Three-dimensional image analytical detection of intussusceptive pillars in murine lung.

A variety of diseases can lead to loss of lung tissue. Currently, this can be treated only symptomatically. In mice, a complete compensatory lung growth within 21 days after resection of the left lung can be observed. Understanding and transferring this concept of compensatory lung growth to humans would greatly improve therapeutic options. Lung growth is always accompanied by a process called angiogenesis forming new capillary blood vessels from preexisting ones. Among the processes during lung growth, the formation of transluminal tissue pillars within the capillary vessels (intussusceptive pillars) is observed. Therefore, pillars can be understood as an indicator for active angiogenesis and microvascular remodelling. Thus, their detection is very valuable when aiming at characterization of compensatory lung growth. In a vascular corrosion cast, these pillars appear as small holes that pierce the vessels. So far, pillars were detected visually only based on 2D images. Our approach relies on high-resolution synchrotron microcomputed tomographic images. With a voxel size of 370 nm we exploit the spatial information provided by this imaging technique and present the first algorithm to semiautomatically detect intussusceptive pillars. An at least semiautomatic detection is essential in lung research, as manual pillar detection is not feasible due to the complexity and size of the 3D structure. Using our algorithm, several thousands of pillars can be detected and subsequently analysed, e.g. regarding their spatial arrangement, size and shape with an acceptable amount of human interaction. In this paper, we apply our novel pillar detection algorithm to compute pillar densities of different specimens. These are prepared such that they show different growing states. Comparing the corresponding pillar densities allows to investigate lung growth over time.

Sensitivity and specificity of ultrasound in spinal trauma in 29 consecutive patients.

PURPOSE: Indication for surgery in spine trauma patients depends on the extent of destruction of the spine. Computer-assisted tomography scan (CAT scan) is not suitable to diagnose type B-injuries. Aim of the study was to investigate whether ultrasound is able to detect destruction of the posterior ligament complex (PLC). METHODS: Twenty-nine patients were included. The results of ultrasound were compared with magnetic resonance imaging (MRI), CAT scan, X-ray, intraoperative findings. Statistical analysis was carried out by an independent observer. RESULTS: In 27 cases both ultrasound and MRI had the same result. In two cases, ultrasound failed to detect ligamentous injury. The sensitivity of ultrasound was 0.82 (CI 0.48-0.98), its specificity: 1. MRI and Ultrasound findings had a strong positive correlation (phi = 0.85, Cohen's kappa: 0.85, with 95 % confidence interval 0.65-1) and a high significance (Fischer's exact test: p < 0.0001). CONCLUSION: Ultrasound may indicate rupture or integrity of PLC in cases where MRI is missing.

The interference of distal humeral plating with the medial and lateral collateral ligaments of the elbow.

INTRODUCTION: The aim of the present study was to determine the anatomical relationship and evaluate the potential interference of today's common distal humerus plates with the medial and lateral collateral ligaments of the elbow. MATERIALS AND METHODS: The elbow joints of 23 embalmed upper extremities were dissected. Three different brands of distal humerus double-plating systems were applied in a standardized fashion. We used a caliper to measure the amount of absolute overlap of the plates on the corresponding collateral ligaments. RESULTS: The data show contact and overlap with the medial and lateral collateral ligaments in all tested medial and lateral plates. The posterolateral and posteromedial plates showed no contact with the ligaments, yet they did contact the posterior joint capsules. The medial plates showed less contact/overlap when compared with the lateral and extended medial plates. CONCLUSION: Based on the present data, we conclude that distal humerus plating using the perpendicular technique with standard-sized medial plates shows the least amount of overlap over the medial and lateral collateral ligaments. The extent of the overlap of the ligaments by the humeral plates is clearly shown in the present study.

In vitro and in vivo characterization of porcine acellular dermal matrix for gingival augmentation procedures.

BACKGROUND AND OBJECTIVE: Recently, porcine acellular dermal matrix (PADM) has been proposed as a possible alternative to autogenous grafts in periodontal plastic surgery. The aim of the present study was to investigate the in vitro responses of four different oral cell lines cultured on a novel PADM. Furthermore, tissue reaction to PADM was evaluated histologically after subcutaneous implantation in mice. MATERIAL AND METHODS: Human gingival fibroblasts (HGF), human osteoblast-like cells, human umbilical vein endothelial cells and human oral keratinocytes (HOK) were cultured and transferred on to the PADM. A tissue culture polystyrene surface served as the control. The viability of all tested cell lines on PADM was measured by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetric assay and PrestoBlue(®) reagent. The ToxiLight(®) assay was performed to analyze the effect of PADM on adenylate kinase release. PADM was implanted into nude mice subcutaneously and subjected to histological analysis after 21 d. RESULTS: Using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetric assays, all tested cell lines cultured on PADM demonstrated a significant increase of viability compared to the control group (each p < 0.001) with the exception of HGF and HOK after 3 d (each p > 0.05). According to the PrestoBlue(®) analysis, all cell lines demonstrated a significant increase of viability compared to the control group at the particular points of measurement after 18 h (HGF p < 0.01; human osteoblast-like cells, human umbilical vein endothelial cells, HOK each p < 0.001). No significant cytotoxic effects of PADM on the tested cell lines could be observed, as assessed by changes in adenylate kinase release. Subcutaneous implantation of PADM into nude mice demonstrated good integration with surrounding tissues and significant revascularization of its collagen structure. CONCLUSION: Overall, the results suggest that PADM is a promising substitute for autogenous soft tissue grafts in periodontal surgery.

Bisphosphonates' antiangiogenic potency in the development of bisphosphonate-associated osteonecrosis of the jaws: influence on microvessel sprouting in an in vivo 3D Matrigel assay.

OBJECTIVES: Bisphosphonate-associated osteonecrosis of the jaw (BP-ONJ) is an adverse side effect of long-term bisphosphonate treatment. One theory of BP-ONJ etiology suggests a negative influence of these agents on angiogenesis and vascularization. This in vivo study analyzed the effects of bisphosphonates on angiogenesis in a 3D Matrigel assay. MATERIALS AND METHODS: Matrigel plugs were implanted into fifty 6-8-week-old female nude mice. Ten animals each were treated either with clodronate, ibandronate, pamidronate, zoledronate, or carrier solution as controls. The microvessel density (MVD), microvessel area (MVA), and microvessel size (MVS) in Matrigel plugs were analyzed after 21 days of treatment by immunohistochemistry and exemplary 3D microvascular corrosion castings. RESULTS: All bisphosphonates induced a statistically significant decrease of MVD (p each <0.001), whereby the nitrogen-containing bisphosphonates (N-BPs) demonstrated a clearly stronger effect than non-nitrogen-containing bisphosphonates (NN-BP) clodronate (control 166, clodronate 99, ibandronate 48, pamidronate 47, zoledronate 35 microvessels/mm(2)). Referring to MVA, similar results could be detected. MVS was significantly increased especially by ibandronate (103 %) compared to control group (p < 0.001). Scanning electron microscope scans of the corrosion castings confirmed these results. CONCLUSIONS: The stronger influence on MVD by N-BPs compared to the NN-BP clodronate may explain for the lack of BP-ONJ after treatment with NN-BPs. CLINICAL RELEVANCE: Ibandronate induced a strong increase of MVS. In combination with the reduced MVD, this could result only in a fractional reduced perfusion which might be an explanation for the lower occurrence of BP-ONJ in patients receiving ibandronate compared to patients receiving pamidronate or zoledronate.

[Risk of injury to children's fingers in power-operated motor vehicle windows]. / Verletzungsrisiko von Kinderfingern in automatischen Scheibenschliessanlagen von Kraftfahrzeugen.

BACKGROUND: This study examines the dimensions of children's fingers and the risk of jam injuries in a 4-mm gap between glass and gasket of power-operated motor vehicle windows. MATERIAL AND METHODS: The diameter of the proximal, middle, and distal phalanx and of the proximal and distal interphalangeal joint of each finger of the right hand of 160 children was measured in a cross-sectional investigation. Six different drawings in cross section of gaskets and glass window panes of current motor vehicle side door windows at a vertical gap of 4 mm were drawn in correct proportion. The larger actual width of the oblique gap between window glass and gasket was measured and related to the diameters of children's fingers. RESULTS: Almost all fingers and joints fit in the largest actual gap of 18 mm between glass and gasket of one seal design. CONCLUSION: The European guideline 74/60/EWG specifications currently pertaining to closing force restriction do not eliminate the risk of potentially serious injury to children's fingers in motor vehicle power windows.

Finger injuries caused by power-operated windows of motor vehicles: an experimental cadaver study.

The aim of this experimental cadaver study was to investigate which kinds of lesions could occur in jam events between the glass and seal entry of power-operated motor vehicle side door windows at two different closing forces. Ten hands of fresh cadaver specimens were used. Three different hand positions chosen to simulate real events in which a finger is jammed between the glass and seal entry of the window of a current motor vehicle were examined. The index, middle, ring, and little finger of each hand were separately jammed both at the proximal and distal interphalangeal joint at closing forces of 300 and 500 N with a constant window glass closing speed of 10 cm/s. Macroscopically visible injuries were documented and radiographs of all fingers were obtained in two standard planes. At a closing force of 300 N, contusion marks of the skin, palmar joint instabilities and superficial skin lesions occurred, whilst at 500 N superficial skin lesions, superficial and deep open crush injuries, and fractures were observed. The results of this study experimentally demonstrate the kinds of finger injuries that could be expected in real jam events between the glass and seal entry in automatic power-operated windows.

Biomechanical and histological evaluation of abdominal wall compliance with intraperitoneal onlay mesh implants in rabbits: a comparison of six different state-of-the-art meshes.

BACKGROUND: An ideal prosthetic mesh for incisional hernia repair should mimic the anisotropic compliance of the abdominal wall, and at lower loads should exhibit higher distensibility without impairment of safety at higher loads. This study evaluated the biomechanical properties of six meshes in a rabbit model. METHODS: New Zealand white rabbits were used for this study. Two meshes of the same brand (Ethicon Physiomesh™, Bard Composix(®) L/P, Gore Dualmesh(®), Bard Sepramesh(®), Ethicon Proceed(®) or Parietex™ Composite) were implanted into each animal for assessment of intra-abdominal hernia repair, with a total of ten meshes per group. Twelve weeks after implantation, the abdominal walls with ingrown meshes were harvested and examined biomechanically with a plunger test. The mesh-tissue compliance was evaluated by the forces exerted at given displacements and also described through a simple mathematical approximation. Abdominal wall samples were collected for histopathology, cell turnover and morphometry. RESULTS: No mesh-related complications were seen. The adhesion score was significantly higher in Bard Composix(®) L/P and Ethicon Proceed(®) meshes. Significant shrinkage was seen in Gore Dualmesh(®) and Parietex™ Composite meshes. Physiomesh™ exhibited the highest compliance during plunger testing, characterized by lower, more physiological reaction forces against tissue displacement than the competitor meshes. In contrast, the safety modulus was comparable in all groups. Histology showed less collagen and less foreign body reaction in the Physiomesh™ samples contributing to patient's comfort. CONCLUSION: In terms of safety, this study showed no superiority of any single mesh. The comfort modulus however differed, being lowest in the newly developed Physiomesh™.

Entrapment of adult fingers between window glass and seal entry of a motor vehicle side door: an experimental study for investigation of the force at the subjective pain threshold.

In modern motor vehicles with automatic power windows, a potential hazard exists for jam events of fingers between the window glass and seal entry. This study determined entrapment forces acting on adult fingers at the subjective maximum pain threshold during entrapment in such windows. The length and the girth of the proximal and distal interphalangeal joints of the triphalangeal fingers of the right hands of 109 participants (60 men, 49 women) were measured; the diameter was calculated from girth, which was assumed to be circular. The automatic power window system of a motor vehicle side door was changed to a mechanical system. During entrapment the force distributed across the four proximal interphalangeal joints (PIPs), and separately on the proximal interphalangeal (iPIP) and then the distal interphalangeal (iDIP) joints of the index finger was measured using a customized force sensor. The maximum bearable entrapment force was 97.2 ± 51.8 N for the PIPs, 43.4 ± 19.9 N for the iPIP, and 36.9 ± 17.8 N for the iDIP. The positive correlation between finger diameter and maximum entrapment force was significant. Particularly with regard to the risk to children's fingers, the 100 N statutory boundary value for closing force of electronic power windows should be reduced.

Antitumor effect of the vascular-disrupting agent ZD6126 in a murine renal cell carcinoma model.

ZD6126 is a vascular-disrupting agent that affects the endothelial tubulin cytoskeleton causing selective occlusion of tumor vasculature and extensive tumor cell necrosis. The present study evaluated the antitumor and antivascular activities of ZD6126 in the clinically relevant murine renal cell carcinoma (RENCA) model and also evaluated biological response to therapy using color Doppler imaging as biomarker. Mice were implanted with RENCA tumor cells (day 0) and established tumors were treated with ZD6126 (100 mg/kg i.p.) or vehicle with repeated intermittent doses on day 10, 14 and 18. ZD6126 treatment led to a significant reduction in tumor size and was associated with extensive tumor necrosis and a reduction in tumor blood flow versus controls. MVD increased with intermittent treatment (day 10, 14 and 18). In an additional study, animals were treated at day 19 and quantitative three-dimensional microvascular corrosion casting was performed to enable detailed assessment of the tumor vascular architecture. Corrosion casting showed that tumor vessel architecture is affected by treatment, whereas pre-existing vessels in control tissues are practically not affected. Inter-vessel and inter-branch distances as well as vessel diameters are influenced by treatment. In conclusion, ZD6126 showed potent antitumor efficacy in the RENCA model and our data suggest that decrease in tumor blood flow may be a useful surrogate marker of treatment effect.

Forces and deformations of the abdominal wall--a mechanical and geometrical approach to the linea alba.

Force-elongation responses of the human abdominal wall in the linea alba region were determined by tensile tests in which the linea alba was seen to exhibit a nonlinear elastic, anisotropic behavior as is frequently observed in soft biological tissues. In addition, the geometry of the abdominal wall was determined, based on MRI data. The geometry can be specified by principal radii of curvature in longitudinal of approximately 470 mm and in the transverse direction of about 200 mm. The determined radii agree with values found in other studies. Mechanical stresses, deformations and abdominal pressures for load cases above 6% elongation can be related using Laplace's formula and our constitutive and geometrical findings. Results from uni- and biaxial tensile tests can thus be compared using this model. Calculations confirm that abdominal pressures of approximately 20 kPa correspond to related biaxial forces of about 3.4N/mm in the transverse and 1.5 N/mm in the longitudinal direction. Young's moduli can be calculated with respect to the uniaxial as well as the biaxial loading. At these physiological loadings, a compliance ratio of about 2:1 between the longitudinal and transversal directions is found. Young's moduli of about 50 kPa occur in transversal direction and of about 20 kPa in longitudinal direction at transverse and longitudinal strains both in the order of 6%. These findings coincide with results from other investigations in which the properties of the abdominal wall have been examined.

Lengths, girths, and diameters of children's fingers from 3 to 10 years of age.

We obtained data on the lengths, girths, and diameters of the fingers of children from 3 to 10 years of age. A total of 160 children (78 girls, 82 boys) were examined in a cross-sectional investigation. The length of each finger of the right hand of every child was measured, as were the girths of the proximal, middle, and distal phalanges, and of the proximal and distal interphalangeal joint. The average length of the thumb was 49 (35-65) mm. The index and ring fingers both averaged 69 (index, 50-88; ring, 42-96) mm in length, while the middle and little fingers averaged 72 (57-100) and 56 (40-74) mm, respectively. Average diameter, calculated from the girth measurement, was 16 (11-22) mm for the thumb, 15 mm for both the index (9-22) and middle (10-21) fingers, 14 (10-20) and 13 (8-19) mm for the ring and little fingers, respectively. The average length of each finger increased by 37% from 3 to 10 years of age, average girth by 24%, and diameter increased by 20%. We observed no differences in length, girth, and diameter between the sexes. The dimensions of children's fingers are relevant to injuries from automatic, power-operated window lifters of motor vehicles because risk of injury to a finger jammed between an ascending window and the seal entry depends upon the diameter of the finger. Additionally, short fingers of young children can be jammed over almost their entire length in the oblique design of a car window seal entry.

Three-dimensional analysis of tumour vascular corrosion casts using stereoimaging and micro-computed tomography.

OBJECTIVE: In order to perform effective translational research for cancer therapy, we need to employ pre-clinical models which reflect the clinical situation. The purpose of this study was to quantitatively compare the vascular architecture of human colorectal cancer and experimental tumour models to determine the suitability of animal models for vascular studies and antivascular therapy. METHODS: In this study we investigated the three-dimensional properties of colonic tumour vasculature in both human clinical tissues (normal mucosa control [n=20], carcinoma [n=20] and adenoma [n=6]) and murine colorectal xenografts (LS147T [n=6] and SW1222 [n=6]). Scanning Electron Microscope Stereoimaging (SEM) and X-ray Micro-Computed Tomography (Micro-CT) methods were employed for 3D analyses of the vascular corrosion casts from these tissues. RESULTS: Morphological measurements showed that there were significant differences in the underlying morphology in the different tissues. Of the studied xenografts, LS147T is more consistently similar to the vascular architecture of the human carcinoma than SW1222. The only reversal of this is for the inter-vessel distance. CONCLUSION: While SEM stereoimaging provided better surface detailed resolution of the corrosion casts, it was complimented by the fully 3D micro-CT method. Comparison made between the xenografts and clinical tumours showed that the LS147T xenografts shared many similarities with the clinical tumour vasculature. This study provides insight into how to select the most suitable pre-clinical models for translational studies of clinical cancer therapy.

Dimensions of the myopectineal orifice: a human cadaver study.

BACKGROUND: In mesh implantations, there needs to be a balance between mass and the size of the meshes. However, the mesh size should allow for adequate prevention of hernia recurrence by sufficient coverage of the entire myopectineal orifice (MPO). AIM AND OBJECTIVE: The aim was to determine the MPO dimensions and several pelvic parameters. METHODS: Thirty-two inguinal regions were assessed in 16 embalmed cadavers. RESULTS: The MPO averaged 7.8 cm (+/-3.0) in width and 6.5 cm (+/-1.9) in height. The weak inguinal area cranially to the inguinal ligament was 4.5 cm (+/-1.7) high. We found significant gender differences: MPOs in males showed the same width and height (7.6 x 7.6 cm), whereas in females, the MPO width was greater than its height (8.1 x 5.3 cm). Noticeable correlations of the MPO parameters to constitutional parameters were found for body size versus the height of the MPO (r = 0.5005) and interspinous distance versus the height of the MPO (r = 0.7653). CONCLUSIONS: A mesh measuring 10 x 8 cm is suitable for both genders: in females, it will cover the whole MPO, including the infraligamental part, whereas in males, the weak inguinal area is preferentially covered.

Angiogenesis and vascular network of teratocarcinoma from embryonic stem cell transplant into seminiferous tubules.

BACKGROUND: Carcinoma in situ (CIS) of the testis is considered to be a precancerous germinal cell lesion, but the precise cellular and molecular mechanisms underlying transformation of CIS into invasive pluripotent cancer cells remain to be elucidated. Moreover, a satisfactory animal model for the experimental study of germinal tumours has not been developed to date. METHODS: We have developed a tumour model that involves the microinjection of green fluorescent protein-labelled embryonic stem (ES) cells (which are functionally equivalent to CIS cells) into syngenic mouse seminiferous tubules, a unique cell microenvironment in which germinal cells mature and CIS arise. To characterise the vascularisation of teratocarcinomas, which arise after cell transplant, we used immunohistochemistry, together with a qualitative and quantitative analysis of scanning electron microscopy images of corrosion casting samples. RESULTS: Embryonic stem cells transplanted into seminiferous tubules did not differentiate into germinal cells, but rather they behaved as invasive embryonal carcinoma (EC) stem cells. The vascular pattern of the experimental teratocarcinomas showed a highly disorganised architecture, and some of the neoplastic capillaries were derived, at least in part, from the original transplanted ES cells. CONCLUSION: The transplantation of pluripotent ES cells into seminiferous tubules efficiently recapitulates the early stages of development of teratocarcinomas. Consequently, this method constitutes a novel in vivo model to study the mechanisms of invasion and progression of experimental germinal tumours.

External stenting with a new polyester mesh reduces neointimal hyperplasia of vein grafts in a sheep model.

OBJECTIVE: External stents placed around vein grafts have demonstrated effectiveness in reducing neointimal hyperplasia by preventing distension of the thin-walled vein grafts when exposed to arterial pressure. However, the ideal stent material has yet to be defined. The following study investigates the short- and long-term effects of an innovative polyester mesh stent designed with optimized adaptation of circumferential compliance. METHODS: Following in vitro definition of the ideal macro-porous polyester stent material, a total of 12 sheep underwent implantation of bilateral carotid artery vein graft bypasses. In six sheep, the short term outcome (four weeks of implantation) was investigated by comparing the newly-designed stent to native veins, micro-porous PTFE stent grafts and metallic Biocompound stents (BCGs). Flow volume and graft diameter were measured prior to explantation. Grafts were evaluated histologically with respect to morphometry and immunoassaying. In the long-term group (6-month implantation time), the polyester stent was compared to native veins. RESULTS: All stents effectively prevented dilatation of the graft. Perfused vessel diameters of the polyester veins were 8.3+/-0.6 mm. BCG as well as PTFE veins showed diameter reduction to 7.4+/-0.7 mm (p<0.05) and 7.8+/-0.4 mm (p<0.05), respectively. Both in the short and long terms, the new polyester stent led to significantly higher reduction of neointimal hyperplasia and luminal encroachment compared to the native vessel. It proved superior to PTFE stenting, while the Biocompound material failed to prove efficacy. CONCLUSIONS: It was demonstrated that the new macro-porous polyester mesh stent reduced neointimal hyperplasia more effectively than other commercially available stents.

Protecting the endothelial integrity of internal thoracic arteries.

BACKGROUND: Previous functional studies on human internal thoracic arteries, comparing the effect of the traditional harvesting method (occlusion with a clip) with a method leaving the artery perfused, revealed considerably impaired endothelial function associated with enhanced contractility after clipping. We have now investigated whether these observations could be correlated (1) with plasma markers of endothelial dysfunction, and (2) with structural changes in the endothelial layer. METHODS: 32 patients were randomly distributed into groups of clipped and perfused arteries. Arterial blood samples were obtained from both the artery and extracorporeal circulation to determine sP-selectin, sE-selectin, sL-selectin, and thrombomodulin using enzyme-linked immunosorbent assay. Arteries from three patients were examined by scanning electron microscopy. RESULTS: Concentrations of sP-selectin and thrombomodulin were significantly higher in plasma from clipped arteries compared to perfused arteries, whereas sE-selectin and sL-selectin concentrations were similar within the groups. Scanning electron microscopy revealed significant structural changes and loss of endothelial cells in clipped arteries. CONCLUSION: Biochemical and structural results support our findings that leaving the internal thoracic artery perfused preserves endothelial function in the arterial graft.

[Angiogenesis VEGF165 gene therapy with AdVEGF- a new delay procedure for flaps]. / Angiogenese-Gentherapie mit AdVEGF.

A regular tissue functioning requires the adequate supply of oxygen and nutrient via blood vessels. The sequences of formation and maturation of vessels are initiated and maintained by different growth factors. The VEGF growth factor plays an exceptional role in these mechanisms. The creation of sublethal ischemia as an angiogenic stimulus known as "Delay" is a well established procedure in plastic surgery, although the underlying molecular biological mechanisms still remain unknown. The important role of VEGF and its regulation depending on oxygen pressure suggest a strong connection between this growth factor and the delay phenomenon. The VEGF concentration in skin and underlying muscle was measured in overdimensioned random pattern flaps on 32 male Sprague-Dawley rats after either VEGF gene therapy or circumcision without elevation of the flap and compared to controls. Additional random pattern flaps were raised seven days post gene therapy or delay. The effect on the flap perfusion was measured postoperatively using Indocyanine green Laser Fluoroscopy and the size of the surviving and necrotic areas of the flaps were analysed. The skin of the random pattern flaps showed both in the Delay group and in the VEGF gene therapy group a significantly elevated VEGF concentration compared to the controls. The underlying rectus abdominis muscle showed no significant differences in VEGF concentration between the groups. The flap perfusion postoperatively was significantly increased solely in the VEGF gene therapy group. The analysis of the surviving area of the flaps showed a significant increase over the controls in the gene therapy group. The Delay procedure results in a significantly and locally raised concentration of the VEGF growth factor. The gene therapeutical use of this growth factor allows us to raise flap perfusion and to reduce necrosis. Both VEGF gene therapy and Delay seem to promote similar mechanisms whereas the gene therapy produced superior results in this setting.

[Microvessel visualisation: applications and limitations]. / Möglichkeiten und Grenzen der Mikrogefässdarstellung.

Vessel systems and the microvascular unit may be studied by a variety of morphological techniques which enable improved structure-function analyses. Two-dimensional light and electron microscopic sectioning techniques with and without specific markers are complemented by 3D vascularisation and perfusion study techniques. Static ex vivo modalities still prevail, however, the future will be dominated by intravital high-resolution techniques.

AdVEGF165 gene transfer increases survival in overdimensioned skin flaps.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a key regulator of angiogenesis. VEGF A also plays an important role in wound healing of the skin by promoting angiogenesis and by stimulating blood vessel growth. Therefore we tested the hypothesis that flap survival could be increased by the preoperative injection of AdVEGF(165). METHODS: We studied the effect of AdVEGF(165) in an overdimensioned ischemic random-pattern-flap model in the rat (n = 50) with a length-to-width ratio of 4 : 1. VEGF cDNA was administered in two concentrations of 5 x 10(8) plaque-forming units (pfU) and 1 x 10(9) pfU using a recombinant adenoviral vector. Recombinant virus was injected subdermally 7, 3 or 0 days prior to flap harvest for the lower concentration and 7 days prior for the higher concentration. Flap survival and necrosis were observed at day 7, the day the animals were sacrificed. RESULTS: Adenoviral gene transfer with VEGF(165) 3 and 7 days before flap harvest showed a significantly increased flap survival of 50% together with a significantly reduced necrosis (p < 0.01). Injection using a titer of 1 x 10(9) pfU 7 days prior to surgery increased flap survival even more, though failing to reach statistical significance compared to the lower concentration. VEGF protein concentration in the injected skin was significantly higher than in controls (p < 0.01). Flap perfusion was increased as well, demonstrated by indocyanine green (ICG) fluoroscopy (p < 0.001). CONCLUSIONS: Our results confirm the important role of VEGF(165) on angiogenesis in ischemic flaps. Indeed by injecting VEGF(165) at 3 to 7 days preoperatively in a concentration of 1 x 10(9) pfU our data show that length-to-width ratio for random-pattern-flaps could be increased from 2 : 1 to 3 : 1 and therefore may allow a wider range of applications of this simple flap technique.