Histopathologic and morphometric evaluation of the skin abnormalities induced by erbium:YAG and carbon dioxide lasers in 10 patients

In the 1960s, carbon dioxide (CO2) laser therapy started to be applied to eliminate wrinkles, actinic scars, and acne because of its capacity of induce intracellular water vaporization. However, recent studies have shown the efficacy of the erbium laser in removing delicate and moderate scars. Furthermore, the postoperative lesions induced by the erbium laser seem to resolve faster and with less erythematous pattern compared with lesions induced by the CO2 laser. The purpose of this study was to determine the immediate pathologic alterations caused by single applications of CO2 and erbium lasers and their association in human skin shreds.Ten white female patients aged 30 to 63 years underwent rhytidectomy, and their respective shreds, which were prepared for excision, were tattooed with the CO2 laser, the erbium laser, or a combination of both in random order and number of applications, before final removal. This project was approved by the local ethical committee. After surgical removal, these tattooed shreds were fixed in 10% buffered formalin and submitted to histopathologic analysis. Morphometric studies demonstrated the normal skin thickness and thickness of the laser-treated area, and their subtraction resulted in the ablation damage values. Residual thermal damage corresponded to the thickness of the affected skin from the most superficial layer of tissue in the laser-treated area down to the deepest dermal area with basophilic degeneration of collagen fibers.Our results showed that two CO2 applications resulted in greater ablation and residual thermal damage when compared with only one CO2 application. The same was true in comparisons of one and two applications of the erbium laser. Both results were statistically significant (p < 0.05). When one isolated erbium and one isolated CO2 application were compared, ablation damage was greater in the former group, although with no statistical significance. One CO2 plus one erbium application compared with one isolated CO2 application showed similar ablation damage but greater residual thermal damage in the latter group (p < 0.05). These observations might contribute to our understanding of the lesions caused in the human skin by erbium and CO2 lasers and eventually help determine the ideal laser combination for the appropriate surgical treatment.     

翠绿宝石激光脱毛350例疗效分析

目的:评价翠绿宝石激光脱毛治疗的疗效。方法:使用波长为755 nm的GentleLASE Plus激光对350例门诊患者的不同部位的体毛进行多次治疗,并按不同部位和治疗次数进行分组,分析其疗效。结果:经过多次治疗后,总体治愈率为83.71%,疗效与不同的部位有关,且与治疗次数呈正相关。其中5例出现暂时性色素沉着。结论:使用GentleLASE Plus激光进行脱毛治疗效果良好,并发症少,是目前较为理想的脱毛治疗方法。     

Side effects and complications of variable-pulsed erbium:yttrium-aluminum-garnet laser skin resurfacing: extended experience with 50 patients

Recent advances in technology have provided laser surgeons with new options for cutaneous laser resurfacing. Despite its popularity, there is limited information on the short-term and long-term side effects and complications of variable-pulsed erbium:yttrium-aluminum-garnet (erbium:YAG) laser skin resurfacing. The purpose of this study was to prospectively evaluate postoperative wound healing, side effects, and complications of multiple-pass, variable-pulsed erbium:YAG laser skin resurfacing for facial photodamage, rhytides, and atrophic scarring. Fifty consecutive patients with facial photodamage, rhytides, or atrophic scarring were treated with a variable-pulsed erbium:YAG laser. Side effects and complications relating to postoperative healing, erythema, and pigmentary changes were tabulated. Patients were evaluated at postoperative days 3 through 7 and at 1, 3, 6, and 12 months after laser skin resurfacing. The average time for reepithelialization was 5.1 days. Prolonged erythema (>1 month) was observed in three patients (6 percent). Transient hyperpigmentation occurred in 20 patients (40 percent), with an average duration of 10.4 weeks. No cases of hypopigmentation or scarring were seen. In summary, a variable-pulsed erbium:YAG laser can safely be used for the treatment of facial photodamage, rhytides, and atrophic scarring. Although more postoperative erythema is seen after variable-pulsed erbium:YAG laser treatment than is usually produced with a short-pulsed erbium:YAG system, the side-effect profile and recovery period after variable-pulsed erbium:YAG laser skin resurfacing still are more favorable than after multiple-pass carbon dioxide laser skin resurfacing.     

Skin contraction with pulsed CO2 and erbium:YAG laser

The purpose of this study was to assess the physical response of skin to laser resurfacing in a real-time, quantitative fashion. The study was designed to assess skin contraction from two opposite standpoints. First, change in tension was measured during laser application while samples were held at constant length. Second, change in length of a sample under no tension was measured during laser treatment. These two disparate analyses represent the two possible extremes of the clinical situation in which skin exists under some tension with some laxity to allow for decrease in length. A custom apparatus with digital interface for skin tension measurements was used to produce single sample tracings of change in skin tension with laser treatment. Length change was measured for individual samples by continuous sonomicrometer readings. Individual sample data were then plotted in a time versus tension/length graph. Skin contracts immediately to a peak level and then relaxes to a sustained plateau level for both CO2 and erbium:YAG lasers. Increased contraction was noted when the beam penetrated into the dermis. Greater peak and plateau contraction is observed after the beam has penetrated into the dermis. Skin contraction varies directly with energy for CO2 and erbium:YAG laser. Findings were similar when skin tension was measured with the sample held at constant length and when length change was measured with the sample under no tension. Char left on the skin after a pass with CO2 laser substantially decreases skin contraction. High-density settings with CO2 laser yield pulse stacking, which effectively irradiates the same portion of tissue with char on it. Skin contraction varies inversely with computer pattern density settings for CO2 laser due to this pulse stacking effect. Density has little effect on skin contraction for the erbium:YAG laser because little char is generated. Histologic analysis identified a zone of coagulated dermis that correlates linearly with skin contraction.     

Flow cytometer in the infrared: inexpensive modifications to a commercial instrument.

BACKGROUND: The application of molecules that fluoresce in the infrared (IR) region to measure cell products would be enhanced by a flow cytometer capable of measuring them. To our knowledge, none exist at this time. Accordingly, we have developed such an instrument. METHODS: A Becton Dickinson LSR flow cytometer was modified to include a small 785-nm IR diode laser the size of a C cell battery with 44-mW output power. The instrument was modified further to accommodate this laser in addition to a 405-nm solid-state laser, a 488-nm air-cooled argon laser, and a 658-nm solid-state laser. Because the IR laser is dangerous to the eye, the laser beams were viewed for optical alignment using a CCD camera and video monitor. An avalanche photodiode was used in place of a photomultiplier tube because its detection sensitivity in the IR region is superior. RESULTS: To assess performance, scatter and fluorescence measurements were made using microspheres that fluoresce in the IR region, and human leukocytes were stained with CD45 biotin followed by a streptavidin conjugated with an IR dye. An avalanche photodiode was 2.3 to 2.8 times more sensitive than a photomultiplier tube for detecting IR fluorescence. Cells stained with CD45 biotin and avidin conjugated with an IR dye could easily be resolved and their fluorescence quantified; there was virtually no autofluorescence. In addition, a lipophilic membrane dye that emits in the IR region was studied. HL60 cells were stained with this dye and they exhibited bright fluorescence intensity. CONCLUSION: A commercial instrument could be modified to accommodate an IR laser for exciting dyes that fluoresce in the IR region. This new capability will extend the range of fluorescence that can be measured by flow cytometry.     

Combined erbium:YAG laser resurfacing and face lifting

Facial aging occurs secondary to gravity-induced tissue ptosis and photoaging. Combined face lifting and carbon dioxide laser resurfacing provides a comprehensive one-stage approach to facial rejuvenation but is condemned by many plastic surgeons due to the nonspecific thermal effects of the laser and risk of skin necrosis. Newer high-energy erbium:YAG lasers allow precise tissue ablation with minimal thermal effect. In this study, various facial rejuvenation techniques were combined with simultaneous erbium:YAG laser resurfacing to assess results and complications. A total of 257 patients from Florida, Melbourne, Australia, and Tel Aviv, Israel, underwent combined erbium:YAG laser resurfacing and surgical facial rejuvenation. Various face-lift methods were used, including endoscopic, deep plane, and subcutaneous. Simultaneous, full-facial laser resurfacing was performed using a variety of erbium:YAG lasers. It was found that combined laser resurfacing and face lifting was successful in greater than 95 percent of patients with minimal morbidity. Two patients (1 percent) (both heavy smokers) developed small areas of skin necrosis that healed with minor pigment changes. Five patients (2 percent) developed synechia that was treated with no residual effect. Two additional patients (1 percent) developed temporary ectropion. There were no other cases of scarring, infection, or cosmetically obvious hypopigmentation. Although larger studies are necessary, it seems that the lack of thermal injury from the erbium:YAG laser makes it possible to safely perform laser resurfacing with surgical facial rejuvenation in nonsmokers. However, the authors caution that familiarity with the nuances of erbium:YAG laser resurfacing be obtained before performing combined laser resurfacing and face lifting.     

Evaluation of six fluorescent protein stains for use in flow microfluorometry.

Flow microfluorometric (FMF) analysis of stained cells has provided protein distribution histograms for large populations of cells. Spectral data and staining protocols were evaluated for six fluorescent protein dyes suggested for staining cells in liquid suspension. The requirements for dyes and/or staining protocol included minimal cell clumping and cell loss, near-optimal dye excitation at existing laser wavelengths, and tenacity of the dye/protein interaction. These criteria were best satisfied by fluorescein isothiocyanate (FITC) and rhodamine B isothiocyanate (RITC). Both fluorescamine and 8-aniline-1-naphthalene sulfonic acid (ANSA) showed potential applicability for use in systems where excitation wavelengths in the ultraviolet range are available. Protein staining with fluorescamine was extremely rapid. Brilliant sulfaflavine and 1-dimethyl-aminonaphthalene-5-sulfonyl chloride (DANSYL) were found unsatisfactory in these studies, since the former dye tended to diffuse from the cells, while the latter induced excessive cell clumping and cell loss. These techniques have application to immunofluorescence analysis and can also be profitably employed in dual-parameter analysis systems in connection with double-staining techniques for simultaneous DNA and protein analysis.     

Photodynamic approaches to elimination and prevention of atherosclerotic changes in the vessels

Photodynamic therapy (PDT) is a method based on a selective accumulation of a photo-sensitive dye in the target cells with their subsequent destruction with the aid of a low-intensity laser radiation of a certain wavelength. It is widely used today in clinical practice, mostly in oncology. This review elucidates possibilities of using the PDT in prophylactics and treatment of cardiovascular pathological conditions, atherosclerosis in particular. Data are cited indicating a successful application of the PDT for preventing the intima hyperplasia following and angioplasty. A possibility of using the PDT for treatment and prophylactics of atherosclerotic alterations in the vessels with due consideration of specifics of formation and cullular composition of the atherosclerotic lesions.     

不同光源对K562细胞ALA-PDT作用的实验研究

目的:利用532 nm脉冲激光、532 nm连续激光和氙灯对K562细胞进行基于5-氨基乙酰丙酸的光动力疗法(ALA-PDT),研究在不同光照条件下细胞抑制率的变化情况,为实现体外ALA-PDT的高效率选择合适的光源。方法:在其他条件相同的情况下,采用不同的光源、不同的光剂量对ALA-PDT组细胞进行辐照,利用O-LYMPUS倒置荧光显微镜和显微镜数码相机系统观察细胞的形态学变化并拍照,利用光学显微镜进行台盼兰拒染法检测细胞的抑制率变化情况。结果:532 nm连续激光和脉冲激光对K562细胞的ALA-PDT抑制率均较低,增加光剂量也不能有效提高ALA-PDT的抑制率;氙灯在功率密度为350 mW/cm2、光照5 min时就能达到最佳的光剂量,此时单纯光照对K562细胞的光损伤作用很小且ALA-PDT效率很高。结论:宽光谱、高功率的氙灯对K562细胞的ALA-PDT效果远优于532 nm激光,对体外ALA-PDT实验比较适用。     

Evaluation of Six Fluorescent Protein Stains for Use in Flow Microfluorometry

Flow microfluorometric (FMF) analysis of stained cells has provided protein distribution histograms for large populations of cells. Spectral data and staining protocols were evaluated for six fluorescent protein dyes suggested for staining cells in liquid suspension. The requirements for dyes and/or staining protocol included minimal cell clumping and cell loss, near-optimal dye excitation at existing laser wavelengths, and tenacity of the dye/protein interaction. These criteria were best satisfied by fluoresoein isothiocyanate (FITC) and rho-damine B isothiocyanate (RITC). Both fluoreseamine and 8-aniline-1-naphthakne sulfonic acid (ANSA) showed potential applicability for use in systems where excitation wavelengths in the ultraviolet range are available. Protein staining with fluorescamine was extremely rapid. Brilliant sulfaflavine and 1-dimethyl-aminonaphthalene-5-sulfonyl chloride (DANSYL) WCR found unsatisfactory in these studies, since the former dye tended to diffuse from the all., while the latter induced excessive all clumping and cell loss. These techniques have application to immunofluoregcence analysis and can also be profitably employed in dual-parameter analysis systems in connection with double-staining techniques for simultaneous DNA and protein analysis.     

Evaluation of Six Fluorescent Protein Stains for Use in Flow Microfluorometry

Flow microfluorometric (FMF) analysis of stained cells has provided protein distribution histograms for large populations of cells. Spectral data and staining protocols were evaluated for six fluorescent protein dyes suggested for staining cells in liquid suspension. The requirements for dyes and/or staining protocol included minimal cell clumping and cell loss, near-optimal dye excitation at existing laser wavelengths, and tenacity of the dye/protein interaction. These criteria were best satisfied by fluoresoein isothiocyanate (FITC) and rho-damine B isothiocyanate (RITC). Both fluoreseamine and 8-aniline-1-naphthakne sulfonic acid (ANSA) showed potential applicability for use in systems where excitation wavelengths in the ultraviolet range are available. Protein staining with fluorescamine was extremely rapid. Brilliant sulfaflavine and 1-dimethyl-aminonaphthalene-5-sulfonyl chloride (DANSYL) WCR found unsatisfactory in these studies, since the former dye tended to diffuse from the all., while the latter induced excessive all clumping and cell loss. These techniques have application to immunofluoregcence analysis and can also be profitably employed in dual-parameter analysis systems in connection with double-staining techniques for simultaneous DNA and protein analysis.     

Cellular Destruction Following Transmyocardial Laser Revascularization (TMR)

Summary During transmyocardial revascularization, cellular destruction of cardiomyocytes occurs as a result of the high-energy laser. However, the features of myocardial cellular destruction are unclear. The present study was undertaken to examine the structural characteristics of cell death in the myocardium following transmyocardial revascularization. Myocardial specimens from 3 male patients who had died within 11 days following laser revascularization were collected within 1 h of death and were analyzed by immunohistochemistry and electron microscopy. For immunohistochemistry, antibodies to pro-apoptotic proteins CPP32 and BAX were used. Immunohistochemical examination demonstrated the presence of cells expressing both CPP32 and BAX along the laser channel. Electron microscopic analysis revealed that the lining surface of laser channels consisted of condensed acellular debris and dead cells. No endothelialization of channels was noted. The lumen of laser channels were surrounded by a rim of acellular debris with several outer concentric rims of cardiomyocytes showing features of cellular destruction. The present study identified features of both necrotic and apoptotic cellular death following laser revascularization.     

Difference in Membrane Repair Capacity Between Cancer Cell Lines and a Normal Cell Line

Electroporation-based treatments and other therapies that permeabilize the plasma membrane have been shown to be more devastating to malignant cells than to normal cells. In this study, we asked if a difference in repair capacity could explain this observed difference in sensitivity. Membrane repair was investigated by disrupting the plasma membrane using laser followed by monitoring fluorescent dye entry over time in seven cancer cell lines, an immortalized cell line, and a normal primary cell line. The kinetics of repair in living cells can be directly recorded using this technique, providing a sensitive index of repair capacity. The normal primary cell line of all tested cell lines exhibited the slowest rate of dye entry after laser disruption and lowest level of dye uptake. Significantly, more rapid dye uptake and a higher total level of dye uptake occurred in six of the seven tested cancer cell lines (p < 0.05) as well as the immortalized cell line (p < 0.001). This difference in sensitivity was also observed when a viability assay was performed one day after plasma membrane permeabilization by electroporation. Viability in the primary normal cell line (98 % viable cells) was higher than in the three tested cancer cell lines (81–88 % viable cells). These data suggest more effective membrane repair in normal, primary cells and supplement previous explanations why electroporation-based therapies and other therapies permeabilizing the plasma membrane are more effective on malignant cells compared to normal cells in cancer treatment.     

Er,Cr:YSGG激光照射对牙根尖封闭效果影响的实验研究

目的:深入探讨在根尖倒充填术中Er,Cr:YSGG激光照射对牙根尖的封闭效果,为此激光在口腔临床上的应用与推广提供重要的理论与实验依据。方法:将56颗新拔除的单根牙随机分成7组,每组8颗牙。去除牙冠,常规进行根管预备、根管充填。将倒充填的牙根尖进行预备,然后银汞合金倒充填。预备方法:1手机切除根尖后,制备成烧瓶状洞形;2手机处理方法同1,然后Er,Cr:YSGG激光于不同能量(1W、2W)及不同方式(喷水或无水)下对洞内壁进行照射。结合微渗漏检测与扫描电镜观察对Er,Cr:YSGG激光照射后的牙根尖封闭效果进行评估。结果:Er,Cr:YSGG激光可有效去除根切面上的玷污层及碎屑;与手机加银汞倒充填组相比,激光照射后的根尖染料微渗漏深度有所减少;但只有1WEr,Cr:YSGG激光于无水条件下照射时,根尖染料微渗漏深度显著减少(P<0.05)。结论:Er,Cr:YSGG激光应用于根尖倒充填术中可减少微渗漏的发生;促进根尖封闭;在特定的能量参数下可获得优于常规银汞倒充填术的治疗效果。因此我们推荐临床上将Er,Cr:YSGG激光照射与常规银汞倒充填术结合使用,这将有助于提高根尖倒充填术治疗的成功率。     

Photo-immunotargeting with haematoporphyrin conjugates activated by a low-power He-Ne laser

Summary A combined method has been developed for selective cytolysis in vitro as well as in vivo using a photosensitizer haematoporphyrin-protein conjugate as the targeting molecule and low-power He-Ne laser (632.8 nm) irradiation in order to activate the sensitizer to its excited, toxic triplet energy state. The specificity of the procedure was demonstrated in vitro by purging a mixed cell population from one component, and in vivo in an animal (nude mice) xenograft tumour model, where human cancer cells were destroyed by the immunotargeting method using monoclonal-antibody-haematoporphyrin (mAb-HP) conjugate (a-PNAr-I mAbs, which bind to the cell surface antigens of gastric cancer cells) and soft laser irradiation. The cell destruction was dependent on the doses of both mAb-HP and He-Ne laser light energy, and occurred only in target cell populations.     

Skin closure with dye-enhanced laser welding and fibrinogen

The topical application of wavelength-specific dye and fibrinogen has been used to enhance laser closure of vascular anastomoses. We compared the closure of skin incisions by two different dye-enhanced, fibrinogen-based laser welding systems [argon laser (power density 4.78 W/cm2) with fluorescein isothiocyanate dye (n = 32) and diode laser (power density 9.55 W/cm2) with indocyanine green dye (n = 32)] with closure by interrupted 5-0 nylon suture (n = 64) and examined tensile strength, hydroxyproline production, histology, and cosmesis. Two 3-cm full-thickness incisions were made on the shaved backs of 64 rats. One incision was closed with suture, whereas the other, after treatment with the appropriate dye, was welded with either argon- or diode-lasered fibrinogen. At postoperative days 5, 10, 15, and 28, the closure sites were harvested and sectioned for analysis. Initially, wounds closed with argon-lasered fibrinogen showed less inflammatory response, greater collagen production (34.61 +/- 0.74 mg/gm), and greater mean peak stress at rupture (64.85 lbs/in2) than those closed with suture (16.42 +/- 3.20 mg/gm, 26.68 lbs/in2) (p less than 0.05). By 15 days, both argon and diode laser closures are superior in strength and collagen production to suture closure (p less than 0.05). At 28 days, diode laser closures (1315.60 lbs/in2) are stronger than suture closures (998.09 lbs/in2), whereas both are stronger than argon laser closures (813.16 lbs/in2) (p less than 0.05). Cosmetically, argon-welded wounds consistently appeared finer and lacked cross-hatched suture scars.(ABSTRACT TRUNCATED AT 250 WORDS)     

The Diatom Staurosirella pinnata for Photoactive Material Production

A native isolate of the colonial benthic diatom Staurosirella pinnata was cultivated for biosilica production. The silicified cell walls (frustules) were used as a source of homogeneous and structurally predictable porous biosilica for dye trapping and random laser applications. This was coupled with the extraction of lipids from biomass showing potential to fabricate photoactive composite materials sustainably. The strain was selected for its ease of growth in culture and harvesting. Biosilica and lipids were obtained at the end of growth in indoor photobioreactors. Frustules were structurally characterized microscopically and their chemistry analyzed with Fourier Transform Infrared Spectroscopy. Frustule capacity of binding laser dyes was evaluated on a set of frustules/Rhodamine B (Rho B) solutions and with respect to silicon dioxide and diatomite by Fluorescence Spectroscopy demonstrating a high affinity for the organic dye. The effect of dye trapping property in conveying Rho B emission to frustules, with enhancement of scattering events, was analyzed on Rho B doped polyacrylamide gels filled or not with frustules. Amplified spontaneous emission was recorded at increasing pump power indicating the onset of a random laser effect in frustule filled gels at lower power threshold compared to unfilled matrices.     

Laser induced cell fusion in combination with optical tweezers: the laser cell fusion trap.

A single-beam gradient force optical trap was combined with a pulsed UV laser microbeam in order to perform laser induced cell fusion. This combination offers the possibility to selectively fuse two single cells without critical chemical or electrical treatment. The optical trap was created by directing a Nd:YAG laser, at a wavelength of 1.06 microns, into a microscope and focusing the laser beam with a high numerical aperture objective. The UV laser microbeam, produced by a nitrogen-pumped dye laser (366 nm), was collinear with the trapping beam. Once inside the trap, two cells could be fused with several pulses of the UV laser microbeam, attenuated to an energy of approximately 1 microJ/pulse in the object plane. This method of laser induced cell fusion should provide increased selectivity and efficiency in generating viable hybrid cells.     

Optical characteristics of a gallium laser plasma

Results are presented from studies of the emission from an erosion gallium laser plasma at a moderate intensity (W=(1–5)×10⁸ W/cm²) of a 1.06-μm laser radiation. It is shown that, under these conditions, the lower excited states of gallium atoms are populated most efficiently. Among the ions, only the most intense GaII lines are observed in the emission spectrum. The populations of GaI and GaII excited states are not related to direct electron excitation, but are determined by the recombination of gallium ions with slow electrons. The recombination times of GaIII and GaII ions in the core of the plasma jet are determined from the waveforms of emission in the GaII and GaI spectral lines and are equal to 10 and 140 ns, respectively. The results obtained are of interest for spectroscopic diagnostics of an erosion plasma produced from gallium-containing layered crystals during the laser deposition of thin films. __________ Translated from Fizika Plazmy, Vol. 27, No. 1, 2001, pp. 85–88. Original Russian Text Copyright ? 2001 by Shuaibov, Shimon, Dashchenko, Shevera, Chuchman.     

Quantification of extracellular polymeric substances in biofilms by confocal laser scanning microscopy

Extracellular polymeric substances (EPS) in a biofilm were quantified by measuring the total cell volume from a 3-D image of the biofilm using confocal laser scanning microscope after staining cells with a fluorescent dye specific for nucleic acids. The EPS content was the difference between the volatile solids in the biofilm and the total cell mass, which could be quantified from the measured cell volume.