Endocultivation: continuous application of rhBMP-2 via mini-osmotic pumps to induce bone formation at extraskeletal sites.

The continuous presence of recombinant human bone morphogenetic protein 2 (rhBMP-2) inside a scaffold may be crucial to the outcome in bone tissue engineering. This study investigated whether the release of the growth factor rhBMP-2 via different continuous application schemes influences histomorphological aspects of the hard and soft tissues induced. Three-dimensionally printed hydroxyapatite scaffolds were implanted into one latissimus dorsi muscle of 42 female Lewis rats. Simultaneously implanted mini-osmotic pumps were used to provide a continuous application of rhBMP-2 over 1, 2, or 4 weeks (total dose 200µg). A reference group received rhBMP-2 at the time of implantation only, and a control group received only block implantation. Bone density and histological examinations were performed after 8 weeks. No significant difference in bone density was found between the groups; however, the blood vessel count differed significantly between the groups receiving continuous treatments and both the control group and simultaneous rhBMP-2 treatment group (P<0.0001). Soft tissue types were distributed differently among the study groups. RhBMP-2 application via mini-osmotic pumps is as suitable for inducing bone formation as a single application at the time of implantation. The time interval over which rhBMP-2 was administered had no impact on the amount of new bone formation, probably due to the study duration and low local concentrations of growth factor.

Endocultivation: the influence of delayed vs. simultaneous application of BMP-2 onto individually formed hydroxyapatite matrices for heterotopic bone induction.

When bone morphogenetic protein (BMP) is delivered to matrices in vivo may affect tissue engineered bone constructs for jaw reconstruction after cancer surgery. This study compared the effects of BMP application at different times after matrix implantation for heterotopic bone induction in a rat model. Hydroxyapatite blocks were implanted unilaterally onto the surface of the latissimus dorsi muscle. A second block was implanted onto the contralateral muscle after 1, 2 or 4 weeks and 200 µg rhBMP-2 was injected into the blocks on both sides. Bone formation and density inside the blocks was analysed by CT and histology. 8 weeks after BMP application increases in bone density within the scaffolds were most pronounced in the simultaneous application group (179 HU). Less pronounced increases were observed for the 1 (65 HU), 2 (58 HU) and 4 (31 HU; p<0.0001) week delay group. Homogeneous bone induction started from the central channel of the blocks. Capillaries and larger vessels were seen in all constructs, samples receiving delayed BMP treatment demonstrated significantly greater neovascularization. Delayed application of BMP was less effective for heterotopic bone formation than simultaneous application. A central channel allows homogeneous bone induction directly from the centre of the blocks.

Comparison of vacuum and conventional wound dressings for full thickness skin grafts in the minipig model.

The aim of this study was to compare vacuum and conventional dressings and to follow revascularization with laser-Doppler spectroscopy and histological evaluations. In six minipigs, full thickness skin grafts were harvested on both sides of the back and transplanted to the contralateral site. One site was covered with a classical cotton dressing, the other with a vacuum dressing. For 10 days, oxygen, haemoglobin, flow and velocity levels were measured by laser-Doppler spectroscopy. Data were analyzed with ANOVA. Histological evaluation included haematoxylin and eosin (H&E) as well as CD31 immunohistochemical staining. Oxygen levels were significantly reduced in the vacuum dressing group compared with the classic dressing group during the first 3 days after transplantation. Haemoglobin levels were slightly, but significantly, higher in the vacuum dressing group over the whole observation period. On the second and third day after transplantation single capillaries were detectable in the histological evaluation. Starting from the fourth day, capillary number increased. Similar results for the classical pressure dressing and the vacuum dressing were observed. The present experimental model provides a standardized and reliable test system for evaluating revascularization of full thickness skin grafts in conjunction with growth factors and other enhancers of revascularization.

Life-threatening bite injury of the lower lip after use of metamizole.

INTRODUCTION: Bite wounds of the oral mucosa heal after eliminating the causative irritant, but there are serious exceptions from the rule. We present the case of a 37-year-old woman with an ulcer of the mucosa of the lower lip, which had been present for 10 days, and leucopenia. DISCUSSION: Agranulocytosis after the use of metamizole is part from leukaemia and lues, a rare reason for non-healing ulcers of the mucosa of the oral cavity without fulminant signs for inflammation. CONCLUSION: As this is a life-threatening disease, medical therapy must begin immediately.

Inducible HSP70 levels in thoracic wound fluid indicate myocardial damage after open heart surgery.

AIM: Heat shock proteins (HSP) act as chaperones and protect cellular homeostasis. They are induced by multiple forms of cell stress. Nothing is known about their rise in the drainage fluid after cardiac and thoracic operations. METHODS: Forty-five patients undergoing open heart surgery or major lung resections were studied. Levels of inducible HSP70 were measured daily in thoracic wound fluid and blood with ELISA tests in addition to multiple conventional blood and wound fluid parameters. Data were analyzed using analysis of covariance and Spearman's rank-correlation coefficient. RESULTS: Inducible HSP70 was detectable in all thoracic fluid samples. Mean HSP70-level after open heart surgery was 64 ng/mL and 40 ng/mL in patients without extracorporeal circulation (P=0.015). Drainage HSP70 levels were correlated with white blood cell counts in wound fluid (P<0.0001) and blood CK-MB levels (P<0.0001). Correlation coefficients between white blood cells in blood and local HSP-levels were explicitly lower (P=0.0023). There was a steady decrease in HSP levels in drainage fluid from day 1 to day 4 (P<0.0001). CONCLUSIONS: Inducible HSP70 is present in postoperative thoracic drainage fluid. Drainage levels are higher in patients after open heart surgery and are correlated with blood CK-MB levels, indicating cardiomyocyte damage. Our data further suggest that extracellular HSP70 is released also by white blood cells. Additional studies are needed to examine the role of extracellular HSP70 in wound healing processes and myocardial damage.

Reaction time in automated kinetic perimetry: effects of stimulus luminance, eccentricity, and movement direction.

PURPOSE: To determine the effects of stimulus eccentricity and luminance level on the reaction time (RT) of young normal volunteers during automated kinetic campimetry. METHODS: We used a specially designed video-campimetric device equipped with a continuous infrared (IR) pupillographic fixation control (Tübingen Computer Campimeter) and recorded reaction times upon presenting horizontally moving small circular stimuli (size 26'; constant angular velocity 2 degrees /s) starting at 16 locations within the central 30 degrees -radius of the visual field. Two different levels of stimulus luminance were used (41.6 cd/m(2) and 110 cd/m(2)), while background luminance was 10 cd/m(2). Each stimulus was presented a total of six times in a randomized order. Subjects were 12 healthy young individuals (aged 21-30 years) with normal ophthalmic examinations. An analysis of variance (ANOVA) was performed on the data. RESULTS: RTs showed considerable inter- and intra-individual variation with individual least squares means (LSM, fitted values of a linear model) ranging from 305 to 454 ms, and residual standard deviation (R.S.D.) 66 ms. Reaction times did not differ significantly as a function of stimulus direction (P>0.6). Higher luminance levels produced significantly reduced reaction times for all stimulus locations and directions (mean reduction: 16 ms; P<0.0001). Reaction times increased with increasing eccentricity, in the mean by 1.8 ms per degree of visual angle, from 365+/-4 ms (S.E.M.) foveally, to 407+/-2 ms at 30 degrees eccentricity; (P<0.0001). CONCLUSIONS: Automated kinetic perimetry should be designed to cope with significant, variable interindividual response characteristics. Other stimulus related factors, such as eccentricity or luminance level, have a significant but comparatively small effect on reaction time within the central 30 degrees -radius visual field in healthy young individuals.