Pub Date : 2011-11-01DOI: 10.1016/j.mla.2011.08.009
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Pub Date : 2011-11-01DOI: 10.1016/j.mla.2011.08.005
Gergely Klinda, Peter Urban, Carsten M. Philipp, H.-Peter Berlien
Objective
This retrospective study was undertaken to examine the efficiency of a 532 nm laser device in the treatment of molluscum contagiosum.
Material and methods
The medical records were analyzed of 32 patients with molluscum contagiosum who had been treated in our clinic with a 532 nm laser (IDAS; Quantel Derma GmbH) in the time period 09/2005 to 04/2011.
Results
Of all the patients treated, a total of 84% required only one treatment, 13% required two and only one patient (3%) needed three laser sessions. The treatment was well-tolerated and only a few of the patients developed side effects, such as hypopigmentation and atrophic skin texture changes.
Conclusion
The efficiency of the 532 nm laser is comparable with the dye laser, but the 532 nm laser is lower in price and more commonly available than the dye laser. We recommend further prospective studies about the 532 nm laser in the treatment of molluscum contagiosum.
Ziel
In dieser retrospektiven Studie wurde die Effektivität eines 532 nm-Lasers (IDAS; Quantel Derma GmbH) in der Behandlung von Dellwarzen (Molluscum contagiosum) untersucht.
Material und Methoden
Es wurden die Behandlungsdokumentationen von 32 Patienten, die in unserer Klinik zwischen 09/2005 und 04/2011 wegen Molluscum contagiosum behandelt wurden, retrospektiv analysiert.
Ergebnisse
84% der Patienten benötigte eine Behandlung, 13% zwei und ein Patient (3%) drei Behandlungen. Die Behandlung wurde gut toleriert, die Komplikationsrate war gering. Die Komplikationen bestanden in Hypopigmentierungen und atrophischen Hautveränderungen.
Schlussfolgerung
Die Effektivität des 532 nm-Lasers ist vergleichbar mit der des Farbstofflasers. Der 532 nm-Laser ist weiter verbreitet und preiswerter als der Farbstofflaser. Wir empfehlen weitere prospektive Studien zur Beurteilung des Stellenwertes des 532 nm-Lasers bei der Behandlung von Molluscum contagiosum.
目的回顾性研究532 nm激光治疗传染性软疣的疗效。材料与方法对32例经532 nm激光治疗的传染性软疣患者的病历进行分析;Quantel Derma GmbH)于2005年9月至2011年4月期间。结果在所有治疗的患者中，84%的患者只需要一次治疗，13%的患者需要两次治疗，只有1例患者(3%)需要三次激光治疗。治疗耐受性良好，只有少数患者出现副作用，如色素减退和皮肤质地萎缩改变。结论532 nm激光器的效率可与染料激光器相媲美，但价格较染料激光器低，使用更为普遍。我们建议对532 nm激光治疗传染性软疣进行进一步的前瞻性研究。激光激光器(IDAS)的研究进展(英文)Quantel Derma GmbH)在der Behandlung von delwarzen(传染性软疣)untersucht。材料与方法:回顾性分析2005年9月至2011年4月在德国克立克医院死亡的32例传染性软疣病例。ergebnis84% der Patienten benötigte eine Behandlung, 13% zwei和ein Patient (3%) drei Behandlungen。死在床上，死在床上，死在床上。黄萎病与色素减退症的关系Hautveränderungen。SchlussfolgerungDie Effektivität des 532nm - lasers - ist vergleichbar mit der des Farbstofflasers。Der 532nm - laser list(激光表):超宽和预宽，也可用于Farbstofflaser。本文对传染性软疣的激光治疗进行了前瞻性研究。
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Pub Date : 2011-11-01DOI: 10.1016/j.mla.2011.08.003
Daniela Göppner , Norma Mechow , Julia Liebscher , Erik Thiel , Gunter Seewald , Harald Gollnick , Carsten M. Philipp , Karl-Heinz Schönborn
Background and objectives
Two-photon microscopy (TPM) generates high definition images from biological samples such as human skin in a non-invasive way. The focus of an ultrashort pulsed laser is scanned through the tissue and the fluorescence photons induced by two-photon excitation of endogeneous fluorophores are collected, measured and displayed as a scanning image. So far, one of the main disadvantages of TPM is the limitation of the scanning area to tenths of a millimeter by the field of view of the focusing optics. Essential pieces of information, such as the differentiation between benign or malign changes in biological tissue, are therefore not captured. Within a BMBF-supported research project “FluoTOM”, a new modified TPM scanning technique was developed to overcome these shortcomings, and pilot tests have been made on benign and malignant samples of human skin ex vivo.
Material and methods
The field of view of the new TPM was extended from the current standard of <500 μm to >5 mm. The spectral band option allows the synchronous recording of one to four images with exact local congruence, using photons from a choice of different spectral windows. These images are finally merged into a false-color presentation for interpretation. The spectral wavelengths can be chosen in the range of 350–650 nm according to a specific diagnostic question. Real-time photography is used as an adjunctive method to locate the scan position with respect to the sample to be examined. Samples under investigation were unstained tissue slides or full paraffin-embedded biopsy/tissue blocks. The images obtained by our new method were subsequently compared with standard H&E histology.
Results
Vertical laser scan images of superior quality from unstained histological slides as well as from the paraffin blocks of different benign and malignant cutaneous samples were made. When directly compared to the routine H&E histology, which is the gold standard, relevant tissue structures and changes in the normal architecture could be clearly identified and confirmed respectively.
Conclusion
A new non-invasive TPM scanning technique has been established that provides high-resolution imaging of biological tissue, such as human skin, in a distortion-free way and with a diagnostic relevant imaging size, thus allowing the assessment of tissue-significant medical issues in greater detail. Current use of our new method ex vivo shows that it is particularly appropriate for diagnosis and research applications of human skin.
Hintergrund
Zwei-Photonen-Mikroskopie (2PM) erzeugt bekanntlich nicht-invasiv hochaufgelöste Bilder aus dem Volumen von biologischem Gewebe. Der Fokus eines ultrakurz gepulsten Lasers führt hierbei zur sogenannten Zweiphotonenanregung endogener Fluorophore, deren Emissionen gesammelt, gemess
背景和目的双光子显微镜(TPM)以非侵入性的方式从生物样品(如人体皮肤)中生成高清晰度图像。将超短脉冲激光的焦点扫描穿过组织，收集、测量内源性荧光团双光子激发引起的荧光光子，并以扫描图像显示。到目前为止，TPM的主要缺点之一是受聚焦光学元件视场的限制，扫描面积仅为十分之一毫米。因此，诸如生物组织中良性或恶性变化的区分等基本信息不能被捕获。在bmbf支持的研究项目“FluoTOM”中，开发了一种新的改进TPM扫描技术来克服这些缺点，并在人体皮肤的良性和恶性样本上进行了中试试验。材料与方法新型TPM的视场由现行标准的500 μm扩展到5 mm。光谱波段选项允许同步记录一到四张具有精确局部一致性的图像，使用来自不同光谱窗口的光子。这些图像最后被合并成一个假彩色的表示，以供解释。光谱波长可根据具体诊断问题在350-650 nm范围内选择。实时摄影被用作辅助方法来定位相对于待检查样品的扫描位置。调查样本为未染色的组织切片或全石蜡包埋活检/组织块。通过我们的新方法获得的图像随后与标准H&E组织学进行比较。结果从未染色的组织学切片和不同良恶性皮肤标本的石蜡块上获得了高质量的垂直激光扫描图像。与常规H&E组织学(金标准)直接比较，可以清楚地识别和确认相关组织结构和正常结构的变化。结论建立了一种新的无创TPM扫描技术，可以以无扭曲的方式提供生物组织(如人体皮肤)的高分辨率成像，并且具有诊断相关的成像尺寸，从而可以更详细地评估组织重要的医学问题。目前我们的新方法在体外的应用表明，它特别适合于人体皮肤的诊断和研究应用。HintergrundZwei-Photonen-Mikroskopie(下午2点)erzeugt bekanntlich nicht-invasiv hochaufgeloste bild来自民主党的羊皮纸书卷冯biologischem Gewebe。德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国Bedingt durch das eingeschränkte Bildfeld der fokussierenden Optik ist die Scanfläche der下午2点，beang auf zehntelmm beschränkt。信息科学，信息科学，信息科学，信息科学，信息科学，信息科学，信息科学，信息科学，信息科学[1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1] [1]材料与方法荷兰技术修饰技术研究中心:Bildfeld von derzeit standardmäßig <500 μm auf > 5mm ausgedeent and die Bildqualität in ihrer Beurteilbarkeit entscheidend gesteigert werden。Eine Bildgebungsoption mit spectre - band - diskriminierung erlaubt es, mehrere exakt deckungsgleiche Bilder mitteles Photonen unterschiedlicher Spektralkanäle synchryzu erzegen, die zur visualinterpretation zu einem Falschfarbenbild fusion werden können。In Abhängigkeit von der spezifischen Fragestellung können die spektralen Fenster In einem Wellenlängenbereich von 350-650 nm ausgewählt werden。在Situs-Foto gewährleistet进行扫描定位和探测。Untersucht wurden ungefärbte Gewebeschnitte oder in Paraffinblöcke eingebettete Biopsie-/Gewebeproben。Die erzielten Bilder wurden anschließend mit herkömmlichen histologischen H&E-Schnitten bez<s:1> glich ihrer Aussagefähigkeit verglichen und bewertet。ErgebnisseHochauflösende Bilder von histologischen Schnitten sowie Paraffinblöcken gut- wie bösartiger Hautveränderungen konnten gewonnen werden。Im Vergleich mit histologischen H&E-Schnitten ließen sich krankheitsrelevant Veränderungen identifizieren。[2]夜间侵入性扫描技术[m] -扫描技术präsentiert, die hochauflösende生物化学学报，[j]。der Haut, in verzerrungsfreier Darstellung and mit diagnostich relevter Bildgröße ermöglicht and so deren Beurteilbarkeit in Hinblick of signante medizinische Fragestellungen entscheidendverbessert。 该技术被证明可以用于诊断和研究皮肤病。
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Pub Date : 2011-11-01DOI: 10.1016/j.mla.2011.09.001
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Pub Date : 2011-08-01DOI: 10.1016/j.mla.2011.05.003
Stefan Andree , Henrike Wilms , Jürgen Helfmann
Background
The skin protects the body against environmental stress, for example contact with biological or chemical substances. The protection must be effective for substances soluble in both polar solvents such as water, and non-polar solvents such as oil. Therefore a specific balance of water and lipids contained in the skin is necessary to maintain the protection against both of these chemicals. Potential applications of assessing the water concentration in skin range from determining the effect of soap-like substances on the water balance in the skin over the efficiency of hydrating cosmetics to detecting water deposition in skin due to heart or kidney diseases.
Objective
To our knowledge, there is currently no accepted in vivo method for quantitative determination of the water content in skin. Our aim was therefore to apply the spatially resolved reflectance (SRR) method to assess in vivo the water concentration in the skin.
Materials and methods
An overview on the advantages and disadvantages of our set-up is presented compared to other SRR set-ups and other (invasive) methods to assess the water concentration in skin. The set-up is described and the procedure for the evaluation of the absorption coefficient, from which the water concentration is determined. The performance of our set-up was examined by using fluid phantoms containing dimethyl sulfoxide (DMSO), water and scattering particles at different concentrations.
Results
The determined water concentration correlates linearly to the expected concentration with a correlation coefficient R2 of 0.977. The derived water concentration, taken from the spatially and spectrally resolved skin measurements presented here, is comparable to the concentration expected from the literature and found to be a realistic figure of 80%.
Conclusion
The optical non-invasive determination of water concentration in the skin is feasible using SRR.
Hintergrund
Die Haut schützt den Körper vor Stress durch Umwelteinflüsse, zum Beispiel durch Kontakt mit biologischen oder chemischen Substanzen. Der Schutz muss sowohl für fett- als auch wasserlösliche Substanzen funktionieren. Daher ist es notwendig, dass sich eine Balance zwischen Wasser und Lipiden in der Haut ausbildet, um gegen beide Arten von Substanzen zu schützen.
Die Bestimmung der Wasserkonzentration der Haut spielt daher eine besondere Rolle, wenn beispielsweise Effekte von Seifen oder ähnlichen Substanzen auf den Wasserhaushalt der Haut oder die Effizienz feuchtigkeitsspendender Kosmetika untersucht oder aber Wassereinlagerungen verursacht durch Herz- oder Nierenkrankheiten bestimmt werden sollen.
Zielsetzung
Zurzeit sind uns keine Verfahren für die nicht-invasive, quantitative In-vivo-Messung des Wassergehaltes bekannt, die
皮肤保护身体免受环境压力，例如与生物或化学物质接触。对于可溶于极性溶剂(如水)和非极性溶剂(如油)的物质，保护必须有效。因此，皮肤中所含的水和脂的特定平衡是必要的，以保持对这两种化学物质的保护。评估皮肤中水分浓度的潜在应用范围包括确定肥皂样物质对皮肤中水分平衡的影响，而不是保湿化妆品的效率，以及检测由于心脏或肾脏疾病导致的皮肤中的水分沉积。目的据我们所知，目前还没有一种公认的体内定量测定皮肤含水量的方法。因此，我们的目的是应用空间分辨反射率(SRR)方法来评估体内皮肤中的水浓度。材料和方法与其他SRR装置和其他(侵入性)方法相比，概述了我们装置的优点和缺点，以评估皮肤中的水分浓度。描述了装置和评估吸收系数的程序，从吸收系数中确定水的浓度。通过使用含有二甲基亚砜(DMSO)、水和不同浓度的散射粒子的流体幻影来检测我们的装置的性能。结果水的测定浓度与期望浓度呈线性相关，相关系数R2为0.977。从空间和光谱分辨率的皮肤测量中得出的水浓度与文献中预期的浓度相当，并且发现这是80%的现实数字。结论SRR法测定皮肤水分浓度是可行的。[参考文献][endnoteref: 1] [endnoteref: 1] [endnoteref: 1] [endnoteref: 1] [endnoteref: 1] [endnoteref: 1]。Der Schutz muss sowohl f r fett- also auch wasserlösliche Substanzen functionieren。在未来的发展中，我们将在未来的发展中，在未来的发展中，在未来的发展中，在未来的发展中，在未来的发展中，我们将在未来的发展中取得更大的进步。德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国德国ZielsetzungZurzeit sind uns keine Verfahren f r死亡夜间侵入性，定量的体内messung des Wassergehaltes bekant, die allgemein anerkant sind。Ziel war es daher, die ortsaufgelöste rkstreuung f r die Ermittelung der wasserkonzation von Haut in vivo anzuwenden。材料基金MethodenZunächst wid ein Überblick ber die Vor- und Nachteile unseres Verfahrens gegen ber anderen ortsaufgelösten Messverfahren und anderen (invasiven) Methoden zur Bestimmung des Wassergehaltes der Haut gegeben。他说:“我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是说，我的意思是我的意思。”研究结果表明:1 . flbe_sigphantome (flbe_sigphantome)， 2 . DMSO, 2 .二氧化钛，3 . Streuer, 3 . Einsatz kamen。ergebnisdie ermittelten Wasserkonzentrationen verhalten sich linear (bestestimmtheitsmasß R2 von 0,977) zur tatsächlichen Wasserkonzentration。Die aus den ortsaufgelösten Hautmessungen ermittelte wasserkonzation beträgt 80%，是在现实主义者Wert ist。ZusammenfassungDie optische，夜间侵入性尿路尿路尿路集中ortsaufgelöste尿路尿路尿路尿路尿路尿路尿路尿路尿路尿路尿路möglich。
{"title":"Feasibility of dermal water content determination by spatially resolved reflectance","authors":"Stefan Andree , Henrike Wilms , Jürgen Helfmann","doi":"10.1016/j.mla.2011.05.003","DOIUrl":"10.1016/j.mla.2011.05.003","url":null,"abstract":"<div><h3>Background</h3><p>The skin protects the body against environmental stress, for example contact with biological or chemical substances. The protection must be effective for substances soluble in both polar solvents such as water, and non-polar solvents such as oil. Therefore a specific balance of water and lipids contained in the skin is necessary to maintain the protection against both of these chemicals. Potential applications of assessing the water concentration in skin range from determining the effect of soap-like substances on the water balance in the skin over the efficiency of hydrating cosmetics to detecting water deposition in skin due to heart or kidney diseases.</p></div><div><h3>Objective</h3><p>To our knowledge, there is currently no accepted <em>in vivo</em> method for quantitative determination of the water content in skin. Our aim was therefore to apply the spatially resolved reflectance (SRR) method to assess <em>in vivo</em> the water concentration in the skin.</p></div><div><h3>Materials and methods</h3><p><span>An overview on the advantages and disadvantages of our set-up is presented compared to other SRR set-ups and other (invasive) methods to assess the water concentration in skin. The set-up is described and the procedure for the evaluation of the absorption coefficient, from which the water concentration is determined. The performance of our set-up was examined by using fluid phantoms containing </span>dimethyl sulfoxide (DMSO), water and scattering particles at different concentrations.</p></div><div><h3>Results</h3><p>The determined water concentration correlates linearly to the expected concentration with a correlation coefficient <em>R</em><sup>2</sup> of 0.977. The derived water concentration, taken from the spatially and spectrally resolved skin measurements presented here, is comparable to the concentration expected from the literature and found to be a realistic figure of 80%.</p></div><div><h3>Conclusion</h3><p>The optical non-invasive determination of water concentration in the skin is feasible using SRR.</p></div><div><h3>Hintergrund</h3><p>Die Haut schützt den Körper vor Stress durch Umwelteinflüsse, zum Beispiel durch Kontakt mit biologischen oder chemischen Substanzen. Der Schutz muss sowohl für fett- als auch wasserlösliche Substanzen funktionieren. Daher ist es notwendig, dass sich eine Balance zwischen Wasser und Lipiden in der Haut ausbildet, um gegen beide Arten von Substanzen zu schützen.</p><p>Die Bestimmung der Wasserkonzentration der Haut spielt daher eine besondere Rolle, wenn beispielsweise Effekte von Seifen oder ähnlichen Substanzen auf den Wasserhaushalt der Haut oder die Effizienz feuchtigkeitsspendender Kosmetika untersucht oder aber Wassereinlagerungen verursacht durch Herz- oder Nierenkrankheiten bestimmt werden sollen.</p></div><div><h3>Zielsetzung</h3><p>Zurzeit sind uns keine Verfahren für die nicht-invasive, quantitative <em>In-vivo</em>-Messung des Wassergehaltes bekannt, die ","PeriodicalId":88584,"journal":{"name":"Medical laser application : international journal for laser treatment and research","volume":"26 3","pages":"Pages 109-118"},"PeriodicalIF":0.0,"publicationDate":"2011-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.mla.2011.05.003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54921108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2011-08-01DOI: 10.1016/j.mla.2011.05.007
H.-Peter Berlien
{"title":"Reply to Prof. Bäumler's letter to the editor entitled “Powerful medical light sources such as lasers or IPL in the hands of laypersons?”","authors":"H.-Peter Berlien","doi":"10.1016/j.mla.2011.05.007","DOIUrl":"10.1016/j.mla.2011.05.007","url":null,"abstract":"","PeriodicalId":88584,"journal":{"name":"Medical laser application : international journal for laser treatment and research","volume":"26 3","pages":"Pages 145-146"},"PeriodicalIF":0.0,"publicationDate":"2011-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.mla.2011.05.007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54921164","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Human bone marrow-derived mesenchymal stem cells (BM-MSCs) are a promising cell source for regenerative medical applications because they can be easily isolated and expanded ex vivo. However, due to the lack of specific cell surface markers, it is difficult to identify whether ex vivo isolated BM-MSC populations are free of stromal fibroblasts. Bright-field microscopical analyses are insufficient to determine fibroblastic contaminations since these two cell types have similar cell morphologies.
Materials and methods
We employed and compared traditional flow cytometric (FACS) analysis, in vitro differentiation assays, and Raman spectroscopy to distinguish between human BM-MSCs and fibroblasts.
Results
We found that FACS analysis, utilizing previously described fibroblast-identifying antibodies, was inadequate in separating stromal fibroblasts from BM-MSCs as over 75% of the BM-MSCs shared these antigens. In vitro differentiation assays revealed that, in contrast to fibroblasts, BM-MSCs could be successfully differentiated into adipocytes, osteoblasts and chondrocytes. Using this method it was possible to discriminate between the two cell types. However, the need for prolonged in vitro culture periods of up to 4 weeks is a major disadvantage of this test method. Raman spectroscopy, a non-contact technique measuring the wavelength and intensity of inelastic scattered light from molecules by employing high-power near-infrared lasers, distinguished ultra-fast between BM-MSCs and fibroblasts (integration time of 100 s/cell).
Conclusion
Based on the results, we conclude that Raman spectroscopy is a suitable tool for the rapid detection of fibroblastic contaminations in BM-MSC cultures.
Hintergrund
Aus dem Knochenmark gewonnene mesenchymale Stammzellen (BM-MSCs) sind eine vielversprechende Zellquelle für Anwendungen innerhalb der regenerativen Medizin. Es ist jedoch schwierig zu bestimmen, ob isolierte BM-MSC-Populationen frei von Fibroblasten oder anderen, die Stammzellen kontaminierenden Zellen sind. Ziel der vorliegenden Studie war es festzustellen, ob sich die Raman-Spektroskopie zur nicht-invasiven, kontaktfreien Unterscheidung humaner BM-MSCs und Fibroblasten eignet.
Material und Methoden
Es wurden verschiedene Verfahren (Durchflusszytometrie, In-vitro-Differenzierungsassays, Raman-Spektroskopie) hinsichtlich der Differenzierbarkeit humaner BM-MSCs und Fibroblasten untersucht und verglichen.
Ergebnisse
Unsere Daten zeigen, dass Hellfeld-mikroskopische Analysen allein nicht ausreichen, um Fremdzellkontaminationen zu bestimmen, da BM-MSCs und Fibroblasten sehr ähnliche Zellmorphologien aufweisen. Auch die traditionelle durchflusszytometrische (FACS)-Analyse unter Verwendung zuvor beschriebener zell-populationsspez
人骨髓间充质干细胞(BM-MSCs)是一种很有希望用于再生医学应用的细胞来源，因为它们易于分离和体外扩增。然而，由于缺乏特异性的细胞表面标记物，很难确定离体分离的BM-MSC群体是否不含间质成纤维细胞。明场显微镜分析不足以确定成纤维细胞污染，因为这两种细胞类型具有相似的细胞形态。材料和方法我们采用传统的流式细胞术(FACS)分析、体外分化分析和拉曼光谱来区分人脑-间充质干细胞和成纤维细胞。结果我们发现，利用先前描述的成纤维细胞鉴定抗体的FACS分析不足以从BM-MSCs中分离间质成纤维细胞，因为超过75%的BM-MSCs共享这些抗原。体外分化实验显示，与成纤维细胞相比，BM-MSCs可以成功分化为脂肪细胞、成骨细胞和软骨细胞。使用这种方法可以区分两种细胞类型。然而，需要延长体外培养时间长达4周是该测试方法的主要缺点。拉曼光谱是一种利用高功率近红外激光测量分子非弹性散射光的波长和强度的非接触技术，可以超快地区分BM-MSCs和成纤维细胞(集成时间为100 s/细胞)。结论拉曼光谱是一种适合于骨髓间充质干细胞培养中成纤维细胞污染快速检测的工具。[中文]:间充质雄性Stammzellen (BM-MSCs)的研究进展。[2][1][1][1][1][1][1][1][1][1][1][1][1][2]。研究结果表明:人骨髓间充质干细胞与成纤维细胞的间充质干细胞具有夜间侵袭性。材料与方法(durchflusszymetrie, in -vitro differenzierungsassays, raman - spektroskie)研究了人骨髓间充质干细胞(bmmscs)和成纤维细胞(Fibroblasten, untersuht and verglichen)之间的差异。ergebnisunsere Daten zeigen, dass hellfield - microskopische Analysen allein nicht ausreichen, um fredmzellkontamationen zuestimmen, da BM-MSCs和Fibroblasten sehr ähnliche Zellmorphologien aufweisen。传统的纤维母细胞及骨髓间充质干细胞(bmmscs)分析。体外差异测定:zeigten, dass BM-MSCs, Gegensatz zu成纤维细胞，富含脂肪蛋白，成骨细胞和软骨蛋白的差异细胞。研究结果表明:1 .体外培养方法:1 .体外培养方法:1 .体外培养方法:Die Raman-Spektroskopie ermöglichte hingegen Die schnelle Differenzierung zwischen BM-MSCs和Fibroblasten。【关键词】纤维母细胞，培养，培养，成纤维细胞。
{"title":"Non-contact discrimination of human bone marrow-derived mesenchymal stem cells and fibroblasts using Raman spectroscopy","authors":"Marieke Pudlas , Daniel Alejandro Carvajal Berrio , Miriam Votteler , Steffen Koch , Sibylle Thude , Heike Walles , Katja Schenke-Layland","doi":"10.1016/j.mla.2011.05.004","DOIUrl":"10.1016/j.mla.2011.05.004","url":null,"abstract":"<div><h3>Background</h3><p>Human bone marrow-derived mesenchymal stem cells (BM-MSCs) are a promising cell source for regenerative medical applications because they can be easily isolated and expanded <em>ex vivo</em>. However, due to the lack of specific cell surface markers, it is difficult to identify whether <em>ex vivo</em> isolated BM-MSC populations are free of stromal fibroblasts. Bright-field microscopical analyses are insufficient to determine fibroblastic contaminations since these two cell types have similar cell morphologies.</p></div><div><h3>Materials and methods</h3><p>We employed and compared traditional flow cytometric (FACS) analysis, <em>in vitro</em> differentiation assays, and Raman spectroscopy to distinguish between human BM-MSCs and fibroblasts.</p></div><div><h3>Results</h3><p>We found that FACS analysis, utilizing previously described fibroblast-identifying antibodies, was inadequate in separating stromal fibroblasts from BM-MSCs as over 75% of the BM-MSCs shared these antigens. <em>In vitro</em> differentiation assays revealed that, in contrast to fibroblasts, BM-MSCs could be successfully differentiated into adipocytes, osteoblasts and chondrocytes. Using this method it was possible to discriminate between the two cell types. However, the need for prolonged <em>in vitro</em> culture periods of up to 4 weeks is a major disadvantage of this test method. Raman spectroscopy, a non-contact technique measuring the wavelength and intensity of inelastic scattered light from molecules by employing high-power near-infrared lasers, distinguished ultra-fast between BM-MSCs and fibroblasts (integration time of 100<!--> <!-->s/cell).</p></div><div><h3>Conclusion</h3><p>Based on the results, we conclude that Raman spectroscopy is a suitable tool for the rapid detection of fibroblastic contaminations in BM-MSC cultures.</p></div><div><h3>Hintergrund</h3><p>Aus dem Knochenmark gewonnene mesenchymale Stammzellen (BM-MSCs) sind eine vielversprechende Zellquelle für Anwendungen innerhalb der regenerativen Medizin. Es ist jedoch schwierig zu bestimmen, ob isolierte BM-MSC-Populationen frei von Fibroblasten oder anderen, die Stammzellen kontaminierenden Zellen sind. Ziel der vorliegenden Studie war es festzustellen, ob sich die Raman-Spektroskopie zur nicht-invasiven, kontaktfreien Unterscheidung humaner BM-MSCs und Fibroblasten eignet.</p></div><div><h3>Material und Methoden</h3><p>Es wurden verschiedene Verfahren (Durchflusszytometrie, <em>In-vitro</em>-Differenzierungsassays, Raman-Spektroskopie) hinsichtlich der Differenzierbarkeit humaner BM-MSCs und Fibroblasten untersucht und verglichen.</p></div><div><h3>Ergebnisse</h3><p>Unsere Daten zeigen, dass Hellfeld-mikroskopische Analysen allein nicht ausreichen, um Fremdzellkontaminationen zu bestimmen, da BM-MSCs und Fibroblasten sehr ähnliche Zellmorphologien aufweisen. Auch die traditionelle durchflusszytometrische (FACS)-Analyse unter Verwendung zuvor beschriebener zell-populationsspez","PeriodicalId":88584,"journal":{"name":"Medical laser application : international journal for laser treatment and research","volume":"26 3","pages":"Pages 119-125"},"PeriodicalIF":0.0,"publicationDate":"2011-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.mla.2011.05.004","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54921119","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2011-08-01DOI: 10.1016/j.mla.2011.05.002
Uwe J. Netz, Jan Toelsner, Uwe Bindig
We report on the development and production of tissue-like optical phantoms for detection and characterization of fluorescent objects close to the tissue surface. The phantoms mimic bulk optical properties such as light scattering and absorption of female breast tissue and human skin. Fluorescence sources are embedded for a given well-defined localization and volume to simulate pathologic lesions that are marked by specific molecular substances labeled with a fluorescent dye. The phantoms were made of epoxy resin with added titanium dioxide and black epoxy color paste for adjustment of the tissue-like scattering and absorbing properties. Indocyanine green was used as a fluorescent dye. The phantoms were evaluated with an experimental set-up for a new method for in vivo laser fluorescence tomography in the frequency domain.
Wir berichten in diesem Artikel über die Entwicklung und Herstellung von Phantomen mit gewebeähnlichen optischen Eigenschaften für die Detektion und Charakterisierung von oberflächennahen Fluoreszenzquellen. Die Phantome simulieren die optischen Parameter bezügliche Lichtstreuung und –absorption von weiblichem Brustgewebe bzw. Hautgewebe. Zur Simulation von pathologischen Gewebearealen, die mit einem spezifischen molekularen Marker belegt sind, an den ein Fluoreszenzfarbstoff gekoppelt ist, wurden in die Phantome Fluoreszenzquellen mit definierter Position und Ausdehnung integriert. Die Phantome bestehen aus Epoxidharz sowie Titandioxid und einer schwarzen Epoxid-Farbpaste zum Einstellen der gewebeähnlichen Streu- und Absorptionseigenschaften. Als Fluoreszenzfarbstoff wurde Indocyaningrün verwendet. Die Phantome wurden mit einem Experimentalaufbau für ein neues Verfahren einer In-vivo-Laserfluoreszenztomografie in der Frequency-Domain evaluiert.
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Pub Date : 2011-08-01DOI: 10.1016/S1615-1615(11)00040-8
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Pub Date : 2011-08-01DOI: 10.1016/j.mla.2011.04.001
Ivy Ndhundhuma , Carmen Hauser , Claudia Scalfi-Happ , Angelika Rück , Rudolf Steiner
Objective
The exposure of a photosensitizer to visible light, preferably laser light in the red region, in the presence of molecular oxygen results in a photochemical reaction described as photodynamic therapy (PDT). Photodynamic therapy is increasingly used for the treatment of skin cancers. The subcellular localization of the photosensitizer has been shown to be a key factor in the outcome of PDT. Mitochondrial localized photosensitizers are able to induce apoptosis very rapidly. Lysosomal localized photosensitizers can elicit either a necrotic or an apoptotic response. The present work contributes to the classification of the subcellular targets and cell death pathway of PDT using hydrophilic aluminum (III) phthalocyanine chloride tetrasulfonate (AlPcS4).
Material and methods
The localization of AlPcS4 in metastatic human melanoma (HT-144) cells was studied by means of fluorescent probes (Rh-123, LysoTracker® and Fluo-3) and a laser scanning microscope (LSM).
Results
The laser sources of the LSM induced a PDT effect with a redistribution of AlPcS4 inside the entire cell, followed by the increased fluorescence of AlPcS4 in the cytoplasm of HT-144 cells.
Conclusion
The results suggest that AlPcS4 is a potential sensitizer for melanoma skin cancer treatments. However, a comparison of its cytotoxicity, uptake and accumulation in normal and cancer cells deserve further investigation.
Hintergrund
Die Bestrahlung eines Photosensibilisators mit sichtbarem Licht, vorzugsweise Laserlicht im roten Spektralbereich, und in Gegenwart von molekularem Sauerstoff führt zu einer photochemischen Reaktion, die als photodynamische Therapie (PDT) bezeichnet wird. Die PDT wird zunehmend für die Behandlung von Hauttumoren eingesetzt. Für den Erfolg der PDT ist die subzelluläre Lokalisation des Photosensibilisators ausschlaggebend. In Mitochondrien lokalisierte Photosensibilisatoren können beschleunigt Apoptose auslösen. In Lysosomen lokalisierte Photosensibilisatoren können entweder Nekrose oder Apoptose hervorrufen. Die vorliegende Arbeit trägt zur Klassifikation subzellulärer Zielstrukturen bei, die über entsprechende Signalwege zum Zelltod führen.
Material und Methoden
Die Lokalisation von hydrophilem, tetra-sulfoniertem Aluminium(III)-Phthalocyanin-Chlorid (AlPcS4) in metastasierenden humanen Melanom-Zellen (HT-144) wurde mit Hilfe von Fluoreszenzmarkern (Rhodamin 123, LysoTracker® und Fluo-3) und dem Laser-Scan-Mikroskop (LSM) untersucht.
Ergebnisse
Die Laserstrahlquellen im LSM erzeugten einen PDT-Effekt, der zu einer Umverteilung des AlPcS4 in den Zellen führte, gefolgt von einem Fluoreszenzanstieg des AlPcS4 im Zytoplasma der HT-144 Zellen.
Zusammenfassung
AlPcS
目的将光敏剂暴露于可见光，最好是红色区域的激光，在存在分子氧的情况下导致称为光动力治疗(PDT)的光化学反应。光动力疗法越来越多地用于治疗皮肤癌。光敏剂的亚细胞定位已被证明是PDT结果的关键因素。线粒体定位光敏剂能够快速诱导细胞凋亡。溶酶体定位光敏剂可引起坏死或凋亡反应。本研究有助于利用亲水性酞菁四磺酸铝(AlPcS4)对PDT的亚细胞靶点和细胞死亡途径进行分类。材料和方法采用荧光探针(Rh-123、LysoTracker®和Fluo-3)和激光扫描显微镜(LSM)研究AlPcS4在转移性人黑色素瘤(HT-144)细胞中的定位。结果LSM激光源诱导HT-144细胞产生PDT效应，使AlPcS4在整个细胞内重新分布，细胞质中AlPcS4的荧光增强。结论AlPcS4是治疗黑色素瘤皮肤癌的潜在增敏剂。然而，其在正常细胞和癌细胞中的细胞毒性、摄取和积累的比较值得进一步研究。光敏剂与光化学反应(photochemischen reakchen)、光动力学治疗(photodynamische Therapie (PDT))、光化学反应(photochemischen reakchen)、光动力学治疗(photodynamische Therapie, PDT)等。Die PDT wind zunememederfendi . [j] Die Behandlung von hautumenenesetz。 r den Erfolg der PDT ist die subzelluläre光敏剂的定位。在线粒体lokalisierte光敏bilisatoren können beschleunigt凋亡auslösen。在溶酶体中，lockalisierte光敏分解蛋白können与Nekrose oder凋亡蛋白结合。Die vorliegende Arbeit trägt zur分类subzellulärer Zielstrukturen bei, Die ber entsprechende Signalwege zum Zelltod f hren。材料与方法铝(III)-酞青素-氯化物(AlPcS4)在转移性人黑色素- zellen (HT-144)中的局部化，荧光标记物(Rhodamin 123, LysoTracker®和Fluo-3)和激光扫描-微孔(LSM)的局部化。ergebnisdie Laserstrahlquellen im LSM erzeeugten einen pdt - effet, der zu einer Umverteilung des AlPcS4 in den Zellen f， gefolgt von einem Fluoreszenzanstieg des AlPcS4 im Zytoplasma der HT-144 Zellen。人黑色素- zellen (HT-144)光敏增效剂;jedoch sind weitere vergleichende Messungen和gesunden zellenendi。
{"title":"Subcellular co-localization of aluminum (III) phthalocyanine chloride tetrasulphonate with fluorescent markers in the human melanoma cell-line HT-144","authors":"Ivy Ndhundhuma , Carmen Hauser , Claudia Scalfi-Happ , Angelika Rück , Rudolf Steiner","doi":"10.1016/j.mla.2011.04.001","DOIUrl":"10.1016/j.mla.2011.04.001","url":null,"abstract":"<div><h3>Objective</h3><p>The exposure of a photosensitizer to visible light, preferably laser light in the red region, in the presence of molecular oxygen results in a photochemical reaction described as photodynamic therapy (PDT). Photodynamic therapy is increasingly used for the treatment of skin cancers. The subcellular localization of the photosensitizer has been shown to be a key factor in the outcome of PDT. Mitochondrial localized photosensitizers are able to induce apoptosis very rapidly. Lysosomal localized photosensitizers can elicit either a necrotic or an apoptotic response. The present work contributes to the classification of the subcellular targets and cell death pathway of PDT using hydrophilic aluminum (III) phthalocyanine chloride tetrasulfonate (AlPcS<sub>4</sub>).</p></div><div><h3>Material and methods</h3><p>The localization of AlPcS<sub>4</sub> in metastatic human melanoma (HT-144) cells was studied by means of fluorescent probes (Rh-123, LysoTracker<sup>®</sup> and Fluo-3) and a laser scanning microscope (LSM).</p></div><div><h3>Results</h3><p>The laser sources of the LSM induced a PDT effect with a redistribution of AlPcS<sub>4</sub> inside the entire cell, followed by the increased fluorescence of AlPcS<sub>4</sub> in the cytoplasm of HT-144 cells.</p></div><div><h3>Conclusion</h3><p>The results suggest that AlPcS<sub>4</sub> is a potential sensitizer for melanoma skin cancer treatments. However, a comparison of its cytotoxicity, uptake and accumulation in normal and cancer cells deserve further investigation.</p></div><div><h3>Hintergrund</h3><p>Die Bestrahlung eines Photosensibilisators mit sichtbarem Licht, vorzugsweise Laserlicht im roten Spektralbereich, und in Gegenwart von molekularem Sauerstoff führt zu einer photochemischen Reaktion, die als photodynamische Therapie (PDT) bezeichnet wird. Die PDT wird zunehmend für die Behandlung von Hauttumoren eingesetzt. Für den Erfolg der PDT ist die subzelluläre Lokalisation des Photosensibilisators ausschlaggebend. In Mitochondrien lokalisierte Photosensibilisatoren können beschleunigt Apoptose auslösen. In Lysosomen lokalisierte Photosensibilisatoren können entweder Nekrose oder Apoptose hervorrufen. Die vorliegende Arbeit trägt zur Klassifikation subzellulärer Zielstrukturen bei, die über entsprechende Signalwege zum Zelltod führen.</p></div><div><h3>Material und Methoden</h3><p>Die Lokalisation von hydrophilem, tetra-sulfoniertem Aluminium(III)-Phthalocyanin-Chlorid (AlPcS<sub>4</sub>) in metastasierenden humanen Melanom-Zellen (HT-144) wurde mit Hilfe von Fluoreszenzmarkern (Rhodamin 123, LysoTracker<sup>®</sup> und Fluo-3) und dem Laser-Scan-Mikroskop (LSM) untersucht.</p></div><div><h3>Ergebnisse</h3><p>Die Laserstrahlquellen im LSM erzeugten einen PDT-Effekt, der zu einer Umverteilung des AlPcS<sub>4</sub> in den Zellen führte, gefolgt von einem Fluoreszenzanstieg des AlPcS<sub>4</sub> im Zytoplasma der HT-144 Zellen.</p></div><div><h3>Zusammenfassung</h3><p>AlPcS<s","PeriodicalId":88584,"journal":{"name":"Medical laser application : international journal for laser treatment and research","volume":"26 3","pages":"Pages 93-100"},"PeriodicalIF":0.0,"publicationDate":"2011-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.mla.2011.04.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54921066","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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