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1.
Biomed Mater Eng ; 34(6): 537-544, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37334576

RESUMO

BACKGROUND: A combination of synthetic porous materials and BMP-2 has been used to promote fracture healing. For bone healing to be successful, it is important to use growth factor delivery systems that enable continuous release of BMP-2 at the fracture site. We previously reported that in situ-formed gels (IFGs) consisting of hyaluronan (HyA)-tyramine (TA), horseradish peroxidase and hydrogen peroxide enhance the bone formation ability of hydroxyapatite (Hap)/BMP-2 composites in a posterior lumbar fusion model. OBJECTIVE: We examined the effectiveness of IFGs-HyA/Hap/BMP-2 composites for facilitating osteogenesis in refractory fracture model mice. METHODS: After establishing the refractory fracture model, animals were either treated at the site of fracture with Hap harboring BMP-2 (Hap/BMP-2) or IFGs-HyA with Hap harboring BMP-2 (IFGs-HyA/Hap/BMP-2) (n = 10 each). Animals that underwent the fracture surgery but did not receive any treatment were considered the control group (n = 10). We determined the extent of bone formation at the fracture site according to findings on micro-computed tomography and histological studies four weeks following treatment. RESULTS: Animals treated with IFGs-HyA/Hap/BMP-2 demonstrated significantly greater bone volume, bone mineral content and bone union than those treated with vehicle or IFG-HyA/Hap alone. CONCLUSIONS: IFGs-HyA/Hap/BMP-2 could be an effective treatment option for refractory fractures.


Assuntos
Durapatita , Ácido Hialurônico , Camundongos , Animais , Microtomografia por Raio-X , Proteína Morfogenética Óssea 2 , Osteogênese , Consolidação da Fratura
2.
Biomed Mater Eng ; 34(1): 67-76, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-35694914

RESUMO

BACKGROUND: Mesenchymal stem cell (MSC)-based therapies offer potential for bone repair. MSC spheroid cultures may harbor enhanced therapeutic potential over MSC monolayers through increased secretion of trophic factors. However, the impact of spheroid size on trophic factor expression is unclear. OBJECTIVE: We investigated the effect of spheroid size on trophic factor-related gene expression. METHODS: KUM10, a murine MSC line was used. RNA-seq was used to screen the transcriptional profiles of MSC monolayer and spheroid cultures. Differentially expressed genes identified in RNA-seq were evaluated by q-PCR in cultures of 5 × 104 (S group), 5 × 105 (M group), 5 × 106 (L group) cells/well. RESULTS: Comparison of expression levels between KUM10 monolayer and spheroid cultures identified 2140 differentially expressed genes, of which 1047 were upregulated and 1093 were downregulated in KUM10 spheroids. Among these, 12 upregulated genes (Bmp2, Fgf9, Fgf18, Ngf, Pdgfa, Pdgfb, Tgfb1, Vegfa, Vegfc, Wnt4, Wnt5a, Wnt10a) were associated with secretory growth factors. Of these, expression of Fgf9, Fgf18, Vegfa and Vegfc was elevated in the L group, and Pdgfb and Tgfb1 was elevated in the S group. CONCLUSIONS: Spheroid size may impact trophic factor expression. Our results will be useful for future studies assessing the utility of MSC spheroids for treating bone injury.


Assuntos
Células-Tronco Mesenquimais , Esferoides Celulares , Camundongos , Animais , Esferoides Celulares/metabolismo , Transcriptoma , Proteínas Proto-Oncogênicas c-sis/metabolismo , Proteínas Proto-Oncogênicas c-sis/farmacologia , Linhagem Celular
3.
J Plast Reconstr Aesthet Surg ; 75(9): 3166-3173, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35868973

RESUMO

INTRODUCTION: Diagnostic imaging modalities to evaluate the three-dimensional distribution of thoracodorsal artery perforators (TDAPs) are lacking. In this study, TDAPs were visualized and characterized using photoacoustic imaging. MATERIAL AND METHODS: In this study, 34 sites in the lateral chest wall of 18 individuals were analyzed. The region extending 5 cm ventral and 5 cm dorsal to the lateral edge of the latissimus dorsi (LD) and 5-15 cm from the posterior axillary fold was scanned using photoacoustic imaging. The largest perforator closest to the edge of the LD was characterized. The location of the stem portion and the orientation of the longest cutaneous branch of the perforator were described. The relationship between the maximal depth of delineation on photoacoustic images and the depth of the deep fascia was assessed. RESULTS: On average, 2.6 perforators (range, 1-5 perforators) were visualized in the region of interest. The distribution of the TDAP stem portion was similar to that in previous studies. Cutaneous branches were preferentially oriented in a medial-caudal direction. The length of delineated cutaneous branches varied (range, 7-78 mm) depending on the thickness of the subcutaneous layer. Vessels under the LD were observed when the subcutaneous layer was thin. CONCLUSION: Photoacoustic imaging can successfully visualize TDAPs in three dimensions. Visualization of TDAPs varied by the thickness of the subcutaneous layer. A thin deep fascia of the LD might be a cause of deep laser penetration.


Assuntos
Retalho Perfurante , Técnicas Fotoacústicas , Artérias , Humanos , Imageamento Tridimensional , Retalho Perfurante/irrigação sanguínea , Retalhos Cirúrgicos/irrigação sanguínea , Tórax
4.
Cureus ; 14(5): e25509, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35663656

RESUMO

Several types of calcium phosphate (CaP) biomaterial carriers have been designed to deliver bone morphogenetic protein-2 (BMP-2) to augment spinal fusion in spinal surgery. Here, we evaluated an in situ-formed hydrogel (IFH) constructed from hyaluronan (IFH-HA) combined with a BMP2/hydroxyapatite (HAP) composite in bone formation in a murine model of posterolateral lumbar fusion (PLF). HAP was submerged in HA-tyramine (TA) polymer solution containing horseradish peroxidase (HRP) and 2 µg BMP-2 (BMP2/HA-TA/HRP solution). H2O2 was added to initiate the curing reaction (BMP-2/IFH-HA). phosphate-buffered saline (PBS) was added to the BMP2/HA-TA/HRP solution (BMP-2/HA-TA) instead of H2O2 to evaluate the effectiveness of the curing reaction. HAP immersed in PBS was used as a control. PLF model mice were randomly assigned to receive one these composites (n = 10 each). X-ray images were taken to assess the bone fusion, and microcomputed tomography analysis was conducted to examine new bone formation at the graft site four weeks following surgery. No evidence of fusion was observed four weeks after surgery in the Control or BMP2/HA-TA group. In contrast, the BMP2/IFH-HA group exhibited newly formed bone between the transverse processes and bone union in coronal sections. Relative to the Control and BMP2/HA-TA groups, the BMP2/IFH-HA group showed significantly greater bone volume. The BMP2/IFH-HA group also showed significantly elevated bone mineral content relative to the BMP2/HA-TA group. A composite comprising BMP2/HAP and IFH-HA, thus, enhanced the new bone formation in a murine model of PLF, suggesting its promise for augmenting spinal fusion.

5.
BMC Neurosci ; 23(1): 37, 2022 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-35725384

RESUMO

BACKGROUND: Autologous vein wrapping (VW) is used in the treatment of recurrent chronic constriction neuropathy and traumatic peripheral nerve injury. However, use of autologous veins is limited by the inability to obtain longer veins of sufficient length for larger sites. Frozen allograft tissue has several advantages, including its availability for large grafts, avoidance of donor-site morbidity, and shorter operation time. Here, we investigated the effect of frozen vein wrapping (FVW) in Wistar rats as a model of sciatic nerve injury. RESULTS: The rats were grouped by treatment as (i) untreated after chronic constriction injury surgery (CCI; control group), (ii) treated with vein wrapping using freshly isolated vein (VW), and (iii) treated with vein wrapping using frozen vein (FVW). Mechanical allodynia was assessed with von Frey filaments on postoperative days (PODs) 1, 3, 5, 7, and 14. Gene expression of HO-1 was evaluated by quantitative polymerase chain reaction (qPCR). The response of heme oxygenase-1 gene, Hmox-1, expression to VW and FVW was assessed by RT-PCR. Both VW and FVW significantly increased withdrawal threshold levels compared to the untreated control group on POD 1, 3, and 5. Both VW and FVW also showed increased HO-1 expression compared to the CCI group. CONCLUSIONS: FVW increased the withdrawal threshold similar to VW in a rat CCI model for short periods. Frozen vein wrapping using vein allograft without donor site morbidity may be an alternative therapeutic option.


Assuntos
Lesões por Esmagamento , Neuropatia Ciática , Animais , Constrição , Constrição Patológica/metabolismo , Modelos Animais de Doenças , Hiperalgesia/metabolismo , Ratos , Ratos Wistar , Nervo Isquiático/lesões , Neuropatia Ciática/metabolismo
6.
Ultrason Imaging ; 44(2-3): 96-104, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-35549598

RESUMO

Photoacoustic (PA) technology can be used for non-invasive imaging of blood vessels. In this paper, we report on our prototype PA imaging system with a newly designed ultrasound sensor and its visualization performance of microvascular in animal. We fabricated an experimental system for animals using a high-frequency sensor. The system has two modes: still image mode by wide scanning and moving image mode by small rotation of sensor array. Optical test target, euthanized mice and rats, and live mice were used as objects. The results of optical test target showed that the spatial resolution was about two times higher than that of our conventional prototype. The image performance in vivo was evaluated in euthanized healthy mice and rats, allowing visualization of detailed blood vessels in the liver and kidneys. In tumor-bearing mice, different results of vascular induction were shown depending on the type of tumor and the method of transplantation. By utilizing the video imaging function, we were able to observe the movement of blood vessels around the tumor. We have demonstrated the feasibility of the system as a less invasive animal experimental device, as it can acquire vascular images in animals in a non-contrast and non-invasive manner.


Assuntos
Neoplasias , Técnicas Fotoacústicas , Animais , Imageamento Tridimensional/métodos , Camundongos , Neoplasias/diagnóstico por imagem , Técnicas Fotoacústicas/métodos , Ratos , Ultrassonografia
7.
Int J Mol Sci ; 23(6)2022 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-35328395

RESUMO

Animal studies suggest that pain-related-molecule upregulation in degenerated intervertebral discs (IVDs) potentially leads to low back pain (LBP). We hypothesized that IVD mechanical stress and axial loading contribute to discogenic LBP's pathomechanism. This study aimed to elucidate the relationships among the clinical findings, radiographical findings, and pain-related-molecule expression in human degenerated IVDs. We harvested degenerated-IVD samples from 35 patients during spinal interbody fusion surgery. Pain-related molecules including tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-6, calcitonin gene-related peptide (CGRP), microsomal prostaglandin E synthase-1 (mPGES1), and nerve growth factor (NGF) were determined. We also recorded preoperative clinical findings including body mass index (BMI), Oswestry Disability Index (ODI), and radiographical findings including the vacuum phenomenon (VP) and spinal instability. Furthermore, we compared pain-related-molecule expression between the VP (-) and (+) groups. BMI was significantly correlated with the ODI, CGRP, and mPGES-1 levels. In the VP (+) group, mPGES-1 levels were significantly higher than in the VP (-) group. Additionally, CGRP and mPGES-1 were significantly correlated. Axial loading and mechanical stress correlated with CGRP and mPGES-1 expression and not with inflammatory cytokine or NGF expression. Therefore, axial loading and mechanical stress upregulate CGRP and mPGES-1 in human degenerated IVDs, potentially leading to chronic discogenic LBP.


Assuntos
Degeneração do Disco Intervertebral , Disco Intervertebral , Dor Lombar , Animais , Índice de Massa Corporal , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Humanos , Interleucina-6/metabolismo , Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/metabolismo , Dor Lombar/etiologia , Fator de Crescimento Neural/metabolismo , Vácuo
8.
Biomed Res Int ; 2021: 7988320, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34337052

RESUMO

Age is a key factor in intervertebral disc (IVD) degeneration; however, the changes that occur in IVDs with age are not fully understood. Tissue-resident macrophages are critical for tissue homeostasis and are regulated by transforming growth factor- (TGF-) ß. We examined changes in the proportion of resident macrophages in young versus aged mice and the role of TGF-ß in regulating resident macrophages in IVDs. IVDs were harvested from 4-month (young) and 18-month-old (aged) C57BL/6J mice. The proportion of macrophages in IVDs was determined using flow cytometry (n = 5 for each time point) and the expression of Cd11b, Cd206, and Tgfb genes, which encode CD11b, CD206, and TGF-ß protein, respectively, using real-time PCR. To study the role of TGF-ß in the polarization of resident macrophages, resident macrophages isolated from IVDs from young and aged mice were treated with recombinant TGF-ß with and without a TGF-ß inhibitor (SB431542). Additionally, SB431542 was intraperitoneally injected into young and aged mice, and Cd206 expression was examined using real-time PCR (n = 10 for each time point). The proportion of CD11b+ and CD11b+ CD206+ cells was significantly reduced in aged versus young mice, as was Cd11b, Cd206, and Tgfb expression. TGF-ß/IL10 stimulation significantly increased the expression of Cd206, an M2 macrophage marker, in disc macrophages from both young and aged mice. Meanwhile, administration of a TGF-ß inhibitor significantly reduced Cd206 expression compared to vehicle control in both groups. Conclusion. Resident macrophages decrease with age in IVDs, which may be associated with the concomitant decrease in TGF-ß. Our findings provide new insight into the mechanisms of age-related IVD pathology.


Assuntos
Envelhecimento/metabolismo , Disco Intervertebral/metabolismo , Lectinas Tipo C/metabolismo , Macrófagos/metabolismo , Lectinas de Ligação a Manose/metabolismo , Receptores de Superfície Celular/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Animais , Biomarcadores/metabolismo , Células Cultivadas , Masculino , Receptor de Manose , Camundongos Endogâmicos C57BL
9.
BMC Musculoskelet Disord ; 22(1): 634, 2021 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-34301215

RESUMO

BACKGROUND: Intervertebral disc (IVD) degeneration is a major cause of low back pain (LBP). Following disc injury, nerve growth factor (NGF) concentrations rise in IVDs, and anti-NGF therapy has been shown to attenuate LBP in humans. Increased levels of tumor necrosis factor-α (TNF-α) and transforming growth factor-ß (TGF-ß) in degenerative IVDs and in in vitro studies suggest that these factors promote NGF production. However, whether these factors regulate NGF in vivo remains unclear. Thus, we studied NGF regulation in a mouse model of IVD injury. METHODS: After inducing IVD injury, we examined mRNA levels of Tnfa, Tgfb, and Ngf in IVDs from control and IVD-injured mice across 7 days. To do this, we used magnetic cell separation to isolate CD11b ( +) (macrophage-rich) and CD11b (-) (IVD cell-rich) cell fractions from injured IVDs. To study the effect of TNF-α on Ngf expression, we examined Ngf expression in injured IVDs from C57BL/6 J and Tnfa-knockout (KO) mice (C57BL/6 J background). To study the effect of TGF-ß on Ngf expression, C57/BL6J mice were given an intraperitoneal injection of either the TGF-ß inhibitor SB431542 or DMSO solution (vehicle) one and two days before harvesting IVDs. RESULTS: mRNA expression of Tnfa, Tgfb, and Ngf was significantly increased in injured IVDs. Tnfa was predominantly expressed in the CD11b ( +) fraction, and Tgfb in the CD11b (-) fraction. Ngf expression was comparable between CD11b ( +) and CD11b (-) fractions, and between wild-type and Tnfa-KO mice at post-injury day (PID) 1, 3, and 7. SB431542 suppressed TGF-ß-mediated Ngf expression and NGF production in vitro. Further, administration of SB431542 significantly reduced Ngf expression in IVDs such that levels were below those observed in vehicle-treated animals at PID3 and PID7. CONCLUSION: A TGF-ß inhibitor reduced Ngf expression in a mouse model of IVD injury, suggesting that TGF-ß may regulate NGF expression in vivo.


Assuntos
Degeneração do Disco Intervertebral , Fator de Crescimento Neural/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Animais , Disco Intervertebral , Degeneração do Disco Intervertebral/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Fator de Crescimento Transformador beta/antagonistas & inibidores
10.
Biomed Mater Eng ; 32(4): 207-215, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33780358

RESUMO

BACKGROUND: An enzymatic crosslinking strategy using hydrogen peroxide and horseradish peroxidase is receiving increasing attention for application with in situ-formed hydrogels (IFHGs). IFHGs may also be ideal carrier materials for bone repair, although their ability to carry bone morphogenetic protein-2 (BMP2) has yet to be examined. OBJECTIVE: We examined the effectiveness of an IFHG made of hyaluronan (IFHG-HA) containing BMP2 for promoting bone formation in a mouse critical size bone defect model. METHODS: C57/BL6J mice received a 2-mm femoral critical-sized bone defect before being randomly assigned to one of the following treatment groups (n = 6): control (no treatment), IFHG-HA only, PBS with BMP2, and IFHG-HA with BMP2. X-ray radiographs were utilized to track new bone formation, and micro-computed tomography and histological examination were performed on new bone formed at the bone defect site two weeks after surgery. RESULTS: Mice treated with PBS with BMP2 and IFHG-HA with BMP2 had greater bone volume (BV) and bone mineral content (BMC) than those receiving control, and successfully achieved consolidation. Mice treated with IFHG-HA with BMP2 had significantly higher BV and BMC than those treated with PBS with BMP2. CONCLUSIONS: IFHG-HA may be an effective carrier for BMP2 to enable delivery for bone defect repair.


Assuntos
Hidrogéis , Osteogênese , Aceleração , Animais , Proteína Morfogenética Óssea 2 , Ácido Hialurônico , Camundongos , Crânio , Microtomografia por Raio-X
11.
J Orthop Res ; 39(8): 1755-1762, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-32856747

RESUMO

Multiple human and animal studies suggest that the upregulation of inflammatory cytokines and other pain-related molecules in degenerated or injured intervertebral discs (IVDs) may cause discogenic low back pain (LBP). We previously reported that macrophages in injured IVD in mice produced inflammatory cytokines, but not other pain-related molecules. CD14 is a monocyte marker expressed mainly by macrophages. The aim of the current study was to evaluate the role of CD14-positive cells in inflammatory cytokine and pain-related molecule expression in human degenerated IVD. IVD samples were harvested from 14 patients, including 10 with lumbar spinal stenosis, four with adult spinal deformity, and one with lumbar disc herniation during spinal interbody fusion surgery. Harvested IVD-derived mononuclear cells were obtained and CD14-positive (+) and CD14-negative (-) cells were separated using CD14 antibody and streptavidin-labeled magnetic beads. Inflammatory cytokines messenger RNA (mRNA) in the CD14(+) and CD14(-) cells, including tumor necrosis factor ɑ (TNFA), in, terleukin-1ß (IL1B) and IL6, were determined using quantitative polymerase chain reaction (qPCR) and their expression levels were compared. To evaluate factors controlling the regulation of pain-related molecules mRNA expression, cultured CD14(-) and CD14(+) cells from IVDs were stimulated with recombinant human TNF-ɑ and IL-1ß and levels of pain-related molecules, including calcitonin gene-related peptide (CGRP) and nerve growth factor (NGF) were determined using qPCR. Levels of TNFA, IL1B, IL6, and NGF in CD14(+) cells were significantly increased compared with those in CD14(-) cells (TNFA, p = 0.006; IL1B, p = .017; IL6, p = .010; NGF, p = .027). Following TNFA stimulation, NGF levels were significantly increased in CD14(-) and CD14(+) cells (CD14(-), p = .003; CD14(+), p < .001) and CGRP was significantly increased in CD14(-) IVD cells (p = .040). Following IL1B stimulation, NGF levels were significantly increased in CD14(-) cells (p = .004). CD14(+) cells had higher TNFA, IL1B, IL6, and NGF expressions than CD14(-) cells in human degenerated IVDs. Additionally, TNFA stimulation promoted the upregulation of NGF and CGRP in CD14(-) cells. These findings suggested that CD14(+) cells directly and indirectly contributed to inflammatory cytokine and pain-related molecule expression in human degenerated IVD. CD14(+) cells might be important in the pathological mechanism of chronic discogenic LBP in humans.


Assuntos
Degeneração do Disco Intervertebral , Disco Intervertebral , Dor Lombar , Animais , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Citocinas/metabolismo , Humanos , Interleucina-6/metabolismo , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/patologia , Dor Lombar/etiologia , Dor Lombar/patologia , Camundongos , Fator de Crescimento Neural/metabolismo , RNA Mensageiro/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
12.
J Orthop Surg Res ; 15(1): 471, 2020 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-33054796

RESUMO

BACKGROUND: Delivery of bone morphogenetic protein-2 (BMP-2) via animal-derived absorbable collagen materials is used for the treatment of large bone defects. However, the administration of bovine proteins to humans is associated with the risk of zoonotic complications. We therefore examined the effect of combining BMP-2 with collagen-like peptides, poly(POG)n, in a critical-sized bone defect mouse model. METHODS: A 2-mm critical-sized bone defect was created in the femur of 9-week-old male C57/BL6J mice. Mice were randomly allocated into one of four treatment groups (n = 6 each): control (no treatment), poly(POG)n only, 0.2 µg, or 2.0 µg BMP-2 with poly(POG)n. New bone formation was monitored using soft X-ray radiographs, and bone formation at the bone defect site was examined using micro-computed tomography and histological examination at 4 weeks after surgery. RESULTS: Administration of 2.0 µg of BMP-2 with poly(POG)n promoted new bone formation and resulted in greater bone volume and bone mineral content than that observed in the control group and successfully achieved consolidation. In contrast, bone formation in all other groups was scarce. CONCLUSIONS: Our findings suggest the potential of BMP-2 with poly(POG)n as a material, free from animal-derived collagen, for the treatment of large bone defects.


Assuntos
Proteína Morfogenética Óssea 2/administração & dosagem , Proteína Morfogenética Óssea 2/farmacologia , Colágeno , Portadores de Fármacos , Fêmur/lesões , Fêmur/fisiopatologia , Osteogênese/efeitos dos fármacos , Fator de Crescimento Transformador beta/administração & dosagem , Fator de Crescimento Transformador beta/farmacologia , Animais , Modelos Animais de Doenças , Fêmur/diagnóstico por imagem , Géis , Masculino , Camundongos Endogâmicos C57BL , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacologia , Microtomografia por Raio-X
13.
J Orthop Surg Res ; 15(1): 426, 2020 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-32948214

RESUMO

BACKGROUND: An enzymatic crosslinking strategy using hydrogen peroxide and horseradish peroxidase is receiving increasing attention for application with in situ-formed hydrogels (IFHs). Several studies have reported the application of IFHs in cell delivery and tissue engineering. IFHs may also be ideal carrier materials for bone repair, although their potential as a carrier for bone morphogenetic protein (BMP)-2 has yet to be examined. Here, we examined the effect of an IFH made of hyaluronic acid (IFH-HA) containing BMP-2 in promoting osteogenesis in a mouse refractory fracture model. METHODS: Immediately following a fracture procedure, animals either received no treatment (control) or an injection of IFH-HA/PBS or IFH-HA containing 2 µg BMP-2 (IFH-HA/BMP-2) into the fracture site (n = 16, each treatment). RESULTS: Fracture sites injected with IFH-HA/BMP-2 showed significantly greater bone volume, bone mineral content, and bone union compared with sites receiving no treatment or treated with IFH-HA/PBS alone (each n = 10). Gene expression levels of osteogenic markers, Alpl, Bglap, and Osx, were significantly raised in the IFH-HA/BMP-2 group compared to the IFH-HA/PBS and control groups (each n = 6). CONCLUSION: IFH-HA/BMP-2 may contribute to the treatment of refractory fractures.


Assuntos
Proteína Morfogenética Óssea 2/administração & dosagem , Proteína Morfogenética Óssea 2/farmacologia , Fraturas do Fêmur/tratamento farmacológico , Fraturas do Fêmur/fisiopatologia , Fêmur/metabolismo , Fêmur/patologia , Hidrogéis/administração & dosagem , Osteogênese/efeitos dos fármacos , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Densidade Óssea , Modelos Animais de Doenças , Fraturas do Fêmur/genética , Fraturas do Fêmur/patologia , Fêmur/fisiopatologia , Expressão Gênica/efeitos dos fármacos , Injeções Intralesionais , Camundongos Endogâmicos C57BL , Tamanho do Órgão , Osteogênese/genética , Fator de Transcrição Sp7/genética , Fator de Transcrição Sp7/metabolismo
14.
Artigo em Japonês | MEDLINE | ID: mdl-32963140

RESUMO

It is important to optimize the exposure dose when conducting interventional radiology, but optimization is difficult for medical centers to achieve independently. In 2005, we administered a questionnaire on the measurement of dose rates and awareness of exposure reduction when performing percutaneous coronary intervention. Ten years later, we conducted a follow-up survey of the same 31 centers to determine the current situation and identify trends. The results of the survey showed that the mean fluoroscopy dose rate decreased to 55% of the 2005 value, from 28.2 to 15.6 mGy/min, and the mean radiography dose rate decreased to 71% of the 2005 value, from 4.2 to 3.0 mGy/s. Dose rates for both fluoroscopy and radiography decreased by 84% of facilities. The results also indicated greater cooperation by physicians compared to 10 years ago. In particular, there was a considerable increase in the exchange of ideas with physicians regarding exposure, suggesting a stronger level of interest in exposure. The overall score for questionnaire items was 33% higher than that in the previous survey. These results show that in the past 10 years, awareness of exposure reduction has improved, and dose optimization has been a major factor in the downward trend in dose rates in radiography and fluoroscopy.


Assuntos
Intervenção Coronária Percutânea , Radiografia Intervencionista , Angiografia Coronária , Fluoroscopia , Seguimentos , Doses de Radiação , Inquéritos e Questionários , Raios X
15.
Cureus ; 12(7): e9331, 2020 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-32714714

RESUMO

The global coronavirus disease 2019 (COVID-19) pandemic has caused several million infections and hundreds of thousands of deaths. A large number of healthcare workers have died as a result of infection with this virus. Therefore, elective surgery was markedly reduced or stopped in our hospital's orthopedic department. The detection of asymptomatic COVID-19-positive patients became key to reducing the infection risk to physicians and staff to allow orthopedic surgery to be performed. A total of 21 patients were scheduled to undergo orthopedic surgery, including elective surgery, in Shonantobu General Hospital, Chigasaki City, Kanagawa, Japan. All 21 patients gave permission to undergo loop-mediated isothermal amplification (LAMP) screening the day before surgery. None of the 21 patients we tested was positive for COVID-19. All patients remained asymptomatic during the two to four weeks of postoperative follow-up. No physicians or medical staff developed COVID-19 symptoms. This was a very small study in a city with a relatively low incidence of COVID-19. We found that LAMP screening was accurate, in terms of its negative predictive value. Larger studies are needed.

16.
J Orthop Res ; 38(8): 1703-1709, 2020 08.
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.


Assuntos
Degeneração do Disco Intervertebral/imunologia , Deslocamento do Disco Intervertebral/imunologia , Macrófagos , Animais , Células da Medula Óssea/metabolismo , Quimiocinas/metabolismo , Proteínas de Fluorescência Verde , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos
17.
Cent Eur J Immunol ; 45(4): 377-381, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33613092

RESUMO

Recent evidence suggests that synovial macrophage activation may be involved in cartilage destruction and pain in osteoarthritis (OA). The macrophage-inducible C-type lectin (Mincle) Clec4e is expressed in macrophages and is regulated in inflammatory conditions. Given that the regulation of Mincle in synovial macrophages has not been elucidated, we investigated the expression and regulation of Mincle in human synovial tissue (ST) harvested from patients with radiographic knee OA during total knee arthroplasty. Immunohistochemical and flow cytometric analyses were used to identify cells with Mincle expression in resected tissues. CD14-positive (CD14+; macrophage-rich cell fraction) and CD14-negative (CD14-; fibroblast-rich cell fraction) cells were extracted from the ST and used to assess MINCLE mRNA expression levels. To determine the role of tumor necrosis factor alpha (TNF-α) in the regulation of MINCLE expression, TNF-α was used to stimulate cultured CD14+ cells. Immunohistochemical staining revealed Mincle-positive cells in the synovial lining layer. Flow cytometric analysis showed that CD45+CD14+ cells were Mincle positive while CD45-/CD14- cells were Mincle negative. MINCLE expression was significantly higher in CD14+ cells than in CD14- cells. Stimulation of cultured CD14+ macrophages with TNF-α significantly increased MINCLE mRNA expression, while stimulation with TNF-α neutralizing antibody significantly decreased expression. That Mincle expression was observed in synovial macrophages and its expression was induced by TNF-α suggests that Mincle might have a key role in synovial inflammation in the osteoarthritic synovium.

18.
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.


Assuntos
Cabeça/cirurgia , Pescoço/cirurgia , Retalho Perfurante/irrigação sanguínea , Técnicas Fotoacústicas , Procedimentos de Cirurgia Plástica/métodos , Cuidados Pré-Operatórios , Cirurgia Assistida por Computador , Adulto , Idoso , Feminino , Cabeça/diagnóstico por imagem , Humanos , Lasers , Masculino , Pessoa de Meia-Idade , Pescoço/diagnóstico por imagem , Estudos Prospectivos , Coxa da Perna/irrigação sanguínea , Coxa da Perna/diagnóstico por imagem , Coxa da Perna/cirurgia , Parede Torácica/cirurgia , Ultrassonografia
19.
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
20.
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
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