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1.
Prostaglandins Other Lipid Mediat ; 151: 106475, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32711127

RESUMO

Better knowledge of the breast tumor microenvironment is required for surgical resection and understanding the processes of tumor development. Raman spectroscopy is a promising tool that can assist in uncovering the molecular basis of disease and provide quantifiable molecular information for diagnosis and treatment evaluation. In this work, eighty-eight frozen breast tissue sections, including forty-four normal and forty-four tumor sections, were mapped in their entirety using a 250-µm-square measurement grid. Two or more smaller regions of interest within each tissue were additionally mapped using a 25 µm-square step size. A deep learning algorithm, convolutional neural network (CNN), was developed to distinguish histopathologic features with-in individual and across multiple tissue sections. Cancerous breast tissue were discriminated from normal breast tissue with 90 % accuracy, 88.8 % sensitivity and 90.8 % specificity with an excellent Area Under the Receiver Operator Curve (AUROC) of 0.96. Features that contributed significantly to the model were identified and used to generate RGB images of the tissue sections. For each grid point (pixel) on a Raman map, color was assigned to intensities at frequencies of 1002 cm-1 (Phenylalanine), 869 cm-1 (Proline, CC stretching of hydroxyproline-collagen assignment, single bond stretching vibrations for the amino acids proline, valine and polysaccharides) and 1309 cm-1 (CH3/CH2 twisting or bending mode of lipids). The Raman images clearly associate with hematoxylin and eosin stained tissue sections and allow clear visualization of boundaries between normal adipose, connective tissue and tumor. We demonstrated that this simple imaging technique allows high-resolution, straightforward molecular interpretation of Raman images. Raman spectroscopy provides rapid, label-free imaging of microscopic features with high accuracy. This method has application as laboratory tool and can assist with intraoperative tissue assessment during Breast Conserving surgery.


Assuntos
Neoplasias da Mama/patologia , Análise Espectral Raman , Microambiente Tumoral , Aprendizado Profundo , Feminino , Humanos
2.
Biomed Mater ; 12(4): 045008, 2017 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-28357996

RESUMO

Few studies have been reported that focus on developing implant surface nanofiber (NF) coating to prevent infection and enhance osseointegration by local drug release. In this study, coaxial doxycycline (Doxy)-doped polycaprolactone/polyvinyl alcohol (PCL/PVA) NFs were directly deposited on a titanium (Ti) implant surface during electrospinning. The interaction of loaded Doxy with both PVA and PCL NFs was characterized by Raman spectroscopy. The bonding strength of Doxy-doped NF coating on Ti implants was confirmed by a stand single-pass scratch test. The improved implant osseointegration by PCL/PVA NF coatings in vivo was confirmed by scanning electron microscopy, histomorphometry and micro computed tomography (µCT) at 2, 4 and 8 weeks after implantation. The bone contact surface (%) changes of the NF coating group (80%) is significantly higher than that of the no NF group (<5%, p < 0.05). Finally, we demonstrated that a Doxy-doped NF coating effectively inhibited bacterial infection and enhanced osseointegration in an infected (Staphylococcus aureus) tibia implantation rat model. Doxy released from NF coating inhibited bacterial growth up to 8 weeks in vivo. The maximal push-in force of the Doxy-NF coating (38 N) is much higher than that of the NF coating group (6.5 N) 8 weeks after implantation (p < 0.05), which was further confirmed by quantitative histological analysis and µCT. These findings indicate that coaxial PCL/PVA NF coating doped with Doxy and/or other drugs have great potential in enhancing implant osseointegration and preventing infection.


Assuntos
Doxiciclina/farmacologia , Osseointegração/efeitos dos fármacos , Poliésteres/química , Álcool de Polivinil/química , Infecções Estafilocócicas/prevenção & controle , Staphylococcus aureus/química , Staphylococcus aureus/efeitos dos fármacos , Tíbia/fisiologia , Titânio/química , Animais , Doxiciclina/química , Nanofibras , Próteses e Implantes , Ratos , Microtomografia por Raio-X
3.
J Neurooncol ; 120(1): 55-62, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25038847

RESUMO

Raman spectroscopy provides a molecular signature of the region being studied. It is ideal for neurosurgical applications because it is non-destructive, label-free, not impacted by water concentration, and can map an entire region of tissue. The objective of this paper is to demonstrate the meaningful spatial molecular information provided by Raman spectroscopy for identification of regions of normal brain, necrosis, diffusely infiltrating glioma and solid glioblastoma (GBM). Five frozen section tissues (1 normal, 1 necrotic, 1 GBM, and 2 infiltrating glioma) were mapped in their entirety using a 300-µm-square step size. Smaller regions of interest were also mapped using a 25-µm step size. The relative concentrations of relevant biomolecules were mapped across all tissues and compared with adjacent hematoxylin and eosin-stained sections, allowing identification of normal, GBM, and necrotic regions. Raman peaks and peak ratios mapped included 1003, 1313, 1431, 1585, and 1659 cm(-1). Tissue maps identified boundaries of grey and white matter, necrosis, GBM, and infiltrating tumor. Complementary information, including relative concentration of lipids, protein, nucleic acid, and hemoglobin, was presented in a manner which can be easily adapted for in vivo tissue mapping. Raman spectroscopy can successfully provide label-free imaging of tissue characteristics with high accuracy. It can be translated to a surgical or laboratory tool for rapid, non-destructive imaging of tumor margins.


Assuntos
Mapeamento Encefálico/métodos , Neoplasias Encefálicas/patologia , Encéfalo/patologia , Glioblastoma/patologia , Glioma/patologia , Imagem Molecular/métodos , Análise Espectral Raman/métodos , Idoso , Estudos de Casos e Controles , Seguimentos , Secções Congeladas , Humanos , Pessoa de Meia-Idade , Necrose , Prognóstico
4.
Cancer Metastasis Rev ; 33(2-3): 673-93, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24510129

RESUMO

There is a need in prostate cancer diagnostics and research for a label-free imaging methodology that is nondestructive, rapid, objective, and uninfluenced by water. Raman spectroscopy provides a molecular signature, which can be scaled from micron-level regions of interest in cells to macroscopic areas of tissue. It can be used for applications ranging from in vivo or in vitro diagnostics to basic science laboratory testing. This work describes the fundamentals of Raman spectroscopy and complementary techniques including surface enhanced Raman scattering, resonance Raman spectroscopy, coherent anti-Stokes Raman spectroscopy, confocal Raman spectroscopy, stimulated Raman scattering, and spatially offset Raman spectroscopy. Clinical applications of Raman spectroscopy to prostate cancer will be discussed, including screening, biopsy, margin assessment, and monitoring of treatment efficacy. Laboratory applications including cell identification, culture monitoring, therapeutics development, and live imaging of cellular processes are discussed. Potential future avenues of research are described, with emphasis on multiplexing Raman spectroscopy with other modalities.


Assuntos
Neoplasias da Próstata/diagnóstico , Análise Espectral Raman/métodos , Biomarcadores Tumorais/metabolismo , Diagnóstico por Imagem , Humanos , Masculino , Metabolômica/métodos , Neoplasias da Próstata/metabolismo , Proteômica/métodos
5.
J Neurooncol ; 116(3): 477-85, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24390405

RESUMO

The need exists for a highly accurate, efficient and inexpensive tool to distinguish normal brain tissue from glioblastoma multiforme (GBM) and necrosis boundaries rapidly, in real-time, in the operating room. Raman spectroscopy provides a unique biochemical signature of a tissue type, with the potential to provide intraoperative identification of tumor and necrosis boundaries. We aimed to develop a database of Raman spectra from normal brain, GBM, and necrosis, and a methodology for distinguishing these pathologies. Raman spectroscopy was used to measure 95 regions from 40 frozen tissue sections using 785 nm excitation wavelength. Review of adjacent hematoxylin and eosin sections confirmed histology of each region. Three regions each of normal grey matter, necrosis, and GBM were selected as a training set. Ten regions were selected as a validation set, with a secondary validation set of tissue regions containing freeze artifact. Grey matter contained higher lipid (1061, 1081 cm(-1)) content, whereas necrosis revealed increased protein and nucleic acid content (1003, 1206, 1239, 1255-1266, 1552 cm(-1)). GBM fell between these two extremes. Discriminant function analysis showed 99.6, 97.8, and 77.5% accuracy in distinguishing tissue types in the training, validation, and validation with freeze artifact datasets, respectively. Decreased classification in the freeze artifact group was due to tissue preparation damage. This study shows the potential of Raman spectroscopy to accurately identify normal brain, necrosis, and GBM as a tool to augment pathologic diagnosis. Future work will develop mapped images of diffuse glioma and neoplastic margins toward development of an intraoperative surgical tool.


Assuntos
Neoplasias Encefálicas/patologia , Encéfalo/patologia , Secções Congeladas , Glioblastoma/patologia , Necrose/patologia , Análise Espectral Raman , Idoso , Mapeamento Encefálico , Análise Discriminante , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Tempo
6.
Pediatr Surg Int ; 29(2): 129-40, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23143035

RESUMO

PURPOSE: Create a Raman spectroscopic database with potential to diagnose cancer and investigate two different diagnostic methodologies. Raman spectroscopy measures the energy of photons scattered inelastically by molecules. These molecular signatures form the basis of identifying complex biomolecules and can be used to differentiate normal from neoplastic tissue. METHODS: 1,352 spectra from 55 specimens were collected from fresh or frozen normal brain, kidney and adrenal gland and their malignancies. Spectra were obtained utilizing a Renishaw Raman microscope (RM1000) at 785 nm excitation wavelength with an exposure time of 10 to 20 s/spectrum over three accumulations. Spectra were preprocessed and discriminant function analysis was used to classify spectra based on pathological gold standard. RESULTS: The results of leave 25 % out training/testing validation were as follows: 94.3 % accuracy for training and 91.5 % for testing adrenal, 95.1 % accuracy for training and 88.9 % for testing group of brain, and 100 % accuracy for kidney training/testing groups when tissue origin was assumed. A generalized database not assuming tissue origin provided 88 % training and 85.5 % testing accuracy. CONCLUSION: A database can be made from Raman spectra to classify and grade normal from cancerous tissue. This database has the potential for real time diagnosis of fresh tissue and can potentially be applied to the operating room in vivo.


Assuntos
Neoplasias das Glândulas Suprarrenais/diagnóstico , Neoplasias Encefálicas/diagnóstico , Bases de Dados Factuais/estatística & dados numéricos , Neoplasias Renais/diagnóstico , Análise Espectral Raman/métodos , Criança , Diagnóstico Diferencial , Análise Discriminante , Hospitais Universitários , Humanos , Reprodutibilidade dos Testes
7.
Pediatr Neurosurg ; 48(2): 109-17, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23154646

RESUMO

PURPOSE: Raman spectroscopy can quickly and accurately diagnose tissue in near real-time. This study evaluated the capacity of Raman spectroscopy to diagnose pediatric brain tumors. EXPERIMENTAL DESIGN: Samples of untreated pediatric medulloblastoma (4 samples and 4 patients), glioma (i.e. astrocytoma, oligodendroglioma, ependymoma, ganglioglioma and other gliomas; 27 samples and 19 patients), and normal brain samples (33 samples and 5 patients) were collected fresh from the operating room or from our frozen tumor bank. Samples were divided and tested using routine pathology and Raman spectroscopy. Twelve Raman spectra were collected per sample. Support vector machine analysis was used to classify spectra using the pathology diagnosis as the gold standard. RESULTS: Normal brain (321 spectra), glioma (246 spectra) and medulloblastoma (82 spectra) were identified with 96.9, 96.7 and 93.9% accuracy, respectively, when compared with each other. High-grade ependymomas (41 spectra) were differentiated from low-grade ependymomas (25 spectra) with 100% sensitivity and 96.0% specificity. Normal brain tissue was distinguished from low-grade glioma (118 spectra) with 91.5% sensitivity and 97.8% specificity. For these analyses, the tissue-level classification was determined to be 100% accurate. CONCLUSION: These results suggest Raman spectroscopy can accurately distinguish pediatric brain neoplasms from normal brain tissue, similar tumor types from each other and high-grade from low-grade tumors.


Assuntos
Neoplasias Encefálicas/diagnóstico , Neoplasias Cerebelares/diagnóstico , Glioma/diagnóstico , Meduloblastoma/diagnóstico , Análise Espectral Raman/métodos , Astrocitoma/diagnóstico , Astrocitoma/patologia , Encéfalo/patologia , Neoplasias Encefálicas/patologia , Neoplasias Cerebelares/patologia , Criança , Diagnóstico Diferencial , Ependimoma/diagnóstico , Ependimoma/patologia , Ganglioglioma/diagnóstico , Ganglioglioma/patologia , Glioma/patologia , Humanos , Meduloblastoma/patologia , Gradação de Tumores , Oligodendroglioma/diagnóstico , Oligodendroglioma/patologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise Espectral Raman/normas , Bancos de Tecidos
8.
Pancreas ; 36(2): e1-8, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18376295

RESUMO

OBJECTIVES: Detection of neoplastic changes using optical spectroscopy has been an active area of research in recent times. Raman spectroscopy is a vibrational spectroscopic technique that can be used to diagnose various tumors with high sensitivity and specificity. We evaluated the ability of Raman spectroscopy to differentiate normal pancreatic tissue from malignant tumors in a mouse model. METHODS: We collected 920 spectra, 475 from 31 normal pancreatic tissue and 445 from 29 tumor nodules using a 785-nm near-infrared laser excitation. Discriminant function analysis was used for classification of normal and tumor samples. RESULTS: Using principal component analysis, we were able to highlight subtle chemical differences in normal and malignant tissue. Using histopathology as the gold standard, Raman analysis gave sensitivities between 91% and 96% and specificities between 88% and 96%. CONCLUSIONS: Raman spectroscopy along with discriminant function analysis is a useful method to detect cancerous changes in the pancreas. Pancreatic tumors were characterized by increased collagen content and decreased DNA, RNA, and lipids components compared with normal pancreatic tissue.


Assuntos
Pâncreas/patologia , Neoplasias Pancreáticas/patologia , Espectroscopia de Luz Próxima ao Infravermelho , Análise Espectral Raman , Animais , Linhagem Celular Tumoral , Colágeno/análise , DNA/análise , Análise Discriminante , Humanos , Lipídeos/análise , Camundongos , Neoplasias Experimentais/patologia , Pâncreas/química , Neoplasias Pancreáticas/química , Valor Preditivo dos Testes , Análise de Componente Principal , RNA/análise , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
9.
Biopolymers ; 89(3): 235-41, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18041066

RESUMO

Raman spectroscopy shows potential in differentiating tumors from normal tissue. We used Raman spectroscopy with near-infrared light excitation to study normal breast tissue and tumors from 11 mice injected with a cancer cell line. Spectra were collected from 17 tumors, 18 samples of adjacent breast tissue and lymph nodes, and 17 tissue samples from the contralateral breast and its adjacent lymph nodes. Discriminant function analysis was used for classification with principal component analysis scores as input data. Tissues were examined by light microscopy following formalin fixation and hematoxylin and eosin staining. Discriminant function analysis and histology agreed on the diagnosis of all contralateral normal, tumor, and mastitis samples, except one tumor which was found to be more similar to normal tissue. Normal tissue adjacent to each tumor was examined as a separate data group called tumor bed. Scattered morphologically suspicious atypical cells not definite for tumor were present in the tumor bed samples. Classification of tumor bed tissue showed that some tumor bed tissues are diagnostically different from normal, tumor, and mastitis tissue. This may reflect malignant molecular alterations prior to morphologic changes, as expected in preneoplastic processes. Raman spectroscopy not only distinguishes tumor from normal breast tissue, but also detects early neoplastic changes prior to definite morphologic alteration.


Assuntos
Biomarcadores Tumorais/análise , Neoplasias da Mama/química , Mama/química , Lesões Pré-Cancerosas/química , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Análise Espectral Raman/métodos , Animais , Biomarcadores Tumorais/química , Mama/patologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Diagnóstico Diferencial , Modelos Animais de Doenças , Feminino , Técnicas Histológicas , Camundongos , Transplante de Neoplasias , Lesões Pré-Cancerosas/diagnóstico , Sensibilidade e Especificidade , Espectroscopia de Luz Próxima ao Infravermelho/instrumentação , Análise Espectral Raman/instrumentação
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