3, 4], with obvious translational effects in a clinical setting and possibly in cancer survival. Physical exercise science is coming of age not only in rehabilitation clinical haematology/oncology, but also as a potential therapeutic tool.

PKCε expression is required during proplatelet formation in murine model

Megakaryocytes (MK) remodel their cytoplasm into long proplatelet extensions to generate platelets [1]. We have previously demonstrated that PKCepsilon expression is strictly regulated during megakaryocytopoiesis (MKpoiesis), and its forced expression in the late phases of MK differentiation impairs platelet production [2,3]. However, our preliminary data suggest that PKCepsilon positive platelets may be released around the acute event of myocardial infarction, affecting their aggregation potential. Primary fetal liver (FL) cells isolated from CD1 pregnant mice are the preferential model to study the platelet formation mechanism [4]. Therefore, here we focused on the mouse PKCepsilon positive model to elucidate the role of PKCepsilon in MK maturation. Our data show that not only PKCepsilon expression increases during mouse MK differentiation, but also its forced down-regulation strongly reduces pro-platelet formation. Therefore, PKCepsilon is strongly required for murine proplatelet production. On the basis of these results and other known model systems, we show that PKCepsilon has a relevant role in the completion of platelet release.

Opposite effects of VWF during megakaryocytic vs erythroid differentiation

Hemopoiesis is formed by stem cells (SC), progenitor cells and short-lived precursors, that continuously interact with the extracellular matrix and the cells in the bone marrow microenvironment. The complexity of hematopoiesis consists in the requirement of a highly regulated progression through different steps of proliferation and maturation, that span from the self-renewal of SC, their commitment, the arrest of proliferation and terminal differentiation of mature, functional elements with a defined lifespan. The downstream progeny of Human SC (HSC) have been characterized, and lineage restricted oligopotent progenitor cells for lymphoid (common lymphoid progenitor, CLP) and myeloid (common myeloid progenitor, CMP), granulocyte-monocyte progenitor (GMP) and megakaryocyte-erithrocyte progenitor (MEP) lineages have been identified. In normal hematopoiesis, cytokine and growth factors (i.e. erythropoietin – EPO – and thrombopoietin – TPO –) are essential for these various functions, and act by binding to their cell-surface receptors and triggering complex cascades of intracellular signaling. We recently found that VWF is able to increase platelet production during TPO-induced MK differentiation, acting as a cytokine. It was also able to restore normal platelet production in TPO-treated CD34 cells obtained from VWD2B patients. Here, we tested the role of VWF both in MK and erythroid differentiation. VWF boosted the TPO-induced differentiation of human CD34-derived MK, as expected. On the contrary, EPO-induced erythroid differentiation of CD34 cells was inhibited by addition of VWF to the culture medium. Our data demonstrate that VWF plays different roles during both erythropoiesis and megakaryocytopoiesis, making it attractive to speculate that VWF could be involved in the control of the MEP commitment. Further experiments are needed to elucidate the effects of VWF on the common precursor.