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1.
J Anat ; 243(2): 274-283, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-36943032

RESUMO

The effects of sex on human facial morphology have been widely documented. Because sexual dimorphism is relevant to a variety of scientific and applied disciplines, it is imperative to have a complete and accurate account of how and where male and female faces differ. We apply a comprehensive facial phenotyping strategy to a large set of existing 3D facial surface images. We investigate facial sexual dimorphism in terms of size, shape, and shape variance. We also assess the ability to correctly assign sex based on shape, both for the whole face and for subregions. We applied a predefined data-driven segmentation to partition the 3D facial surfaces of 2446 adults into 63 hierarchically linked regions, ranging from global (whole face) to highly localized subparts. Each facial region was then analyzed with spatially dense geometric morphometrics. To describe the major modes of shape variation, principal components analysis was applied to the Procrustes aligned 3D points comprising each of the 63 facial regions. Both nonparametric and permutation-based statistics were then used to quantify the facial size and shape differences and visualizations were generated. Males were significantly larger than females for all 63 facial regions. Statistically significant sex differences in shape were also seen in all regions and the effects tended to be more pronounced for the upper lip and forehead, with more subtle changes emerging as the facial regions became more granular. Males also showed greater levels of shape variance, with the largest effect observed for the central forehead. Classification accuracy was highest for the full face (97%), while most facial regions showed an accuracy of 75% or greater. In summary, sex differences in both size and shape were present across every part of the face. By breaking the face into subparts, some shape differences emerged that were not apparent when analyzing the face as a whole. The increase in facial shape variance suggests possible evolutionary origins and may offer insights for understanding congenital facial malformations. Our classification results indicate that a high degree of accuracy is possible with only parts of the face, which may have implications for biometrics applications.


Assuntos
Face , Lábio , Adulto , Humanos , Feminino , Masculino , Face/anatomia & histologia , Lábio/anatomia & histologia , Imageamento Tridimensional/métodos , Caracteres Sexuais
2.
J Genet Couns ; 2023 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-37632295

RESUMO

Genomic technologies are now utilized for the genetic diagnosis of vascular anomalies. This provides the opportunity for genetic counselors to make a significant contribution to patient care for this complex disease. The aim of this study was to explore Australian healthcare professionals' perspectives on the relatively recent integration of molecular diagnostic testing for vascular anomalies, with or without genetic counseling support. Nine semi-structured interviews were conducted with Australian healthcare professionals involved in the provision of care for individuals with vascular anomalies. Thematic analysis identified six themes: (1) Molecular diagnosis is beneficial; (2) psychosocial needs can motivate families to pursue a molecular diagnosis; (3) molecular genetic testing for vascular anomalies is complex; (4) genetic service provision is not a one size fits all; (5) a client-centered approach for genetic service provision can go a long way; and (6) the value of genetic counselors. Based on our findings, implementation of a vascular anomalies genetic diagnostic program inclusive of genetic counseling may be challenging, yet such programs are likely to benefit both patients and their families, as well as healthcare professionals. As this paradigm shift unfolds, genetic counselors have an opportunity to contribute to the vascular anomaly field by educating healthcare professionals and patients, by participating in multidisciplinary clinics to support complex cases and by raising awareness regarding their practice and potential contributions.

3.
J Pediatr Orthop ; 41(1): e67-e73, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32815867

RESUMO

BACKGROUND: Intramuscular venous malformations, often erroneously called "intramuscular hemangiomas," present to pediatric orthopaedic surgeons either as a differential diagnosis of tumor or as a cause of muscle pain. Treatment options include injection sclerotherapy or surgery. There is some literature to indicate that sclerotherapy can reduce pain, but little evidence on the effectiveness of surgery. The primary aim of this study was to evaluate the efficacy of surgical resection for intramuscular venous malformations, with a secondary aim to evaluate the natural history and presentation of intramuscular venous malformations to improve clinician understanding of this condition. METHODS: A retrospective chart analysis was performed of cases identified from a vascular anomalies database from January 2004 and December 2018. Primary outcome was change in preoperative and postoperative pain. Natural history of the lesion was assessed, including age when the lesion was first noticed, when it became painful, and when it required treatment. RESULTS: Fifty-four cases were identified in the study period of which 40 underwent surgery. Pain improved in 36 of 39 patients (92.3%) who had pain before surgery and 29 (74.4%) were pain free after surgery. All 13 patients who required whole muscle excision to resect the lesion experienced an improvement in pain and 10 (76.9%) were pain free. A history of previous intervention, with surgery or sclerotherapy showed a trend towards less successful surgical outcomes. Patients presented across a wide age range from infancy to adulthood, but the most common presentation was pain with exercise between 9 and 16 years of age, with presence of a mass in about half of cases. CONCLUSIONS: Surgery, when performed by a surgeon with appropriate experience, is an effective first-line treatment for painful intramuscular venous malformations, offering pain relief in the majority of cases. Magnetic resonance imaging and ultrasound are diagnostic in most cases. The majority of lesions are resectable, meaning they can be removed with a margin leaving a functional limb. Sometimes resection of a whole muscle is required. LEVEL OF EVIDENCE: Level IV-case series.


Assuntos
Músculo Esquelético/irrigação sanguínea , Escleroterapia , Malformações Vasculares , Procedimentos Cirúrgicos Vasculares , Veias , Adolescente , Criança , Feminino , Humanos , Imageamento por Ressonância Magnética/métodos , Masculino , Dor/etiologia , Dor/cirurgia , Manejo da Dor , Estudos Retrospectivos , Escleroterapia/efeitos adversos , Escleroterapia/métodos , Resultado do Tratamento , Malformações Vasculares/diagnóstico por imagem , Malformações Vasculares/fisiopatologia , Malformações Vasculares/cirurgia , Procedimentos Cirúrgicos Vasculares/efeitos adversos , Procedimentos Cirúrgicos Vasculares/métodos , Veias/anormalidades , Veias/cirurgia
4.
Int Wound J ; 14(6): 967-972, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28251817

RESUMO

Haemangioma of infancy, a benign tumour of blood vessels, is the most common tumour of infancy. Ulceration, the most common complication, presents a unique wound care challenge. A retrospective audit of medical records of children with haemangioma of infancy who presented to the Royal Children's Hospital, Melbourne, Australia, between January 2000 and December 2014 was undertaken with an aim to examine wound management of ulcerated haemangioma of infancy. In total, 535 hospital medical records were identified as suitable, of which 352 were randomly selected and audited, of which 84 patients had ulcerated haemangioma of infancy, and 62 were subject to wound management. Of these, 35 were successfully managed by wound dressings, 9 were not fully healed at the time of last review, and 18 were referred for surgical excision. Patients attended an average of five outpatient visits, and the average time from presentation to documented healing was 105 days. There were a total of 225 episodes of wound dressing, for which there was a documented follow-up appointment at which healing could be assessed. Although a wide range of dressings were used, there was no clear pattern of benefit of one dressing over another. Wounds were less likely to be healed after the use of a silver-impregnated dressing. Pain was poorly documented. Clinical assessment of whether wounds were infected was of no help in planning treatment. There is considerable variability in the management of this difficult wound group, and further prospective studies are required.


Assuntos
Bandagens , Hemangioma/complicações , Cicatrização/fisiologia , Ferimentos e Lesões/etiologia , Ferimentos e Lesões/terapia , Austrália , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Estudos Prospectivos , Estudos Retrospectivos
5.
Angiogenesis ; 17(1): 1-15, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23884796

RESUMO

Human lymphatic vascular malformations (LMs), also known as cystic hygromas or lymphangioma, consist of multiple lymphatic endothelial cell-lined lymph-containing cysts. No animal model of this disease exists. To develop a mouse xenograft model of human LM, CD34(Neg)CD31(Pos) LM lymphatic endothelial cells (LM-LEC) were isolated from surgical specimens and compared to foreskin CD34(Neg)CD31(Pos) lymphatic endothelial cells (LECs). Cells were implanted into a mouse tissue engineering model for 1, 2 and 4 weeks. In vitro LM-LECs showed increased proliferation and survival under starvation conditions (P < 0.0005 at 48 h, two-way ANOVA), increased migration (P < 0.001, two-way ANOVA) and formed fewer (P = 0.029, independent samples t test), shorter tubes (P = 0.029, independent samples t test) than foreskin LECs. In vivo LM-LECs implanted into a Matrigel™-containing mouse chamber model assembled to develop vessels with dilated cystic lumens lined with flat endothelium, morphology similar to that of clinical LMs. Human foreskin LECs failed to survive implantation. In LM-LEC implanted chambers the percent volume of podoplanin(Pos) vessels was 1.18 ± 2.24 % at 1 week, 6.34 ± 2.68 % at 2 weeks and increasing to 7.67 ± 3.60 % at 4 weeks. In conclusion, the significantly increased proliferation, migration, resistance to apoptosis and decreased tubulogenesis of LM-LECs observed in vitro is likely to account for their survival and assembly into stable LM-like structures when implanted into a mouse vascularised chamber model. This in vivo xenograft model will provide the basis of future studies of LM biology and testing of potential pharmacological interventions for patients with lymphatic malformations.


Assuntos
Proliferação de Células , Separação Celular , Células Endoteliais , Sobrevivência de Enxerto , Vasos Linfáticos , Animais , Antígenos CD34/metabolismo , Sobrevivência Celular , Criança , Pré-Escolar , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Células Endoteliais/transplante , Feminino , Xenoenxertos , Humanos , Lactente , Vasos Linfáticos/anormalidades , Vasos Linfáticos/metabolismo , Vasos Linfáticos/patologia , Masculino , Camundongos , Camundongos SCID , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Fatores de Tempo , Engenharia Tecidual/métodos
6.
Artif Organs ; 38(4): 352-61, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23981068

RESUMO

We have designed a laboratory extracorporeal normothermic blood perfusion system for whole organs (e.g., kidney) that achieves pulsatile flow, low levels of hemolysis, and a blood priming volume of 60 mL or less. Using this uniquely designed extracorporeal circuit, we have achieved perfusion of two isolated ex vivo constructs. In the first experiment, we successfully perfused a rabbit epigastric flap based on the femoral vessels. In the second experiment, we were able to perfuse the isolated rabbit kidney for 48 h (range for all kidneys was 12-48 h) with excellent urine output, normal arterial blood gasses at 24 h, and normal ex vivo kidney histology at the conclusion of the experiments. These parameters have not been achieved before with any known or previously published laboratory extracorporeal circuits. The study has implications for prolonged organ perfusion prior to transplantation and for tissue engineering of vascularized tissues, such as by the perfusion of decellularized organs.


Assuntos
Circulação Extracorpórea/instrumentação , Animais , Desenho de Equipamento , Circulação Extracorpórea/métodos , Rim/irrigação sanguínea , Fluxo Pulsátil , Coelhos
8.
J Paediatr Child Health ; 48(10): 902-6, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22897120

RESUMO

INTRODUCTION: Propranolol has recently emerged as an effective drug treatment for infantile haemangiomas. The side effect profile of the drug and the safety of administering propranolol in outpatient settings in this age group remain uncertain. We report our experience with 200 infants and children prescribed propranolol to treat infantile haemangiomas, including 37 patients considered to have a poor response to treatment. METHOD: Patients were prescribed propranolol (1 mg/kg/dose bd) as outpatients at the Vascular Anomalies Service at the Royal Children's Hospital, Melbourne. RESULTS: The median age at commencement was 4 months (range 5 days-7 years). Twenty patients were older than 12 months at commencement. The median duration of treatment was 8 months. About 80% of treated haemangiomas were on the face. Approximately 50% of patients were considered to have an excellent response, 30% to have a good response and 20% to have a poor response. All segmental facial haemangiomas responded well. In contrast, 25% of focal facial haemangiomas responded poorly. Sleep disturbance was the most common side effect. Gross motor abnormalities including delayed walking were observed in 13 patients. CONCLUSION: Propranolol appears to be an effective treatment for infantile haemangiomas, particularly large segmental facial lesions. A poor response was seen in 20% of patients. Treatment has been provided in an outpatient setting without major complications and with excellent parental compliance. The side effect profile appears to be favourable, but further follow-up is required to identify unexpected long-term side effects.


Assuntos
Assistência Ambulatorial , Antineoplásicos/uso terapêutico , Hemangioma/tratamento farmacológico , Propranolol/uso terapêutico , Neoplasias Cutâneas/tratamento farmacológico , Administração Oral , Criança , Pré-Escolar , Esquema de Medicação , Neoplasias Faciais/tratamento farmacológico , Feminino , Seguimentos , Humanos , Lactente , Recém-Nascido , Masculino , Resultado do Tratamento
9.
J Vasc Surg ; 53(2): 435-44, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21030196

RESUMO

OBJECTIVES: Dense angiogenic sprouting occurs from arteriovenous loops (AVLs) incorporating autologous vein grafts inserted into empty plastic chambers in vivo. The purpose of this study was to determine if angiogenesis from the AVL was limited by substituting an "off the shelf" cold-stored allograft vein instead of an autologous vein. METHODS: Four Sprague Dawley rat groups (two AVL configurations × two chamber types) were established for both 2-week and 6-week harvest. Control AVLs were autologous femoral vein grafts harvested from the left femoral vein that were surgically inserted between the cut femoral artery and vein on the right side. Experimental "allograft" AVLs were rat femoral veins cold-stored (4°C, sterile) for 4 to 7 weeks and then microsurgically interposed between the right femoral artery and vein of an unrelated rat. The two AVL types were inserted in one of two plastic chamber types--smooth or perforated. At harvest, the AVL constructs were checked for patency, weighed, their volume determined, and histology undertaken. Morphometric assessment of percent and absolute volume of major tissue components (including blood vessels) at 6 weeks was completed. RESULTS: There were no significant differences between autograft and allograft groups in construct weight, volume, or morphology at 2 or 6 weeks. No statistical differences occurred in the percent or absolute vascular volume of AVLs incorporating a cold-stored allograft vs autologous vein grafts at 6 weeks regardless of the chamber type. However, perforated chambers caused significant increases in construct weight (P = .015), volume (P = .006), and percent and absolute connective tissue volume at 6 weeks (P = .001) compared to smooth chamber constructs, regardless of the graft type. CONCLUSION: Cold-stored small-caliber allografts interposed in AVLs do not inhibit microcirculatory development and can be used in composite tissue engineering.


Assuntos
Bioprótese , Implante de Prótese Vascular/instrumentação , Prótese Vascular , Artéria Femoral/cirurgia , Veia Femoral/transplante , Neovascularização Fisiológica , Engenharia Tecidual , Enxerto Vascular , Animais , Proliferação de Células , Temperatura Baixa , Artéria Femoral/patologia , Artéria Femoral/fisiopatologia , Veia Femoral/patologia , Veia Femoral/fisiopatologia , Masculino , Microcirculação , Desenho de Prótese , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Preservação de Tecido , Transplante Autólogo , Transplante Homólogo , Grau de Desobstrução Vascular
10.
Artigo em Inglês | MEDLINE | ID: mdl-34649968

RESUMO

Nevus sebaceous syndrome (NSS) is a rare, multisystem neurocutaneous disorder, characterized by a congenital nevus, and may include brain malformations such as hemimegalencephaly or focal cortical dysplasia, ocular, and skeletal features. It has been associated with several eponyms including Schimmelpenning and Jadassohn. The isolated skin lesion, nevus sebaceous, is associated with postzygotic variants in HRAS or KRAS in all individuals studied. The RAS proteins encode a family of GTPases that form part of the RAS/MAPK signaling pathway, which is critical for cell cycle regulation and differentiation during development. We studied an individual with nevus sebaceous syndrome with an extensive nevus sebaceous, epilepsy, intellectual disability, and hippocampal sclerosis without pathological evidence of a brain malformation. We used high-depth gene panel sequencing and droplet digital polymerase chain reaction (PCR) to detect and quantify RAS/MAPK gene variants in nevus sebaceous and temporal lobe tissue collected during plastic and epilepsy surgery, respectively. A mosaic KRAS c.34G > T; p.(Gly12Cys) variant, also known as G12C, was detected in nevus sebaceous tissue at 25% variant allele fraction (VAF), at the residue most commonly substituted in KRAS Targeted droplet digital PCR validated the variant and quantified the mosaicism in other tissues. The variant was detected at 33% in temporal lobe tissue but was absent from blood and healthy skin. We provide molecular confirmation of the clinical diagnosis of NSS. Our data extends the histopathological spectrum of KRAS G12C mosaicism beyond nevus sebaceous to involve brain tissue and, more specifically, hippocampal sclerosis.


Assuntos
Nevo , Proteínas Proto-Oncogênicas p21(ras) , Encéfalo , Humanos , Recidiva Local de Neoplasia , Proteínas ras
11.
Sci Rep ; 11(1): 12175, 2021 06 09.
Artigo em Inglês | MEDLINE | ID: mdl-34108542

RESUMO

Craniofacial dysmorphism is associated with thousands of genetic and environmental disorders. Delineation of salient facial characteristics can guide clinicians towards a correct clinical diagnosis and understanding the pathogenesis of the disorder. Abnormal facial shape might require craniofacial surgical intervention, with the restoration of normal shape an important surgical outcome. Facial anthropometric growth curves or standards of single inter-landmark measurements have traditionally supported assessments of normal and abnormal facial shape, for both clinical and research applications. However, these fail to capture the full complexity of facial shape. With the increasing availability of 3D photographs, methods of assessment that take advantage of the rich information contained in such images are needed. In this article we derive and present open-source three-dimensional (3D) growth curves of the human face. These are sequences of age and sex-specific expected 3D facial shapes and statistical models of the variation around the expected shape, derived from 5443 3D images. We demonstrate the use of these growth curves for assessing patients and show that they identify normal and abnormal facial morphology independent from age-specific facial features. 3D growth curves can facilitate use of state-of-the-art 3D facial shape assessment by the broader clinical and biomedical research community. This advance in phenotype description will support clinical diagnosis and the understanding of disease pathogenesis including genotype-phenotype relations.


Assuntos
Anormalidades Múltiplas/patologia , Anormalidades Craniofaciais/patologia , Face/patologia , Imageamento Tridimensional/métodos , Modelos Estatísticos , Atrofia Muscular/patologia , Anormalidades Múltiplas/genética , Anormalidades Múltiplas/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antropometria , Estudos de Casos e Controles , Criança , Pré-Escolar , Anormalidades Craniofaciais/genética , Anormalidades Craniofaciais/metabolismo , Face/anormalidades , Feminino , Seguimentos , Gráficos de Crescimento , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Atrofia Muscular/genética , Atrofia Muscular/metabolismo , Fenótipo , Prognóstico , Adulto Jovem
12.
Plast Reconstr Surg Glob Open ; 8(5): e2847, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-33154878

RESUMO

Three-dimensional (3D) photography is becoming widely used in plastic surgery. It provides an accurate and reproducible record of the facial surface anatomy and could be a versatile tool for treatment planning and assessment. However, the existing software tools available for the assessment of 3D facial imaging often give highly misleading results. The goal of this special topic article is to give clinicians an insight into methods of 3D image assessment and explain the reasons why results may be misleading. We point toward the advantages of an alternative approach using "nonrigid surface registration" for the comparison of pre- and postsurgical images. This approach is compared with the regular rigid surface registration, and this is illustrated by the assessment of a child with Crouzon syndrome before and after LeFort III osteotomy and distraction. Findings of the standard method imply that changes have occurred that are anatomically not possible, whereas the alternative approach indicates realistic changes. Furthermore, we demonstrate an exciting capacity of 3D image analysis to construct reference populations of normal head size and shape. These can be used to assess the parts of the head that are normal and abnormal pre- and posttreatment of the same child. We conclude that, while 3D image analysis has great potential in surgical assessment, existing software does not always give an adequate assessment. Collaboration among surgeons and engineering and computer science specialists should be encouraged. This way, more comprehensive and accurate techniques in patient assessment and surgical planning can be developed and applied in clinical practice.

13.
J Pediatr Surg ; 53(4): 798-801, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28599969

RESUMO

BACKGROUND: Infection in low flow malformations is difficult to diagnose and treat. Initial presentation can be followed by cycles of recurrent infection lasting several years. The optimal duration of antibiotic therapy to prevent recurrence of infection has not been established. METHODS: All cases of infection in low flow malformations at the Royal Children's Hospital over a ten-year period were reviewed. Clinical markers of infection and duration of initial antibiotic treatment were correlated with the development of recurrent episodes of infection. RESULTS: Twenty-one patients met criteria for inclusion. Nineteen were diagnosed as lymphatic malformations and two as venous malformations. The majority of patients (13 or 62%) received a prolonged course of six weeks or more of antibiotics. Eleven (52%) patients went on to have recurrent infections, but these were significantly less likely to be in those treated with a long course of antibiotics (Fisher's exact test, p=0.026). In only 12 of 21 cases could a bacterium be grown. Elevated CRP was the most consistent abnormal laboratory finding in infection. CONCLUSIONS: Longer courses of antibiotics reduce the risk of recurrent infection in low-flow vascular malformations. We recommend an antibiotic course of three months or more at the initial presentation of infection in a low flow malformation. Elevated CRP is the most sensitive test for diagnosis of infection in low-flow malformations. TYPE OF STUDY: Treatment study. LEVEL OF EVIDENCE: III.


Assuntos
Antibacterianos/administração & dosagem , Infecções Cardiovasculares/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Positivas/tratamento farmacológico , Anormalidades Linfáticas/complicações , Malformações Vasculares/complicações , Adolescente , Antibacterianos/uso terapêutico , Infecções Cardiovasculares/etiologia , Infecções Cardiovasculares/prevenção & controle , Criança , Pré-Escolar , Esquema de Medicação , Feminino , Seguimentos , Infecções por Bactérias Gram-Negativas/etiologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Infecções por Bactérias Gram-Positivas/etiologia , Infecções por Bactérias Gram-Positivas/prevenção & controle , Humanos , Lactente , Recém-Nascido , Masculino , Recidiva , Prevenção Secundária
14.
Sci Rep ; 8(1): 4771, 2018 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-29556038

RESUMO

Many disorders present with characteristic abnormalities of the craniofacial complex. Precise descriptions of how and when these abnormalities emerge and change during childhood and adolescence can inform our understanding of their underlying pathology and facilitate diagnosis from craniofacial shape. In this paper we develop a framework for analysing how anatomical differences between populations emerge and change over time, and for binary group classification that adapts to the age of each participant. As a proxy for a disease-control comparison we use a database of 3D photographs of normally developing boys and girls to examine emerging sex-differences. Essentially we define 3D craniofacial 'growth curves' for each sex. Differences in the forehead, upper lip, chin and nose emerge primarily from different growth rates between the groups, whereas differences in the buccal region involve different growth directions. Differences in the forehead, buccal region and chin are evident before puberty, challenging the view that sex differences result from pubertal hormone levels. Classification accuracy was best for older children. This paper represents a significant methodological advance for the study of facial differences between growing populations and comprehensively describes developing craniofacial sex differences.


Assuntos
Desenvolvimento Maxilofacial , Caracteres Sexuais , Crânio/crescimento & desenvolvimento , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Modelos Estatísticos , Análise de Regressão
15.
Tissue Eng ; 13(4): 673-81, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17335401

RESUMO

A novel method of spontaneous generation of new adipose tissue from an existing fat flap is described. A defined volume of fat flap based on the superficial inferior epigastric vascular pedicle in the rat was elevated and inset into a hollow plastic chamber implanted subcutaneously in the groin of the rat. The chamber walls were either perforated or solid and the chambers either contained poly(D,L-lactic-co-glycolic acid) (PLGA) sponge matrix or not. The contents were analyzed after being in situ for 6 weeks. The total volume of the flap tissue in all groups except the control groups, where the flap was not inserted into the chambers, increased significantly, especially in the perforated chambers (0.08 +/- 0.007 mL baseline compared to 1.2 +/- 0.08 mL in the intact ones). Volume analysis of individual component tissues within the flaps revealed that the adipocyte volume increased and was at a maximum in the chambers without PLGA, where it expanded from 0.04 +/- 0.003 mL at insertion to 0.5 +/- 0.08 mL (1250% increase) in the perforated chambers and to 0.16 +/- 0.03 mL (400% increase) in the intact chambers. Addition of PLGA scaffolds resulted in less fat growth. Histomorphometric analysis rather than simple hypertrophy documented an increased number of adipocytes. The new tissue was highly vascularized and no fat necrosis or atypical changes were observed.


Assuntos
Tecido Adiposo/anatomia & histologia , Tecido Adiposo/crescimento & desenvolvimento , Retalhos Cirúrgicos/patologia , Retalhos Cirúrgicos/fisiologia , Engenharia Tecidual/instrumentação , Engenharia Tecidual/métodos , Tecido Adiposo/irrigação sanguínea , Animais , Desenho de Equipamento , Análise de Falha de Equipamento , Masculino , Tamanho do Órgão , Ratos , Ratos Sprague-Dawley , Técnicas de Cultura de Tecidos/instrumentação , Técnicas de Cultura de Tecidos/métodos
16.
ANZ J Surg ; 77(3): 118-20, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17305981

RESUMO

BACKGROUND: Obesity is recognized as a risk factor for wound healing problems in surgery, but the reason for this increased risk is unclear. It is now recognized that truncal obesity, as measured by either the waist circumference or the waist-hip ratio (WHR), is a strong risk factor for an array of metabolic problems termed 'metabolic syndrome'. It was suggested that increased WHR would be a sensitive marker of obesity-related wound healing impairment and therefore would predict skin graft failure. METHOD: Consecutive patients undergoing full-thickness skin graft of the head and neck region in a private practice setting were enrolled prospectively. The body mass index and WHR were recorded at the time of surgery and skin graft take was assessed at 1-2 weeks. RESULTS: There was a strong correlation between increased WHR and graft failure; all four patients in the study having a WHR greater than 1 showed partial or total graft loss compared with only one patient having a 10% loss in the remainder (r = -0.76, P = 0.002). CONCLUSION: There is a strong correlation between truncal obesity as measured by the WHR and skin graft failure. WHR or waist circumference should be considered as a preoperative assessment tool in identifying patients at risk of surgical complications.


Assuntos
Síndrome Metabólica/diagnóstico , Transplante de Pele , Relação Cintura-Quadril , Cicatrização , Adulto , Idoso , Idoso de 80 Anos ou mais , Humanos , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Prospectivos , Falha de Tratamento
17.
Cell Transplant ; 15(4): 319-24, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16898225

RESUMO

As in vivo tissue engineering of complex tissues and organs progresses, there is a need for an independently vascularized, alterable, and recoverable model. Current models of islet cell transplantation (release into the portal venous system, placement under the renal capsule, and microencapsulation) lack these qualities. We have developed a model of angiogenesis and spontaneous tissue generation in the rat that lends itself as a potential platform for tissue engineering. In this experiment, we examined the effectiveness of such a model in addressing some of the shortcomings of endocrine pancreatic transplantation. An arteriovenous loop was created in the groins of five adult inbred Sprague-Dawley rats, and placed within polycarbonate chambers. Isolated pancreatic islet cell clusters were placed within the chambers, suspended in a matrix of Matrigel. The chambers were recovered at 3 weeks, and the newly generated tissue was processed for histologic and immunohistochemical analysis. By 3 weeks, spontaneous generation of angiogenesis and collagen matrix and deposition of a collagen matrix was observed. Surviving islet cells were identified by histology and their viability was confirmed via immunohistochemistry for insulin and glucagon. This study demonstrates the ability to maintain viability and functionality of transplanted islet cells on a tissue-engineered platform with an independent vascular supply. The model provides the ability to alter the graft environment via matrix substitution, cellular coculture, and administration of growth factors. The transplanted tissues are recoverable without animal sacrifice and are microsurgically transferable. This model may provide an in vivo culture platform for the study of islet transplantation.


Assuntos
Diabetes Mellitus Experimental/cirurgia , Transplante das Ilhotas Pancreáticas/métodos , Ilhotas Pancreáticas/fisiologia , Engenharia Tecidual/métodos , Animais , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/fisiopatologia , Glucagon/metabolismo , Sobrevivência de Enxerto/fisiologia , Insulina/metabolismo , Ilhotas Pancreáticas/irrigação sanguínea , Ilhotas Pancreáticas/citologia , Masculino , Neovascularização Fisiológica/fisiologia , Projetos Piloto , Ratos , Ratos Sprague-Dawley , Transplante Homólogo
18.
Stem Cell Reports ; 7(3): 518-526, 2016 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-27594589

RESUMO

The ability to reliably express fluorescent reporters or other genes of interest is important for using human pluripotent stem cells (hPSCs) as a platform for investigating cell fates and gene function. We describe a simple expression system, designated GAPTrap (GT), in which reporter genes, including GFP, mCherry, mTagBFP2, luc2, Gluc, and lacZ are inserted into the GAPDH locus in hPSCs. Independent clones harboring variations of the GT vectors expressed remarkably consistent levels of the reporter gene. Differentiation experiments showed that reporter expression was reliably maintained in hematopoietic cells, cardiac mesoderm, definitive endoderm, and ventral midbrain dopaminergic neurons. Similarly, analysis of teratomas derived from GT-lacZ hPSCs showed that ß-galactosidase expression was maintained in a spectrum of cell types representing derivatives of the three germ layers. Thus, the GAPTrap vectors represent a robust and straightforward tagging system that enables indelible labeling of PSCs and their differentiated derivatives.


Assuntos
Expressão Gênica , Genes Reporter , Vetores Genéticos/genética , Células-Tronco Pluripotentes/metabolismo , Transgenes , Sistemas CRISPR-Cas , Diferenciação Celular , Linhagem Celular , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Humanos , Células-Tronco Pluripotentes/citologia , Nucleases dos Efetores Semelhantes a Ativadores de Transcrição
19.
J Vis Exp ; (99): e52691, 2015 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-25992474

RESUMO

Lymphatic system disorders such as primary lymphedema, lymphatic malformations and lymphatic tumors are rare conditions that cause significant morbidity but little is known about their biology. Isolating highly pure human lymphatic endothelial cells (LECs) from diseased and healthy tissue would facilitate studies of the lymphatic endothelium at genetic, molecular and cellular levels. It is anticipated that these investigations may reveal targets for new therapies that may change the clinical management of these conditions. A protocol describing the isolation of human foreskin LECs and lymphatic malformation lymphatic endothelial cells (LM LECs) is presented. To obtain a single cell suspension tissue was minced and enzymatically treated using dispase II and collagenase II. The resulting single cell suspension was then labelled with antibodies to cluster of differentiation (CD) markers CD34, CD31, Vascular Endothelial Growth Factor-3 (VEGFR-3) and PODOPLANIN. Stained viable cells were sorted on a fluorescently activated cell sorter (FACS) to separate the CD34(Low)CD31(Pos)VEGFR-3(Pos)PODOPLANIN(Pos) LM LEC population from other endothelial and non-endothelial cells. The sorted LM LECs were cultured and expanded on fibronectin-coated flasks for further experimental use.


Assuntos
Células Endoteliais/citologia , Citometria de Fluxo/métodos , Humanos , Linfadenite/patologia , Linfedema/patologia , Metaloproteinase 8 da Matriz/química
20.
Plast Reconstr Surg ; 113(1): 260-9, 2004 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-14707645

RESUMO

The authors previously described a model of tissue engineering in rats that involves the insertion of a vascular pedicle and matrix material into a semirigid closed chamber, which is buried subcutaneously. The purpose of this study was to develop a comparable model in mice, which could enable genetic mutants to be used to more extensively study the mechanisms of the angiogenesis, matrix production, and cellular migration and differentiation that occur in these models. A model that involves placing a split silicone tube around blood vessels in the mouse groin was developed and was demonstrated to successfully induce the formation of new vascularized tissue. Two vessel configurations, namely, a flow-through pedicle (n = 18 for three time points) and a ligated vascular pedicle (n = 18), were compared. The suitability of chambers constructed from either polycarbonate or silicone and the effects of incorporating either Matrigel equivalent (n = 18) or poly(DL-lactic-co-glycolic acid) (n = 18) on angiogenesis and tissue production were also tested. Empty chambers, chambers with vessels only, and chambers with matrix only served as control chambers. The results demonstrated that a flow-through type of vascular pedicle, rather than a ligated pedicle, was more reliable in terms of patency, angiogenesis, and tissue production, as were silicone chambers, compared with polycarbonate chambers. Marked angiogenesis occurred with both types of extracellular matrix scaffolds, and there was evidence that native cells could migrate into and survive within the added matrix, generating a vascularized three-dimensional construct. When Matrigel was used as the matrix, the chambers filled with adipose tissue, creating a highly vascularized fat flap. In some cases, new breast-like acini and duct tissue appeared within the fat. When poly(dl-lactic-co-glycolic acid) was used, the chambers filled with granulation and fibrous tissue but no fat or breast tissue was observed. No significant amount of tissue was generated in the control chambers. Operative times were short (25 minutes), and two chambers could be inserted into each mouse. In summary, the authors have developed an in vivo murine model for studying angiogenesis and tissue-engineering applications that is technically simple and quick to establish, has a high patency rate, and is well tolerated by the animals.


Assuntos
Modelos Animais , Retalhos Cirúrgicos/irrigação sanguínea , Engenharia Tecidual/métodos , Animais , Materiais Biocompatíveis , Colágeno , Combinação de Medicamentos , Artéria Femoral , Veia Femoral , Ácido Láctico , Laminina , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neovascularização Fisiológica , Ácido Poliglicólico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Polímeros , Proteoglicanas , Grau de Desobstrução Vascular
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