Nonlinear optical microscopy
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AIM: Clubfoot is a congenital deformity affecting the musculoskeletal system, resulting in contracted and stiff tissue in the medial part of the foot. Minoxidil (MXD) has an inhibitory effect on lysyl hydroxylase, which influences the quality of extracellular matrix crosslinking, and could therefore be used to reduce the stiffness and to improve the flexibility of the tissue. We assessed the in vitro antifibrotic effects of minoxidil on clubfoot-derived cells. METHODS: Cell viability and proliferation were quantified by xCELLigence, MTS, and LIVE/DEAD assays. The amount of collagen I deposited into the extracellular matrix was quantified using immunofluorescence with subsequent image segmentation analysis, hydroxyproline assay, and Second Harmonic Generation imaging. Extracellular matrix contraction was studied in a 3D model of cell-populated collagen gel lattices. RESULTS: MXD concentrations of 0.25, 0.5, and 0.75 mM inhibited the cell proliferation in a concentration-dependent manner without causing a cytotoxic effect. Exposure to ≥0.5 mM MXD resulted in a decrease in collagen type I accumulation after 8 and 21 days in culture. Changes in collagen fiber assembly were observed by immunofluorescence microscopy and nonlinear optical microscopy (second harmonic generation). MXD also inhibited the contraction of cell-populated collagen lattices (0.5 mM by 22%; 0.75 mM by 28%). CONCLUSIONS: Minoxidil exerts an in vitro inhibitory effect on the cell proliferation, collagen accumulation, and extracellular matrix contraction processes that are associated with clubfoot fibrosis. This study provides important preliminary results demonstrating the potential relevance of MXD for adjuvant pharmacological therapy in standard treatment of relapsed clubfoot.
- MeSH
- kolagen typu I MeSH
- kolagen MeSH
- konzervativní terapie MeSH
- lidé MeSH
- minoxidil farmakologie MeSH
- pes equinovarus * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
... The Goldman equation (resting membrane voltage) 80 -- 3.4.3 Nonlinear thermodynamics and dissipative ... ... Skorpíková) 149 -- 8.2 Optical properties of the eye 154 -- 8.2.1 The structure of the eyeball and the ... ... optical microscopes 220 -- 11.6.1.4 Optical scanning microscopes 221 -- 11.6.2 Electron microscopy 223 ... ... -- 11.6.2.1 Transmission electron microscopy 223 -- 11.6.2.2 Scanning electron microscopy (SEM) 224 ... ... -- 11.6.3 Acoustic microscopy 225 -- 11.7 Bone densitometry (I. ...
2nd rev. ed. 325 s. : il., tab. ; 30 cm
- Konspekt
- Biochemie. Molekulární biologie. Biofyzika
- NLK Obory
- fyzika, biofyzika
- NLK Publikační typ
- učebnice vysokých škol
This paper highlights some of the key technologies of using two innovative molecular imaging modalites, magnetic resonance imaging (MRI) and nonlinear optical microscopy, for imaging intravenously injected ultra small paramagnetic iron oxide nanoparticles cross linked with antibodies (CLUSPIO) in the amyotrophic lateral sclerosis (ALS) experimental model in vivo or ex vivo, respectively. Intensive efforts have been made in investigating the causes of abnormalities in lipid metabolism, monitored in some neurodegenerative disorders systems. It has been shown that an abnormal accumulation of some common lipids in motor nerve cells may play a critical role in the development of amyotrophic lateral sclerosis. The presented experiments were performed on brain specimens from the transgenic rat model expressing multiple copies of mutated (G93A) human SOD-1 gene, after CD4+ lymphocytes were magnetically labeled with i.v.i. CLUSPIO antibodies. In vivo MRI revealed marked signal intensity enhancements in specific pathological regions of the ALS rat brain as compared to the wild type. Surface-enhanced coherent anti-Stokes Raman scattering (SECARS) microscopy indicated cellular interactions based on lipids association to anti-CD4 CLUSPIO
- MeSH
- amyotrofická laterální skleróza diagnóza etiologie MeSH
- antigeny CD4 diagnostické užití metabolismus MeSH
- experimenty na zvířatech MeSH
- financování organizované MeSH
- lymfocyty fyziologie metabolismus patologie MeSH
- magnetická rezonanční tomografie metody přístrojové vybavení využití MeSH
- metabolismus lipidů MeSH
- mikroskopie metody přístrojové vybavení využití MeSH
- molekulární zobrazování metody využití MeSH
- nanočástice diagnostické užití MeSH
- neurodegenerativní nemoci diagnóza etiologie MeSH
- potkani Sprague-Dawley MeSH
- potkani transgenní MeSH
- Ramanova spektroskopie metody MeSH
- sloučeniny železa diagnostické užití MeSH
- statistika jako téma MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
We investigated possibilities of the combination of the one- and two-photon excitation microscopy for examination of the experimental melanoma tissue in vivo, in mice under general anesthesia, and ex vivo on freshly harvested specimens. Our aim was to obtain sufficiently informative images of unstained tumor tissues and their modifications after hyperthermia treatment. The mouse experimental melanoma structure was studied and compared with normal tissue from the same animal by using confocal and nonlinear microscopy techniques based on (i) one-photon excitation (1PE) fluorescence, (ii) 1PE reflectance, (iii) second harmonic generation imaging, and (iv) two-photon excitation autofluorescence. We checked different spectral conditions and other settings of image acquisition, as well as combinations of the above imaging modalities, to fully exploit the potential of these techniques in the evaluation of treated and untreated cancer tissue morphology. Our approach enabled to reveal the collagen fiber network in relation with the other tissues, and to identify invasive tumor cells. It also proved to be useful for the examination of interrelationships between functional and morphological aspects based on optical properties of the tissues, especially in studies of changes between the tumor and control tissue, as well as changes induced by physical treatments, e.g., delivery of microwave hyperthermia treatment. These differences were also evaluated quantitatively, when we found out that the maximum Euler-Poincare characteristic reflects well the melanoma morphological structure. The results showed that the proposed investigative approach could be suitable also for a direct evaluation of tissue modifications induced by clinical interventions. (c) 2009 Wiley-Liss, Inc.
- MeSH
- indukovaná hypertermie MeSH
- melanom experimentální patologie radioterapie MeSH
- mikroskopie MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
... The Goldman equation (resting membrane voltage) 72 -- 3.4.3 Nonlinear thermodynamics and dissipative ... ... Skorpíková) 141 -- 8.2 Optical properties of the eye 146 -- 8.2.1 The structure of the eyeball and the ... ... optical microscopes 212 -- 11.6.1.4 Optical scanning microscopes 213 -- 11.6.2 Electron microscopy 215 ... ... -- 11.6.2.1 Transmission electron microscopy 216 -- 11.6.2.2 Scanning electron microscopy (SEM) 216 ... ... -- 11.6.3 Acoustic microscopy 217 -- 11.7 Bone densitometry (I. ...
1. vyd. viii, 317 s. : il. ; 30 cm
