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1.
J Orthop Res ; 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31965590

RESUMO

Macrophages, particularly M1 macrophages, produce proinflammatory cytokines and contribute to the degenerative process in injured intervertebral discs (IVDs). We previously showed that macrophages in both intact and injured IVDs increased following IVD injury. Resident macrophages and macrophages recruited from the peripheral blood have distinct roles in tissue. However, it remains to be determined whether increased macrophages derive from resident or recruited macrophages. We investigated the origin of M1 macrophages in injured IVDs using green fluorescent protein (GFP) transgenic bone marrow chimeric mice. The M1 macrophage marker, CD86, increased in both disc-derived resident macrophages and bone marrow-derived macrophages (BMMs) after lipopolysaccharide/interferon γ stimulation in vitro. Following IVD injury, the proportion of cells positive for the CD86 ligand, the F4/80 antigen, and the surface glycoprotein CD11b (CD86+ CD11b+ F4/80+) significantly increased in GFP+ populations at days 3, 7, and 14. In contrast, CD86+ CD11b+ F4/80+ cells in GFP- populations significantly increased on day 3, and thereafter decreased on days 7 and 14. The proportion of CD86+ CD11b+ F4/80+ cells in the GFP+ populations was significantly higher than that in the GFP- populations at days 1, 3, 7, and 14. Monocyte chemoattractant protein-1 expression in disc-derived macrophages, but not in BMMs, increased following interleukin-1ß stimulation. Our results suggest M1 macrophages following IVD injury originate from recruited macrophages. Resident macrophages may behave differently in IVD injury. The role of resident macrophages needs to be clarified. Further investigation is needed.

2.
J Biomed Mater Res B Appl Biomater ; 108(2): 326-332, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31016841

RESUMO

Several nerve conduits have been investigated for their potential as alternative sources of autografts for bridging neural gaps. However, autologous nerve transplants remain the most effective for nerve repair. We examined clinically approved nerve conduits containing collagen and polyglycolic acid (PGA-c) combined with collagen-binding basic fibroblast growth factor (bFGF) containing a polycystic kidney disease (PKD) domain and collagen binding domain (CBD) (bFGF-PKD-CBD) in a rat 15-mm sciatic nerve critical-size defect model. The treatment groups were: PGA-c immersed in phosphate-buffered saline (PBS) (PGA-c/PBS group), bFGF (PGA-c/bFGF group), or bFGF-PKD-CBD (PGA-c/bFGF-PKD-CBD group), and no treatment (Defect group). Gait and histological analyses were performed. Four weeks after treatment, the recovery rate of the paw print area was significantly greater in the PGA-c/bFGFPKD-CBD group than the PGA-c/PBS and PGA-c/bFGF groups. Mean intensity of paw prints was significantly greater in the PGA-c/bFGF-PKD-CBD group than the PGA-c/PBS and Defect groups. Swing time was significantly greater in the PGA-c/PBS, PGA-c/bFGF, and PGA-c/bFGF-PKD-CBD groups than the Defect group. At 8 weeks, all three parameters were significantly greater in the PGA-c/PBS, PGA-c/bFGF, and PGA-c/bFGF-PKD-CBD groups than the Defect group. Regenerated myelinated fibers were observed in 7/8 (87.5%) rats in the PGA-c/bFGF-PKD-CBD group after 8 weeks, and in 1/8 (12.5%) and 3/8 (37.5%) rats in the PGA-c/PBS and PGA-c/bFGF groups, respectively. PGA-c/bFGF-PKD-CBD composites may be promising biomaterials for promoting functional recovery of long-distance peripheral nerve defects in clinical practice.

3.
Microsurgery ; 40(3): 324-330, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31713920

RESUMO

BACKGROUND: Photoacoustic tomography (PAT) is a noninvasive vascular imaging modality that uses near-infrared pulse laser beams and ultrasound (US) to visualize vessels. We previously demonstrated the utility of PAT for visualizing anterolateral thigh (ALT) perforators in a clinical study of 10 thighs in 5 healthy adults. Evaluation of the correlation between PAT and US findings showed that PAT had comparable diagnostic potential but was superior in visualizing subcutaneous microvessels; however, there was no comparison with intraoperative findings. In this study, we used a newly developed technique to transfer a PAT image to a body-attachable transparent sheet to compare PAT and intraoperative findings. METHODS: Eight patients were recruited in this prospective study. Patient age ranged from 32 to 79 years (average 60). Seven ALT flaps were applied in head and neck reconstruction. One flap was elevated in chest wall reconstruction. Each PAT scan of an 18 cm × 13.5 cm region took approximately 5 min. Acquired data were processed three-dimensionally using a novel imaging software program. Perforator vessel data from PAT imaging were traced and corrected for projection onto medical film sheets. The correlation between the perforator stem portions predicted by PAT and the intraoperative findings at the level of the fascia-penetrating points was evaluated, and distal branching patterns were analyzed. RESULTS: PAT imaging showed 16 perforators in 8 thighs. Intraoperative surgical findings revealed that all the perforator penetrating points at the deep fascia level matched the PAT findings within 10 mm. None of the eight ALT flaps demonstrated postoperative complications. The perforator complexes were classified as type I in three cases (19%), type II in eight cases (50%), and type III in five cases (31%). CONCLUSIONS: PAT imaging matched the intraoperative findings within 10 mm. Preoperative vascular evaluation allows for the creation of a vascular map for facilitating ALT flap surgeries.

4.
Biomed Res Int ; 2019: 6959056, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31662989

RESUMO

Background: Previous studies suggest the presence of an association of vascular endothelial growth factor (VEGF) with osteoarthritis (OA) severity and pain in patients with knee OA. VEGF expression in human synovial fibroblasts (SFs) is induced by transforming growth factor-beta (TGFß). However, the signaling pathway governing TGFß-mediated regulation of VEGF in SFs has not been identified. Methods: OA patients who underwent total knee arthroplasty had their synovial tissue (SYT) extracted and the constituent SFs cultured. The cells were stimulated with culture medium (control), human recombinant TGFß (hrTGFß), hrTGFß + ALK5 inhibitor SB505124, hrTGFß + transforming growth factor activating kinase 1 (TAK1) inhibitor (5Z)-7-oxozeaenol, or hrTGFß + p38 inhibitor SB203580 for 6 h. VEGF mRNA expression in SFs was examined using real-time polymerase chain reaction and VEGF protein production in the cell supernatant was examined using enzyme-linked immunosorbent assay. Additionally, phosphorylated levels of SMAD2 and p38 were examined using western blotting. Results: ALK5 (SB505124) and TAK1 (5Z-oxozeaenol) inhibitors completely suppressed TGFß-induced VEGF mRNA expression and VEGF protein production. Both SB505124 and 5Z-oxozeaenol also suppressed SMAD2 and p38 phosphorylation. The p38 inhibitor (SB203580) partially inhibited TGFß-mediated VEGF mRNA and VEGF protein production. Conclusion: TGFß-mediated regulation of VEGF expression and VEGF protein production in the SYT of OA patients occurs through both the canonical and noncanonical pathway.


Assuntos
Fibroblastos/metabolismo , Osteoartrite do Joelho/metabolismo , Transdução de Sinais/fisiologia , Membrana Sinovial/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Células Cultivadas , Fibroblastos/efeitos dos fármacos , Humanos , Imidazóis/farmacologia , MAP Quinase Quinase Quinases/metabolismo , Osteoartrite do Joelho/tratamento farmacológico , Piridinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Proteína Smad2/metabolismo , Membrana Sinovial/efeitos dos fármacos , Zearalenona/análogos & derivados , Zearalenona/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
5.
Diabetes Metab Syndr Obes ; 12: 1151-1159, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31406471

RESUMO

Purpose: While research has identified diabetes mellitus (DM) as a risk factor for knee osteoarthritis (KOA), the underlying mechanisms are not fully understood. Studies suggest that Toll-like receptor 4 (TLR4) expression is elevated in osteoarthritic lesions of OA patients and in target tissues of insulin resistance such as adipose tissue and skeletal muscle in patients with DM. TLR4 is associated with inflammation and catabolic response via regulation of matrix metalloproteases (MMPs). We hypothesized that TLR4 and MMP expression may be increased in the synovial tissue (SYN) of KOA patients with diabetic pathology. We therefore investigated TLR and MMP expression in the SYN of KOA patients with and without high haemoglobin A1c concentrations. Patients and methods: A total of 171 patients radiographically diagnosed with KOA were grouped based on their HbA1c concentration (HbA1c ≥6.5 and HbA1c <6.5). We used real-time polymerase chain reaction to compare the expression of TLRs (TLR2, TLR4) and MMPs (MMP2, MMP3, MMP9 and MMP13) in patients' SYN between the two groups. MMP13 regulation by the TLR4 ligand, lipopolysaccharide (LPS), in SYN cells was examined in culture by stimulating SYN cells with LPS or vehicle (culture medium) for 24 h. Results: The expression of TLR4 and MMP13 in the HbA1c ≥6.5 group was significantly elevated compared to that in the HbA1c <6.5 group. In contrast, TLR2, MMP2, MMP3 and MMP9 expression levels were similar between the groups. MMP13 mRNA and MMP13 protein levels in SYN cells were significantly higher following stimulation with LPS compared to vehicle. Conclusions: TLR4 and MMP13 expression were elevated in the synovium of osteoarthritis patients with high HbA1c concentrations. Our results may provide insight into the pathology of OA patients with DM.

6.
Artigo em Japonês | MEDLINE | ID: mdl-31327775

RESUMO

PURPOSE: We conducted a multicenter study to investigate the current status of difference between the actual values at the patient entrance reference point (PERP) and display air kerma. METHODS: We exposure dose and fluoroscopy dose were measured by 32 apparatuses at 32 member institutions of the Japanese Society of Circulation Imaging Technology (CITEC) under unified conditions, and the actual measured values and display air kerma were compared. We entrance doses during fluoroscopy and imaging were measured at the PERP, with focus detector distance (FDD) 110 cm, a copper plate of 3.5 mm in thickness adhered to the front face of flat panel detector (FPD) as absorber, field-of-view (FOV) 18 cm, and the frame rate of 15 f/s, excluding the bed. Display air kerma were recorded at the same time. JIS (Z 4751-2-43: 2012) specify "The reference air kerma rate and the cumulative reference air kerma shall not deviate from their respective display air kerma by more than ±35% over the range of 6 mGy/min and 100 mGy to the maximum value." The number of apparatuses display air kerma deviated from this condition and its percentage were obtained. RESULTS: The mean difference percentage between actual measured values and display air kerma in 32 apparatuses was approximately 15.6%, with some apparatuses showing substantially different display air kerma. CONCLUSION: In order to estimate patients' skin exposure dose from display air kerma more accurately, it is necessary to perform calibration of the apparatus by regular dose measurement or convert values.


Assuntos
Raios X , Calibragem , Fluoroscopia , Humanos , Doses de Radiação , Radiografia , Inquéritos e Questionários
7.
Diabetes Metab Syndr Obes ; 12: 377-382, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31114272

RESUMO

Purpose: While research suggests that obesity is a risk factor for knee osteoarthritis (KOA), the mechanisms are not fully understood. Mast cell (MC) numbers are increased in the osteoarthritic synovium and in the adipose tissue of obese individuals. We hypothesized that MC numbers are increased in the synovium of obese KOA patients. This study investigated MC marker and MC-generated cytokine/growth factor expression in the synovium of obese KOA patients. Patients and methods: Patients radiographically diagnosed with KOA (male: 38, female: 132) were allocated to three groups based on their body mass index (BMI): normal (<25 kg/m2), overweight (25-29.99 kg/m2) and obese (≥30 kg/m2), according to the World Health Organization BMI classification. We used real-time polymerase chain reaction to compare the expression of MC markers (CD117, CD203c) and growth factors/cytokines (FGF2, VEGFA, TNFA, and IL8) in patients' synovium among the groups. Results: CD117 expression was significantly higher in the obese group than in the normal and overweight groups. CD203c and FGF2 expression were higher in the obese group than in the normal group. FGF2 expression levels were significantly correlated with those of CD117 (ρ=0.487) and CD203c (ρ=0.751). Conclusion: MC markers CD117 and CD203c, and FGF2 were highly expressed in the synovium of obese KOA patients. Further investigations are needed to reveal the role of MCs in the relationship between obesity and osteoarthritis pathology.

8.
J Orthop Res ; 37(10): 2258-2263, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31115924

RESUMO

Autologous vein wrapping is used to treat recurrent chronic constriction neuropathy and traumatic peripheral nerve injury. However, its use is restricted due to the inability to obtain sufficiently long veins for larger grafts. We previously reported that vein-derived basic fibroblast growth factor (bFGF) promotes heme oxygenase-1 (HO-1), which reduces allodynia via its anti-inflammatory properties. To mimic vein wrapping, we developed a collagen sheet impregnated with bFGF. Chronic constriction injury (CCI) was induced in male Wistar rats as a model of sciatic nerve injury, and the rats were divided into three groups: (i) untreated after CCI surgery (control group), (ii) treated with a collagen sheet wrap impregnated with phosphate-buffered saline (PBS/CS group), and (iii) treated with a collagen sheet wrap impregnated with bFGF (bFGF/CS group). Pain behavior (von Frey test) was evaluated on postoperative days (PODs) 1, 5, 7, and 14. Quantitative polymerase chain reaction was conducted on sciatic nerve RNA to quantify HO-1 gene, Hmox1, expression. Enzyme-linked immunosorbent assay were used to determine HO-1 protein levels on POD 1. von Frey testing showed significantly greater pain hypersensitivity in the control and PBS/CS groups than the bFGF/CS group. In the bFGF/CS group, Hmox1 messenger RNA and HO-1 protein levels were significantly increased in the sciatic nerve compared with the control and PBS/CS groups on PODs 1 and 5 and POD 1, respectively. The bFGF/CS group showed decreased allodynia and HO-1 induction, as observed with vein wrapping. Therefore, local application of bFGF may be an alternative treatment strategy for compressive neuropathy and peripheral nerve trauma in clinical settings. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2258-2263, 2019.


Assuntos
Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Hiperalgesia/terapia , Traumatismos dos Nervos Periféricos/terapia , Neuropatia Ciática/terapia , Animais , Colágeno , Portadores de Fármacos , Avaliação Pré-Clínica de Medicamentos , Heme Oxigenase (Desciclizante)/metabolismo , Distribuição Aleatória , Ratos , Nervo Isquiático/metabolismo , Suínos
9.
BMC Musculoskelet Disord ; 20(1): 199, 2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31077169

RESUMO

BACKGROUND: Chronic inflammation with aging contributes to sarcopenia. Previous studies have suggested that the accumulation of adipose tissue in skeletal muscle, referred to as intermuscular adipose tissue (IMAT), increases with age and is associated with inflammation. However, the mechanism governing ectopic inflammation in skeletal muscle due to aging is not fully understood. Leptin, an adipocytokine derived from adipose tissue, is an important mediator of inflammatory processes. We examined changes in leptin levels with age and whether leptin contributes to ectopic inflammation. METHODS: To evaluate ectopic inflammation in skeletal muscle, we measured alterations to the expression of inflammatory cytokine genes (Il1b, Il6, and Tnfa) and muscle break down-related gene (MuRF1 and Atrogin1) in the quadricep muscles of young (10 weeks) and aged (48 weeks) female rats using quantitative reverse-transcription polymerase chain reaction (Q-RT-PCR). Histological examination was performed to identify the extent of IMAT. Leptin mRNA and leptin protein expression were examined using Q-RT-PCR and enzyme-linked immunosorbent assay, respectively. The effect of leptin on the mRNA expression of Il1b, Il6, and Tnfa in quadricep muscle-derived cells was also examined by stimulating the cells with 0 (control), 1, or 10 µg/mL rat recombinant leptin using Q-RT-PCR. RESULTS: Aged rats had significantly higher Il6, MuRF1, and Atrogin1 but not Il1b and Tnfa, expression and greater levels of IMAT in their quadricep muscles than young rats. Aged rats also had significantly higher leptin expression and leptin protein concentration in their quadricep muscles than young rats. The addition of exogenous leptin to quadricep muscle-derived cells significantly increased the gene expression of Il1b and Il6 but not Tnfa. CONCLUSIONS: Our results suggest that elevated leptin levels due to aging cause ectopic inflammation through IL-6 in the skeletal muscle of aged rats.


Assuntos
Tecido Adiposo/metabolismo , Envelhecimento/metabolismo , Interleucina-6/metabolismo , Leptina/metabolismo , Músculo Esquelético/metabolismo , Tecido Adiposo/imunologia , Envelhecimento/imunologia , Animais , Modelos Animais de Doenças , Feminino , Interleucina-6/imunologia , Modelos Animais , Músculo Esquelético/imunologia , Ratos , Ratos Sprague-Dawley , Sarcopenia/imunologia
10.
BMC Musculoskelet Disord ; 20(1): 204, 2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31077183

RESUMO

BACKGROUND: Nerve growth factor (NGF) contributes to pain in knee osteoarthritis (KOA) patients. Transforming growth factor-beta (TGF-ß) stimulates NGF expression in chondrocytes from KOA patients. However, the correlation between synovial TGF-ß and NGF levels has not been sufficiently studied in human KOA patients. Further, the mechanism governing NGF regulation by TGF-ß in synovial cells is unclear. METHODS: During total knee arthroplasty, we extracted the synovial tissue (SYT) of 107 subjects with unilateral Kellgren/Lawrence grade 3-4 KOA confirmed by radiography. We examined the distribution of TGF-ß and NGF using immunohistochemistry, and analyzed the relationship between NGF and TGFB mRNA levels. Cultured synovial cells extracted from SYT were exposed to culture medium (control), human recombinant TGF-ß (rhTGF-ß), rhTGF-ß + ALK5 inhibitor SB505124, rhTGF-ß + transforming growth factor activating kinase 1 (TAK1) inhibitor (5Z)-7-oxozeaenol, or rhTGF-ß + p38 inhibitor SB203580 for 30 min, 6 h and 24 h. NGF mRNA expressed by the cultured cells and NGF protein levels in the cell supernatant were detected by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Phosphorylation of p38 was evaluated by western blotting. RESULTS: NGF mRNA levels were positively correlated with those of TGFB. Cells expressing TGF-ß and NGF protein were observed in the lining layer of SYT. TGF-ß stimulated increased NGF mRNA expression and NGF protein production. The ALK5 inhibitor completely suppressed the TGF-ß-mediated increase in NGF expression and NGF production in synovial cells. ALK5, TAK1 and p38 inhibitors inhibited the TGF-ß-induced phosphorylation of p38, and TAK1 and p38 inhibitors partially inhibited the TGF-ß-mediated increase in NGF expression and NGF production in synovial cells. CONCLUSION: TGF-ß regulates NGF production via the TGF-ß/ALK5 signaling pathway in osteoarthritic synovium. This effect may partially occur through inhibition of the TAK1/p38 pathway in the SYT of KOA patients.


Assuntos
Artralgia/patologia , Fator de Crescimento Neural/metabolismo , Osteoartrite do Joelho/complicações , Membrana Sinovial/patologia , Fator de Crescimento Transformador beta1/metabolismo , Idoso , Artralgia/etiologia , Células Cultivadas , Feminino , Humanos , Masculino , Osteoartrite do Joelho/patologia , Cultura Primária de Células , Membrana Sinovial/citologia , Sinoviócitos/metabolismo
11.
J Hand Surg Eur Vol ; 44(2): 187-195, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30335597

RESUMO

This study aimed to characterize in vivo human digital arteries in three-dimensions using photoacoustic tomography in order to understand the specific mechanism underlying arterial deformation associated with movement of the proximal interphalangeal joint. Three-dimensional morphological data were obtained on the radialis indicis artery (radial artery of the index finger) at different angles of the joint. The association between increased curvature of the deformation and the anatomical region was assessed. Characteristic morphological deformations in areas of major deformation were determined. The deformation of the artery was characterized by three consecutive curves in juxta-articular regions, which were particularly noticeable when the joint was flexed at an angle of ≥ 60°. The change in the curvature of the deformation during 30°-90° of flexion was lower in middle-aged individuals than in young individuals. Better understanding of the mechanism underlying deformation of the digital arteries may contribute to advancements in flap procedures and rehabilitation strategies after digital artery repair.


Assuntos
Artérias/diagnóstico por imagem , Articulações dos Dedos/fisiologia , Amplitude de Movimento Articular/fisiologia , Adulto , Humanos , Imageamento Tridimensional , Pessoa de Meia-Idade , Técnicas Fotoacústicas , Tomografia , Adulto Jovem
12.
PLoS One ; 13(11): e0207661, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30444907

RESUMO

We aimed to describe the development of an inference model for computer-aided diagnosis of lung nodules that could provide valid reasoning for any inferences, thereby improving the interpretability and performance of the system. An automatic construction method was used that considered explanation adequacy and inference accuracy. In addition, we evaluated the usefulness of prior experts' (radiologists') knowledge while constructing the models. In total, 179 patients with lung nodules were included and divided into 79 and 100 cases for training and test data, respectively. F-measure and accuracy were used to assess explanation adequacy and inference accuracy, respectively. For F-measure, reasons were defined as proper subsets of Evidence that had a strong influence on the inference result. The inference models were automatically constructed using the Bayesian network and Markov chain Monte Carlo methods, selecting only those models that met the predefined criteria. During model constructions, we examined the effect of including radiologist's knowledge in the initial Bayesian network models. Performance of the best models in terms of F-measure, accuracy, and evaluation metric were as follows: 0.411, 72.0%, and 0.566, respectively, with prior knowledge, and 0.274, 65.0%, and 0.462, respectively, without prior knowledge. The best models with prior knowledge were then subjectively and independently evaluated by two radiologists using a 5-point scale, with 5, 3, and 1 representing beneficial, appropriate, and detrimental, respectively. The average scores by the two radiologists were 3.97 and 3.76 for the test data, indicating that the proposed computer-aided diagnosis system was acceptable to them. In conclusion, the proposed method incorporating radiologists' knowledge could help in eliminating radiologists' distrust of computer-aided diagnosis and improving its performance.


Assuntos
Diagnóstico por Computador/métodos , Neoplasias Pulmonares/diagnóstico por imagem , Interpretação de Imagem Radiográfica Assistida por Computador/métodos , Algoritmos , Teorema de Bayes , Humanos , Neoplasias Pulmonares/patologia , Cadeias de Markov , Modelos Teóricos , Método de Monte Carlo , Variações Dependentes do Observador
13.
Sci Rep ; 8(1): 14930, 2018 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-30297721

RESUMO

Photoacoustic (PA) imaging (PAI) has been shown to be a promising tool for non-invasive blood vessel imaging. A PAI system comprising a hemispherical detector array (HDA) has been reported previously as a method providing high morphological reproducibility. However, further improvements in diagnostic capability will require improving the image quality of PAI and fusing functional and morphological imaging. Our newly developed PAI system prototype not only enhances the PA image resolution but also acquires ultrasonic (US) B-mode images at continuous positions in the same coordinate axes. In addition, the pulse-to-pulse alternating laser irradiation shortens the measurement time difference between two wavelengths. We scanned extremities and breasts in an imaging region 140 mm in diameter and obtained 3D-PA images of fine blood vessels, including arterioles and venules. We could estimate whether a vessel was an artery or a vein by using the S-factor obtained from the PA images at two wavelengths, which corresponds approximately to the haemoglobin oxygen saturation. Furthermore, we observed tumour-related blood vessels around breast tumours with unprecedented resolution. In the future, clinical studies with our new PAI system will help to elucidate various mechanisms of vascular-associated diseases and events.


Assuntos
Arteríolas/diagnóstico por imagem , Técnicas Fotoacústicas/instrumentação , Tomografia/instrumentação , Vênulas/diagnóstico por imagem , Algoritmos , Desenho de Equipamento , Feminino , Humanos , Técnicas Fotoacústicas/métodos , Tomografia/métodos
14.
Lipids Health Dis ; 17(1): 215, 2018 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-30205824

RESUMO

BACKGROUND: The infrapatellar fat pad (IFP) is implicated in knee osteoarthritis (KOA). Calcitonin gene-related peptide (CGRP), a vasoactive neuropeptide expressed in joint tissues and synovial tissues (ST), was recently found to be associated with KOA progression and pain. CGRP is expressed in the IFPs of human KOA patients; however, its regulation has not been elucidated. METHODS: IFPs and STs were harvested from 138 KOA patients during total knee replacement (TKR) and analyzed for CGRP, cycloxygenase-2 (COX-2), and microsomal prostaglandin E synthase-1 (mPGES-1) expression using real-time polymerase chain reaction (PCR). To investigate CGRP regulation by prostaglandin E2 (PGE2), adipocytes (Ad) and the stromal vascular fraction (SVF) were harvested from IFPs using collagenase. Synovial cells (SYC) were also harvested from ST and stimulated with vehicle (serum-free culture medium), PGE2, or CGRP. RESULTS: CGRP, COX-2, and mPGES-1 expression levels were significantly higher in IFPs than STs. PGE2 stimulation increased CGRP expression in Ad, the SVF, and SYC; however, CGRP expression was significantly higher in PGE2-stimulated SVF than PGE2-stimulated SYC. CGRP stimulation had no effect on COX-2 or mPGES-1 expression. CONCLUSIONS: CGRP expression in the IFP of KOA patients is regulated by the COX-2/mPGES-1/PGE2 pathway.


Assuntos
Tecido Adiposo/metabolismo , Peptídeo Relacionado com Gene de Calcitonina/genética , Osteoartrite do Joelho/genética , Tecido Adiposo/patologia , Idoso , Idoso de 80 Anos ou mais , Ciclo-Oxigenase 2/genética , Dinoprostona/genética , Regulação da Expressão Gênica , Humanos , Osteoartrite do Joelho/patologia , Prostaglandina-E Sintases/genética , Transdução de Sinais/genética
15.
Biomed Res Int ; 2018: 8393194, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29770338

RESUMO

Basic fibroblast growth factor 2 (bFGF) accelerates bone formation during fracture healing. Because the efficacy of bFGF decreases rapidly following its diffusion from fracture sites, however, repeated dosing is required to ensure a sustained therapeutic effect. We previously developed a fusion protein comprising bFGF, a polycystic kidney disease domain (PKD; s2b), and collagen-binding domain (CBD; s3) sourced from the Clostridium histolyticum class II collagenase, ColH, and reported that the combination of this fusion protein with a collagen-like peptide, poly(Pro-Hyp-Gly)10, induced mesenchymal cell proliferation and callus formation at fracture sites. In addition, C. histolyticum produces class I collagenase (ColG) with tandem CBDs (s3a and s3b) at the C-terminus. We therefore hypothesized that a bFGF fusion protein containing ColG-derived tandem CBDs (s3a and s3b) would show enhanced collagen-binding activity, leading to improved bone formation. Here, we examined the binding affinity of four collagen anchors derived from the two clostridial collagenases to H-Gly-Pro-Arg-Gly-(Pro-Hyp-Gly)12-NH2, a collagenous peptide, by surface plasmon resonance and found that tandem CBDs (s3a-s3b) have the highest affinity for the collagenous peptide. We also constructed four fusion proteins consisting of bFGF and s3 (bFGF-s3), s2b-s3b (bFGF-s2b-s3), s3b (bFGF-s3b), and s3a-s3b (bFGF-s3a-s3b) and compared their biological activities to those of a previous fusion construct (bFGF-s2b-s3) using a cell proliferation assay in vitro and a mouse femoral fracture model in vivo. Among these CB-bFGFs, bFGF-s3a-s3b showed the highest capacity to induce mesenchymal cell proliferation and callus formation in the mice fracture model. The poly(Pro-Hyp-Gly)10/bFGF-s3a-s3b construct may therefore have the potential to promote bone formation in clinical settings.


Assuntos
Clostridium histolyticum/metabolismo , Colágeno/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , Colagenase Microbiana/metabolismo , Osteogênese/fisiologia , Animais , Fraturas do Fêmur/metabolismo , Consolidação da Fratura/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Peptídeos/metabolismo , Ligação Proteica , Ratos , Ratos Wistar
16.
Plast Reconstr Surg ; 141(5): 1288-1292, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29697630

RESUMO

The distal branching pattern of perforators is associated with thin anterolateral thigh flap failure. The purpose of this study was to investigate the feasibility of using photoacoustic tomography as a diagnostic imaging modality to identify anterolateral thigh perforators and their branching patterns in the subcutaneous layer. Ten thighs in five healthy men were studied. The anterolateral aspect of the midthigh was examined using photoacoustic tomography. The correlation between photoacoustic tomography and ultrasound findings was evaluated. To determine the detectability of photoacoustic tomography by depth, the depth of vessels in the stem portion was compared to the depth of the deep fascia measured by ultrasound. Branching patterns of vessels in the adipose and suprafascial layers were evaluated by three-dimensional observation. A total of 18 perforators were visualized by photoacoustic tomography. Photoacoustic tomography and ultrasound had comparable diagnostic potential for the detection of perforators. Photoacoustic tomography visualized microvessels in the subcutaneous layer, especially those in oblique or horizontal orientations. The estimated mean depth of visualized vessels was 9 mm; the maximum depth was 13 mm. There was a strong correlation between the depth of visualized vessels in the stem portion and the depth of the deep fascia. Three-dimensional observation of photoacoustic tomographic images showed the branching morphology of perforators. This study showed the applicability of photoacoustic tomography to identification of the branching patterns of anterolateral thigh perforators in vivo, although limited visualization of subfascial vessels is a technical issue. The authors believe that photoacoustic tomography has the potential to be a new imaging modality for thin anterolateral thigh flap surgery. CLINICAL QUESTION/LEVEL OF EVIDENCE: Diagnostic, IV.


Assuntos
Retalho Perfurante/irrigação sanguínea , Técnicas Fotoacústicas/métodos , Coxa da Perna/irrigação sanguínea , Tomografia/métodos , Adulto , Artérias/diagnóstico por imagem , Estudos de Viabilidade , Voluntários Saudáveis , Humanos , Masculino , Microvasos/diagnóstico por imagem , Retalho Perfurante/efeitos adversos , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/prevenção & controle , Procedimentos Cirúrgicos Reconstrutivos/efeitos adversos , Procedimentos Cirúrgicos Reconstrutivos/métodos , Coxa da Perna/diagnóstico por imagem , Ultrassonografia , Veias/diagnóstico por imagem
17.
J Orthop Res ; 2018 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-29508435

RESUMO

Upregulation of inflammatory cytokines and various growth factors is a significant contributor to discogenic low back pain. The aim of this study was to investigate possible regulation of pain-related molecules by macrophages and the role of macrophage-derived molecules in injured intervertebral disc (IVD)s. C57BL/6J mice were used in this study. We characterized the expression profiles of genes for tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, nerve growth factor (NGF), and vascular endothelial growth factor (VEGF) in both intact and injured IVDs. We examined whether macrophage depletion, induced by systemic injection of clodronate-laden liposomes, affected the expression of these molecules in injured IVDs. The effect of TNF-alpha on cultured F4/80-CD11b-cells in injured IVDs was investigated. Expression of TNF-alpha and IL-1beta was significantly increased in injured IVDs, but decreased by macrophage depletion. Expression of NGF and VEGF was also significantly increased, but by contrast was not decreased by macrophage depletion. TNF-alpha treatment of F4/80-cells from injured IVDs upregulated NGF, VEGF, cyclooxygenase (COX)-2, and microsomal prostaglandin E synthase-1 (mPGES1). IVD injury upregulated inflammatory cytokines and various growth factors. Macrophages in the injured IVDs produced inflammatory cytokines, but not growth factors. Macrophage-derived inflammatory cytokines regulate growth factors and pain-related molecules. These findings demonstrate further complexity in the pathogenesis of discogenic pain. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

18.
F1000Res ; 7: 1813, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30854189

RESUMO

Background : A breast-specific photoacoustic imaging (PAI) system prototype equipped with a hemispherical detector array (HDA) has been reported as a promising system configuration for providing high morphological reproducibility for vascular structures in living bodies. Methods : To image the vasculature of human limbs, a newly designed PAI system prototype (PAI-05) with an HDA with a higher density sensor arrangement was developed. The basic device configuration mimicked that of a previously reported breast-specific PAI system. A new imaging table and a holding tray for imaging a subject's limb were adopted. Results : The device's performance was verified using a phantom. Contrast of 8.5 was obtained at a depth of 2 cm, and the viewing angle reached up to 70 degrees, showing sufficient performance for limb imaging. An arbitrary wavelength was set, and a reasonable PA signal intensity dependent on the wavelength was obtained. To prove the concept of imaging human limbs, various parts of the subject were scanned. High-quality still images of a living human with a wider size than that previously reported were obtained by scanning within the horizontal plane and averaging the images. The maximum field of view (FOV) was 270 mm × 180 mm. Even in movie mode, one-shot 3D volumetric data were obtained in an FOV range of 20 mm in diameter, which is larger than values in previous reports. By continuously acquiring these images, we were able to produce motion pictures. Conclusion : We developed a PAI prototype system equipped with an HDA suitable for imaging limbs. As a result, the subject could be scanned over a wide range while in a more comfortable position, and high-quality still images and motion pictures could be obtained.


Assuntos
Sistemas Computacionais , Extremidades/diagnóstico por imagem , Imageamento Tridimensional , Técnicas Fotoacústicas , Humanos , Lasers , Imagens de Fantasmas , Postura
19.
Biomed Res Int ; 2017: 4371460, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28761877

RESUMO

Cell-based regenerative therapy has the potential to repair bone injuries or large defects that are recalcitrant to conventional treatment methods, including drugs and surgery. Here, we developed a multilayered cell-based bone formation system using cells coated with fibronectin-gelatin (FN-G) nanofilms. The multilayered mesenchymal cells (MLMCs) were formed after two days of culture and were shown to express higher levels of BMP-2 and VEGF compared to monolayer cultures of MCs. The MLMCs were used as a graft material in combination with a fusion protein consisting of basic fibroblast growth factor (bFGF), polycystic kidney disease (PKD) domain, and the collagen-binding domain (CBD) of Clostridium histolyticum collagenase. In femur sites grafted with the MLMCs, significantly higher levels of callus volume and bone mineral content were observed compared to the sham controls. The callus volume and bone mineral content were further increased in femur sites grafted with bFGF-PKD-CBD/MLMCs. Taken together, these results suggest that bFGF-PKD-CBD/MLMCs, which can be simply and rapidly generated in vitro, have the potential to promote bone repair when grafted into large defect sites.


Assuntos
Fêmur/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Osteogênese/fisiologia , Animais , Proteína Morfogenética Óssea 2/metabolismo , Calo Ósseo/metabolismo , Osteócitos/metabolismo , Ratos , Ratos Sprague-Dawley
20.
J Orthop Surg Res ; 12(1): 65, 2017 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-28431586

RESUMO

BACKGROUND: The infrapatellar fat pad (IPFP) has been implicated as a possible source of osteoarthritis (OA) development and knee pain due to the production of inflammatory mediators and the existence of nerve fibers within this structure. Calcitonin gene-related peptide (CGRP) is a vasodilatory neuropeptide that is localized to joint tissues and has recently been implicated in the development of knee OA and OA pain. To date, however, the expression levels of CGRP in the IPFP of human knee OA patients have not been examined. METHODS: IFFP and synovial (SYN) tissues were harvested from 100 individuals with radiographic knee OA (unilateral Kellgren/Lawrence [K/L] grades 2-4) during total knee arthroplasty and subjected to immunohistochemical analysis for CGRP localization. In addition, the messenger RNA (mRNA) expression levels of CGRP and cyclooxygenase-2 (COX-2) in the collected tissues were evaluated and compared using real-time PCR analysis of total RNA extracts. CGRP and COX-2 mRNA expression were also compared among individuals with K/L grades 2-4. RESULTS: CGRP-positive cells were detected in the capillaries within the IPFP and lining layer of SYN tissue. The expression levels of CGRP in the IPFP were positively correlated with COX-2 and were significantly higher than those in SYN tissue. CGRP expression in tissue from the KL4 group was twofold higher than that from the KL2 group. CONCLUSIONS: The IPFP of knee OA patients produces relatively high levels of CGRP, which may be regulated by COX-2 at the transcriptional level. Further studies are needed to determine if CGRP levels are directly linked to OA pathology.


Assuntos
Tecido Adiposo/metabolismo , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Osteoartrite do Joelho/metabolismo , Membrana Sinovial/metabolismo , Idoso , Idoso de 80 Anos ou mais , Ciclo-Oxigenase 2/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
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