In vitro and in vivo tissue repair with laser-activated chitosan adhesive

BACKGROUND AND OBJECTIVES: Sutures are currently the gold standard for wound closure but they are still unable to seal tissue and may induce scarring or inflammation. Biocompatible glues, based on polysaccharides such as chitosan, are a possible alternative to conventional wound closure. In this study, the adhesion of laser-activated chitosan films is investigated in vitro and in vivo. In particular we examine the effect of varying the laser power, as well as adding a natural cross-linker (genipin) to the adhesive composition. STUDY DESIGN/MATERIALS AND METHODS: Flexible and insoluble strips of chitosan films (surface area approximately 34 mm(2), thickness approximately 20 microm) were bonded to sheep intestine using several laser powers (0, 80, 120, and 160 mW) at 808-nm wavelength. The strength of repaired tissue was tested by a calibrated tensiometer to select the best power. A natural cross-linker (genipin) was also added to the film and the tissue repair strength compared with the strength of plain films. The adhesive was also bonded in vivo to the sciatic nerve of rats and the thermal damage induced by the laser assessed 4 days post-operatively. RESULTS: Chitosan adhesives successfully repaired intestine tissue, attaining a maximum repair strength of 14.7+/-4.3 kPa (n = 30) at the laser power of 120 mW. The chitosan-genipin films achieved lower repair strength (9.1+/-2.9 kPa). The laser caused partial demyelination of axons at the site of operation, but the myelinated axons retained a normal morphology proximally and distally. CONCLUSIONS: The chitosan adhesive effectively bonded to tissue causing only localized thermal damage in vivo, when the appropriate laser parameters were selected.     

Carotid artery anastomosis with albumin solder and near infrared lasers: a comparative study.

BACKGROUND AND OBJECTIVE: Laser tissue-welding has been used for anastomosis of carotid arteries. During welding, thermal injury sustained by the vessel walls should be minimized to prevent thrombosis. Two different types of lasers were used and effects on tissue damage were studied in vitro and in vivo. STUDY DESIGN/MATERIALS AND METHODS: End-to-end anastomosis of dog carotid arteries (n = 10) was performed by using a human albumin solder (HAS) in conjunction with Nd:YAG or diode lasers (lambda = 1.32 microm and 1.9 microm, respectively). The arteries were evaluated for patency and evidence of histologic injury after 21 days. Another group of arteries was laser soldered in vitro to measure the intimal and adventitial temperatures by using thermocouples. RESULTS: The arteries repaired with the diode laser sustained significantly less thermal damage than those repaired with Nd:YAG laser, both in vitro and in vivo. In particular, the intimal temperature was significantly lower (P < 0.05) for the diode than for the Nd:YAG repairs (approximately 35 degrees C and approximately 50 degrees C, respectively). In the latter group, the patency rate was 75%, but thrombosis occurred in 75% of the specimens at 21 days. All diode anastomoses were patent and thrombosis developed in only 17% of the arteries. CONCLUSION: Use of the diode laser and albumin solders may provide a means to successfully repair carotid arteries with minimal thermal damage.     

An investigation into the inhibitory effect of ultraviolet radiation on Trichophyton rubrum

Fungal infection of nails, onychomycosis, is predominantly caused by Trichophyton rubrum. This infection is an important public health concern due to its persistent nature and high recurrence rates. Alternative treatments are urgently required. One such alternative is phototherapy involving the action of photothermal or photochemical processes. The aim of this novel study was to assess which wavelengths within the ultraviolet (UV) spectrum were inhibitory and equally important nail transmissible. Initial irradiations of T. rubrum spore suspensions were carried out using a tunable wavelength lamp system (fluence ≤3.1 J/cm²) at wavelengths between 280 and 400 nm (UVC to UVA) to evaluate which wavelengths prevented fungal growth. Light-emitting diodes (LEDs) of defined wavelengths were subsequently chosen with a view to evaluate and potentially implement this technology as a low-cost “in-home” treatment. Our experiments demonstrated that exposure at 280 nm using an LED with a fluence as low as 0.5 J/cm² was inhibitory, i.e., no growth following a 2-week incubation (p?<?0.05; one-way ANOVA), while exposure to longer wavelengths was not. A key requirement for the use of phototherapy in the treatment of onychomycosis is that it must be nail transmissible. Our results indicate that the treatment with UVC is not feasible given that there is no overlap between the antifungal activity observed at 280 nm and transmission through the nail plate. However, a potential indirect application of this technology could be the decontamination of reservoirs of infection such as the shoes of infected individuals, thus preventing reinfection.     

Prognostic value of serum IL-10 and soluble IL-2 receptor levels in aggressive non-Hodgkin's lymphoma

Summary We investigated the prognostic significance of interleukin-10 (IL-10) and soluble interleuckin-2 receptor (sIl-2r) levles in the pretreatment serum of 105 individuals with newly-diagnosed aggressive non-Hodgkin's lymphoma (NHL). Commercially available enzyem-linked immunoassay kits were used for cytokine and receptor measurements. Detectable levels of IL-10 were found in 42 (40%) patients at diagnosis, with no correlation with clinico-haematological parameters, but in no control samples (P < 0.001).
Pretreatment concentrations of sIL-2r were markedly increased in individuals with NHL when compared to controls (2614 ± 893 U/ml v 219 ± 65U/ml, P < 0.001), patients with stage III/IV presenting higher values than those with stage II disase (3885 ± 1196U/ml v 1732 ± 646U/ml, P < 0.001). No single parameter was associated with the achiveement of complete remission, but the combination of elevated IL-10 and of sIL-2r greater than 3000U/ml selected a subset of patients with a high probability of failing induction therapy (P < 0.001). Lifetable analysis also indicated thatj patients with these characteristics have a significantly shorter event-free survival. In a multivariate analysis the combination of IL-10 with sIL-2r was found to have greater predictive strength than the combination of IL-10 with β₂-micro-globulin. We conclude that IL-10 and sIL-2r measurements can be expected to improve existing methods of risk assignment in aggressive NHL.     

Albumin-genipin solder for laser tissue repair

BACKGROUND: Laser tissue soldering (LTS) is an alternative technique to suturing for tissue repair that avoids foreign body reaction and provides immediate sealing of the wound. One of the major drawbacks of LTS, however, is the weak tensile strength of the solder welds when compared to sutures. In this study, a crosslinking agent of low cytotoxicity was investigated for its ability to enhance the bond strength of albumin solders with sheep intestine. STUDY DESIGN/MATERIALS AND METHODS: Solder strips were welded onto rectangular sections of sheep small intestine using a diode laser. The laser delivered in continuous mode a power of 170 +/- 10 mW at lambda = 808 nm, through a multimode optical fiber (core size = 200 microm) to achieve a dose of 10.8 +/- 0.5 J/mg. The solder thickness and surface area were kept constant throughout the experiment (thickness = 0.15 +/- 0.01 mm, area = 12 +/- 1.2 mm2). The solder was composed of 62% bovine serum albumin (BSA), 0.38% genipin, 0.25% indocyanin green dye (IG), and water. Tissue welding was also performed with a BSA solder without genipin, as a control group. The repaired tissue was tested for tensile strength by a calibrated tensiometer. Murine fibroblasts were also cultured in extracted media from heat-denatured genipin solder to assess cell growth inhibition in a 48 hours period. RESULTS: The tensile strength of the genipin solder was doubled that of the BSA solder (0.21 +/- 0.04 N and 0.11 +/- 0.04 N, respectively; P = 10(-15) unpaired t-test, N = 30). Media extracted from crosslinked genipin solder showed negligible toxicity to fibroblast cells under the culture conditions examined here. CONCLUSION: Addition of a chemical crosslinking agent, such as genipin, significantly increased the tensile strength of adhesive-tissue bonds. A proposed mechanism for this enhanced bond strength is the synergistic action of mechanical adhesion with chemical crosslinking by genipin.     

Diode‐pumped fiber lasers: A new clinical tool?

BACKGROUND AND OBJECTIVE: Diode-pumped fiber lasers are a compact and an efficient source of high power laser radiation. These laser systems have found wide recognition in the area of lasers as a result of these very practical characteristics and are now becoming important tools for a large number of applications. In this review, we outline the basic physics of fiber lasers and illustrate how a number of clinical procedures would benefit from their employment. STUDY DESIGN/MATERIALS AND METHODS: The pump mechanisms, the relevant pump and laser transitions between the energy levels, and the main properties of the output from fiber lasers will be briefly reviewed. The main types of high power fiber lasers that have been demonstrated will be examined along with some recent medical applications that have used these lasers. We will also provide a general review of some important medical specialties, highlighting why these fields would gain from the introduction of the fiber laser. RESULTS/CONCLUSION: It is established that while the fiber laser is still a new form of laser device and hence not commercially available in a wide sense, a number of important medical procedures will benefit from its general introduction into medicine. With the number of medical and surgical applications requiring high power laser radiation steadily increasing, the demand for more efficient and compact laser systems providing this capacity will grow commensurately. The high power fiber laser is one system that looks like a promising modality to meet this need.     

Self-expandable chitosan stent: design and preparation

Stents are largely used in surgical procedures to relieve pathological obstructions. The purpose of the present study was to design and prepare a biocompatible stent with a self-expandable mechanism. Thin films were prepared from deacetylated chitosan (4% w/v) dissolved in acetic acid solution (2% v/v). The chitosan films were tested by a calibrated tensiometer to measure the Young's module (E). The films were used to manufacture stents by pulling and winding them around a cylindrical rod in a helical fashion. Thirteen stents (diameter = 0.5 +/- 0.05 mm, length approximately 4 mm) were inserted into the vas deferens of wistar rats. Upon stent insertion, the vasal anastomosis was achieved with a laser-soldering technique. The animals were sacrificied 8 weeks later. The stress test showed that the chitosan film was elastic (maximum strain = 105% +/- 6%, E = 0.7655 +/- 0.0288 Mpa). The stents self-expanded by releasing their elastic energy. All the stents but one remained open inside the vasa despite high incidence of sperm granuloma. A biocompatible and self-expandable stent with a helical design is proposed.     

Bone marrow segmentation in leukemia using diffusion and T (2) weighted echo planar magnetic resonance imaging

Magnetic resonance images of leukemic bone marrow were acquired over large regions of the pelvis and lower abdomen with minimal interference from overlying tissues using diffusion and T(2) weighted echo planar imaging. Data acquisition times were on the order of 1 min for scanning volumes of up to 25 l at a spatial resolution of 31 microl. A survey of 21 patients with leukemia and eight healthy adult volunteers was undertaken to determine the magnitude of the observed effect and its dependence upon specific pathologies. The acquisition methods yielded high-quality segmentation of leukemic bone marrow prior to therapy in seven of seven patients with acute lymphocytic leukemia, chronic lymphocytic leukemia or chronic myelogenous leukemia, and who had hypercellular (>95%) bone marrow at the time of the study. The quality of the segmentation was sufficient to allow the use of maximum intensity projection images which afforded a convenient evaluation of both intra- and extramedullary disease. The measured signal-to-noise ratios agreed with a theoretical estimate that accounted for the percentage cellularity, T(2) relaxation time of water, and self-diffusion coefficient of water in iliac bone marrow. In addition, the mean signal-to-noise ratios from iliac marrow were strongly dependent upon the time after the initiation of chemotherapeutic regimens, implying that the methods may be useful for therapeutic monitoring.     

Laser-Activated Solid Protein Solder for Nerve Repair: In Vitro Studies of Tensile Strength and Solder/Tissue Temperature

. Laser-activated solid protein solder strips have been developed for peripheral nerve repair. Indocyanine green dye added to the solder strongly absorbs diode wavelengths (∼800 nm) and causes localised heating and coagulation of the albumin protein solder. The protein strengthens the tissue join, particularly during the acute healing phase postoperative, while shielding the underlying axons from excessive thermal damage. In this investigation of the solid protein solder technique for nerve repair, the effect of laser irradiance on weld strength and solder and tissue temperature were studied. The tensile strength of repaired nerves rose steadily with increased irradiance reaching a maximum of 105±10 N/cm² at 12.7 W/cm². At higher irradiances, tensile strength fell. The maximum temperature reached at the solder surface and at the solder/nerve interface, measured using a non-contact fibre optic radiometer and thermocouple, respectively, also rose steadily with laser irradiance. At 12.7 W/cm², the temperatures reached at the surface and at the interface were 88±5°C and 71±4°C, respectively. This in vitro investigation demonstrates the feasibility of the laser-activated solid protein solder strips for peripheral nerve repair. The laser irradiance and the corresponding solder surface temperature for optimal tensile strength have been identified. Paper received 20 May 1998; accepted following revision 17 February 1999.