Paolo Matteini

Photothermally-induced disordered patterns of corneal collagen revealed by SHG imaging.

The loss of organization of the corneal collagen lattice induced by photothermal effects was analyzed by using second-harmonic generation (SHG) imaging. Porcine cornea samples were treated with low-power laser irradiation in order to get localized areas of tissue disorganization. The disorder induced within the irradiated area of corneal stroma was quantified by means of Discrete Fourier Transform, auto-correlation and entropy analyses of the SHG images. Polarization modulated SHG measurements allowed to probe the changes in the structural anisotropy of sub-micron hierarchical levels of the stromal collagen. Our results emphasize the great potential of the SHG imaging to detect subtle modifications in the collagen assembly. The proposed analytical methods may be used to track several genetic, pathologic, accidental or surgical-induced disorder states of biological tissues.

Chitosan Films Doped with Gold Nanorods as Laser-Activatable Hybrid Bioadhesives

Chitosan is a linear chain polysaccharide comprising glucosamine and N -acetyl glucosamine residues joined together by β -1,4-glycosidic bonds. It is derived as an acids-soluble material by deacetylation of chitin, which is commonly found in the outer skeleton of crustaceans and in the cell walls of fungi. A variety of fundamental properties such as excellent biocompatibility and biodegradability make chitosan a very attractive material for biomedical applications including wound dressing, tissue engineering and drug delivery. [ 1 , 2 ] The peculiar tendency of chitosan to form films with high mechanical strength, good elasticity and rather slow biodegradation is among the principal reasons why this material is gaining a primary role in medical technology. [ 2–4 ] Examples of applications of chitosan films are the localized chemotherapy based on the local delivery and sustained release of antineoplastic agents after cancer removal [ 5 ] and the tissue repair by closing chronic, accidental, and surgical wounds. [ 4 , 6 ] In order to enable these applications, the chitosan films need to be secured to the target, which may be achieved by suturing. [ 7 ] However, in many cases, suturing may be hardly feasible such as for poorly accessible or delicate body regions (e.g., intracranial, myocardial, etc.). In addition metals and synthetic materials in use for implantation can interfere with the functional rehabilitation of the site due to foreign-body reactions, causing infl ammation and abnormal wound healing. [ 8 ]

Gold nanorods as new nanochromophores for photothermal therapies

Results and perspectives on the biomedical exploitation of gold nanorods with plasmon resonances in the near infrared window are reported. The authors describe experimental studies of laser-activated nanoparticles in the direct welding of connective tissues, which may become a valuable technology in biomedicine. In particular, colloidal gold nanorods excited by diode laser radiation at 810 nm were used to mediate functional photothermal effects and weld eyes lens capsules and arteries. The preparation of biopolymeric matrices including gold nanorods is also described, as well as preliminary tests for their application in the closure of wounds in vessels and tendons. Finally, the use of these nanoparticles for future applications in the diagnosis, imaging and therapy of cancer is discussed.

Photothermal effects in connective tissues mediated by laser-activated gold nanorods

We report a study on the application of laser-activated nanoparticles in the direct welding of connective tissues, which may become a valuable technology in biomedicine. We use colloidal gold nanorods as new near-infrared chromophores to mediate functional photothermal effects in the eye lens capsules. Samples obtained ex vivo from porcine eyes are treated to simulate heterotransplants with 810-nm diode laser radiation in association with a stain of gold nanorods of aspect ratio approximately 4. This stain is applied at the interface between a patch of capsule from a donor eye and the capsule of a recipient eye. Then, by administration of laser pulses of 40 msec and approximately 100-140 J/cm(2), we achieved the local denaturation of the endogenous collagen filaments, which reveals that the treated area reached temperatures above 50 degrees C. The thermal damage is confined within 50-70 mum in a radial distance from the irradiated area.

Structural Behavior of Highly Concentrated Hyaluronan

When investigated under high concentration conditions, hyaluronan (HA) solutions in physiological saline are shown to generate stable superstructures. An abrupt change in the rheological properties observed on increasing the temperature suggests the breaking of certain cooperative bonds. The thermal disruption of the HA superstructure is accompanied by a sharp transition from a long- to a restricted-connectivity water structuring, which is interpreted as a concurrent transition from a stable to a temporary polymer network. The intermolecular associations are considered to be originated by hydrophobic interactions between the nonpolar groups of the polymer backbones.

Laser tissue welding in ophthalmic surgery

Laser welding of ocular tissues is an alternative technique or adjunct to conventional suturing in ophthalmic surgery. It is based on the photothermal interaction of laser light with the main components of the extracellular matrix of connective tissues. The advantages of the welding procedure with respect to standard suturing and stapling are reduced operation times, lesser inflammation, faster healing and increased ability to induce tissue regeneration. The procedure we set up is based on the use of an infrared diode laser in association with the topical application of the chromophore Indocyanine Green. Laser light may be delivered either continuously or in pulses, thus identifying two different techniques that have been applied clinically in various types of transplants of the cornea.

In vivo carotid artery closure by laser activation of hyaluronan-embedded gold nanorods

We prove the first application of near-infrared-absorbing gold nanorods (GNRs) for in vivo laser closure of a rabbit carotid artery. GNRs are first functionalized with a biopolymeric shell and then embedded in hyaluronan, which gives a stabilized and handy laser-activable formulation. Four rabbits undergo closure of a 3-mm longitudinal incision performed on the carotid artery by means of a 810-nm diode laser in conjunction with the topical application of the GNRs composite. An effective surgery is obtained by using a 40-W/cm(2) laser power density. The histological and electron microscopy evaluation after a 30-day follow-up demonstrates complete healing of the treated arteries with full re-endothelization at the site of GNRs application. The absence of microgranuloma formation and/or dystrophic calcification is evidence that no host reaction to nanoparticles interspersed through the vascular tissue occurred. The observation of a reshaping and associated blue shift of the NIR absorption band of GNRs after laser treatment supports the occurrence of a self-terminating process, and thus of additional safety of the minimally invasive laser procedure. This study underlines the feasibility of using GNRs for in vivo laser soldering applications, which represents a step forward toward the introduction of nanotechnology-based therapies in minimally invasive clinical practices.

SERS Detection of Amyloid Oligomers on Metallorganic-Decorated Plasmonic Beads

Protein misfolded proteins are among the most toxic endogenous species of macromolecules. These chemical entities are responsible for neurodegenerative disorders such as Alzheimers, Parkinsons, Creutzfeldt-Jakobs and different non-neurophatic amyloidosis. Notably, these oligomers show a combination of marked heterogeneity and low abundance in body fluids, which have prevented a reliable detection by immunological methods so far. Herein we exploit the selectivity of proteins to react with metallic ions and the sensitivity of surface-enhanced Raman spectroscopy (SERS) toward small electronic changes in coordination compounds to design and engineer a reliable optical sensor for protein misfolded oligomers. Our strategy relies on the functionalization of Au nanoparticle-decorated polystyrene beads with an effective metallorganic Raman chemoreceptor, composed by Al(3+) ions coordinated to 4-mercaptobenzoic acid (MBA) with high Raman cross-section, that selectively binds aberrant protein oligomers. The mechanical deformations of the MBA phenyl ring upon complexation with the oligomeric species are registered in its SERS spectrum and can be quantitatively correlated with the concentration of the target biomolecule. The SERS platform used here appears promising for future implementation of diagnostic tools of aberrant species associated with protein deposition diseases, including those with a strong social and economic impact, such as Alzheimers and Parkinsons diseases.

Thermal Transitions of Fibrillar Collagen Unveiled by Second-Harmonic Generation Microscopy of Corneal Stroma

The thermal transitions of fibrillar collagen are investigated with second-harmonic generation polarization anisotropy microscopy. Second-harmonic generation images and polarization anisotropy profiles of corneal stroma heated in the 35-80°C range are analyzed by means of a theoretical model that is suitable to probe principal intramolecular and interfibrillar parameters of immediate physiological interest. Our results depict the tissue modification with temperature as the interplay of three destructuration stages at different hierarchical levels of collagen assembly including its tertiary structure and interfibrillar alignment, thus supporting and extending previous findings. This method holds the promise of a quantitative inspection of fundamental biophysical and biochemical processes and may find future applications in real-time and postsurgical functional imaging of collagen-rich tissues subjected to thermal treatments.

Emerging concepts of laser-activated nanoparticles for tissue bonding.

We report recent achievements and future perspectives of minimally invasive bonding of biological tissues triggered by laser light. In particular, we review new advancements in the biomedical exploitation of near-infrared absorbing gold nanoparticles as an original solution for the photothermal closure of surgical incisions. Advanced concepts of laser tissue bonding involving the application of hybrid nanocomposites obtained by inclusion of nanochromophores into biopolymer scaffolds are also introduced. The perspectives of tissue bonding are discussed in the following aspects: (1) tissue bonding with highly-stabilized nanochromophores, (2) enhanced tissue bonding with patterned nanocomposites, (3) real-time monitoring of temperature distributions, (4) tracking of tissue regeneration based on the optical resonances of gold nanoparticles.