V. N. Bagratashvili

Laser shaping of composite cartilage grafts

INTRODUCTION The restoration of malformations of different etiology in the head and neck area continue to be a problem to the reconstructive surgeon. Many of these problems are created by the destruction of cartilage. The success of the reconstructive effort very often depends on the selection of a composite cartilage graft of proper size, shape, and thickness, which has to replace the missing cartilage. Despite the best of surgeons intentions, the postoperative result is not always satisfactory due to the difficulty in obtaining a cartilage of the proper shape. Using a carbon dioxide laser, composite cartilage samples 0.4 to 1 mm thick taken from rabbits ears were irradiated. METHODS Rabbit ear cartilage with covering epithelium was used. The thickness of the composite graft measured 0.4 to 1 mm. Specimens were reshaped, treated with a carbon dioxide laser, then immersed in saline. CONCLUSION It was observed that it was possible to change the shape of the cartilage, which then had the tendency to retain its new form for several days. Thicker composite grafts retained the new shape more satisfactorily. The significance of this experiment for future corrective surgery in various parts of the head and neck area is evident. We anticipate that this technique may be useful to mold grafted cartilage for use in complex reconstructions such as nasal, auricular, and tracheal deformities.

Speckle-contrast monitoring of tissue thermal modification

Measurements of the contrast value of time-averaged speckle-modulated images of cartilage tissue are used to study tissue thermal modification in the case of laser-light treatment. This modification is related to thermally induced internal stress relaxation in the matrix of the treated tissue. The specific feature of the evolution of time-averaged speckle contrast with a change in the current temperature of modified collagen tissue is the typical looplike form of the contrast-temperature dependencies associated with irreversible changes in tissue structure and correlated with changes in the tissue diffuse transmittance and the tissue internal stress mentioned by other researchers.

Osteoblast growth on titanium foils coated with hydroxyapatite by pulsed laser ablation.

Pulsed laser ablation is a new method for deposition of thin layers of hydroxyapatite (HA) on to biomaterial surfaces. In this paper, we report activity and morphology of osteoblasts grown on HA surfaces fabricated using different laser conditions. Two sets of films were deposited from dense HA targets, at three different laser fluences: 3, 6 and 9 Jcm(-2). One set of the surfaces was annealed at 575 degrees C to increase the crystallinity of the deposited films. Primary human osteoblasts were seeded onto the material surfaces and cytoskeletal actin organisation was examined using confocal laser scanning microscopy. The annealed surfaces supported greater cell attachment and more defined cytoskeletal actin organisation. Cell activity, measured using the alamar Blue assay, was also found to be significantly higher on the annealed samples. In addition, our results show distinct trends that correlate with the laser fluence used for deposition. The cell activity increases with increasing fluence. This pattern was repeated for alkaline phosphatase production by the cells. Differences in cell spreading were apparent which were correlated with the fluence used to deposit the HA. The optimum surface for initial attachment and spreading of osteoblasts was one of the HA films deposited using 9 J cm(-2) laser fluence and subsequently annealed at 575 degrees C.

Physical, chemical, and biological characterization of pulsed laser deposited and plasma sputtered hydroxyapatite thin films on titanium alloy

The physical, chemical, and biological properties of pulsed laser deposited (PLD) and plasma sputtered (PS) hydroxyapatite (HA) coatings were compared. Human osteoblast-like cell responses to these coatings in vitro were assayed for proliferation and phenotypic expression. PS coatings formed smooth and continuous thin films that followed the contours of the substrate surface. PLD coatings consisted of numerous spheroidal micro- and macroparticles. The crystallinity of all coatings was quantified by comparison with the HA target used for both the PS and PLD processes. The XRD and FTIR results indicated that unannealed PLD coatings deposited at room temperature had X-ray spectra consistent with an amorphous structure and were found to dissolve after only a few hours in saline solution. Annealing at 400 degrees C increased the crystallinity (87-98%), which resulted in improved stability and cell activity. The PS coatings showed greater chemical stability than the unannealed PLD coatings and contained an approximate 15% crystalline phase, increasing to 65% postannealing. Cell proliferation and alkaline phosphatase production were significantly higher on unannealed PS specimens than the other coating treatments. There may be benefits in engineering the presence of a minor percentage of a microcrystalline phase in an amorphous or nanometer scale polycrystalline HA structure.

Biocompatibility and osteogenic potential of human fetal femur-derived cells on surface selective laser sintered scaffolds

For optimal bone regeneration, scaffolds need to fit anatomically into the requisite bone defects and, ideally, augment cell growth and differentiation. In this study we evaluated novel computationally designed surface selective laser sintering (SSLS) scaffolds for their biocompatibility as templates, in vitro and in vivo, for human fetal femur-derived cell viability, growth and osteogenesis. Fetal femur-derived cells were successfully cultured on SSLS-poly(d,l)-lactic acid (SSLS-PLA) scaffolds expressing alkaline phosphatase activity after 7days. Cell proliferation, ingrowth, Alcian blue/Sirius red and type I collagen positive staining of matrix deposition were observed for fetal femur-derived cells cultured on SSLS-PLA scaffolds in vitro and in vivo. SSLS-PLA scaffolds and SSLS-PLA scaffolds seeded with fetal femur-derived cells implanted into a murine critical-sized femur segmental defect model aided the regeneration of the bone defect. SSLS techniques allow fabrication of biocompatible/biodegradable scaffolds, computationally designed to fit any defect, providing a template for cell osteogenesis in vitro and in vivo.

Laser shaping of cartilage

The carbon dioxide laser has been used for the first time to change the cartilages shape. After the laser irradiation the cartilage has the tendency to retain its new form. Different types of laser modified cartilage structures were studied. The inferred physical mechanism for cartilage shaping using the stresses relaxation process is presented. The clinical significance of the results for corrective laser surgery is discussed.

Atomic force microscopic study of the surface morphology of apatite films deposited by pulsed laser ablation

Atomic force microscopy (AFM) has been used to study the surface morphology of apatite films deposited on metallic and polyethylene substrates by laser ablation using KrF and transversely excited atmospheric CO2 lasers. The films are found to consist of a smooth apatite coating with macroparticles scattered on the surface. A wide variety of macroparticles, differing in size, shape and roughness, were found and analysed employing the high spatial resolution of AFM (< 1 nm). We have investigated the correlation between the apatite film morphology and the deposition conditions. Of particular importance are laser fluence, gas pressure, the nature of the target and the substrate temperature. We have explained these dependencies on the basis of a theoretical model which includes evaporation and a cluster-type laser ablation mechanism.

Stress relaxation and cartilage shaping under laser radiation

The problem of a purposeful change of the shape of cartilage is of great importance for otolaryngology, orthopaedics, and plastic surgery. In 1992 we have found a possibility of controlled shaping of cartilage under moderate laser heating. This paper presents new results in studies of that phenomenon. We have measured temperature and stress in a tissue undergoing to irradiation with a Holmium laser. Study of cartilage structure allowed us to find conditions for laser shaping without pronounced alterations in the structure of matrix.

SERS substrates formed by gold nanorods deposited on colloidal silica films

We describe a new approach to the fabrication of surface-enhanced Raman scattering (SERS) substrates using gold nanorod (GNR) nanopowders to prepare concentrated GNR sols, followed by their deposition on an opal-like photonic crystal (OPC) film formed on a silicon wafer. For comparative experiments, we also prepared GNR assemblies on plain silicon wafers. GNR-OPC substrates combine the increased specific surface, owing to the multilayer silicon nanosphere structure, and various spatial GNR configurations, including those with possible plasmonic hot spots. We demonstrate here the existence of the optimal OPC thickness and GNR deposition density for the maximal SERS effect. All other things being equal, the analytical integral SERS enhancement of the GNR-OPC substrates is higher than that of the thick, randomly oriented GNR assemblies on plain silicon wafers. Several ways to further optimize the strategy suggested are discussed.

Fabrication of polymer scaffolds for tissue engineering using surface selective laser sintering

A new approach to the fabrication of individual implants and scaffolds for tissue engineering—surface selective laser sintering (SSLS)—is proposed and realized. In contrast to the conventional selective laser sintering, the SSLS method makes it possible to sinter polymer microparticles and melt the near-surface layer rather than the microparticle as a whole. The effect of the laser radiation parameters and the structure and composition of the raw products on the structure and properties of the biomaterials sintered by the laser radiation is analyzed. This approach makes possible both the application of thermally unstable polymers (e.g., polylactides or polylactoglycolides) and the fabrication of scaffolds with incorporated bioactive proteins. The results obtained yield a regular physical basis for a new technology of the fabrication of various polymer scaffolds that represent important materials and elements of modern tissue engineering. The flexibility of the SSLS method is especially important at the stage of investigation of the cell and tissue responses needed for the optimization of the material for a specific application in tissue engineering.