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1.
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
2.
Clin Sci (Lond) ; 133(4): 583-595, 2019 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-30777884

RESUMO

Recent reports, including ours, have indicated that microRNA (miR)-33 located within the intron of sterol regulatory element binding protein (SREBP) 2 controls cholesterol homeostasis and can be a potential therapeutic target for the treatment of atherosclerosis. Here, we show that SPAST, which encodes a microtubule-severing protein called SPASTIN, was a novel target gene of miR-33 in human. Actually, the miR-33 binding site in the SPAST 3'-UTR is conserved not in mice but in mid to large mammals, and it is impossible to clarify the role of miR-33 on SPAST in mice. We demonstrated that inhibition of miR-33a, a major form of miR-33 in human neurons, via locked nucleic acid (LNA)-anti-miR ameliorated the pathological phenotype in hereditary spastic paraplegia (HSP)-SPG4 patient induced pluripotent stem cell (iPSC)-derived cortical neurons. Thus, miR-33a can be a potential therapeutic target for the treatment of HSP-SPG4.


Assuntos
Terapia Genética/métodos , Células-Tronco Pluripotentes Induzidas/metabolismo , MicroRNAs/genética , Células-Tronco Neurais/metabolismo , Neuritos/metabolismo , Oligonucleotídeos/genética , Paraplegia Espástica Hereditária/terapia , Espastina/genética , Regiões 3' não Traduzidas , Sítios de Ligação , Células Cultivadas , Regulação da Expressão Gênica , Humanos , Células-Tronco Pluripotentes Induzidas/patologia , MicroRNAs/antagonistas & inibidores , MicroRNAs/metabolismo , Células-Tronco Neurais/patologia , Neuritos/patologia , Neurogênese , Oligonucleotídeos/metabolismo , Fenótipo , Paraplegia Espástica Hereditária/genética , Paraplegia Espástica Hereditária/metabolismo , Paraplegia Espástica Hereditária/patologia , Espastina/metabolismo
3.
Cleft Palate Craniofac J ; 56(8): 1026-1037, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30773047

RESUMO

BACKGROUND: Congenital midfacial hypoplasia often requires intensive treatments and is a typical condition for the Binder phenotype and syndromic craniosynostosis. The growth trait of the midfacial skeleton during the early fetal period has been assumed to be critical for such an anomaly. However, previous embryological studies using 2-dimensional analyses and specimens during the late fetal period have not been sufficient to reveal it. OBJECTIVE: To understand the morphogenesis of the midfacial skeleton in the early fetal period via 3-dimensional quantification of the growth trait and investigation of the developmental association between the growth centers and midface. METHODS: Magnetic resonance images were obtained from 60 human fetuses during the early fetal period. Three-dimensional shape changes in the craniofacial skeleton along growth were quantified and visualized using geometric morphometrics. Subsequently, the degree of development was computed. Furthermore, the developmental association between the growth centers and the midfacial skeleton was statistically investigated and visualized. RESULTS: The zygoma expanded drastically in the anterolateral dimension, and the lateral part of the maxilla developed forward until approximately 13 weeks of gestation. The growth centers such as the nasal septum and anterior portion of the sphenoid were highly associated with the forward growth of the midfacial skeleton (RV = 0.589; P < .001). CONCLUSIONS: The development of the midface, especially of the zygoma, before 13 weeks of gestation played an essential role in the midfacial development. Moreover, the growth centers had a strong association with midfacial forward growth before birth.


Assuntos
Craniossinostoses , Face , Desenvolvimento Fetal , Maxila , Desenvolvimento Maxilofacial , Face/embriologia , Feminino , Humanos , Maxila/embriologia , Maxila/crescimento & desenvolvimento , Morfogênese , Gravidez , Zigoma
4.
J Surg Res ; 221: 173-182, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29229125

RESUMO

BACKGROUND: We previously developed collagen/gelatin sponges (CGS) able to sustain and release basic fibroblast growth factor (bFGF) and reported that this CGS impregnated with bFGF promoted dermis-like tissue formation. We herein confirmed the single-sustained release of hepatocyte growth factor (HGF) and the dual sustained release of HGF and bFGF from CGSs, and explored its efficacy using a murine model of skin defects. MATERIALS AND METHODS: The sustained release of HGF alone and both HGF and bFGF from CGSs were evaluated in vitro. CGSs (8 mm in diameter) impregnated with normal saline solution (NSS) (NSS group), HGF solution (10 or 50 µg/cm2) (HGF-L or HGF-H group), bFGF solution (7 µg/cm2) (bFGF group), or HGF (10 µg/cm2) and bFGF (7 µg/cm2) solution (HGF + bFGF group) were implanted into full-thickness skin defects on the backs of mice. The wound area, neoepithelium length, dermis-like tissue formation and newly formed capillaries were evaluated. RESULTS: The single release of HGF and the dual release of HGF and bFGF from CGSs were confirmed. At week 1, the wound closure and neoepithelium length were promoted in the HGF-L group compared with the NSS group. At week 2, the wound closure, neoepithelium length, dermis-like tissue formation and newly formed capillary formation were promoted in the bFGF and HGF + bFGF groups compared with the NSS and HGF-H groups. Newly formed capillary formation was superior in the HGF + bFGF group compared with the bFGF group. CONCLUSIONS: The dual release of HGF and bFGF from CGS was a promising treatment for full-thickness skin defects.


Assuntos
Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Fator de Crescimento de Hepatócito/administração & dosagem , Alicerces Teciduais , Cicatrização/efeitos dos fármacos , Animais , Colágeno , Gelatina , Camundongos Endogâmicos C57BL , Suínos
5.
Int J Mol Sci ; 19(5)2018 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-29695130

RESUMO

Keloids occur after failure of the wound healing process; inflammation persists, and various treatments are ineffective. Keloid pathogenesis is still unclear. We have previously analysed the gene expression profiles in keloid tissue and found that HtrA1 was markedly up-regulated in the keloid lesions. HtrA1 is a serine protease suggested to play a role in the pathogenesis of various diseases, including age-related macular degeneration and osteoarthritis, by modulating extracellular matrix or cell surface proteins. We analysed HtrA1 localization and its role in keloid pathogenesis. Thirty keloid patients and twelve unrelated patients were enrolled for in situ hybridization, immunohistochemical, western blot, and cell proliferation analyses. Fibroblast-like cells expressed more HtrA1 in active keloid lesions than in surrounding lesions. The proportion of HtrA1-positive cells in keloids was significantly higher than that in normal skin, and HtrA1 protein was up-regulated relative to normal skin. Silencing HtrA1 gene expression significantly suppressed cell proliferation. HtrA1 was highly expressed in keloid tissues, and the suppression of the HtrA1 gene inhibited the proliferation of keloid-derived fibroblasts. HtrA1 may promote keloid development by accelerating cell proliferation and remodelling keloid-specific extracellular matrix or cell surface molecules. HtrA1 is suggested to have an important role in keloid pathogenesis.


Assuntos
Regulação da Expressão Gênica , Serina Peptidase 1 de Requerimento de Alta Temperatura A/genética , Queloide/genética , Queloide/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Biópsia , Proliferação de Células , Células Cultivadas , Feminino , Fibroblastos/metabolismo , Técnicas de Silenciamento de Genes , Humanos , Imuno-Histoquímica , Queloide/metabolismo , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/genética , Pele/metabolismo , Pele/patologia , Regulação para Cima , Adulto Jovem
6.
Prenat Diagn ; 37(9): 907-915, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28675493

RESUMO

OBJECTIVES: Disturbance of the development of the nasal septum in the early prenatal period causes congenital facial anomalies characterized by a flat nose and defects of the anterior nasal spine (ANS), such as Binder phenotype. The present research aimed to assess the development of the nasal septum and the ANS with growth in the early prenatal period. METHODS: Magnetic resonance images were obtained from 56 specimens. Mid-sagittal images were analyzed by using geometric morphometrics for the development of the nasal septum, and angle analysis was performed for the development of the ANS. Additionally, we calculated and visualized the ontogenetic allometry of the nasal septum. RESULTS: Our results showed that the nasal septum changed shape in the anteroposterior direction in smaller specimens, while it maintained an almost isometric shape in larger specimens. Furthermore, mathematical evidence revealed that the maturation periods of the shapes of the ANS and the nasal septum were around 12 and 14 weeks of gestation, respectively. CONCLUSION: The anteroposterior development of the nasal septum is specific until 14 weeks of gestation, and it is important for nasal protrusion and the development of the ANS. Therefore, the disturbance of such development could induce low nasal deformity, including Binder phenotype. © 2017 John Wiley & Sons, Ltd.


Assuntos
Imageamento por Ressonância Magnética , Septo Nasal/embriologia , Nariz/anormalidades , Feminino , Idade Gestacional , Humanos , Fenótipo , Gravidez
7.
Ann Plast Surg ; 78(6): 651-658, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28230648

RESUMO

INTRODUCTION: As the take rate of cultured epidermal autografts in burn wound treatment is variable, widely expanded meshed auto skin grafts are often used in combination with cultured epidermal autograft to increase the take rate and achieve definitive wound coverage. However, a long time (3-4 weeks) required to prepare a cultured epidermis sheet is a disadvantage. Allogeneic cultured epidermis can be prepared in advance and cryopreserved to be used in combination with auto meshed skin grafts for treating third-degree burns. Nevertheless, the human cultured epidermis (hCE) has not been proved to accelerate wound healing after meshed skin grafting. Here, we investigated the effect of hCE on wound healing in a rat model of meshed skin grafting. MATERIALS AND METHODS: Human cultured epidermis was prepared from human neonatal foreskin and assessed by the release of growth factors into the culture medium using enzyme-linked immunosorbent assay. Skin wounds were inflicted on male F344 rats and treated by the application of widely meshed (6:1 ratio) autogenous skin grafts with or without hCE (n = 8 rats per group). Wound area, neoepithelium length, granulation tissue formation, and neovascularization were evaluated on day 7 postgrafting. RESULTS: Human cultured epidermis secreted IL-1α, Basic fibroblast growth factor, platelet-derived growth factor-AA, TGF-α, TGF-ß1, and vascular endothelial growth factor in vitro. In rats, hCE accelerated wound closure (P = 0.003), neoepithelium growth (P = 0.019), and granulation tissue formation (P = 0.043), and increased the number of capillaries (P = 0.0003) and gross neovascularization area (P = 0.008) compared with the control group. CONCLUSIONS: The application of hCE with meshed grafts promoted wound closure, possibly via secretion of growth factors critical for cell proliferation and migration, suggesting that hCE can enhance the healing effect of widely expanded skin autografts.


Assuntos
Queimaduras/cirurgia , Células Epidérmicas , Tecido de Granulação/citologia , Reepitelização/fisiologia , Transplante de Pele/métodos , Animais , Autoenxertos , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Humanos , Masculino , Ratos , Ratos Endogâmicos F344 , Cicatrização/fisiologia
8.
J Craniofac Surg ; 28(5): 1302-1304, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28570398

RESUMO

The authors performed distraction osteogenesis using The Maxillary Distractor System (SYNTHES) to maxillary hypoplasia patient with cleft lip palate, and consequently improved the aesthetic complexion of the patient. Velopharyngeal insufficiency developed after bone elongation; the authors improved the insufficiency with conservative therapies such as articulatory training using the bulb attached palatal lift prosthesis. The authors were successful and accepted postoperative speech outcome.


Assuntos
Fenda Labial/cirurgia , Fissura Palatina/cirurgia , Osteogênese por Distração , Retrognatismo/cirurgia , Insuficiência Velofaríngea/terapia , Estética , Humanos , Masculino , Má Oclusão/cirurgia , Osteogênese por Distração/efeitos adversos , Insuficiência Velofaríngea/etiologia , Adulto Jovem
9.
Dev Dyn ; 245(2): 123-31, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26509917

RESUMO

BACKGROUND: After palatoplasty, incomplete velopharyngeal closure in speech articulation sometimes persists, despite restoration of deglutition function. The levator veli palatini (LVP) is believed to be significantly involved with velopharyngeal function in articulation; however, the development and innervation of LVP remain obscure. The development of LVP in human embryos and fetuses has not been systematically analyzed using the Carnegie stage (CS) to standardize documentation of development. RESULTS: The anlage of LVP starts to develop at CS 21 beneath the aperture of the auditory tube (AT) to the pharynx. At CS 23, LVP runs along AT over its full length, as evidenced by three-dimensional image reconstruction. In the fetal stage, the lesser palatine nerve (LPN) is in contact with LVP. CONCLUSIONS: The positional relationship between LVP and AT three-dimensionally, suggesting that LVP might be derived from the second branchial arch. Based on histological evidence, we hypothesize that motor components from the facial nerve may run along LPN, believed to be purely sensory. The multiple innervation of LVP by LPN and pharyngeal plexus may explain the postpalatoplasty discrepancy between the partial impairment in articulation vs. the functional restoration of deglutition. That is, the contribution of LPN is greater in articulation than in deglutition.


Assuntos
Fissura Palatina/patologia , Músculos Palatinos/embriologia , Palato/embriologia , Feminino , Humanos , Imageamento Tridimensional , Imageamento por Ressonância Magnética , Masculino , Músculos Palatinos/inervação , Músculos Palatinos/patologia
10.
Cells Tissues Organs ; 201(3): 170-9, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27002537

RESUMO

High hydrostatic pressure (HHP) technology is a physical method for inactivating tissue. We reported that nevus specimens were inactivated after HHP at 200 MPa and that the inactivated nevus could be used as autologous dermis for covering skin defects. In this study, we verified the inactivation of nevus specimens using a newly developed portable HHP device which will be used in a clinical trial. Nevus tissue specimens were obtained from 5 patients (mean age 7.2 years, range 1-19). We cultured fibroblasts and nevus cells from the tissue specimens and then evaluated their inactivation after HHP at 200 MPa by confirming the attachment of the suspensions and by the live/dead staining of the suspensions, through the dissociation of the cells on chamber slides and by the live/dead staining of the remaining cells. The cells were also quantitatively evaluated by WST-8 assay. We then confirmed the inactivation of the nevus specimens after HHP using explant culture. Our results indicated that fibroblasts and nevus cells were inactivated after HHP at 200 MPa, with the exception of a small percentage of green-colored cells, which reflected the remaining activity of the cellular esterases after HHP. No cells migrated from the nevus specimens after HHP at 200 MPa. We verified the inactivation of fibroblasts and nevus cells cultured from nevus specimens, and in the nevus samples themselves after pressurization at 200 MPa using this device. This device could be used in clinical trials for giant congenital melanocytic nevi and may thus become useful in various medical fields.


Assuntos
Fibroblastos/patologia , Nevo Pigmentado/patologia , Nevo Pigmentado/terapia , Neoplasias Cutâneas/patologia , Neoplasias Cutâneas/terapia , Pele/patologia , Adolescente , Sobrevivência Celular , Células Cultivadas , Criança , Pré-Escolar , Fibroblastos/citologia , Humanos , Pressão Hidrostática , Lactente , Adulto Jovem
11.
J Surg Res ; 201(2): 378-87, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27020822

RESUMO

BACKGROUND: Gelatin has been used as a material sustaining the release of basic fibroblast growth factor (bFGF), which promotes fibroblast proliferation and capillary formation and accelerates wound healing. In the application of these materials, bFGF is impregnated immediately before application, and it is difficult to conform the shape to the wound. In this study, we prepared a pliable and plastic gelatin gel sheet (GGS) that sustains bFGF and conforms to the shape of the wound as a result of cross-linking just before application. In addition, we examined the sustained release profile of bFGF from GGS and its effect on wound healing in murine skin defects. MATERIALS AND METHODS: A 13-wt% gelatin solution was mixed with bFGF before cross-linking with 1% glutaraldehyde solution. GGSs impregnated with 7 µg/cm(2) of bFGF were incubated in phosphate-buffered saline and collagenase solution, and GGS degradation and bFGF release were evaluated. In the murine experiments, GGSs treated without bFGF and GGSs impregnated with 1, 3.5, 7, or 14 µg/cm(2) of bFGF were applied to full-thickness skin defects created on the backs of C57BL/6JJcl mice, and the wound closure, epithelial length, extent of granulation tissue and capillary formation were compared. RESULTS: bFGF was released according to the degradation of GGS in phosphate-buffered saline, and the remaining bFGF was released in collagenase solution. In the animal studies, epithelialization was accelerated in the GGSs treated with 1 and 3.5 µg/cm(2) of bFGF, and granulation tissue formation and angiogenesis were promoted based on the amount of bFGF impregnated into the GGS. CONCLUSIONS: GGS impregnated with bFGF is capable of sustaining the release of bFGF, with consequent accelerated epithelialization, granulation tissue formation, and angiogenesis in vivo. GGS is a novel and promising wound dressing that sustains bFGF and can be adapted to the shape of various wounds in the treatment of both acute and chronic wounds.


Assuntos
Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Cicatrização/efeitos dos fármacos , Animais , Preparações de Ação Retardada , Avaliação Pré-Clínica de Medicamentos , Elasticidade , Gelatina , Masculino , Camundongos Endogâmicos C57BL , Pele/patologia , Ferimentos e Lesões/patologia
12.
J Surg Res ; 201(2): 446-54, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27020831

RESUMO

BACKGROUND: Epidermal growth factor (EGF) plays an important role in wound healing. However, EGF must be applied daily due to rapid inactivation in vivo. We investigated the sustained release of EGF from gelatin gel sheets (GGSs) and the efficacy of GGSs impregnated with EGF for promoting wound healing. MATERIALS AND METHODS: GGSs impregnated with EGF were prepared by cross-linking via glutaraldehyde to gelatin solution containing EGF. The sustained release of EGF and the bioactivity of released EGF were evaluated. Then, three kinds of GGSs containing NSS (normal saline solution; NSS group), 2.5 µg of EGF (EGF-L group), or 25 µg of EGF (EGF-H group) were applied to full-thickness skin defects created on the backs of mice. The wounds covered with polyurethane film without GGS were used as a control (PUF group). The wound area, neoepithelium length, regenerated granulation tissue, and newly formed capillaries were evaluated. RESULTS: EGF was sustained and released from GGS as it degraded. The bioactivity of released EGF was confirmed. EGF-L group promoted the neoepithelium length, regenerated granulation tissue, and newly formed capillaries compared with those in the PUF and NSS groups. The area of regenerated granulation tissue in the NSS group (week 1: 2.6 + 0.2 mm(2), week 2: 2.8 + 0.3 mm(2)) was larger than that in the PUF group (week 1: 0.6 + 0.1 mm(2), week 2: 1.0 + 0.1 mm(2)). The area of newly formed capillaries in the EGF-L group (9967 + 1903 µm(2)) was larger than that of the EGF-H group (3485 + 1050 µm(2)). CONCLUSIONS: GGSs impregnated with EGF-L showed promising results regarding wound healing.


Assuntos
Fator de Crescimento Epidérmico/administração & dosagem , Cicatrização/efeitos dos fármacos , Animais , Criança , Preparações de Ação Retardada , Feminino , Gelatina , Glutaral , Tecido de Granulação/irrigação sanguínea , Humanos , Masculino , Camundongos Endogâmicos C57BL
13.
J Artif Organs ; 19(2): 167-74, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26497310

RESUMO

Nicotine has been reported to prolong the wound healing; however, we showed that the topical application of 10(-4) M nicotine promoted murine wound healing. The objective of this study was to explore the wound healing effects of nicotine in combination with collagen scaffold using skin defects in rabbit. Three full-thickness skin defects 8 mm in diameter were made on the rabbit auricle. Artificial dermis was applied to the defects, and 10 µl of nicotine solution (10(-5), 10(-4), and10(-3) M), bFGF solution (0.5 µg/10 µl), and both bFGF and 10(-4) M nicotine solutions were injected into the artificial dermis once daily for 7 days. Rabbits were sacrificed on day 10, 15, or 20, and the wound healing process was evaluated. bFGF was superior in the formation of the dermis-like tissue and capillaries. In nicotine groups, the epithelial length and the dermis-like tissue formations in the 10(-4) M group were superior, in contrast, those were inhibited in the 10(-3) M group. The synergistic effect of bFGF and 10(-4) M nicotine was not confirmed. This study suggests that the topical application of 10(-4) M nicotine promoted wound healing in rabbit, but the effect was not apparent compared with murine models.


Assuntos
Estimulantes Ganglionares/administração & dosagem , Nicotina/administração & dosagem , Pele Artificial , Pele/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Administração Tópica , Animais , Colágeno/administração & dosagem , Avaliação Pré-Clínica de Medicamentos , Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Masculino , Camundongos , Coelhos , Pele/irrigação sanguínea , Alicerces Teciduais
14.
Proc Natl Acad Sci U S A ; 110(8): 2852-7, 2013 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-23382201

RESUMO

Elastic fiber assembly requires deposition of elastin monomers onto microfibrils, the mechanism of which is incompletely understood. Here we show that latent TGF-ß binding protein 4 (LTBP-4) potentiates formation of elastic fibers through interacting with fibulin-5, a tropoelastin-binding protein necessary for elastogenesis. Decreased expression of LTBP-4 in human dermal fibroblast cells by siRNA treatment abolished the linear deposition of fibulin-5 and tropoelastin on microfibrils. It is notable that the addition of recombinant LTBP-4 to cell culture medium promoted elastin deposition on microfibrils without changing the expression of elastic fiber components. This elastogenic property of LTBP-4 is independent of bound TGF-ß because TGF-ß-free recombinant LTBP-4 was as potent an elastogenic inducer as TGF-ß-bound recombinant LTBP-4. Without LTBP-4, fibulin-5 and tropoelastin deposition was discontinuous and punctate in vitro and in vivo. These data suggest a unique function for LTBP-4 during elastic fibrogenesis, making it a potential therapeutic target for elastic fiber regeneration.


Assuntos
Proteínas da Matriz Extracelular/metabolismo , Proteínas de Ligação a TGF-beta Latente/fisiologia , Proteínas Recombinantes/metabolismo , Animais , Células HEK293 , Humanos , Proteínas de Ligação a TGF-beta Latente/metabolismo , Camundongos , Camundongos Knockout , Ligação Proteica , Interferência de RNA
15.
Skeletal Radiol ; 45(4): 541-7, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26497541

RESUMO

OBJECTIVE: To describe a technique and its clinical applications of three-dimensional ultrasonography to type VI and VII radial polydactyly for identification of potential muscular anomalies. MATERIALS AND METHODS: Ultrasonographic examinations were performed at an out-patient department without sedation or an operative room prior to surgery. The palm was scanned in the transverse direction using a 18-MHz linear transducer under speed regulation at 3 mm/s. Sequential images acquired at 0.2 mm intervals were converted into volume data. After validation of the technique, patients with a radial polydactyly in association with triphalangism (type VII) or with polydactylies of metacarpal duplication (type VI) were included for the examination. RESULTS: Five hands of five patients, one with type VI and four with type VII, were included the study. All the patients were male and the ages at examination ranged from 7 months to 2 years. Of the five patients, four examinations were performed at an out-patient department without sedation and one was under anesthesia just prior to surgery. The muscular abnormalities identified were mal-positions of the thenar muscles in a type VI case and a deficiency of the abductor pollicis brevis muscle in a type VII case with a delta phalanx in the ulnar part. CONCLUSION: Three-dimensional ultrasound technique could be an aid to plan strategies in radial polydactyly if intrinsic muscular anomalies are suspected to be involved.


Assuntos
Dedos/anormalidades , Imageamento Tridimensional/métodos , Músculo Esquelético/diagnóstico por imagem , Polidactilia/diagnóstico por imagem , Ultrassonografia/métodos , Humanos , Interpretação de Imagem Assistida por Computador , Lactente , Masculino
16.
Ann Plast Surg ; 76(6): 652-8, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27176561

RESUMO

OBJECTIVE: The manual application of hot water or hot metal to an animal's skin surface is often used to prepare burn wound models. However, manual burn creation is subject to human variability. We developed a new device that can control the temperature, time, and pressure of contact to produce precise and reproducible animal burn wounds and investigated the conditions required to prepare various burn wounds using our new device. METHODS: We prepared burn wounds on F344 rats using 3 contact times 2, 4, and 10 seconds using a stamp heated to 80°C. We observed the wound-healing process macroscopically and histologically and evaluated the burn depth using a laser speckle contrast-imaging device, which evaluated the blood flow of the wound. RESULTS: The changes in the burned area over time, tissue perfusion of the burn wounds, histological evaluation of the burn depth by hematoxylin-eosin and azocarmine and aniline blue staining, and the epithelialization rate (the ratio of the epithelialized area to the wound length) were evaluated on histological sections. Results indicated that the burn wounds prepared with contact times of 2, 4, and 10 seconds corresponded to superficial dermal burns, deep dermal burns, and full-thickness burns, respectively. CONCLUSIONS: We demonstrated that partial- and full-thickness burn wounds can be precisely and reproducibly created with our new automated burning device.


Assuntos
Queimaduras/etiologia , Equipamentos e Provisões Elétricas , Temperatura Alta/efeitos adversos , Modelos Animais , Ratos Endogâmicos F344/lesões , Pele/lesões , Animais , Queimaduras/diagnóstico por imagem , Queimaduras/patologia , Masculino , Ratos , Pele/irrigação sanguínea , Pele/diagnóstico por imagem , Pele/patologia , Fatores de Tempo
17.
Ann Otol Rhinol Laryngol ; 124(2): 116-25, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25115594

RESUMO

OBJECTIVE: Treatment of vocal fold scarring remains challenging. We have previously reported the therapeutic effects of local injection of basic fibroblast growth factor (bFGF) in animal models and humans. A novel collagen/gelatin sponge (CGS) is capable of sustained release of bFGF, which compensates for its quick absorption in vivo, avoiding multiple injections. This study aimed to evaluate the biocompatibility and efficacy of the CGS in rat vocal fold fibroblasts prior to human trials. METHODS: Fibroblasts extracted from Sprague-Dawley rat vocal folds were seeded onto a CGS and then cultivated with bFGF at concentrations of 0, 10, and 100 ng/mL. Vocal fold fibroblast morphology, adhesion, proliferation, and gene expression were measured under these 3-dimensional conditions. RESULTS: Cells adhered to the CGS from day 1. Although no significant differences in cell morphology were detected, cell proliferation was accelerated by bFGF administration. Expression of endogenous bFGF and hepatocyte growth factor was significantly up-regulated at 10 ng/mL bFGF. The expression of procollagen I and procollagen III was significantly suppressed, whereas HAS-1 and HAS-2 were up-regulated at 10 and 100 ng/mL bFGF. CONCLUSION: The collagen/gelatin sponge is biocompatible with vocal fold fibroblasts and may be useful as a bFGF drug delivery system for the treatment of scarred vocal folds.


Assuntos
Fenômenos Fisiológicos Celulares/efeitos dos fármacos , Colágeno/farmacologia , Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Fibroblastos/metabolismo , Esponja de Gelatina Absorvível/farmacologia , Gelatina/farmacologia , Prega Vocal/patologia , Animais , Materiais Biocompatíveis/farmacologia , Cicatriz , Relação Dose-Resposta a Droga , Sistemas de Liberação de Medicamentos , Fibroblastos/patologia , Substâncias de Crescimento/administração & dosagem , Humanos , Teste de Materiais/métodos , Ratos , Ratos Sprague-Dawley , Alicerces Teciduais , Distúrbios da Voz/tratamento farmacológico
18.
J Foot Ankle Surg ; 54(6): 1119-23, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25998477

RESUMO

In toe desyndactyly, a dorsal or plantar commissural flap, combined with skin grafts, will ensure an acceptable result. However, the parallel unsightly scars in the longitudinal direction on the dorsum of the toes will sometimes fail to satisfy the patient's and/or the parents' aesthetic expectations. To address this issue, we developed a technique using a transversely oriented transposition flap for web reconstruction, which can spare the dorsal interdigital skin maximally to shift the dorsal scars plantarly such that they become inconspicuous. The design of the flap is simple and uncomplicated surgically. Moreover, the donor site morbidity is minimal, owing to the good healing potential of the transverse scars. This technique could be an alternative in web reconstruction of toe desyndactyly, especially in cases with high cosmetic priority.


Assuntos
Retalhos Cirúrgicos , Sindactilia/cirurgia , Dedos do Pé/anormalidades , Dedos do Pé/cirurgia , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Procedimentos de Cirurgia Plástica , Transplante de Pele
19.
J Artif Organs ; 17(4): 352-7, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25028148

RESUMO

A bilayered artificial dermis (AD) composed of an upper silicone sheet and a lower collagen sponge has been widely applied for skin defects. After application, fibroblasts and capillaries infiltrate the AD and the collagen sponge is replaced by host dermal tissue within a few weeks. However, this delay and the high incidence of infection are concerns regarding the use of AD in the treatment of chronic ulcers. In this study, we compared the neovascularization of conventional AD seeded with autologous fibroblasts (cultured dermis: CD) and collagen/gelatin sponge (CGS), which is a novel artificial dermis capable of sustained release of basic fibroblast growth factor (bFGF) after application using laser Doppler imaging (LDI). CD (n = 5) and CGS impregnated with bFGF (n = 6) were applied to diabetic foot ulcers after debridement. Perfusion units (PUs) were measured just after, and 1, 2 and 3 weeks after application, and complete healing rates within 16 weeks were compared. No significant differences in PUs were seen 1, 2 and 3 weeks after application and in healing rates within 16 weeks between the two groups. This study suggested that CD and CGS treatments were effective, but there were no significant differences between them in the treatment of diabetic ulcers .


Assuntos
Pé Diabético/terapia , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Neovascularização Fisiológica/fisiologia , Pele Artificial , Cicatrização/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Pé Diabético/diagnóstico por imagem , Feminino , Fibroblastos/citologia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do Tratamento , Ultrassonografia
20.
Ann Plast Surg ; 72(1): 84-8, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23241770

RESUMO

BACKGROUND: A bilayered artificial dermis is widely applied for skin defects. Its collagen sponge is biodegraded and replaced with dermis-like tissue after application. There is no reliable method for quantitatively evaluating the blood flow of artificial dermis. In this study, we used laser Doppler imaging to evaluate the perfusion of artificial dermis. MATERIALS AND METHODS: Twelve patients treated with artificial dermis and secondary skin grafting were included. We measured the perfusion unit just after application of artificial dermis, 1 week after, and before skin grafting. RESULTS: Secondary skin grafts of 6 patients took completely, and the others showed partial necrosis. Laser Doppler imaging could detect blood flow in the artificial dermis, and a significant difference was observed in perfusion units between the "complete take" group and "partial necrosis" group before grafting (P < 0.05). CONCLUSIONS: Laser Doppler imaging could be a useful and noninvasive technique for the evaluation of blood flow to the artificial dermis before grafting.


Assuntos
Fluxometria por Laser-Doppler , Neovascularização Fisiológica , Transplante de Pele/métodos , Pele Artificial , Pele/irrigação sanguínea , Adolescente , Adulto , Idoso , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Pele/diagnóstico por imagem , Ultrassonografia
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