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Infrared irradiation of skin for the development of non-invasive health monitoring technologies 红外照射皮肤为发展无创健康监测技术
Pub Date : 2015-06-19 DOI: 10.1117/12.2182345
Hisham Abdussamad Abbas, G. Triplett
Infrared radiation was employed to study the optical transmission properties of pigskin and the factors that influence transmission at room temperature. The skin samples from the forehead of piglets were irradiated using an infrared-pulsed source by varying the beam properties such as optical power, power density, duty cycle, as well as sample thickness. Because infrared radiation in select instances can penetrate through thick-fleshy skin more easily than visible radiation, temperature fluctuations observed within the skin samples stemming from exposure-dependent absorption revealed interesting transmission properties and the limits of optical exposure. Pigskin was selected for this study since its structure most closely resembles that of human skin. Furthermore, the pulsed beam technique compared to continuous operation offers more precise control of heat generation within the skin. Through this effort, the correlated pulsed-beam parameters that influence infrared transmission were identified and varied to minimize the internal absorption losses through the dermis layers. The two most significant parameters that reduce absorption losses were frequency and duty cycle of the pulsed beam. Using the Bouger-Beer-Lambert Law, the absorption coefficient from empirical data is approximated, while accepting that the absorption coefficient is neither uniform nor linear. Given that the optical source used in this study was single mode, the infrared spectra obtained from irradiated samples also reveal characteristics of the skin structure. Realization of appropriate sample conditions and exposure parameters that reduce light attenuation within the skin and sample degradation could give way to novel non-invasive measuring techniques for health monitoring purposes.
采用红外辐射法研究了猪皮在室温下的光传输特性及影响光传输的因素。使用红外脉冲源照射仔猪额头皮肤样品,通过改变光束特性,如光功率、功率密度、占空比以及样品厚度。由于红外辐射在特定情况下比可见光辐射更容易穿透厚肉皮肤,因此在皮肤样品中观察到的由暴露依赖性吸收引起的温度波动揭示了有趣的传输特性和光学暴露的限制。之所以选择猪皮作为研究对象,是因为猪皮的结构与人类皮肤最为相似。此外,与连续操作相比,脉冲束技术可以更精确地控制皮肤内的热量产生。通过这项工作,确定了影响红外传输的相关脉冲光束参数,并对其进行了调整,以最大限度地减少通过真皮层的内部吸收损失。减少吸收损失的两个最重要的参数是脉冲光束的频率和占空比。利用布格-比尔-朗伯定律,在接受吸收系数既非均匀也非线性的情况下,对经验数据的吸收系数进行了近似。由于本研究使用的光源为单模,因此从辐照样品中获得的红外光谱也揭示了皮肤结构的特征。实现适当的样品条件和暴露参数,减少皮肤内的光衰减和样品降解,可以为用于健康监测目的的新型非侵入性测量技术让路。
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引用次数: 1
Innovative parameters obtained for digital analysis of microscopic images to evaluate in vitro hemorheological action of anesthetics 获得显微图像数字分析的创新参数,以评估麻醉剂的体外血液流变学作用
Pub Date : 2015-06-19 DOI: 10.1117/12.2180791
Analía I. Alet, Sabrina Basso, M. Delannoy, N. Alet, M. D'Arrigo, H. Castellini, B. Riquelme
Drugs used during anesthesia could enhance microvascular flow disturbance, not only for their systemic cardiovascular actions but also by a direct effect on the microcirculation and in particular on hemorheology. This is particularly important in high-risk surgical patients such as those with vascular disease (diabetes, hypertension, etc.). Therefore, in this work we propose a set of innovative parameters obtained by digital analysis of microscopic images to study the in vitro hemorheological effect of propofol and vecuronium on red blood cell from type 2 diabetic patients compared to healthy donors. Obtained innovative parameters allow quantifying alterations in erythrocyte aggregation, which can increase the in vivo risk of microcapillary obstruction.
麻醉过程中使用的药物不仅可以增强微血管流动的紊乱,还可以直接影响微循环,特别是血液流变学。这对于高危手术患者,如血管疾病患者(糖尿病、高血压等)尤为重要。因此,在这项工作中,我们提出了一套创新的参数,通过显微镜图像的数字分析获得,以研究异丙酚和维库溴铵对2型糖尿病患者红细胞的体外血液流变学影响,并与健康献血者进行比较。获得的创新参数可以量化红细胞聚集的变化,这可以增加体内微血管阻塞的风险。
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引用次数: 4
Combined phosphorescence-holographic approach for singlet oxygen detection in biological media 生物介质中单线态氧检测的联合磷光-全息方法
Pub Date : 2015-06-19 DOI: 10.1117/12.2180909
I. Semenova, A. Belashov, D. M. Beltukova, N. Petrov, O. Vasyutinskii
The paper presents a novel combined approach aimed to detect and monitor singlet oxygen molecules in biological specimens by means of the simultaneous recording and monitoring of their deactivation dynamics in the two complementary channels: radiative and nonradiative. The approach involves both the direct registration of phosphorescence at the wavelength of about 1270 nm caused by radiative relaxation of excited singlet oxygen molecules and holographic recording of thermal disturbances in the medium produced by their nonradiative relaxation. The data provides a complete set of information on singlet oxygen location and dynamics in the medium. The approach was validated in the case study of photosensitized generation of singlet oxygen in onion cell structures.
本文提出了一种新的组合方法,旨在通过同时记录和监测其在两个互补通道:辐射和非辐射中的失活动力学来检测和监测生物标本中的单线态氧分子。该方法包括直接记录由激发单线态氧分子的辐射弛豫引起的约1270 nm波长的磷光,以及由它们的非辐射弛豫引起的介质热扰动的全息记录。该数据提供了一套完整的单线态氧在介质中的位置和动力学信息。该方法在洋葱细胞结构中光敏生成单线态氧的案例研究中得到了验证。
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引用次数: 0
Raman spectroscopic analysis of oral squamous cell carcinoma and oral dysplasia in the high-wavenumber region 高波数区口腔鳞状细胞癌和口腔发育不良的拉曼光谱分析
Pub Date : 2015-06-19 DOI: 10.1117/12.2180996
L. F. C. S. Carvalho, F. Bonnier, K. O'Callaghan, J. O’Sullivan, S. Flint, Lázaro P. M. Neto, C. A. T. Soto, L. dos Santos, A. Martin, H. Byrne, F. Lyng
Raman spectroscopy can provide a molecular-level signature of the biochemical composition and structure of cells with excellent spatial resolution and could be useful to monitor changes in composition for early stage and non-invasive cancer diagnosis, both ex-vivo and in vivo. In particular, the fingerprint spectral region (400–1,800 cm-1) has been shown to be very promising for optical biopsy purposes. However, limitations to discrimination of dysplastic and inflammatory processes based on the fingerprint region still persist. In addition, the Raman spectral signal of dysplastic cells is one important source of misdiagnosis of normal versus pathological tissues. The high wavenumber region (2,800–3,600 cm-1) provides more specific information based on N-H, O-H and C-H vibrations and can be used to identify the subtle changes which could be important for discrimination of samples. In this study, we demonstrate the potential of the highwavenumber spectral region by collecting Raman spectra of nucleoli, nucleus and cytoplasm from oral epithelial cancer (SCC-4) and dysplastic (DOK) cell lines and from normal oral epithelial primary cells, in vitro, which were then analyzed by area under the curve as a method to discriminate the spectra. In this region, we will show the discriminatory potential of the CH vibrational modes of nucleic acids, proteins and lipids. This technique demonstrated more efficient discrimination than the fingerprint region when we compared the cell cultures.
拉曼光谱可以提供分子水平的细胞生化组成和结构特征,具有良好的空间分辨率,可用于监测早期和非侵袭性癌症诊断,无论是离体还是体内。特别是,指纹光谱区域(400-1,800 cm-1)已被证明是非常有前途的光学活检目的。然而,基于指纹区鉴别发育不良和炎症过程的局限性仍然存在。此外,发育不良细胞的拉曼光谱信号是正常与病理组织误诊的重要来源之一。高波数区域(2,800-3,600 cm-1)提供了基于N-H, O-H和C-H振动的更具体的信息,可用于识别可能对样品区分很重要的细微变化。在这项研究中,我们通过收集口腔上皮癌(SCC-4)和发育异常(DOK)细胞系以及正常口腔上皮原代细胞的核核、细胞核和细胞质的拉曼光谱,在体外证明了高波数光谱区域的潜力,然后通过曲线下面积分析作为鉴别光谱的方法。在这个区域,我们将展示核酸、蛋白质和脂类的CH振动模式的区别潜力。在细胞培养对比中,该技术比指纹区识别效率更高。
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引用次数: 4
Optical fluorescence spectroscopy to detect hepatic necrosis after normothermic ischemia: animal model 荧光光谱法检测常温缺血后肝坏死的动物模型
Pub Date : 2015-06-19 DOI: 10.1117/12.2180897
R. A. Romano, J. D. Vollet-Filho, S. Pratavieira, J. L. Fernández, C. Kurachi, V. Bagnato, O. Castro-e-Silva, A. Sankarankutty
Liver transplantation is a well-established treatment for liver failure. However, the success of the transplantation procedure depends on liver graft conditions. The tissue function evaluation during the several transplantation stages is relevant, in particular during the organ harvesting, when a decision is made concerning the viability of the graft. Optical fluorescence spectroscopy is a good option because it is a noninvasive and fast technique. A partial normothermic hepatic ischemia was performed in rat livers, with a vascular occlusion of both median and left lateral lobes, allowing circulation only for the right lateral lobe and the caudate lobe. Fluorescence spectra under excitation at 532 nm (doubled frequency Nd:YAG laser) were collected using a portable spectrometer (USB2000, Ocean Optics, USA). The fluorescence emission was collected before vascular occlusion, after ischemia, and 24 hours after reperfusion. A morphometric histology analysis was performed as the gold standard evaluation ─ liver samples were analyzed, and the percentage of necrotic tissue was obtained. The results showed that changes in the fluorescence emission after ischemia can be correlated with the amount of necrosis evaluated by a morphometric analysis, the Pearson correlation coefficient of the generated model was 0.90 and the root mean square error was around 20%. In this context, the laser-induced fluorescence spectroscopy technique after normothermic ischemia showed to be a fast and efficient method to differentiate ischemic injury from viable tissues.
肝移植是一种公认的治疗肝衰竭的方法。然而,移植手术的成功取决于肝移植的条件。在几个移植阶段的组织功能评估是相关的,特别是在器官采集期间,当决定移植物的生存能力时。光学荧光光谱是一个很好的选择,因为它是一种无创和快速的技术。在大鼠肝脏中进行部分常温肝缺血,中间和左外侧叶血管闭塞,只允许右外侧叶和尾状叶循环。使用便携式光谱仪(USB2000, Ocean Optics, USA)采集532 nm(双频Nd:YAG激光)激发下的荧光光谱。在血管闭塞前、缺血后和再灌注后24小时采集荧光发射。进行形态计量学组织学分析作为金标准评估──分析肝脏样本,并获得坏死组织的百分比。结果表明,荧光发射在缺血后的变化可与形态学分析评估的坏死程度相关,生成的模型的Pearson相关系数为0.90,均方根误差在20%左右。在此背景下,常温缺血后激光诱导荧光光谱技术是一种快速有效的区分缺血损伤和活组织的方法。
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引用次数: 0
Assembly and characterization of a fluorescence lifetime spectroscopy system for skin lesions diagnostic 用于皮肤病变诊断的荧光寿命光谱系统的组装和表征
Pub Date : 2015-06-19 DOI: 10.1117/12.2180599
Marcelo Saito Nogueira, Ramon Gabriel Texiera Rosa, S. Pratavieira, Camila de Paula D´Almeida, C. Kurachi
The fluorescence spectra and fluorescence lifetime analysis in biological tissues has been presented as a technique of a great potential for tissue characterization for diagnostic purposes. The objective of this study is to assemble and characterize a fluorescence lifetime spectroscopy system for diagnostic of clinically similar skin lesions in vivo. The fluorescence lifetime measurements were performed using the Time Correlated Single Photon Counting (Becker & Hickl, Berlin, Germany) technique. Two lasers, one emitting at 378 nm and another at 445 nm, are used for excitation with 20, 50 and 80 MHz repetition rate. A bifurcated optical fiber probe conducts the excitation light to the sample, the collected light is transmitted through bandpass filters and delivered to a hybrid photomultiplier tube detector. The fluorescence spectra were obtained by using a portable spectrometer (Ocean Optics USB-2000-FLG) with the same excitation sources. An instrument response function of about 300 ps was obtained and the spectrum and fluorescence lifetime of a standard fluorescent molecule (Rhodamine 6G) was measured for the calibration of the system ((4.1 ± 0.3) ns). The assembled system was considered robust, well calibrated and will be used for clinical measurements of skin lesions.
生物组织中的荧光光谱和荧光寿命分析已被认为是一种具有巨大潜力的用于诊断目的的组织表征技术。本研究的目的是组装和表征荧光寿命光谱系统诊断临床相似的皮肤病变在体内。荧光寿命测量使用时间相关单光子计数(Becker & Hickl,柏林,德国)技术进行。使用两个激光器,一个发射波长为378nm,另一个发射波长为445nm,分别以20mhz、50mhz和80mhz的重复频率进行激发。分岔光纤探头将激发光传导至样品,采集到的光通过带通滤波器传输至混合光电倍增管探测器。荧光光谱采用相同激发源的便携式光谱仪(Ocean Optics USB-2000-FLG)获得。获得了约300 ps的仪器响应函数,并测量了标准荧光分子(罗丹明6G)的光谱和荧光寿命((4.1±0.3)ns),用于系统校准。组装的系统被认为是健壮的,校准良好,将用于皮肤病变的临床测量。
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引用次数: 22
A simple dental caries detection system using full spectrum of laser-induced fluorescence 一个简单的龋检测系统,使用全光谱激光诱导荧光
Pub Date : 2015-06-19 DOI: 10.1117/12.2180777
Renata Maciel Rocha-Cabral, F. Mendes, E. Maldonado, D. Zezell
Objectives: to develop an apparatus for the detection of early caries lesions in enamel using the full extent of the tooth fluorescence spectrum, through the integration of a laser diode, fiber optics, filters and one portable spectrometer connected to a computer, all commercially available; to evaluate the developed device in clinical and laboratory tests, and compare its performance with commercial equipment. Methods: clinical examinations were performed in patients with indication for exodontics of premolars. After examinations, the patients underwent surgery and the teeth were stored individually. The optical measurements were repeated approximately two months after extraction, on the same sites previously examined, then histological analysis was carried out. Results: the spectral detector has presented high specificity and moderate sensitivity when applied to differentiate between healthy and damaged tissues, with no significant differences from the performance of the commercial equipment. The developed device is able to detect initial damages in enamel, with depth of approximately 300 μm. Conclusions: we successfully demonstrated the development of a simple and portable system based in laser-induced fluorescence for caries detection, assembled from common commercial parts. As the spectral detector acquires a complete recording of the spectrum from each tissue, it is possible to use it for monitoring developments of caries lesions.
目标:开发一种仪器,通过集成激光二极管、光纤、滤光片和连接到计算机的便携式光谱仪,利用牙齿荧光光谱的全部范围来检测牙釉质的早期龋齿病变,所有这些都是市售的;在临床和实验室测试中评估所开发的设备,并将其性能与商用设备进行比较。方法:对有适应证的前磨牙患者进行临床检查。检查后,患者接受手术治疗,牙齿单独存放。提取后大约两个月,在先前检查的相同部位重复光学测量,然后进行组织学分析。结果:光谱检测仪用于区分健康组织和受损组织具有高特异性和中等灵敏度,与商用设备性能无显著差异。该设备能够检测牙釉质的初始损伤,深度约为300 μm。结论:我们成功地展示了一种基于激光诱导荧光的简单便携式龋检测系统的开发,该系统由常见的商业部件组装而成。由于光谱检测器从每个组织中获得完整的光谱记录,因此可以使用它来监测龋齿病变的发展。
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引用次数: 4
Onychomycosis diagnosis using fluorescence and infrared imaging systems 荧光和红外成像系统诊断甲真菌病
Pub Date : 2015-06-19 DOI: 10.1117/12.2180998
A. P. da Silva, T. C. Fortunato, M. D. Stringasci, C. Kurachi, V. Bagnato, N. Inada
Onychomycosis is a common disease of the nail plate, constituting approximately half of all cases of nail infection. Onychomycosis diagnosis is challenging because it is hard to distinguish from other diseases of the nail lamina such as psoriasis, lichen ruber or eczematous nails. The existing methods of diagnostics so far consist of clinical and laboratory analysis, such as: Direct Mycological examination and culture, PCR and histopathology with PAS staining. However, they all share certain disadvantages in terms of sensitivity and specificity, time delay, or cost. This study aimed to evaluate the use of infrared and fluorescence imaging as new non-invasive diagnostic tools in patients with suspected onychomycosis, and compare them with established techniques. For fluorescence analysis, a Clinical Evince (MM Optics®) was used, which consists of an optical assembly with UV LED light source wavelength 400 nm ± 10 nm and the maximum light intensity: 40 mW/cm2 ± 20%. For infrared analysis, a Fluke® Camera FKL model Ti400 was used. Patients with onychomycosis and control group were analyzed for comparison. The fluorescence images were processed using MATLAB® routines, and infrared images were analyzed using the SmartView® 3.6 software analysis provided by the company Fluke®. The results demonstrated that both infrared and fluorescence could be complementary to diagnose different types of onychomycosis lesions. The simplicity of operation, quick response and non-invasive assessment of the nail patients in real time, are important factors to be consider for an implementation.
甲真菌病是甲板的一种常见疾病,约占所有甲感染病例的一半。甲真菌病的诊断是具有挑战性的,因为它很难与其他疾病的甲层,如牛皮癣,橡胶苔藓或湿疹指甲区分。现有的诊断方法包括临床和实验室分析,如:直接真菌学检查和培养,PCR和PAS染色的组织病理学。然而,它们在灵敏度和特异性、时间延迟或成本方面都有一定的缺点。本研究旨在评估红外和荧光成像作为新的无创诊断工具在疑似甲癣患者中的应用,并将其与现有技术进行比较。荧光分析使用临床仪器(MM Optics®),该仪器由紫外LED光源波长为400 nm±10 nm的光学组件组成,最大光强为40 mW/cm2±20%。红外分析使用Fluke®相机FKL型号Ti400。甲真菌病患者与对照组进行比较分析。使用MATLAB®例程处理荧光图像,使用Fluke®公司提供的SmartView®3.6软件分析红外图像。结果表明,红外与荧光在不同类型的甲癣病变诊断中具有互补作用。手术简单、反应迅速、对甲患者进行实时无创评估是实施该手术需要考虑的重要因素。
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引用次数: 0
Laser speckle contrast imaging of blood flow from anesthetized mice: correcting drifts in measurements due to breathing movements 麻醉小鼠血流的激光散斑对比成像:校正呼吸运动引起的测量漂移
Pub Date : 2015-06-19 DOI: 10.1117/12.2087063
G. Nogueira, Márcio A. C. Ribeiro, Juliane C Campos, J. Ferreira
Background: Laser speckle contrast imaging allows non-invasive assessment of cutaneous blood flow. Although the technique is attractive to measure a quantity related to the skin blood flow (SBF) in anesthetized animal models, movements from breathing can mask the SBF signal. As a consequence, the measurement is overestimated because a variable amount of a DC component due to the breathing movements is added to the SBF signal. Objective: To evaluate a method for estimating the background level of the SBF signal, rejecting artefacts from breathing. Methods: A baseline correction method used for accurate DNA sequencing was evaluated, based on estimating the background level of a signal in small temporal sliding-windows. The method was applied to evaluate a mouse model of hindlimb ischemia. SBF signals from hindlimbs of anesthetized C57BL/6 mice (n=13) were registered. The mean SBF (Fi and Fc from ischemic and control hindlimbs) were computed from the registers and from the corresponding estimated background levels (Fib and Fcb from ischemic and control hindlimbs). Results: The mean values of the percentages (a measure of ischemia) MI = (Fi/Fc).100 and MIb = (Fib/Fcb).100 were computed to be 30±4% and 23±3% respectively (mean ± SE). Evidences of statistical differences between both, ischemic and control hindlimbs, were obtained (p<0.05, paired student-t). The mean error [(MI-MIb)/MIb].100 obtained was 45±14% (mean±SE). Conclusion: The recovery of a corrupted SBF signal by breathing artefacts is feasible, allowing more accurate measurements.
背景:激光散斑对比成像可以对皮肤血流进行无创评估。尽管该技术在麻醉动物模型中测量与皮肤血流量(SBF)相关的数量很有吸引力,但呼吸运动可以掩盖SBF信号。结果,测量结果被高估了,因为呼吸运动引起的直流分量的可变量被添加到SBF信号中。目的:探讨一种排除呼吸伪影的SBF信号背景电平估计方法。方法:基于在小时间滑动窗口中估计信号的背景电平,评估用于精确DNA测序的基线校正方法。应用该方法对小鼠后肢缺血模型进行评价。记录麻醉C57BL/6小鼠后肢SBF信号(n=13)。平均SBF(来自缺血后肢和对照后肢的Fi和Fc)从注册表和相应的估计背景水平(来自缺血后肢和对照后肢的Fib和Fcb)计算。结果:心肌缺血百分数MI = (Fi/Fc)的平均值。100和MIb = (Fib/Fcb)。100例分别为30±4%和23±3% (mean±SE)。缺血后肢与对照组后肢之间的差异有统计学意义(p<0.05,配对student-t)。平均误差[(MI-MIb)/MIb]。100例为45±14%(平均值±SE)。结论:通过呼吸伪影恢复损坏的SBF信号是可行的,可以实现更精确的测量。
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引用次数: 0
Effect of laser phototherapy on human alveolar bone repair: micro tomographic and histomorphometrical analysis 激光光疗对人牙槽骨修复的影响:显微层析和组织形态学分析
Pub Date : 2015-06-19 DOI: 10.1117/12.2181041
M. Romão, M. Marques, A. Cortes, A. Horliana, M. S. Moreira, C. Lascala
The immediate dental implant placement in the molars region is critical, because of the high amount of bone loss and the discrepancy between the alveolar crest thickness and the dental implant platform. Laser phototherapy (LPT) improves bone repair thus could accelerate the implant placement. Twenty patients were selected for the study. Ten patients were submitted to LPT with GaAlAs diode laser (808nm) during molar extraction, immediately after, 24h, 48h, 72h, 96h and 7 days. The irradiations were applied in contact and punctual mode (100mW, 0.04cm2, 0.75J/cm2, 30s per point, 3J per point). The control group (n=10) received the same treatment; however with the power of the laser off. Forty days later samples of the tissue formed inside the sockets were obtained for further microtomography (microCTs) and histomorphometry analyses. Data were compared by the Student t test, whereas those from the different microCT parameters were compared by the Pearson correlation test (p<0.05). The relative bone volume, as well as area was significantly higher (p<0.001) in the lased than the control group. In the control group there were negative correlations between number and thickness, and between number and separation of trabecula (p<0.01). Between thickness and separation of trabecula the correlation was positive (p<0.01). The laser group showed significant negative correlation between the number and the thickness of trabecula (p<0.01). LPT accelerated bone repair. By the Pearson correlation test it was possible to infer that the lased group presented a more homogeneous trabecular configuration, which would allow earlier dental implant placement.
由于牙槽嵴厚度与种植体平台之间存在差异,因此在磨牙区域立即放置种植体是至关重要的。激光光疗(LPT)可以改善骨修复,从而加快种植体的植入。20名患者被选为研究对象。10例患者分别于拔牙时、拔牙后、24h、48h、72h、96h和7d接受GaAlAs二极管激光(808nm) LPT。辐照采用接触和定时模式(100mW, 0.04cm2, 0.75J/cm2, 30s /点,3J /点)。对照组(n=10)给予相同治疗;然而,随着激光的功率关闭。40天后,获得窝内形成的组织样本,用于进一步的显微断层扫描(microct)和组织形态学分析。数据比较采用Student t检验,不同显微ct参数的数据比较采用Pearson相关检验(p<0.05)。实验组相对骨体积和骨面积均显著高于对照组(p<0.001)。对照组小梁数目与厚度、数目与分离度呈负相关(p<0.01)。小梁厚度与分离程度呈正相关(p<0.01)。激光组小梁数量与厚度呈显著负相关(p<0.01)。LPT加速骨修复。通过Pearson相关检验,可以推断出激光组呈现出更均匀的小梁结构,这将允许更早地种植牙。
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引用次数: 0
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Biophotonics South America
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