Patient- and physician-related risk factors for hyperkalaemia in potassium-increasing drug-drug interactions.

PURPOSE: Hyperkalaemia due to potassium-increasing drug-drug interactions (DDIs) is a clinically important adverse drug event. The purpose of this study was to identify patient- and physician-related risk factors for the development of hyperkalaemia. METHODS: The risk for adult patients hospitalised in the University Hospital Zurich between 1 December 2009 and 31 December 2011 of developing hyperkalaemia was correlated with patient characteristics, number, type and duration of potassium-increasing DDIs and frequency of serum potassium monitoring. RESULTS: The 76,467 patients included in this study were prescribed 8,413 potentially severe potassium-increasing DDIs. Patient-related characteristics associated with the development of hyperkalaemia were pulmonary allograft [relative risk (RR) 5.1; p < 0.0001), impaired renal function (RR 2.7; p < 0.0001), diabetes mellitus (RR 1.6; p = 0.002) and female gender (RR 1.5; p = 0.007). Risk factors associated with medication were number of concurrently administered potassium-increasing drugs (RR 3.3 per additional drug; p < 0.0001) and longer duration of the DDI (RR 4.9 for duration ≥6 days; p < 0.0001). Physician-related factors associated with the development of hyperkalaemia were undetermined or elevated serum potassium level before treatment initiation (RR 2.2; p < 0.001) and infrequent monitoring of serum potassium during a DDI (interval >48 h: RR 1.6; p < 0.01). CONCLUSION: Strategies for reducing the risk of hyperkalaemia during potassium-increasing DDIs should consider both patient- and physician-related risk factors.

Injectable living marrow stromal cell-based autologous tissue engineered heart valves: first experiences with a one-step intervention in primates.

AIMS: A living heart valve with regeneration capacity based on autologous cells and minimally invasive implantation technology would represent a substantial improvement upon contemporary heart valve prostheses. This study investigates the feasibility of injectable, marrow stromal cell-based, autologous, living tissue engineered heart valves (TEHV) generated and implanted in a one-step intervention in non-human primates. METHODS AND RESULTS: Trileaflet heart valves were fabricated from non-woven biodegradable synthetic composite scaffolds and integrated into self-expanding nitinol stents. During the same intervention autologous bone marrow-derived mononuclear cells were harvested, seeded onto the scaffold matrix, and implanted transapically as pulmonary valve replacements into non-human primates (n = 6). The transapical implantations were successful in all animals and the overall procedure time from cell harvest to TEHV implantation was 118 ± 17 min. In vivo functionality assessed by echocardiography revealed preserved valvular structures and adequate functionality up to 4 weeks post implantation. Substantial cellular remodelling and in-growth into the scaffold materials resulted in layered, endothelialized tissues as visualized by histology and immunohistochemistry. Biomechanical analysis showed non-linear stress-strain curves of the leaflets, indicating replacement of the initial biodegradable matrix by living tissue. CONCLUSION: Here, we provide a novel concept demonstrating that heart valve tissue engineering based on a minimally invasive technique for both cell harvest and valve delivery as a one-step intervention is feasible in non-human primates. This innovative approach may overcome the limitations of contemporary surgical and interventional bioprosthetic heart valve prostheses.

Clinical decision support for monitoring drug-drug-interactions and potassium-increasing drug combinations: need for specific alerts.

Computer-triggered reminders alerting physicians on every potentially harmful drug-drug-interaction (DDI) induce alert fatigue due to frequent messages of limited clinical relevance. On demand DDI-checks, however, are not commonly used by physicians. Optimal strategies for sustained quality assurance have to consider patients' risk factors and focus on the most significant DDIs only. An approach is proposed based on the analysis of concurrent prescription of potassium-sparing diuretics and potassium supplements (CPPP), which are the most frequent DDIs classified as contraindicated. Although the frequency of monitoring potassium serum levels declined during prolonged periods of CPPP, the likelihood of observing a hyperkalaemia increased. The median treatment period of CPPP was 3.3 days, whereas hyperkalaemia occurred after a median observation time of 4.5 days of CPPP. Thus, computer-triggered reminders for ordering potassium serum levels may be indicated if monitoring has been discontinued after 48h of CPPP.

Prenatally fabricated autologous human living heart valves based on amniotic fluid derived progenitor cells as single cell source.

BACKGROUND: A novel concept providing prenatally tissue engineered human autologous heart valves based on routinely obtained fetal amniotic fluid progenitors as single cell source is introduced. METHODS AND RESULTS: Fetal human amniotic progenitors were isolated from routinely sampled amniotic fluid and sorted using CD133 magnetic beads. After expansion and differentiation, cell phenotypes of CD133- and CD133+ cells were analyzed by immunohistochemistry and flowcytometry. After characterization, CD133- derived cells were seeded onto heart valve leaflet scaffolds (n=18) fabricated from rapidly biodegradable polymers, conditioned in a pulse duplicator system, and subsequently coated with CD133+ derived cells. After in vitro maturation, opening and closing behavior of leaflets was investigated. Neo-tissues were analyzed by histology, immunohistochemistry, and scanning electron microscopy (SEM). Extracellular matrix (ECM) elements and cell numbers were quantified biochemically. Mechanical properties were assessed by tensile testing. CD133- derived cells demonstrated characteristics of mesenchymal progenitors expressing CD44 and CD105. Differentiated CD133+ cells showed features of functional endothelial cells by eNOS and CD141 expression. Engineered heart valve leaflets demonstrated endothelialized tissue formation with production of ECM elements (GAG 80%, HYP 5%, cell number 100% of native values). SEM showed intact endothelial surfaces. Opening and closing behavior was sufficient under half of systemic conditions. CONCLUSIONS: The use of amniotic fluid as single cell source is a promising low-risk approach enabling the prenatal fabrication of heart valves ready to use at birth. These living replacements with the potential of growth, remodeling, and regeneration may realize the early repair of congenital malformations.

Endothelial nitric oxide synthase gene transfer inhibits human smooth muscle cell migration via inhibition of Rho A.

Smooth muscle cell (SMC) migration contributes to vascular remodeling. Nitric oxide (NO) produced via endothelial NO synthase (eNOS) inhibits SMC migration. This study analyzes signal transduction mechanisms of SMC migration targeted by NO. SMCs were cultured from human saphenous veins, and cell migration was studied using Boyden chambers. PDGF-BB (0.1 to 10 ng/ml) stimulated SMC migration in a concentration-dependent manner, which was inhibited by adenoviral-mediated overexpression of eNOS and by the NO donor diethylentriamine NONOate (DETANO, 10 to 10 mol/L). NO release was enhanced in eNOS-transduced SMCs, and L-NAME blunted the effect of eNOS overexpression on migration. PDGF-BB (10 ng/ml) activated Rho A, which was inhibited by the overexpression of eNOS by DETANO and by 8 bromo-cGMP. The inhibitory effect of DETANO on Rho A activity was prevented by the cGMP-dependant kinase inhibitor. Furthermore, inhibition of Rho A by C3 exoenzyme and inhibition of ROCK by Y-27632 diminished cell migration stimulated by PDGF-BB. Finally, in the cells overexpressing constitutively active ROCK mutant (CAT), DETANO failed to prevent PDGF-BB-induced SMC migration. In conclusion, NO inhibits human SMC migration via blockade of the Rho A pathway.

Cryopreserved amniotic fluid-derived cells: a lifelong autologous fetal stem cell source for heart valve tissue engineering.

BACKGROUND AND AIM OF THE STUDY: Fetal stem cells represent a promising cell source for heart valve tissue engineering. In particular, amniotic fluid-derived cells (AFDC) have been shown to lead to autologous fetal-like heart valve tissues in vitro for pediatric application. In order to expand the versatility of these cells also for adult application, cryopreserved AFDC were investigated as a potential life-long available cell source for heart valve tissue engineering. METHODS: Human AFDC were isolated using CD133 magnetic beads, and then differentiated and analyzed. After expansion of CD133- as well as CD133+ cells up to passage 7, a part of the cells was cryopreserved. After four months, the cells were re-cultured and phenotyped by flow cytometry and immunohistochemistry, including expression of CD44, CD105, CD90, CD34, CD31, CD141, eNOS and vWF, and compared to their non-cryopreserved counterparts. The stem cell potential was investigated in differentiation assays. The viability of cryopreserved AFDC for heart valve tissue engineering was assessed by creating heart valve leaflets in vitro. RESULTS: After cryopreservation, amniotic fluid-derived CD133- and CD133+ cells retained their stem cell-like phenotype, expressing mainly CD44, CD90 and CD105. This staining pattern was comparable to that of their non-cryopreserved counterparts. Moreover, CD133- cells demonstrated differentiation potential into osteoblast-like and adipocyte-like cells. CD133+ cells showed characteristics of endothelial-like cells by eNOS, CD141 and beginning vWF expression. When used for the fabrication of heart valve leaflets, cryopreserved CD133- cells produced extracellular matrix elements comparable to their non-cryopreserved counterparts. Moreover, the resulting tissues showed a cellular layered tissue formation covered by functional endothelia. The mechanical properties were similar to those of tissues fabricated from non-cryopreserved cells. CONCLUSION: The study results suggest that the use of cell bank technology fetal amniotic fluid-derived stem cells might represent a life-long available autologous cell source for heart valve tissue engineering, and also for adult application.

Assessment of coronary sinus anatomy between normal and insufficient mitral valves by multi-slice computertomography for mitral annuloplasty device implantation.

INTRODUCTION: Latest techniques enable positioning of devices into the coronary sinus (CS) for mitral valve (MV) annuloplasty. We evaluate the feasibility of non-invasive assessment to determine CS anatomy and its relation to MV annulus and coronary arteries by multi-slice CT (MSCT) in normal and insufficient MV. METHODS: Fifty patients (33 males, 17 females, age 67+/-11 years) were studied retrospectively by 64-MSCT scans for anatomical criteria regarding CS and its relation to MV annulus and circumflex artery (CX). We included 24 patients with severe mitral insufficiency and 26 with no MV disease. Diameter of MV, of proximal and distal ostium of CS, length and volume of CS, angle between anterior interventricular vein (AIV) and CS, caliber change of CX before, under/over and after CS were analysed. Different anatomical correlations were demonstrated: distance of MV annulus to CS, CX to CS. RESULTS: Diameter of proximal CS ostium was significantly larger in insufficient MV compared to normal MV (11+/-2.8 mm vs 9.9+/-2.5 mm; p<0.024). CS was significantly longer in patients with insufficient MV (125.4+/-17 mm vs 108.9+/-18 mm; p<0.003) with also significant differences in volume of CS (p<0.039). Significant difference in annulus diameter, 46.1+/-6mm (insufficient MV) versus 39.5+/-7.5 mm, p<0.004 was observed. Angle CS-AIV was 103.5+/-29 degrees (range 52 degrees -144 degrees ) in insufficient valves versus 118.2+/-24.5 degrees (range 73 degrees -166 degrees ) in normal valves with a tendency to higher angles in normal valves (p=0.06). Distance of MV annulus to CS measured 16+/-4.1/14.2+/-3.6 mm (insufficient/normal MV) without significant difference between groups. In 15 patients CX ran under CS. Eighty-four percent of these patients (13/15) show a decrease in CS caliber in the area of intersection. In 14 patients CS ran over and in one patient the diameter of the CS at intersecting region was smaller. In 16 patients no direct point of contact was visible, in five patients CX to CS positioning was not evaluable. CONCLUSION: There is a significant anatomic difference between normal and insufficient MV, which might be the basis for any interventional approaches through the CS. Exact measurements of all structures and its anatomic correlations are possible with MSCT, which allows pre-interventional planning.

Living autologous heart valves engineered from human prenatally harvested progenitors.

BACKGROUND: Heart valve tissue engineering is a promising strategy to overcome the lack of autologous growing replacements, particularly for the repair of congenital malformations. Here, we present a novel concept using human prenatal progenitor cells as new and exclusive cell source to generate autologous implants ready for use at birth. METHODS AND RESULTS: Human fetal mesenchymal progenitors were isolated from routinely sampled prenatal chorionic villus specimens and expanded in vitro. A portion was cryopreserved. After phenotyping and genotyping, cells were seeded onto synthetic biodegradable leaflet scaffolds (n=12) and conditioned in a bioreactor. After 21 days, leaflets were endothelialized with umbilical cord blood-derived endothelial progenitor cells and conditioned for additional 7 days. Resulting tissues were analyzed by histology, immunohistochemistry, biochemistry (amounts of extracellular matrix, DNA), mechanical testing, and scanning electron microscopy (SEM) and were compared with native neonatal heart valve leaflets. Fresh and cryopreserved cells showed comparable myofibroblast-like phenotypes. Genotyping confirmed their fetal origin. Neo-tissues exhibited organization, cell phenotypes, extracellular matrix production, and DNA content comparable to their native counterparts. Leaflet surfaces were covered with functional endothelia. SEM showed cellular distribution throughout the polymer and smooth surfaces. Mechanical profiles approximated those of native heart valves. CONCLUSIONS: Prenatal fetal progenitors obtained from routine chorionic villus sampling were successfully used as an exclusive, new cell source for the engineering of living heart valve leaflets. This concept may enable autologous replacements with growth potential ready for use at birth. Combined with the use of cell banking technology, this approach may be applied also for postnatal applications.

Functional growth in tissue-engineered living, vascular grafts: follow-up at 100 weeks in a large animal model.

BACKGROUND: Living autologous vascular grafts with the capacity for regeneration and growth may overcome the limitations of contemporary artificial prostheses. Particularly in congenital cardiovascular surgery, there is an unmet medical need for growing replacement materials. Here we investigate growth capacity of tissue-engineered living pulmonary arteries in a growing lamb model. METHODS AND RESULTS: Vascular grafts fabricated from biodegradable scaffolds (ID 18+/-l mm) were sequentially seeded with vascular cells. The seeded constructs were grown in vitro for 21 days using biomimetic conditions. Thereafter, these tissue-engineered vascular grafts (TEVGs) were surgically implanted as main pulmonary artery replacements in 14 lambs using cardiopulmonary bypass and followed up for < or = 100 weeks. The animals more than doubled their body weight during the 2-year period. The TEVG showed good functional performance demonstrated by regular echocardiography at 20, 50, 80, and 100 weeks and computed tomography-angiography. In particular, there was no evidence of thrombus, calcification, stenosis, suture dehiscence, or aneurysm. There was a significant increase in diameter by 30% and length by 45%. Histology showed tissue formation reminiscent of native artery. Biochemical analysis revealed cellularity and proteoglycans and increased collagen contents in all of the groups, analogous to those of native vessels. The mechanical profiles of the TEVG showed stronger but less elastic tissue properties than native pulmonary arteries. CONCLUSIONS: This study provides evidence of growth in living, functional pulmonary arteries engineered from vascular cells in a full growth animal model.

Autologous human tissue-engineered heart valves: prospects for systemic application.

BACKGROUND: Tissue engineering represents a promising approach for the development of living heart valve replacements. In vivo animal studies of tissue-engineered autologous heart valves have focused on pulmonary valve replacements, leaving the challenge to tissue engineer heart valves suitable for systemic application using human cells. METHODS AND RESULTS: Tissue-engineered human heart valves were analyzed up to 4 weeks and conditioning using bioreactors was compared with static culturing. Tissue formation and mechanical properties increased with time and when using conditioning. Organization of the tissue, in terms of anisotropic properties, increased when conditioning was dynamic in nature. Exposure of the valves to physiological aortic valve flow demonstrated proper opening motion. Closure dynamics were suboptimal, most likely caused by the lower degree of anisotropy when compared with native aortic valve leaflets. CONCLUSIONS: This study presents autologous tissue-engineered heart valves based on human saphenous vein cells and a rapid degrading synthetic scaffold. Tissue properties and mechanical behavior might allow for use as living aortic valve replacements.

Geometric models of the aortic and pulmonary roots: suggestions for the Ross procedure.

OBJECTIVE: To discuss geometric factors, which may influence long-term results relating to homograft competence following the Ross procedure, we describe the 3D morphology of the pulmonary and aortic roots. MATERIALS: Measurements were made on 25 human aortic and pulmonary roots. Inter-commissural distances and the heights of the sinuses were measured. For geometrical reconstruction the three commissures and their vertical projections at the root base were used as reference points. RESULTS: In the pulmonary root, the three inter-commissural distances were of similar dimensions (17.9+/-1.6mm, 17.5+/-1.4mm and 18.6+/-1.5mm). In the aortic root, the right inter-commissural distance was greatest (18.8+/-1.9mm), followed by the non-coronary (17.4+/-2.0mm) and left coronary sinus commissures (15.2+/-1.9mm). The mean height of the left pulmonary sinus was greatest (20+/-1.7mm) followed by the anterior (17.5+/-1.4mm) and right pulmonary sinus (18+/-1.66mm). In the aortic root, the height of the right coronary sinus was the greatest (19.4+/-1.9mm) followed by the heights of the non-coronary (17.7+/-1.8mm) and left coronary sinus (17.4+/-1.4mm). Measured differences between parameters determine the tilt angle and direction of the root vector. The tilt angle in the pulmonary root averaged 16.26 degrees , respectively; for the aortic roots, it was 5.47 degrees . CONCLUSIONS: Herein we suggest that the left pulmonary sinus is best implanted in the position of the right coronary sinus, the anterior pulmonary in the position of the non-coronary sinus and the right pulmonary sinus in the position of the left coronary sinus. In this way, the direction of the pulmonary root vector will be parallel to that of the aortic root vector.

Activation of human microvascular endothelial cells with TNF-alpha and hypoxia/reoxygenation enhances NK-cell adhesion, but not NK-Cytotoxicity.

BACKGROUND: Ischemia/reperfusion injury (I/R) and cellular rejection in solid organ transplantation are characterized by adhesion molecule up-regulation on the graft endothelium, a prerequisite for leukocyte recruitment. The contribution of NK cells to I/R and allograft rejection is not well understood. The aim of the present study was to investigate allogeneic interactions between human NK cells and microvascular endothelial cells (MVEC) with special regard to the differential impact of TNF-alpha and hypoxia/reoxygenation in an in vitro model of I/R. METHODS: MVEC were stimulated in vitro for 8 h with TNF-alpha, exposed to hypoxia (1% O2), hypoxia/reoxygenation, and combinations thereof in a hypoxia chamber. Cell surface expression of adhesion molecules on MVEC was analyzed by flow cytometry, and adhesion molecule shedding by ELISA. NK cell adhesion on MVEC was determined under shear stress, and NK cytotoxicity using Cr-release assays. RESULTS: Surface expression of ICAM-1, VCAM-1, and E-/P-selectin on MVEC was up-regulated by TNF-alpha but unaffected by hypoxia/reoxygenation in the absence of TNF-alpha. ICAM-1 expression was further increased by a combination of TNF-alpha and hypoxia/reoxygenation, whereas TNF-alpha-induced E-/P-selectin expression was strongly reversed by hypoxia/reoxygenation. NK cell adhesion increased after exposing MVEC to TNF-alpha and hypoxia/reoxygenation. Susceptibility of MVEC to NK cytotoxicity was enhanced by TNF-alpha and slighty reduced by hypoxia/reoxygenation. CONCLUSIONS: Endothelial activation with TNF-alpha, but not hypoxia/reoxygenation, induced NK cytotoxicity whereas the combination thereof induced the strongest NK cell adhesion. Our findings suggesting a role for NK cells in allograft responses support the development of anti-inflammatory treatment strategies to prevent I/R.

Engineering of biologically active living heart valve leaflets using human umbilical cord-derived progenitor cells.

This study demonstrates the engineering of biologically active heart valve leaflets using prenatally available human umbilical cord-derived progenitor cells as the only cell source. Wharton's Jelly-derived cells and umbilical cord blood-derived endothelial progenitor cells were subsequently seeded on biodegradable scaffolds and cultured in a biomimetic system under biochemical or mechanical stimulation or both. Depending on the stimulation, leaflets showed mature layered tissue formation with functional endothelia and extracellular matrix production comparable with that of native tissues. This demonstrates the feasibility of heart valve leaflet fabrication from prenatal umbilical cord-derived progenitor cells as a further step in overcoming the lack of living autologous replacements with growth and regeneration potential for the repair of congenital malformation.

Clinical anatomy of the atrioventricular node artery.

BACKGROUND AND AIM OF THE STUDY: The study aim was to describe the topographical relationship of the atrioventricular (AV) node artery and mitral valve annulus fibrosus with regard to AV node dysfunction following mitral valve replacement or ring annuloplasty. METHODS: The anatomy of the AV node artery was analyzed in 55 human hearts without previous pathological alterations. Selective coronary angiograms were performed to identify the AV node origin. Run-off of the AV node artery and its topographical relationship to the mitral valve attachment was analyzed in dry-dissected hearts. The position of the AV node was verified by histological sectioning. RESULTS: The AV node artery originated from the right coronary artery in 73% of examined cases, and from the left coronary artery in 27% of cases. The left AV node artery was closely related to the mitral valve attachment, especially at the area of the left proximal part of the posterior leaflet. CONCLUSION: These morphological data were compared to clinical reports emphasizing the postoperative incidence of AV block after mitral valve implantation and ring annuloplasty. The occurrence of early postoperative AV node block ranged from 20% to 37%. By comparing the present data with available literature, it can be stated that there is a high risk of intraoperative damage to the left AV node artery during manipulation of the mitral valve annulus fibrosus.

Does retrograde cerebral perfusion via superior vena cava cannulation protect the brain?

OBJECTIVE: The retrograde cerebral perfusion via cannulation of the superior vena cava is a widespread method for optimising protection of the brain during hypothermic circulatory arrest. METHODS: In 14 cadavers (8 females, 6 males) of the local department of pathology, an examination was performed to check the competence of the valves of the internal jugular veins. After a complete preparation of the superior vena cava, the innominate vein and both internal jugular veins, ligating all side branches, a retrograde perfusion on 7 cadavers was installed, documenting flow and pressure of each internal jugular vein (IJV) in vitro. Afterwards, the veins were opened and their valves inspected. RESULTS: In all 14 cadavers, anatomically and functionally competent valves on the right proximal IJV were found. Only 1/14 cadaver had no valve in the left proximal IJV. Additional rudimentary and incompetent valves could be identified in 1/14 cadaver on the distal right IJV, and in 2/14 cadavers on the left IJV. Retrograde flow measurement of 7/14 cadavers revealed 0 ml/min in 4/7 cadavers, 6 ml/min in 1/7, 340 ml/min in 1/7 and 2500 ml/min in 1/7 cadaver. CONCLUSIONS: As a rule, anatomically and functionally competent valves in the proximal IJV are present. In human beings, they obstruct the direct retrograde inlet to the intracranial venous system, which suggests an unbalanced and unreliable perfusion of the brain. Therefore, retrograde cerebral perfusion by cannulating the superior vena cava may help flushing out embolism and supporting 'the cold jacket' of the brain. However, its effect of retrograde backflow cannot be a sign of adequate cerebral perfusion.

Coronary artery imaging with 64-slice computed tomography from cardiac surgical perspective.

INTRODUCTION: 64-Slice computed tomography (CT) has been introduced with high expectations. This study illustrates the value of 64-slice CT for the diagnosis of significant coronary artery stenoses when images are analysed by cardiovascular surgeons. METHODS: Fifty patients (39 males, 11 females) underwent invasive coronary angiography and 64-slice CT. In these patients, 40 had coronary artery disease and 10 patients had valvular disease. Evaluation of right coronary artery (RCA), left main (LM), left anterior descending artery (LAD), diagonal branch 1 (D1), circumflex branch (CX), and 1st marginal branch was performed by two cardiovascular surgeons. All vessels with a diameter >/=1.5 mm were analysed and a lumen restriction of >50% was considered a significant stenosis. CT image quality was classified as excellent, reduced but still diagnostic, and not assessable. Invasive coronary angiography was taken as gold standard for calculations of diagnostic accuracy. RESULTS: Mean heart rate during CT scan was 65+/-11 beats per minute (bpm). Image quality of 92% (506/550) of all segments was rated as excellent, 5% (27/550) were rated as being of reduced quality but still diagnostic, and 3% (17/550) were considered not assessable. The sensitivity for diagnosing a significant stenosis with CT when including all reliably evaluated segments was 93% (106/114), specificity was 97% (381/392), positive predictive value was 91% (106/117), and negative predictive value was 98% (381/389). CONCLUSION: 64-Slice CT provides a high diagnostic accuracy in assessing significant coronary artery stenosis. Nevertheless, still exist some disadvantages such as strong vessel wall calcifications reducing the reliability for image interpretation. At the moment, 64-slice CT should be used as a complementary imaging modality to invasive coronary angiography.

Cbfa-1 (Runx-2) and osteocalcin expression by human osteoblasts in heparin osteoporosis in vitro.

Heparin may cause adverse effects on bone formation following long-term application. The exact pathomechanism is unclear, but in vitro data suggest an impaired osteoblast function. The transcription axis of Cbfa-1 (Runx-2) and osteocalcin is crucial in maintaining an equilibrium of bone formation and resorption in vivo. We used a human osteoblast cell culture model to further investigate the effect of heparin (low-molecular-weight heparin, dalteparin) on the expression of these two regulators of osteoblast differentiation. At high doses, dalteparin caused a significant inhibition of both osteocalcin and Cbfa-1 expression in vitro. Our data support the hypothesis of a direct inhibition of osteoblast function underlying heparin osteoporosis.

Sixteen-channel multidetector row computed tomography versus coronary angiography in a surgical view.

BACKGROUND: Invasive coronary angiography (ICA) is the gold standard for the diagnosis of coronary artery disease and also for imaging procedures for preoperative planning of coronary artery bypass grafting (CABG). Sixteen-multidetector row computed tomography (MDCT) represents an alternative depiction of coronary vessels. METHODS: Preoperative exams included ICA and MDCT in 50 patients. Two blinded surgical readers independently investigated both diagnostic modalities regarding location, severity, and morphology of the stenoses. The right coronary artery, left anterior descending branch, and circumflex branch--each divided in 3 sections--and the left main artery with a diameter (3) 1.5 mm were rated in both procedures, and the percentage of complete evaluations by MDCT was assessed. RESULTS: Heart rate was 72 +/- 8 bpm. Forty-six percent of patients received a complete MDCT evaluation, and 54% received an incomplete MDCT evaluation. In 62% of these incompletely examined patients, 1 branch was not completely analyzable, in 31% 2 branches; and in 7% all 3 branches. In total, 9% of all segments were incompletely assessed. Investigators detected coronary stenoses in complete evaluations with a sensitivity of 94% and a specificity of 95%. Positive predictive value was 87% and negative predictive value was 98%. Plaque classification in soft and hard plaques was possible. CONCLUSION: Sixteen-MDCT is not a viable alternative diagnostic tool at present. However, although the percentage of incomplete evaluated patients is more then 50%, only 9% of all segments were incompletely assessable. If this technology can be further improved, especially its software, it will become a valid diagnostic tool for coronary artery disease.

Tissue engineering of functional trileaflet heart valves from human marrow stromal cells.

BACKGROUND: We previously demonstrated the successful tissue engineering and implantation of functioning autologous heart valves based on vascular-derived cells. Human marrow stromal cells (MSC) exhibit the potential to differentiate into multiple cell-lineages and can be easily obtained clinically. The feasibility of creating tissue engineered heart valves (TEHV) from MSC as an alternative cell source, and the impact of a biomimetic in vitro environment on tissue differentiation was investigated. METHODS AND RESULTS: Human MSC were isolated, expanded in culture, and characterized by flow-cytometry and immunohistochemistry. Trileaflet heart valves fabricated from rapidly bioabsorbable polymers were seeded with MSC and grown in vitro in a pulsatile-flow-bioreactor. Morphological characterization included histology and electron microscopy (EM). Extracellular matrix (ECM)-formation was analyzed by immunohistochemistry, ECM protein content (collagen, glycosaminoglycan) and cell proliferation (DNA) were biochemically quantified. Biomechanical evaluation was performed using Instron(TM). In all valves synchronous opening and closing was observed in the bioreactor. Flow-cytometry of MSC pre-seeding was positive for ASMA, vimentin, negative for CD 31, LDL, CD 14. Histology of the TEHV-leaflets demonstrated viable tissue and ECM formation. EM demonstrated cell elements typical of viable, secretionally active myofibroblasts (actin/myosin filaments, collagen fibrils, elastin) and confluent, homogenous tissue surfaces. Collagen types I, III, ASMA, and vimentin were detected in the TEHV-leaflets. Mechanical properties of the TEHV-leaflets were comparable to native tissue. CONCLUSION: Generation of functional TEHV from human MSC was feasible utilizing a biomimetic in vitro environment. The neo-tissue showed morphological features and mechanical properties of human native-heart-valve tissue. The human MSC demonstrated characteristics of myofibroblast differentiation.

Geometrical model of the pulmonary root.

BACKGROUND AND AIM OF THE STUDY: The renaissance of the Ross procedure has attracted much attention to the clinical anatomy of the pulmonary valve. Within the reviewed literature which emphasizes the morphology of the pulmonary root, no detailed anatomy has been reported. The study aim was to describe the cardiosurgical-orientated anatomy of the pulmonary valve. METHODS: Morphometric measurements were made on 20 fixed normal pulmonary roots. The roots were fixed under diastolic pressure (5 mmHg) with closed leaflets. The distances between the commissures, and between the intervalvular triangles, were measured. The heights of the sinuses and of the intervalvular triangles were also defined. RESULTS: The mean (+/- SD) of each parameter was calculated. The distances of the right, left and anterior sinus commissures were approximately similar, as were the distances of the intervalvular triangles at the root base. The mean height of the left coronary sinus was maximal, followed by the right and anterior pulmonary sinuses. The left intervalvular triangle was highest, followed by the posterior and right intervalvular triangles. The difference between the height of the left sinus and right intervalvular triangle indicated a tilt angle of 16.26 degrees between the root base and sinotubular junction. CONCLUSION: Based on these measured parameters, a spatial geometrical model was constructed which clearly described the asymmetrical structure of the pulmonary root.