Vesicular Messages from Dental Biofilms for Neutrophils.

The encounter between dental biofilm and neutrophils in periodontitis remains elusive, although it apparently plays a crucial role in the periodontal pathology and constitutes a key topic of periodontology. Dental biofilm and neutrophils were isolated from orally healthy persons and patients with periodontitis. We investigated biofilm and its particle-shedding phenomenon with electron microscopy and nanoparticle tracking analysis (NTA); biofilm shedding-neutrophil interactions were examined ex vivo with epi-fluorescence microscopy. For this purpose, we used acellular dental biofilm shedding, purified lipopolysaccharide (LPS), and phorbol 12-myristate 13-acetate (PMA) as activators, and the interleukin 8 receptor beta (CXCR2) inhibitor and the anti-interleukin 8 receptor alpha (CXCR1) antibody as modulators. The shedding of acellular dental biofilms overwhelmingly consists of bacterial extracellular vesicles (BEVs). The latter induced the moderate formation of neutrophil extracellular traps (NETs) in orally healthy subjects and a strong formation in patients with periodontitis. A CXCR2 inhibitor and an anti-CXCR1 antibody had a minor effect on NET formation. Neutrophils from patients with periodontitis exhibited NET hyper-responsiveness. BEVs were stronger inducers of NET formation than purified LPS and PMA. A plateau of neutrophil responsiveness is reached above the age of 40 years, indicating the abrupt switch of maladaptive trained immunity (TI) into the activated modus. Our results suggest that dental biofilms consist of and disseminate immense amounts of outer membrane vesicles (OMVs), which initiate NET formation via a non-canonical cytosolic LPS/caspase-4/11/Gasdermin D pathway. This modus of NET formation is independent of neutrophil elastase (NE), myeloperoxidase (MPO), peptidylarginine deiminase 4 (PAD4), and toll-like receptors (TLR). In periodontitis, the hyper-responsiveness of neutrophils to BEVs and the increased NET formation appear to be a consequence of TI.

Breaking the Gingival Barrier in Periodontitis.

The break of the epithelial barrier of gingiva has been a subject of minor interest, albeit playing a key role in periodontal pathology, transitory bacteraemia, and subsequent systemic low-grade inflammation (LGI). The significance of mechanically induced bacterial translocation in gingiva (e.g., via mastication and teeth brushing) has been disregarded despite the accumulated knowledge of mechanical force effects on tight junctions (TJs) and subsequent pathology in other epithelial tissues. Transitory bacteraemia is observed as a rule in gingival inflammation, but is rarely observed in clinically healthy gingiva. This implies that TJs of inflamed gingiva deteriorate, e.g., via a surplus of lipopolysaccharide (LPS), bacterial proteases, toxins, Oncostatin M (OSM), and neutrophil proteases. The inflammation-deteriorated gingival TJs rupture when exposed to physiological mechanical forces. This rupture is characterised by bacteraemia during and briefly after mastication and teeth brushing, i.e., it appears to be a dynamic process of short duration, endowed with quick repair mechanisms. In this review, we consider the bacterial, immune, and mechanical factors responsible for the increased permeability and break of the epithelial barrier of inflamed gingiva and the subsequent translocation of both viable bacteria and bacterial LPS during physiological mechanical forces, such as mastication and teeth brushing.

Connection between Periodontitis-Induced Low-Grade Endotoxemia and Systemic Diseases: Neutrophils as Protagonists and Targets.

Periodontitis is considered a promoter of many systemic diseases, but the signaling pathways of this interconnection remain elusive. Recently, it became evident that certain microbial challenges promote a heightened response of myeloid cell populations to subsequent infections either with the same or other pathogens. This phenomenon involves changes in the cell epigenetic and transcription, and is referred to as ''trained immunity''. It acts via modulation of hematopoietic stem and progenitor cells (HSPCs). A main modulation driver is the sustained, persistent low-level transmission of lipopolysaccharide from the periodontal pocket into the peripheral blood. Subsequently, the neutrophil phenotype changes and neutrophils become hyper-responsive and prone to boosted formation of neutrophil extracellular traps (NET). Cytotoxic neutrophil proteases and histones are responsible for ulcer formations on the pocket epithelium, which foster bacteremia and endoxemia. The latter promote systemic low-grade inflammation (SLGI), a precondition for many systemic diseases and some of them, e.g., atherosclerosis, diabetes etc., can be triggered by SLGI alone. Either reverting the polarized neutrophils back to the homeostatic state or attenuation of neutrophil hyper-responsiveness in periodontitis might be an approach to diminish or even to prevent systemic diseases.

Determinate growth is predominant and likely ancestral in squamate reptiles.

Body growth is typically thought to be indeterminate in ectothermic vertebrates. Indeed, until recently, this growth pattern was considered to be ubiquitous in ectotherms. Our recent observations of a complete growth plate cartilage (GPC) resorption, a reliable indicator of arrested skeletal growth, in many species of lizards clearly reject the ubiquity of indeterminate growth in reptiles and raise the question about the ancestral state of the growth pattern. Using X-ray micro-computed tomography (µCT), here we examined GPCs of long bones in three basally branching clades of squamate reptiles, namely in Gekkota, Scincoidea and Lacertoidea. A complete loss of GPC, indicating skeletal growth arrest, was the predominant finding. Using a dataset of 164 species representing all major clades of lizards and the tuataras, we traced the evolution of determinate growth on the phylogenetic tree of Lepidosauria. The reconstruction of character states suggests that determinate growth is ancestral for the squamate reptiles (Squamata) and remains common in the majority of lizard lineages, while extended (potentially indeterminate) adult growth evolved several times within squamates. Although traditionally associated with endotherms, determinate growth is coupled with ectothermy in this lineage. These findings combined with existing literature suggest that determinate growth predominates in both extant and extinct amniotes.

Comparison of four near-infrared spectroscopy devices shows that they are only suitable for monitoring cerebral oxygenation trends in preterm infants.

AIM: Measuring cerebral oxygenation using near-infrared spectroscopy (NIRS) has taken on an increasingly important role in the field of neonatology. Several companies have already developed commercial devices, and more publications are reporting absolute boundary values or percentiles for neonates. We compared four commercially used devices to discover whether they provided consistent results in the same patients. METHODS: We recruited nine preterm infants and tested them for 2 h, using sensors from two different devices. The measurements were carried out six times on each child, so that all four devices were compared with each other. A total of 54 measurements were conducted. The following devices were compared: the NIRO 200 (Hamamatsu Photonics K.K), the INVOS 5100c (Somanetics), the Fore-Sight (CAS Med.) and the SenSmart X-100 (NONIN). RESULTS: The cerebral tissue oxygenation data yielded by the individual devices differed significantly from each other, ranging from a minimum difference of 2.93% to a maximum difference of 12.66%. CONCLUSION: The commercially available NIRS devices showed highly significant differences in local cerebral tissue oxygenation levels, to the extent that the industry cannot agree on uniform and reproducible standards. Therefore, NIRS should only be used for trend measurements in preterm infants.

Characterization of the vasa vasorum in the human great saphenous vein: a scanning electron microscopy and 3D-morphometry study using vascular corrosion casts.

The vasa vasorum (VV) of explanted segments of the human great saphenous vein (Vena saphena magna; HGSV), harvested during dissection for coronary bypass grafts or diseased vein segments from the "Salzburger Landesklinikum," were studied by scanning electron microscopy and three-dimensional morphometry of microvascular corrosion casts. The main objective of this study was to examine the VV's structural arrangement in order to find the most vital segments of the HGSV and in turn to improve the results of coronary bypass surgeries. The study presents a meticulous analysis of the whole microvascular system of the VV of the HGSV and its three-dimensional arrangement. It is one of the first studies yielding detailed quantitative data on geometry of the VV of the HGSV. A detailed insight into different vascular parameters such as vessel diameter, interbranching, intervascular distances, and branching angles at different levels of the VV's angioarchitecture and in different parts of the HGSV in health and disease is given. Further, the geometry of bifurcations was examined in order to compute the physiological optimality principles of this delicate vascular system based on its construction, maintenance, and function.

Ocular microvasculature in adult Xenopus laevis: Scanning electron microscopy of vascular casts.

The microvascular anatomy of choriocapillaris, iris, ciliary body, and superficial vascular hyaloid system of eyes was studied in the permanent aquatic Xenopus laevis by scanning electron microscopy of vascular casts and was compared with that published in two semiaquatic ranid species (Rana esculenta and Rana temporaria), and the urodelian species Triturus criststus carnifex. Results showed that the choriocapillaris in Xenopus consisted of a dense meshwork of wide capillaries displaying polygonal arrays at the scleral side with venules leaving the centers and arterioles supplied from the periphery. The choriocapillaris lacked the multilayered capillary meshwork described in ranids. Iris and ciliary body were supplied by nasal and temporal branches of the iridial artery, which either originated with a common stem from the hyaloid artery or arose as individual vessels from the proximal portions of the semicircular nasal and temporal branches of the hyaloid artery. These branches ran in the pupillary margin and supplied the two-dimensional capillary network of the iris, as well as the three-dimensional network of the ciliary body. Iris and ciliary body drained via parallel running vasa recta into the choriocapillaris. The superficial vascular hyaloid bed (system) was supplied by the hyaloid artery. This artery coursed along the scleral surface of the ventrotemporal choriocapillaris toward the ora serrata, where it bifurcated into a temporal and a nasal semicircular branch. Seven to 10 arterial meridional twigs arose from these branches and supplied the superficial hyaloid capillary bed. Capillaries drained into branches of the hyaloid vein, which ascended toward the ora serrata, where the hyaloid vein joined the temporal branch of the ciliary vein.

Hypertrophic cardiomyopathy is characterized by alterations of the mitochondrial calcium uniporter complex proteins: insights from patients with aortic valve stenosis versus hypertrophic obstructive cardiomyopathy.

Introduction: Hypertrophies of the cardiac septum are caused either by aortic valve stenosis (AVS) or by congenital hypertrophic obstructive cardiomyopathy (HOCM). As they induce cardiac remodeling, these cardiac pathologies may promote an arrhythmogenic substrate with associated malignant ventricular arrhythmias and may lead to heart failure. While altered calcium (Ca2+) handling seems to be a key player in the pathogenesis, the role of mitochondrial calcium handling was not investigated in these patients to date. Methods: To investigate this issue, cardiac septal samples were collected from patients undergoing myectomy during cardiac surgery for excessive septal hypertrophy and/or aortic valve replacement, caused by AVS and HOCM. Septal specimens were matched with cardiac tissue obtained from post-mortem controls without cardiac diseases (Ctrl). Results and discussion: Patient characteristics and most of the echocardiographic parameters did not differ between AVS and HOCM. Most notably, the interventricular septum thickness, diastolic (IVSd), was the greatest in HOCM patients. Histological and molecular analyses showed a trend towards higher fibrotic burden in both pathologies, when compared to Ctrl. Most notably, the mitochondrial Ca2+ uniporter (MCU) complex associated proteins were altered in both pathologies of left ventricular hypertrophy (LVH). On the one hand, the expression pattern of the MCU complex subunits MCU and MICU1 were shown to be markedly increased, especially in AVS. On the other hand, PRMT-1, UCP-2, and UCP-3 declined with hypertrophy. These conditions were associated with an increase in the expression patterns of the Ca2+ uptaking ion channel SERCA2a in AVS (p = 0.0013), though not in HOCM, compared to healthy tissue. Our data obtained from human specimen from AVS or HOCM indicates major alterations in the expression of the mitochondrial calcium uniporter complex and associated proteins. Thus, in cardiac septal hypertrophies, besides modifications of cytosolic calcium handling, impaired mitochondrial uptake might be a key player in disease progression.

Introduction of an alternative standardized radiographic measurement method to evaluate volar angulation in subcapital fractures of the 5th metacarpal.

OBJECTIVE: The purpose of the present study was to compare the intra- and interobserver reliability of two different measurement methods for volar angulation of the 5th metacarpal (MC) in an attempt to establish a new standard measurement method to reduce interobserver discrepancies for therapeutic decisions. MATERIALS AND METHODS: Twenty patients with subcapital fractures of the 5th MC were radiologically investigated. Imaging consisted of a radiographs in antero-posterior and precise lateral view in addition to a CT scan of the 5th MC. Measurement of volar angulation was accomplished using the conventional and the shaft articular surface (SAS) method. The measurements of five investigators were exported to a spreadsheet for statistical analysis to evaluate the intra-and interobserver reliability. RESULTS: The conventional technique showed large differences among the investigators and poor interobserver reliability (W = 0.328 and 0.307) both at injury (p = 0.001) and at follow-up (p = 0.189). The intraobserver concordance of all investigators showed better results with the SAS than with the conventional technique. With the SAS technique, no statistically significant difference among the investigators could be detected at either the time of injury (p = 0.418) or at follow-up (p = 0.526) with excellent interobserver reliability (W = 0.051 and W = 0.041). Evaluation of volar angulation at follow-up using CT scans did not show any statistically significant difference between the techniques with better correlation among the observers with the SAS technique (p = 0.838). CONCLUSIONS: The interobserver correlation of volar angulation with lateral radiographs using the conventional technique was insufficient. Therefore, we recommend the use of the novel SAS technique as standardized measurement method which showed higher accuracy and interobserver reliability in order to facilitate the choice of adequate treatment option. CLINICAL RELEVANCE: A CT scan should be performed in cases with ambivalent treatment options. LEVEL OF EVIDENCE: Evidence-Based Medicine Level II.

Microvascular anatomy of the non-lobulated liver of adult Xenopus laevis: A scanning electron microscopic study of vascular casts.

The microvascular anatomy of the non-lobulated liver of adult Xenopus laevis was studied by scanning electron microscopy of vascular corrosion casts. Hepatic portal veins and hepatic arteries entered hepatic lobes at the hiluses, hepatic veins left at these sites. Intraparenchymal, hepatic portal veins branched up to 10 times before terminal portal venules supplied liver sinusoids. Hepatic arteries closely followed portal vessels. Arteriolar side branches formed anastomoses with close by portal venules (arteriolar-portal anastomoses; APAs), liver sinusoids (arteriolar-sinusoidal anastomoses; ASAs), and peribiliary plexus vessels. Distally, hepatic arteries anastomosed with terminal portal venules having >100 µm in diameter. Liver sinusoids formed a dense three-dimensional network displaying signs of non-sprouting and sprouting angiogenesis evidenced by "holes" and blind ending tapering cast vascular structures (sprouts), respectively. Sinusoids drained via efferent hepatic veins. Right and left hepatic veins drained into the posterior caval vein. Locally, a dense honeycomb-like 3D-meshwork of resin structures was found around terminal portal venules and hepatic arteries. These networks were fed by hepatic arterioles and drained into adjacent terminal portal venules. As their morphologies differed significantly from sinusoids and they were found at sites where diffuse lymphoid tissue is described, we are convinced that they represent the vasculature of diffuse lymphoid tissue areas. Frequencies and diameter ratios of hepatic portal venules versus hepatic arterioles anastomosing with the former (APAs) implicate that the arterial supply contributes to the oxygenation of parenchymal and stromal cells rather than to a significant increase in blood flow towards hepatic sinusoids.

Periodontitis-Derived Dark-NETs in Severe Covid-19.

The frequent severe COVID-19 course in patients with periodontitis suggests a link of the aetiopathogenesis of both diseases. The formation of intravascular neutrophil extracellular traps (NETs) is crucial to the pathogenesis of severe COVID-19. Periodontitis is characterised by an increased level of circulating NETs, a propensity for increased NET formation, delayed NET clearance and low-grade endotoxemia (LGE). The latter has an enormous impact on innate immunity and susceptibility to infection with SARS-CoV-2. LPS binds the SARS-CoV-2 spike protein and this complex, which is more active than unbound LPS, precipitates massive NET formation. Thus, circulating NET formation is the common denominator in both COVID-19 and periodontitis and other diseases with low-grade endotoxemia like diabetes, obesity and cardiovascular diseases (CVD) also increase the risk to develop severe COVID-19. Here we discuss the role of propensity for increased NET formation, DNase I deficiency and low-grade endotoxaemia in periodontitis as aggravating factors for the severe course of COVID-19 and possible strategies for the diminution of increased levels of circulating periodontitis-derived NETs in COVID-19 with periodontitis comorbidity.

Biodegradable Poly(D-L-lactide-co-glycolide) (PLGA)-Infiltrated Bioactive Glass (CAR12N) Scaffolds Maintain Mesenchymal Stem Cell Chondrogenesis for Cartilage Tissue Engineering.

Regeneration of articular cartilage remains challenging. The aim of this study was to increase the stability of pure bioactive glass (BG) scaffolds by means of solvent phase polymer infiltration and to maintain cell adherence on the glass struts. Therefore, BG scaffolds either pure or enhanced with three different amounts of poly(D-L-lactide-co-glycolide) (PLGA) were characterized in detail. Scaffolds were seeded with primary porcine articular chondrocytes (pACs) and human mesenchymal stem cells (hMSCs) in a dynamic long-term culture (35 days). Light microscopy evaluations showed that PLGA was detectable in every region of the scaffold. Porosity was greater than 70%. The biomechanical stability was increased by polymer infiltration. PLGA infiltration did not result in a decrease in viability of both cell types, but increased DNA and sulfated glycosaminoglycan (sGAG) contents of hMSCs-colonized scaffolds. Successful chondrogenesis of hMSC-colonized scaffolds was demonstrated by immunocytochemical staining of collagen type II, cartilage proteoglycans and the transcription factor SOX9. PLGA-infiltrated scaffolds showed a higher relative expression of cartilage related genes not only of pAC-, but also of hMSC-colonized scaffolds in comparison to the pure BG. Based on the novel data, our recommendation is BG scaffolds with single infiltrated PLGA for cartilage tissue engineering.

Arthroscopic all-inside meniscal repair--Does the meniscus heal? A clinical and radiological follow-up examination to verify meniscal healing using a 3-T MRI.

OBJECTIVE: The purpose of this study was to correlate clinical and radiological results using a 3-T MRI to verify meniscal healing after arthroscopic all-inside meniscus repair. MATERIALS AND METHODS: We selected 27 patients (14 men and 13 women) with an average age of 31 ± 9 years and retrospective clinical examinations and radiological assessments using a 3-T MRI after all-inside arthroscopic meniscal repair were conducted. Repair of the medial meniscus was performed in 19 patients and of the lateral meniscus in eight. In 17 patients (63%), we performed concomitant anterior cruciate ligament reconstruction. The mean follow-up period was 4.5 ± 1.7 years. The Lysholm score and Tegner activity index were used for clinical evaluation. Four grades were used to classify the radiological signal alterations within the meniscus: central globular (grade 1); linear horizontal or band-like (grade 2); intrameniscal alterations and linear signal alterations communicating with the articular surface (grade 3); and complex tears (grade 4). RESULTS: At follow-up, the average Lysholm score was 76 ± 15 points, with ten of the patients placed in group 6 based on the Tegner activity index. MRI examinations revealed no signal alteration in three patients, grade 1 in 0, grade 2 in five, grade 3 in 13, and grade 4 in six. The MRI findings correlated positively with the clinical scores in 21 patients (78%). CONCLUSIONS: Correlation of clinical and radiological examination was performed using 3-T MRI. In spite of satisfactory clinical outcomes at follow-up, a radiological signal alteration may still be visible on MRI, which was believed to be scar tissue, but could not be proven definitively. Imaging with a 3-Tesla MRI after meniscal suture surgery provides good but no definitive reliability on meniscus healing and therefore gives no advantage compared to 1.5-T MRI, with good clinical outcome using an all-inside arthroscopic meniscal repair. CLINICAL RELEVANCE: 3T-MRI can not substitute diagnostic arthroscopy in patients with persistent complaints after arthroscopic all-inside meniscal repair.

Microvascular anatomy of the urinary bladder in the adult African clawed toad, Xenopus laevis: A scanning electron microscope study of vascular casts.

We studied urinary bladders of adult male and female Xenopus laevis using light microscopy of stained tissue sections and scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Results showed that bilaterally a vesical artery branched off the femoral artery. At the dorso-lateral serosal surface of the body of the bladder each artery splitted within a short distance into up to five smaller arteries that supplied body and neck regions. Arteries gave off short and long terminal arterioles, which fed the mucosal capillary meshwork. Long terminal arterioles followed dimensional changes of the bladder, while short ones anchored the capillary network to the arterial system. Capillary mesh sizes and shapes varied according to the filling state of the urinary bladder. In the highly to moderately distended (filled) bladder, capillaries were rather straight or undulated only slightly, in the contracted (emptied) bladder they undulated strongly and lay side by side. Postcapillary venules formed by two equally sized capillaries or from capillaries, which serially drained into a small postcapillary venule. Vesical venules formed a large dorsal vesical and a varying number of smaller lateral and ventral vesical veins. The dorsal vesical vein drained either directly or via the posterior hemorrhoidal vein into the common pelvic vein. Lateral and ventral vesical veins also drained into the latter. The vascular patterns found were discussed in respect to the bladder spatial movements during distention (filling) and relaxation (emptying). Furthermore, it was hypothesized that an extensively filled bladder could compress the overlaying abdominal vein forcing part of the blood otherwise drained towards the liver to be detoured via the renal portal veins to the kidneys.

3D morphometric study of domestic fowl glomerulus-A scanning electron microscope study of vascular corrosion casts.

Microvascularization of domestic fowl kidneys was studied using scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Two types of nephrons, mammalian-type (MT) and reptilian-type (RT) nephrons and their glomerular structure were analysed quantitatively by 3D morphometry. A significant difference in shape and size between the MT and RT glomeruli was found. The mean diameter of the RT glomeruli was about 56 µm, while that of MT glomeruli was significantly larger, namely about 80 µm. The afferent arterioles in mammalian-type glomeruli usually bifurcated into two lobular branches and formed a complex glomerular capillary network with numerous loops. Reptilian-type glomeruli consisted of a single capillary forming few loops and leaving the glomerulus as efferent arteriole. Diameters of afferent and efferent arteriolar replicas were similar in all three kidney divisions of MT and RT nephrons. The absence of the interconnecting branches between the MT nephron capillaries at the gross inspection suggests that the mammalian-type nephron glomeruli, although more complex than the reptilian type, are not equivalent to those in mammalian kidneys.

Neutrophils Orchestrate the Periodontal Pocket.

The subgingival biofilm attached to tooth surfaces triggers and maintains periodontitis. Previously, late-onset periodontitis has been considered a consequence of dysbiosis and a resultant polymicrobial disruption of host homeostasis. However, a multitude of studies did not show "healthy" oral microbiota pattern, but a high diversity depending on culture, diets, regional differences, age, social state etc. These findings relativise the aetiological role of the dysbiosis in periodontitis. Furthermore, many late-onset periodontitis traits cannot be explained by dysbiosis; e.g. age-relatedness, attenuation by anti-ageing therapy, neutrophil hyper-responsiveness, and microbiota shifting by dysregulated immunity, yet point to the crucial role of dysregulated immunity and neutrophils in particular. Furthermore, patients with neutropenia and neutrophil defects inevitably develop early-onset periodontitis. Intra-gingivally injecting lipopolysaccharide (LPS) alone causes an exaggerated neutrophil response sufficient to precipitate experimental periodontitis. Vice versa to the surplus of LPS, the increased neutrophil responsiveness characteristic for late-onset periodontitis can effectuate gingiva damage likewise. The exaggerated neutrophil extracellular trap (NET) response in late-onset periodontitis is blameable for damage of gingival barrier, its penetration by bacteria and pathogen-associated molecular patterns (PAMPs) as well as stimulation of Th17 cells, resulting in further neutrophil activation. This identifies the dysregulated immunity as the main contributor to periodontal disease.

Highly porous novel chondro-instructive bioactive glass scaffolds tailored for cartilage tissue engineering.

Cartilage injuries remain challenging since the regenerative capacity of cartilage is extremely low. The aim was to design a novel type of bioactive glass (BG) scaffold with suitable topology that allows the formation of cartilage-specific extracellular matrix (ECM) after colonization with chondrogenic cells for cartilage repair. Highly porous scaffolds with interconnecting pores consisting of 100 % BG were manufactured using a melting, milling, sintering and leaching technique. Scaffolds were colonized with porcine articular chondrocytes (pAC) and undifferentiated human mesenchymal stromal cells (hMSC) for up to 35 days. Scaffolds displayed high cytocompatibility with no major pH shift. Scanning electron microscopy revealed the intimate pAC-scaffold interaction with typical cell morphology. After 14 days MSCs formed cell clusters but still expressed cartilage markers. Both cell types showed aggrecan, SOX9 gene and protein expression, cartilage proteoglycan and sulfated glycosaminoglycan synthesis for the whole culture time. Despite type II collagen gene expression could not anymore be detected at day 35, protein synthesis was visualized for both cell types during the whole culturing period, increasing in pAC and declining after day 14 in hMSC cultures. The novel BG scaffold was stable, cytocompatible and cartilage-specific protein synthesis indicated maintenance of pAC's differentiated phenotype and chondro-instructive effects on hMSCs.

Renal microvasculature in the adult pipid frog, Xenopus laevis: A scanning electron microscope study of vascular corrosion casts.

We studied the opisthonephric (mesonephric) kidneys of adult male and female Xenopus laevis using scanning electron microscopy (SEM) of vascular corrosion casts and light microscopy of paraplast embedded tissue sections. Both techniques displayed glomeruli from ventral to mid-dorsal regions of the kidneys with single glomeruli located dorsally close beneath the renal capsule. Glomeruli in general were fed by a single afferent arteriole and drained via a single thinner efferent arteriole into peritubular vessels. Light microscopy and SEM of vascular corrosion casts revealed sphincters at the origins of afferent arterioles, which arose closely, spaced from their parent renal arteries. The second source of renal blood supply via renal portal veins varied interindividually in branching patterns with vessels showing up to five branching orders before they became peritubular vessels. Main trunks and their first- and second-order branches revealed clear longish endothelial cell nuclei imprint patterns oriented parallel to the vessels longitudinal axis, a pattern characteristic for arteries. Peritubular vessels had irregular contours and were never seen as clear cylindrical structures. They ran rather parallel, anastomosed with neighbors and changed into renal venules and veins, which finally emptied into the ventrally located posterior caval vein. A third source of blood supply of the peritubular vessels by straight terminal portions of renal arteries (vasa recta) was not found.

Microvascular anatomy of ovary and oviduct in the adult African Clawed Toad (Xenopus laevis DAUDIN, 1802)-Histomorphology and scanning electron microscopy of vascular corrosion casts.

Ovaries and oviducts of the adult African Clawed Toad (Xenopus laevis DAUDIN, 1802) were studied by light microscopy (LM) of paraplast embedded tissue sections and scanning electron microscopy (SEM) of vascular corrosion casts (VCCs). Histomorphology revealed that ovarian vessels located in the thecal layers. Ovarian and interlobar arteries displayed a horse-shoe shaped longitudinally running bundle of vascular smooth muscle cells. Follicular blood vessels showed flattened profiles, which were confirmed by scanning electron microscopy in vascular corrosion casts. The flattened profiles obviously led to high intravasal pressures, which locally prevented filling of the follicular capillary bed. Oviduct arteries pierced the fibrous stroma surrounding the oviduct mucosa. In the pars convoluta, the mucosa consisted of a ciliated simple columnar epithelium and tubular oviduct glands that opened between ciliated epithelial cells into the oviduct lumen. Oviduct arteries branched at the basolateral surfaces of tubular glands. After a short tangential course, arterioles branched into capillaries which ran radially between oviduct glands towards the subepithelium. Anastomoses at different heights connected capillaries of neighbouring glands. Subepithelially, capillaries ran longitudinally and undulated. Postcapillary venules radiated centrifugally towards the stroma to finally drain into oviduct veins located in the stroma. Oviduct vascular densities clearly reflected non-ovulatory and ovulatory states.