Biomedical subjects

L Thiele

Publications and source records attributed to L Thiele.

4 recordsLinked to original sources

PubMed2001

Evaluation of particle uptake in human blood monocyte-derived cells in vitro. Does phagocytosis activity of dendritic cells measure up with macrophages?

This work focuses on microparticles as potential antigen delivery systems to target professional antigen-presenting cells. Surface modified polystyrene microparticles were administered to human-derived macrophages (MPhis) and dendritic cells (DCs) in vitro to evaluate the phagocytosis activity of each cell type. To discriminate between internalised particles and those closely attached to the outside of the cells, particle internalisation was verified by confocal laser scanning microscopy. Especially positively charged particles tend to stick to the outer cell membrane and may lead to false positive results when measured by conventional microscopy. In contrast, fluorescence microscopy in combination with an extracellular fluorescence quenching agent (trypan blue) allows the unequivocal assessment of particle uptake for screening purposes. For this assay, the fluorescent label needs to be in direct contact to the quenching agent and cannot be localised inside the particle core. Different types of microparticles varying in size, surface-material and zeta potential resulted in vast differences regarding their uptake by MPhis and DCs as well as the maturation of DCs. Negatively-charged carboxylated and bovine serum albumin-coated particles were phagocytosed by MPhis to a relatively small extent. Interestingly, phagocytosis of these particles was still significantly lower in DCs while positively charged poly-L-lysine (PLL) coated particles induced high phagocytosis activity in both cell types. By comparing our results with literature data, we conclude that phagocytosis activity of DCs and MPhis largely depends on particle size and surface charge and is also influenced by the character of bulk and coating material. PLL can be directed to DCs and MPhis with comparable efficiency and, in addition, induce maturation of DCs.

Antigens, CD↗

PubMed2001

Hydrophilic poly(DL-lactide-co-glycolide) microspheres for the delivery of DNA to human-derived macrophages and dendritic cells.

Biodegradable poly(lactide-co-glycolide) (PLGA) microspheres have a proven track record for drug delivery and are suggested to be ideal carrier systems to target therapeutics into phagocytic cells such as macrophages (MPhis) and dendritic cells (DCs). Microspheres prepared by spray-drying from different PLGA-type polymers were evaluated regarding their effect on phagocytosis, intracellular degradation and viability of human-derived macrophages MPhis and DCs. Even the microspheres prepared from the most hydrophilic polymer RG502H, were efficiently phagocytosed by primary human MPhis and DCs. Interestingly, uptake of PLGA microspheres by DCs as potent immune modulator cells was almost as efficient as uptake by the highly phagocytic MPhis. Phagocytosed microspheres remained inside the cells until decay with none of the microsphere preparations induced significant apoptosis or necrotic cell death. Acidic pH and the phagosomal environment inside the cells enhanced microsphere decay and release of encapsulated material. Degradation of microspheres consisting of the most hydrophilic PLGA polymer RG502H occurred in a reasonable time frame of less than 2 weeks ensuring the release of encapsulated drug during the life span of the cells. To explore important technical and biological aspects of DNA microencapsulation, we have studied DNA loading and in vitro DNA release of microspheres from different PLGA type polymers. Hydrophobicity and molecular weight of the PLGA polymers had profound influence on both the encapsulation efficiency of DNA and its release kinetics in vitro: the hydrophilic polymers showed higher encapsulation efficiency and faster release of intact DNA compared to the hydrophobic ones. These results suggest that microspheres from the PLGA polymer RG502H have improved characteristics for DNA delivery to human MPhis and DCs.

Cells, Cultured↗

PubMed2001

The effect of working tip angulation on root substance removal using Er:YAG laser radiation: an in vitro study.

OBJECTIVES: The present investigation attempted to determine the amount of cementum and/or dentin removal with Er:YAG laser radiation, dependent on the angulation of a specially-developed application tip. MATERIAL AND METHODS: Subject of the study were 150 extracted incisors, canines, premolars, and molars. Source of laser radiation was an Er:YAG laser device emitting pulsed infrared radiation at a wavelength of 2.94 microm with a pulse duration of 250 micros and a pulse repetition rate of 10 pps. The samples of the 3 study groups were irradiated with 380 laser pulses at radiation energies of 60 mJ (group A), 100 mJ (group B), or 180 mJ (group C). In each group, 10 samples were treated at working tip angulations of 15 degrees , 30 degrees , 45 degrees , 60 degrees , and 90 degrees. The substance removal was determined 3-dimensionally using a newly developed laser scanning system (100,000 surface points per sample, accuracy 5 microm) and a special image-analysing software (Match 3D). Statistical analysis was completed with ANOVA followed by multiple comparisons using the Scheffé-test and with linear regression analysis according to Pearson-Bravais (p < 0.05). RESULTS: Strong dependence of substance removal, both determined as maximum depth of the defects (0.5% quantil) as well as defect volume, on the angulation of the working tip was shown. At 60 mJ, the depth of the defects was 41.39 (+/- 32.55) microm at an angulation of 15 degrees and that at 90 degrees was 181.39 (+/- 74.42) microm (R2= 0.921). For the radiation energy at 100 mJ, the depth of the defects ranged from 51.96 (+/- 26.86) microm at 15 degrees to 306.64 (+/- 62.44) microm at 90 degrees (R2 = 0.983). Choosing radiation energies at 180 mJ, the depth of the defects ranged from 64.73 (+/- 27.73) microm at 15 degrees to 639.89 (+/- 47.28) microm at 90 degrees , on average (R2 = 0.853). CONCLUSIONS: The results of the present study provide clear evidence that besides the physical radiation parameters, also the parameters of clinical handling, in particular the angulation of the application tip, has a strong influence on the amount of root substance removal using Er:YAG laser radiation.

Aluminum Silicates↗

PubMed2000

Comparison of multiobjective evolutionary algorithms: empirical results.

In this paper, we provide a systematic comparison of various evolutionary approaches to multiobjective optimization using six carefully chosen test functions. Each test function involves a particular feature that is known to cause difficulty in the evolutionary optimization process, mainly in converging to the Pareto-optimal front (e.g., multimodality and deception). By investigating these different problem features separately, it is possible to predict the kind of problems to which a certain technique is or is not well suited. However, in contrast to what was suspected beforehand, the experimental results indicate a hierarchy of the algorithms under consideration. Furthermore, the emerging effects are evidence that the suggested test functions provide sufficient complexity to compare multiobjective optimizers. Finally, elitism is shown to be an important factor for improving evolutionary multiobjective search.

Algorithms↗