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BACKGROUND: The use of progressive tension sutures (PTSs) for drain-free abdominoplasty closure has been well published in the esthetic literature and found to be effective at reducing abdominal drainage without a worse complication profile. This research aims to evaluate the outcomes of incorporating PTS into donor-site closures for abdominal-based breast reconstruction. METHODS: A retrospective chart review was performed evaluating patients at the University of Chicago Medicine, who underwent autologous breast reconstruction and either PTS closure or standard abdominal closure between 2018 and 2021. RESULTS: A total of 100 patients were included with 50 patients receiving PTS closure and 50 with traditional abdominal closures. Patient demographics, including age (p = 0.82), body mass index (BMI; p = 0.17), diabetes (p = 1.00), tobacco use (p = 0.15), and chemotherapy (p = 1.00) did not significantly differ. Total drain output over the first 72 h was significantly lower in the PTS group compared with the standard closure (SC) group (p = 0.00005). Mean duration of drain placement was shorter by 2 days in the PTS group, but this did not reach significance (p = 0.08). Overall complication rates were lower in the PTS group (p = 0.03), however, no difference was appreciated when separately assessing for seromas (p = 1.00), hematomas (p = 1.00), wound dehiscence (p = 0.58), or surgical-site infections (p = 1.00). More abdominal revisions were observed in the SC group, however, this did not reach significance (p = 0.15) CONCLUSION: The use of PTS with placement of a single drain is a safe option for donor-site closure, and is a compelling technique for patients undergoing abdominal-based breast reconstruction.
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Abdominoplastia , Mamoplastia , Abdominoplastia/métodos , Humanos , Mamoplastia/efeitos adversos , Mamoplastia/métodos , Complicações Pós-Operatórias , Estudos Retrospectivos , Seroma/etiologia , Técnicas de SuturaRESUMO
BACKGROUND: Decreasing length of stay benefits patients and hospital systems alike. This should be accomplished safely without negatively impacting patient outcomes. The authors hypothesize that in the United States, the average length of stay for patients undergoing microsurgical breast reconstruction has decreased since 2012 without a concurrent increase in complication and readmission rates. METHODS: The authors identified female patients who underwent microvascular breast reconstruction (CPT 19364) from the 2012 to 2018 National Surgical Quality Improvement Program database. Trends in complication and readmission rates and length of stay were examined over 7 years. Multivariable logistic regression models and Mann-Kendall trend tests were used to evaluate associations between length of stay and complication and readmission rates. RESULTS: A total of 10,465 cases were identified. The number of autologous microvascular breast reconstruction procedures performed increased annually between 2012 and 2018. Length of stay decreased significantly from 2012 to 2018 (from 4.47 days to 3.90 days) (p < 0.01). Minor and major complication rates remained constant, although major complications showed a decreasing trend (from 27 percent to 21 percent) (p = 0.07). Thirty-day readmission, surgical-site infection, and wound dehiscence rates remained consistent over the study period, whereas rates of blood transfusion or bleeding decreased (p = 0.02). CONCLUSIONS: Using a national sample from 2012 to 2018, the authors observed a significant decrease in length of stay for patients undergoing microvascular breast reconstruction without a concurrent increase in complication and readmission rates. Current efforts to reduce length of stay have been successful without increasing complication or readmission rates. CLINICAL QUESTION/LEVEL OF EVIDENCE: Risk, III.
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Tempo de Internação/tendências , Mamoplastia/métodos , Microvasos/cirurgia , Adulto , Bases de Dados Factuais , Feminino , Humanos , Pessoa de Meia-Idade , Melhoria de Qualidade , Estados Unidos , Procedimentos Cirúrgicos Vasculares/métodosRESUMO
PURPOSE: Cessation of elective surgery during COVID-19 was partly driven by concern for consumption of hospital resources required by critically ill patients. We aim to determine the extent of resource utilization by elective outpatient surgery to assist in ensuring future resource conservation decisions are data driven. METHODS: The study utilized a retrospective cohort gathered from the American College of Surgeons National Surgical Quality Improvement Program database. Participants were adult patients who underwent elective or non-elective surgery between 2017 and 2018. Outcomes included patient characteristics and post-operative outcomes for elective and non-elective surgeries. Post-operative outcomes were used as a surrogate for the consumption of hospital resources. RESULTS: A total of 1,558,938 (79.8%) elective and 393,339 (20.2%) non-elective surgeries were identified. Elective surgery patients were more likely to be outpatient status, have an ASA class < 3, and exhibited lower rates of prolonged ventilation, 30-day reoperation, and 30-day readmissions, and averaged 5 days less of inpatient stay. Elective outpatient surgery (vs. elective inpatient surgery) averaged shorter operative times and exhibited lower rates of readmissions (2.1% vs. 5.5%; p < 0.001), reoperations (1.1% vs. 2.8%; p < 0.001), prolonged ventilation (0.0% vs. 0.3%; p < 0.001), and 30-day mortality (0.1% vs. 0.5%; p < 0.001) and accounted for 30.2% of the overall relative value units ($339,815,038). CONCLUSION: We evaluated utilization of hospital resources by patients undergoing elective outpatient surgery by identifying surgeries performed in 2017-2018 then stratifying them by outpatient status. Elective outpatient surgeries consumed negligible amounts of hospital resources and should not be considered a threat to resources in the setting of high demand by critically ill COVID-19 patients.
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COVID-19 , Complicações Pós-Operatórias , Adulto , COVID-19/epidemiologia , Procedimentos Cirúrgicos Eletivos , Humanos , Tempo de Internação , Readmissão do Paciente , Complicações Pós-Operatórias/epidemiologia , Estudos RetrospectivosRESUMO
BACKGROUND: The rates, severity and consequences of hand and wrist injuries sustained by National Collegiate Athletic Association athletes are not well characterized. This study describes the epidemiology of hand and wrist injuries among collegiate athletes competing in different divisions. METHODS: The National Collegiate Athletic Association Injury Surveillance Program (NCAA-ISP) was accessed from 2004 to 2015 for the following sports: baseball, basketball, football, ice hockey, lacrosse, soccer, wrestling, field hockey, gymnastics, softball and volleyball. The data were used to identify all hand and wrist injuries, the specific injury diagnosis, mean time loss of activity following injury, and need for surgery following injury. These were then stratified by gender. Descriptive statistics were performed to examine the association between sports, event type and division. Student's t test was used to calculate p-values for independent variables. Chi-Square test was used to calculate odds ratio. P < 0.05 was considered significant. RESULTS: 103,098 hand and wrist injuries were reported in in the studied NCAA sports from 2004 to 2015. Male athletes sustained 72,423 injuries (6.01/10,000 athlete exposure) and female athletes sustained 30,675 injuries (4.13/10,000 athlete exposure). Division I athletes sustained significantly more injuries compared to divisions II and III. Overall, 3.78% of hand and wrist injuries required surgical intervention. A significantly higher percentage of division I athletes (both male and female) underwent surgical intervention compared to divisions II and III. The mean time lost due to hand and wrist injury was 7.14 days for all athletes. Division I athletes missed the fewest days due to injury at 6.29 days though this was not significant. CONCLUSIONS: Hand and wrist injuries are common among collegiate athletes. Division I athletes sustain higher rates of injuries and higher surgical intervention rates, while tending to miss fewer days due to injury. Improved characterization of divisional differences in hand and wrist injuries can assist injury management and prevention.
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BACKGROUND: In a surgical field, where surgeons are, "sometimes wrong, but never in doubt," lack of confidence can have detrimental effects on career advancement. In other fields there is evidence that a gap exists between women and men in the amount of confidence they display, and that confidence is a proxy for success. METHODS: This study used the General Self Efficacy Scale and Rosenberg Self-Esteem Scale confidence surveys to assess self confidence amongst female trainees and attending plastic surgeons, to search for baseline characteristics associated with higher confidence scores. RESULTS: Of the 73 participants, protective factors associated with increased female plastic surgeon confidence include age, parity, more advanced academic status, and mentorship. CONCLUSIONS: In order to matriculate into a surgical training program, there must be a measure of confidence and resiliency, but further work needs to be done to identify and address gender gaps in training and early academic careers.
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Equidade de Gênero , Médicas/psicologia , Autoimagem , Cirurgiões/psicologia , Cirurgia Plástica , Logro , Adulto , Escolha da Profissão , Mobilidade Ocupacional , Estudos Transversais , Feminino , Humanos , Internato e Residência , Masculino , Mentores/psicologia , Pessoa de Meia-Idade , Testes Psicológicos , Autoeficácia , Fatores Sexuais , Cirurgiões/educação , Cirurgia Plástica/educação , Inquéritos e Questionários , Estados UnidosRESUMO
Robust, reliable, and reproducible closure of lumbosacral myelomeningocele defects remains a challenge. In infants with lumbosacral myelomeningocele defects, multiple methods of soft tissue coverage have been described. These include various cutaneous, fascial, and muscle flaps and grafts. This is done with relative ease when ample soft tissue is present but becomes extremely difficult for large and distally located defects. We present here our closure technique of lumbosacral myelomeningocele defects in newborns, with associated short- and medium-term outcomes. We demonstrate the anatomy of this technique with fresh cadaver dissection and present a review of demographic and outcome data of 12 consecutive patients treated with this method from June 2014 to August 2019. No major intra- or postoperative complications have been encountered, with a mean follow-up of 22.2 months and median follow up of 18 months. After the neurosurgical repair of lumbosacral myelomeningocele, bilateral composite fascial flaps composed of thoracolumbar and gluteus maximus fascia are elevated in continuity. The paraspinous muscle flaps are then elevated, disinserted distally, and medialized to provide complete muscular coverage of the dural repair. The bilateral composite fascial flaps are medialized and closed over the deep paraspinous muscle flap repair. Two patients experienced areas of small, superficial skin necrosis, one of which healed by secondary intention and the other by debridement and full-thickness skin grafting. Use of bilateral paraspinous muscle flaps and bilateral composite fascial flaps composed of thoracolumbar and gluteus maximus fascia provides robust coverage of lumbosacral defects following myelomeningocele repair in infants.
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INTRODUCTION: Expanding surgical capacity is very difficult in resource-constrained countries. Financial input and additional physical operating room space are needed. The surgical patient volume exists, but the lack of operating room time causes postponement of cases. Hand surgery is particularly important as it improves patient function and allows for a timely return to the workforce after injury. Some hand surgery cases may be performed under local anesthesia with a very basic instrument set in a procedure room. This arrangement eliminates the need and financial burden of an equipped operating room and the need for an anesthesiologist. We hypothesized that performing hand surgery in a simple procedure room by a surgeon with knowledge of adequate local anesthesia could increase hand surgery capacity significantly in a low-income country. METHODS: This technique has been instituted at Komfo Anokye Teaching Hospital in Kumasi, Ghana, with the use of a single procedure room that was previously used for storage. A surgeon trained in performing wide-awake local anesthesia no tourniquet technique visited Komfo Anokye Teaching Hospital and provided lectures about hand surgery under local anesthesia and evidence-based sterility for 6 years. The number and type of cases performed in the procedure room were recorded for the first 11 months after it opened in 2017. RESULTS: For 11 months, use of this room had increased surgical capacity by 33 cases per month. Patient ages ranged from 2 months to 65 years. There have been 358 total cases performed, 240 of which were hand cases. This included washout of hand wounds (n = 87), tendon repair (n = 54) including a single tendon transfer, fracture pinning (n = 33), amputations (n = 24), trigger finger repair (n = 10), nerve repair (n = 6), congenital hand surgery (n = 4), and other (n = 22). Cost savings per case in the procedure room ranges from 500 cedis (US $100) to 2000 cedis (US $400). CONCLUSIONS: The simple procedure room runs more efficiently and is less costly compared with the main operating rooms. Although the case volume has increased surgical capacity significantly, costs to the hospital and patient have decreased. The hospital is reimbursed in a timely fashion for the procedures directly by the patient using this technique. The creation of a single procedure room for wide-awake local anesthesia no tourniquet hand surgery has helped address the issues of inadequate operating room space, time, and expense in resource-constrained Ghana.
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Anestesia Local , Anestésicos Locais , Gana , Mãos/cirurgia , Hospitais , Humanos , Lactente , Salas Cirúrgicas , Pacientes AmbulatoriaisRESUMO
BACKGROUND: Bilateral mastectomy rates are increasing in the United States. The abdomen is the most common harvest site for autologous reconstruction. Nationwide data were examined to determine differences in hospital charges, length of stay (LOS), and early postoperative complications following immediate bilateral pedicled transverse rectus abdominis myocutaneous (pTRAM), free TRAM (fTRAM), deep inferior epigastric perforator (DIEP), and superficial inferior epigastric artery (SIEA) perforator flaps and were compared with unilateral reconstruction. METHODS: Patients who underwent immediate bilateral breast reconstruction using a single method of abdominally based reconstruction were identified using the 2009 to 2014 Nationwide Inpatient Sample Database. Outcomes included total hospital charges, LOS, and immediate postoperative complications. RESULTS: We identified 13,348 cases of bilateral mastectomy with a single type of immediate bilateral autologous flap reconstruction. The majority were bilateral DIEP flaps. Mean total cost for bilateral pTRAM, fTRAM, DIEP, and SIEA flaps was US $21,886.80, US $28,839.40, US $30,051.30, and US $33,784.90, respectively (p < 0.0001). Mean LOS for bilateral pTRAM, fTRAM, DIEP, and SIEA was 4.3, 4.9, 4.5, and 5.4 days, respectively (p = 0.0002), and hematoma rates were 1.93, 2.61, 3.68, and 16.59%, respectively, (p = 0.0001), whereas return to the operating room for vascular anastomosis revision was 0, 1.63, 1.99, and 19.07%, respectively (p < 0.0001). Cost is less for unilateral pTRAM, fTRAM, and DIEP flaps (p < 0.0001). LOS is shorter for unilateral fTRAM versus bilateral (p < 0.0001). No differences were appreciated between unilateral and bilateral hematoma and reoperation rates for any reconstruction (p > 0.1). CONCLUSION: Immediate complication rates were higher in bilateral free flaps compared with bilateral pedicled flaps. pTRAM and fTRAM flap reconstructions are still performed frequently with acceptable immediate results without considering long-term morbidity, aesthetics, and abdominal muscle function. Bilateral SIEA free flaps were associated with significantly higher total cost, LOS, and complication rates compared with other groups. Complications were similar between unilateral and bilateral reconstruction procedures. While cost is significantly greater for bilateral procedures compared with unilateral pTRAM, fTRAM, and DIEP flaps, it is not doubled.
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Neoplasias da Mama/cirurgia , Mamoplastia/métodos , Retalho Miocutâneo/transplante , Reto do Abdome/transplante , Estudos Transversais , Feminino , Preços Hospitalares/estatística & dados numéricos , Humanos , Tempo de Internação/estatística & dados numéricos , Mastectomia , Pessoa de Meia-Idade , Complicações Pós-Operatórias , Estudos Retrospectivos , Transplante Autólogo , Estados UnidosRESUMO
BACKGROUND: Much of the morbidity associated with short bowel syndrome (SBS) is attributed to effects of decreased enteral nutrition and administration of total parenteral nutrition (TPN). We hypothesized that acute SBS alone has significant effects on gene expression beyond epithelial proliferation, and tested this in a zebrafish SBS model. METHODS: In a model of SBS in zebrafish (laparotomy, proximal stoma, distal ligation, n = 29) or sham (laparotomy alone, n = 28) surgery, RNA-Seq was performed after 2 weeks. The proximal intestine was harvested and RNA isolated. The three samples from each group with the highest amount of RNA were spiked with external RNA controls consortium (ERCC) controls, sequenced and aligned to reference genome with gene ontology (GO) enrichment analysis performed. Gene expression of ctnnb1, ccnb1, ccnd1, cyp7a1a, dkk3, ifng1-2, igf2a, il1b, lef1, nos2b, saa1, stat3, tnfa and wnt5a were confirmed to be elevated in SBS by RT-qPCR. RESULTS: RNA-seq analysis identified 1346 significantly upregulated genes and 678 significantly downregulated genes in SBS zebrafish intestine compared to sham with Ingenuity analysis. The upregulated genes were involved in cell proliferation, acute phase response signaling, innate and adaptive immunity, bile acid regulation, production of nitric oxide and reactive oxygen species, cellular barrier and coagulation. The downregulated genes were involved in folate synthesis, gluconeogenesis, glycogenolysis, fatty-acid oxidation and activation and drug and steroid metabolism. RT-qPCR confirmed gene expression differences from RNA-Sequencing. CONCLUSION: Changes of gene expression after 2 weeks of SBS indicate complex and extensive alterations of multiple pathways, some previously implicated as effects of TPN. The systemic sequelae of SBS alone are significant and indicate multiple targets for investigating future therapies.
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Ácidos e Sais Biliares/metabolismo , Expressão Gênica , Sistema Imunitário/imunologia , Sistema Imunitário/metabolismo , Síndrome do Intestino Curto/etiologia , Síndrome do Intestino Curto/metabolismo , Animais , Proliferação de Células , Análise por Conglomerados , Modelos Animais de Doenças , Perfilação da Expressão Gênica , Inflamação/genética , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/patologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Análise de Sequência de RNA , Síndrome do Intestino Curto/patologia , Peixe-ZebraRESUMO
BACKGROUND: Signaling by fibroblast growth factor is critical for epithelial proliferation, differentiation, and the development of many organs, including the intestine. Fibroblast growth factor 10 and fibroblast growth factor 2c are upregulated after massive bowel resection during intestinal adaptation. This pathway is conserved highly. We hypothesized that inhibition of fibroblast growth factor signaling would impair intestinal adaptation in the zebrafish model of short bowel syndrome and allow insight into the negative regulation of this pathway. METHODS: Short bowel syndrome equivalent to a high jejunostomy was generated in adult male hsp70:dnfgfr1-GFP zebrafish, wildtype fish exposed to tyrosine-kinase inhibitor, and wildtype fish in absence of tyrosine-kinase inhibitor. Heat shock in hsp70:dnfgfr1-GFP fish decreases fgf 1 expression. Parameters including weight, proliferation, and differentiation were evaluated after harvest in experimental and control groups. RESULTS: Although short bowel syndrome zebrafish lost more weight relative to sham zebrafish in both groups, heat shock fish with short bowel syndrome lost more weight compared with non-heat shock fish with short bowel syndrome. In the non-heat shock controls, the villus epithelial perimeter increased in short bowel syndrome compared with sham fish, but this did not occur in heat shock fish. Non-heat shock fish with short bowel syndrome fish had significantly increased Bromodeoxyuridine(+) proliferative cells per hemivillus compared with non-heat shock-sham, while heat shock-short bowel syndrome had a more substantial increase in Bromodeoxyuridine(+) cells compared with HS-sham. Non-heat shock-short bowel syndrome demonstrated a significantly increased percentage of Alcian blue(+) goblet cells per hemivillus compared with non-heat shock-sham, while the heat shock-short bowel syndrome demonstrated decreased Alcian blue(+) cells compared with non-heat shock-short bowel syndrome. In contrast, SU5402 inhibited epithelial proliferation while increasing weight loss. CONCLUSION: Inhibition of fibroblast growth factor-1 signaling in short bowel syndrome decreases epithelial adaptation, increases Bromodeoxyuridine-labeled cells at 2 weeks, and exacerbates weight loss while decreasing epithelial goblet cells.
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Proliferação de Células/fisiologia , Enterócitos/fisiologia , Fator 1 de Crescimento de Fibroblastos/fisiologia , Síndrome do Intestino Curto/patologia , Redução de Peso/fisiologia , Animais , Modelos Animais de Doenças , Intestino Delgado/metabolismo , Intestino Delgado/patologia , Masculino , Síndrome do Intestino Curto/etiologia , Síndrome do Intestino Curto/metabolismo , Transdução de Sinais/fisiologia , Peixe-ZebraRESUMO
Background: A modified Appleby procedure for pancreatic body tumors relies upon collateral vessels maintaining blood flow to the proper hepatic artery (PHA) through the pancreaticoduodenal arcade (PDA) off of the superior mesenteric artery (SMA). Compression of the celiac axis by the median arcuate ligament (MAL) promotes the expansion of collateral vessels without preoperative intervention. Case Presentation: A 51-year-old male with asymptomatic compression of the celiac artery presented with new onset insulin-dependent diabetes mellitus. He underwent imaging that demonstrated a locally advanced pancreatic body tumor that encased the superior mesenteric vein and portal vein confluence and involved the common hepatic artery. He had an adequate response to neoadjuvant FOLFIRINOX chemotherapy and underwent an uncomplicated modified Appleby procedure with a margin negative resection. Hepatic blood flow was adequate through the PHA as a result of collateralization of blood flow through the PDA off the SMA. The enhanced collateralization appeared to have occurred secondary to compression of the celiac axis by the MAL. Conclusions: Herein we present a unique case in which improved collateral blood flow through the PDA and the gastroduodenal artery to the PHA occurred due to celiac artery compression by the MAL. This vascular anomaly fortuitously improved the ability to achieve an R0 resection of a locally advanced pancreatic adenocarcinoma of the body of the pancreas by a modified Appleby procedure.
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BACKGROUND: In short bowel syndrome, luminal factors influence adaptation in which the truncated intestine increases villus lengths and crypt depths to increase nutrient absorption. No study has evaluated the effect of adaptation within the distal intestine after intestinal separation. We evaluated multiple conditions, including Igf1r inhibition, in proximal and distal segments after intestinal resection to evaluate the epithelial effects of the absence of mechanoluminal stimulation. METHODS: Short bowel syndrome was created in adult male zebrafish by performing a proximal stoma with ligation of the distal intestine. These zebrafish with short bowel syndrome were compared to sham-operated zebrafish. Groups were treated with the Igf1r inhibitor NVP-AEW541, DMSO, a vehicle control, or water for 2 weeks. Proximal and distal intestine were analyzed by hematoxylin and eosin for villus epithelial circumference, inner epithelial perimeter, and circumference. We evaluated BrdU+ cells, including costaining for ß-catenin, and the microbiome was evaluated for changes. Reverse transcription quantitative polymerase chain reaction was performed for ß-catenin, CyclinD1, Sox9a, Sox9b, and c-Myc. RESULTS: Proximal intestine demonstrated significantly increased adaptation compared to sham-operated proximal intestine, whereas the distal intestine showed no adaptation in the absence of luminal flow. Addition of the Igf1r inhibitor resulted in decreased adaption in the distal intestine but an increase in distal proliferative cells and proximal ß-catenin expression. While some proximal proliferative cells in short bowel syndrome colocalized ß-catenin and BrdU, the distal proliferative cells did not co-stain for ß-catenin. Sox9a increased in the distal limb after division but not after inhibition with the Igf1r inhibitor. There was no difference in alpha diversity or species richness of the microbiome between all groups. CONCLUSION: Luminal flow in conjunction with short bowel syndrome significantly increases intestinal adaption within the proximal intestine in which proliferative cells contain ß-catenin. Addition of an Igf1r inhibitor decreases adaptation in both proximal and distal limbs while increasing distal proliferative cells that do not colocalize ß-catenin. Igf1r inhibition abrogates the increase in distal Sox9a expression that otherwise occurs in short bowel syndrome. Mechanoluminal flow is an important stimulus for intestinal adaptation.
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Intestino Delgado/efeitos dos fármacos , Intestino Delgado/cirurgia , Pirimidinas/antagonistas & inibidores , Pirróis/antagonistas & inibidores , Síndrome do Intestino Curto/patologia , Adaptação Fisiológica/efeitos dos fármacos , Animais , Biomarcadores/metabolismo , Biópsia por Agulha , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Imuno-Histoquímica , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Masculino , Pirimidinas/farmacologia , Pirróis/farmacologia , Distribuição Aleatória , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Sensibilidade e Especificidade , Síndrome do Intestino Curto/tratamento farmacológico , Síndrome do Intestino Curto/cirurgia , Peixe-Zebra , beta Catenina/metabolismoRESUMO
BACKGROUND: Vascular endothelial growth factor (VEGF) is a highly conserved, master regulatory molecule required for endothelial cell proliferation, organization, migration and branching morphogenesis. Podocoryne carnea and drosophila, which lack endothelial cells and a vascular system, express VEGF homologs, indicating potential roles beyond angiogenesis and vasculogenesis. The role of VEGF in the development and homeostasis of the postnatal small intestine is unknown. We hypothesized regulating VEGF bioavailability in the postnatal small intestine would exhibit effects beyond the vasculature and influence epithelial cell stem/progenitor populations. METHODS: VEGF mutant mice were created that overexpressed VEGF in the brush border of epithelium via the villin promotor following doxycycline treatment. To decrease VEGF bioavailability, sFlt-1 mutant mice were generated that overexpressed the soluble VEGF receptor sFlt-1 upon doxycycline administration in the intestinal epithelium. Mice were analyzed after 21 days of doxycycline administration. RESULTS: Increased VEGF expression was confirmed by RT-qPCR and ELISA in the intestine of the VEGF mutants compared to littermates. The VEGF mutant duodenum demonstrated increased angiogenesis and vascular leak as compared to littermate controls. The VEGF mutant duodenum revealed taller villi and increased Ki-67-positive cells in the transit-amplifying zone with reduced Lgr5 expression. The duodenum of sFlt-1 mutants revealed shorter villi and longer crypts with reduced proliferation in the transit-amplifying zone, reduced expression of Dll1, Bmp4 and VE-cadherin, and increased expression of Sox9 and EphB2. CONCLUSIONS: Manipulating VEGF bioavailability leads to profound effects on not only the intestinal vasculature, but epithelial stem and progenitor cells in the intestinal crypt. Elucidation of the crosstalk between VEGF signaling in the vasculature, mesenchyme and epithelial stem/progenitor cell populations may direct future cell therapies for intestinal dysfunction or disease.
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Duodeno/metabolismo , Células-Tronco/metabolismo , Fator A de Crescimento do Endotélio Vascular/fisiologia , Focos de Criptas Aberrantes/genética , Animais , Animais Lactentes , Síndrome de Vazamento Capilar/genética , Divisão Celular , Células Cultivadas , Doxiciclina/farmacologia , Duodeno/irrigação sanguínea , Duodeno/citologia , Duodeno/crescimento & desenvolvimento , Regulação da Expressão Gênica , Processamento de Imagem Assistida por Computador , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Camundongos Transgênicos , Microvilosidades/ultraestrutura , Neovascularização Fisiológica/genética , Organoides , Proteínas Recombinantes de Fusão/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição SOX9/biossíntese , Fatores de Transcrição SOX9/genética , Células-Tronco/citologia , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fator A de Crescimento do Endotélio Vascular/deficiência , Fator A de Crescimento do Endotélio Vascular/genética , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/biossíntese , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
PURPOSE: Tissue-engineered colon (TEC) might potentially replace absent or injured large intestine, but the enteric nervous system (ENS), a key component, has not been investigated. In various enteric neuropathic diseases in which the TEC is derived from aganglionic donor colon, the resulting construct might also be aganglionic, limiting tissue engineering applications in conditions such as Hirschsprung disease (HD). We hypothesized that TEC might contain a diverse population of enteric neuronal subtypes, and that aganglionic TEC can be populated by neurons and glia when supplemented with ENS progenitor cells in the form of neurospheres. MATERIALS AND METHODS: Human and murine organoid units (OU) and multicellular clusters containing epithelium and mesenchyme were isolated from both mouse and human donor tissues, including from normally innervated and aganglionic colon. The OU were seeded onto a biodegradable scaffold and implanted within a host mouse, resulting in the growth of TEC. Aganglionic murine and human OU were supplemented with cultured neurospheres to populate the absent ENS not provided by the OU to rescue the HD phenotype. RESULTS: TEC demonstrated abundant smooth muscle and clusters of neurons and glia beneath the epithelium and deeper within the mesenchyme. Motor and afferent neuronal subtypes were identified in TEC. Aganglionic OU formed TEC with absent neural elements, but neurons and glia were abundant when aganglionic OU were supplemented with ENS progenitor cells. CONCLUSION: Murine and human TEC contain key components of the ENS that were not previously identified, including glia, neurons, and fundamental neuronal subtypes. TEC derived from aganglionic colon can be populated with neurons and glia when supplemented with neurospheres. Combining tissue engineering and cellular replacement therapies represents a new strategy for treating enteric neuropathies, particularly HD.
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Colo , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo , Engenharia Tecidual/métodos , Animais , Técnicas de Cultura de Células , Colo/citologia , Colo/inervação , Colo/metabolismo , Humanos , Camundongos , Camundongos KnockoutRESUMO
Candida biofilm formation is common during infection and environmental growth. We tested the impacts of three biocides (ethanol [EtOH], H(2)O(2), and sodium dodecyl sulfate) on Candida albicans, C. parapsilosis, and C. glabrata biofilms. Higher concentrations of the biocides were required for efficacy against biofilms than for efficacy against planktonic controls. A combination study with two biocides (EtOH and H(2)O(2)) and fluconazole demonstrated that the combination had enhanced efficacy.