Martin Wehner

Laser‐mediated fixation of collagen‐based scaffolds to dermal wounds

Collagen scaffolds are popular for the reconstitution of dermal equivalents. Usually, these scaffolds are fixed with sutures or staples and in many cases these devices have to be removed in a second procedure. Laser‐mediated tissue welding in a wet environment is a potential alternative for collagen scaffold fixation and may be advantageous to suture, staple, and tissue glue fixation.

Morphological effects of nanosecond- and femtosecond-pulsed laser ablation on human middle ear ossicles

We evaluate the feasibility of nanosecond-pulsed and femtosecond-pulsed lasers for otologic surgery. The outcome parameters are cutting precision (in micrometers), ablation rate (in micrometers per second), scanning speed (in millimeters per second), and morphological effects on human middle ear ossicles. We examine single-spot ablations by a nanosecond-pulsed, frequency-tripled Nd:YAG laser (355 nm, beam diameter 10 microm, pulse rate 2 kHz, power 250 mW) on isolated human mallei. A similar system (355 nm, beam diameter 20 microm, pulse rate 10 kHz, power 160-1500 mW) and a femtosecond-pulsed CrLi:SAF-Laser (850 nm, pulse duration 100 fs, pulse energy 40 microJ, beam diameter 36 microm, pulse rate 1 kHz) are coupled to a scanner to perform bone surface ablation over a defined area. In our setups 1 and 2, marginal carbonization is visible in all single-spot ablations of 1-s exposures and longer: With an exposure time of 0.5 s, precise cutting margins without carbonization are observed. Cooling with saline solution result is in no carbonization at 1500 mW and a scan speed of 500 mms. Our third setup shows no carbonization but greater cutting precision, although the ablation volume is lower. Nanosecond- and femtosecond-pulsed laser systems bear the potential to increase cutting precision in otologic surgery.

Laser processing of components for polymer mircofluidic and optoelectronic products

Miniaturization is one of the keywords for the production of customer oriented and highly integrated consumer products like mobile phones, portables and other products from the daily life and there are some first silicon made products like pressure sensors, acceleration sensors and micro fluidic components, which are built in automobiles, washing machines and medical products. However, not all applications can be covered with this material, because of the limitations in lateral and 3-dimensional structuring, the mechanical behavior, the functionality and the costs of silicon. Therefore other materials, like polymers have been selected as suitable candidates for cost effective mass products. This holds especially for medical and optical applications, where the properties of selected polymers, like biocompatibility, inert chemical behavior and high transparency can be used. For this material laser micro processing offers appropriate solutions for structuring as well as for packaging with high flexibility, material variety, structure size, processing speed and easy integration into existing fabrication plants. The paper presents recent results and industrial applications of laser micro processing for polymer micro fluidic devices, like micro analysis systems, micro reactors and medical micro implants, where excimer radiation is used for lateral structuring and diode lasers have used for joining and packaging. Similar technologies have been applied to polymer waveguides to produce passive optoelectronic components for high speed interconnection with surface roughness less than 20 nm and low attenuation. The paper also reviews the technical and economical limitations and the potential of the technology for other micro products.

Surface Characterization and Short-term Adhesion to Zirconia after Ultra-short Pulsed Laser Irradiation

PURPOSE To evaluate the suitability of an ultra-short pulsed laser (USPL) to treat zirconia ceramic surfaces and increase their adhesion to dual-curing resin cement. MATERIALS AND METHODS Twenty 10 × 10 × 5 mm³ blocks were prepared from a zirconia ceramic (Y-TZP). The specimens were polished and randomly assigned to four groups (n = 5) which received the following surface treatments: sandblasting (SB) with Al₂O₃ particles and silica coating (SC) with SiO₂ particles as positive controls; two groups received USPL irradiation, one with 10 scan repetitions (L10) and the other with 20 (L20). Laser irradiation was performed at 1030 nm, 2.3 J/cm², 6 ps pulse duration. The ceramic blocks were duplicated in composite resin and cemented with a dual-curing resin cement. Half of the blocks were then stored in water (37°C) for 24 h and the other half for 1 month. At each time, 40 to 60 sticks per group were subjected to microtensile bond strength testing. Data were analyzed statistically using the Kruskal-Wallis test (α = 0.05). RESULTS Laser-treated zirconia presented statistically significantly higher roughness than did SB and SC. After 24 h, the highest bond strength means (MPa) were achieved by L10 (42.3 ± 10.8) and L20 (37.9 ± 14.4), and both of them were statistically significantly higher than SB (22.0 ± 5.3) and SC (20.8 ± 7.1) (p < 0.05). After 1 month of storage, L10- and L20-treated zirconia still showed significantly higher bond strengths than did SB- and SC-treated zirconia (p < 0.05). CONCLUSION USPL irradiation significantly increases bond strength of zirconia ceramic to dual-curing resin cement and might be an alternative for improving adhesion to this material.

Advantages and drawbacks of Thiol-ene based resins for 3D-printing

The technology of 3D printing is conquering the world and awakens the interest of many users in the most varying of applications. New formulation approaches for photo-sensitive thiol-ene resins in combination with various printing technologies, like stereolithography (SLA), projection based printing/digital light processing (DLP) or two-photon polymerization (TPP) are presented. Thiol-ene polymerizations are known for its fast and quantitative reaction and to form highly homogeneous polymer networks. As the resins are locally and temporally photo-curable the polymerization type is very promising for 3D-printing. By using suitable wavelengths, photoinitiator-free fabrication is feasible for single- and two photon induced polymerization. In this paper divinyl ethers of polyethylene glycols in combination with star-shaped tetrathiols were used to design a simple test-system for photo-curable thiol-ene resins. In order to control and improve curing depth and lateral resolution in 3D-polymerization processes, either additives in chemical formulation or process parameters can be changed. The achieved curing depth and resolution limits depend on the applied fabrication method. While two-/multiphoton induced lithography offers the possibility of micron- to sub-micron resolution it lacks in built-up speed. Hence single-photon polymerization is a fast alternative with optimization potential in sub-10-micron resolution. Absorber- and initiator free compositions were developed in order to avoid aging, yellowing and toxicity of resulting products. They can be cured with UV-laser radiation below 300 nm. The development at Fraunhofer ILT is focusing on new applications in the field of medical products and implants, technical products with respect to mechanical properties or optical properties of 3D-printed objects. Recent process results with model system (polyethylene glycol divinylether/ Pentaerithrytol tetrakis (3-mercaptopropionat), Raman measurements of polymer conversion and surface modifications using bifunctional crosslinkers are presented with advantages, drawbacks and a general outlook.

New stereolithographic resin providing functional surfaces for biocompatible three-dimensional printing:

Stereolithography is one of the most promising technologies for the production of tailored implants. Within this study, we show the results of a new resin formulation for three-dimensional printing which is also useful for subsequent surface functionalization. The class of materials is based on monomers containing either thiol or alkene groups. By irradiation of the monomers at a wavelength of 266 nm, we demonstrated an initiator-free stereolithographic process based on thiol-ene click chemistry. Specimens made from this material have successfully been tested for biocompatibility. Using Fourier-transform infrared spectrometry and fluorescent staining, we are able to show that off-stoichiometric amounts of functional groups in the monomers allow us to produce scaffolds with functional surfaces. We established a new protocol to demonstrate the opportunity to functionalize the surface by copper-catalyzed azide-alkyne cycloaddition chemistry. Finally, we demonstrate a three-dimensional bioprinting concept for the production of potentially biocompatible polymers with thiol-functionalized surfaces usable for subsequent functionalization.

Two-wavelength approach for control of coagulation depth during laser tissue soldering

In laser tissue soldering (LTS) protein solutions are used for closing of incisions or fixation of wound dressings. During coagulation and thermal denaturation of the protein solutions their morphology changes significantly such that light is strongly scattered. When scattering becomes major component extinction increases and the optical penetration depth shrinks which could lead to unsufficient coagulation and bonding. For adaption of extinction during coagulation we are investigating a two-wavelength approach. A strongly absorbed laser wavelength (1540 nm) and weakly absorbed wavelength (980 nm) can be applied simultaneously. Simulation of beam propagation is performed in natural and coagulated state of the solder. The model describes a three-layer system consisting of membrane, solder and phantom. The optical properties are determined by spectrometric measurements both in natural and coagulated state. The absorption coefficient μa, scattering coefficient μs and anisotropy factor γ are determined by numerical analysis from the spectrometric data. Beam propagation is simulated for 980 nm and 1540 nm radiation with ZEMAX® software based on the Monte Carlo method. For both wavelengths the beginning of the process with a clear solder layer, and the final state characterized by a coagulated solder layer are examined. The optical penetration depth depends mainly on the optical properties of the solder, which change in the course of coagulation process. The coagulation depth can be varied between 1.5 mm to 3.5 mm by changing the proportion of both laser sources. This leads to concepts for minimizing heat input while maintaining a constant coagulation depth.

Light-Weight Optical Sensor for Standoff Detection of Fluorescent Biosensors

Cell-based biosensors can be modified to express fluorescent proteins when in contact with trace substances like TNT. Stand-off detection of activated fluorescent biosensors becomes feasible by optical sensors which shine excitation light on the biosensor and collects the fluorescence light. We propose a low cost, light-weight sensor solution which consists of a LED illuminator, a large aperture light collector and a solid state detector. The irradiance level and the detection efficiency are expected to be sufficient for short range detection.

Laser-mediated perforation of plant cells

The functional analysis of plant cells at the cellular and subcellular levels requires novel technologies for the directed manipulation of individual cells. Lasers are increasingly exploited for the manipulation of plant cells, enabling the study of biological processes on a subcellular scale including transformation to generate genetically modified plants. In our setup either a picosecond laser operating at 1064 nm wavelength or a continuous wave laser diode emitting at 405 nm are coupled into an inverse microscope. The beams are focused to a spot size of about 1.5 μm and the tobacco cell protoplasts are irradiated. Optoporation is achieved when targeting the laser focal spot at the outermost edge of the plasma membrane. In case of the picosecond laser a single pulse with energy of about 0.4 μJ was sufficient to perforate the plasma membrane enabling the uptake of dye or DNA from the surrounding medium into the cytosol. When the ultraviolet laser diode at a power level of 17 mW is employed an irradiation time of 200 - 500 milliseconds is necessary to enable the uptake of macromolecules. In the presence of an EYFP encoding plasmid with a C-terminal peroxisomal signal sequence in the surrounding medium transient transformation of tobacco protoplasts could be achieved in up to 2% of the optoporated cells. Single cell perforation using this novel optoporation method shows that isolated plant cells can be permeabilized without direct manipulation. This is a valuable procedure for cell-specific applications, particularly where the import of specific molecules into plant cells is required for functional analysis.

Microsurgical anastomosis of sperm duct by laser tissue soldering

Connection of small vessels is usually done by suturing which is very cumbersome. Laser tissue soldering can circumvent that obstacle if a handy procedure can be defined. Our principle approach consists of a bioresorbable hollow stent with an expected degradation time of 3 weeks in combination with laser soldering. The stent is to be fed into the vessel to stabilize both ends and should allow percolation immediately after joining. The stents are made of Poly(D,L-lactid-co-glycolid) and solder is prepared from bovine serum albumin (BSA) doped with Indocyanine green (ICG) as chromophore to increase the absorption of laser light. After insertion, solder is applied onto the outer surface of the vessel and coagulated by laser radiation. The wavelength of 810 nm of a diode laser fits favorably to absorption properties of tissue and solder such that heating up of tissue is limited to prevent from necrosis and wound healing complications. In our study the preparation of stents, the consistency and doping of solder, a beam delivery instrument and the irradiation conditions are worked out. In-vitro tests are carried out on sperm ducts of Sprague-Dowlae (SD) rats. Different irradiation conditions are investigated and a micro-optical system consisting of a lens and a reflecting prism to ensure simultaneous irradiation of front and back side of the vessels tested. Under these conditions, the short-term rupture strength of laser anastomosis revealed as high as those achieved by suturing.