RESUMEN

BACKGROUND: Linear hypertrophic scar is a common surgical problem that can be difficult to manage, especially for the median sternotomy scar. Botulinum toxin type A (BTA) is widely used in cosmetic surgery and has been shown to improve scar quality recently. The aim of this study was to evaluate the efficacy of BTA injected in the early postoperative of median sternotomy on preventing scar formation. METHODS: In this prospective randomized controlled trial, 19 consecutive patients who underwent median sternotomy were enrolled. The median sternotomy wound in each patient was divided into the upper half and the lower half. Both halves of the wound were randomized to receive the treatment with either BTA or normal saline. At 6-month follow-up, scars were assessed using the Vancouver Scar Scale, scar widths were measured, and patients were asked to evaluate their overall satisfaction. RESULTS: Seventeen patients with median sternotomy wounds completed the entire study. At 6-month follow-up, the mean Vancouver Scar Scale score for the BTA-treated group was 3.44 ± 1.68 and for the normal saline control group was 6.29 ± 2.39, and there was a statistically significant difference between the two groups (P < 0.05). There were also significant improvements in scar width and patient satisfaction for the BTA-treated halves of the wounds (P < 0.05). CONCLUSIONS: The study demonstrates that early postoperative BTA injection can decrease scar formation and reduce scar width in median sternotomy wounds, and the overall appearance is more satisfactory. LEVEL OF EVIDENCE I: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Asunto(s)

Toxinas Botulínicas Tipo A/administración & dosificación , Cicatriz/prevención & control , Esternotomía/métodos , Cicatrización de Heridas/efectos de los fármacos , Adulto , China , Método Doble Ciego , Estética , Femenino , Estudios de Seguimiento , Humanos , Inyecciones Intralesiones , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Valores de Referencia , Medición de Riesgo , Esternotomía/efectos adversos , Factores de Tiempo , Resultado del Tratamiento

RESUMEN

Acute kidney injury (AKI) is a common complication after severe burns. Melatonin has been reported to protect against multiple organ injuries by increasing the expression of SIRT1, a silent information regulator that regulates stress responses, inflammation, cellular senescence and apoptosis. This study aimed to investigate the protective effects of melatonin on renal tissues of burned rats and the role of SIRT1 involving the effects. Rat severely burned model was established, with or without the administration of melatonin and SIRT1 inhibitor. The renal function and histological manifestations were determined to evaluate the severity of kidney injury. The levels of acetylated-p53 (Ac-p53), acetylated-p65 (Ac-p65), NF-κB, acetylated-forkhead box O1 (Ac-FoxO1), Bcl-2 and Bax were analyzed to study the underlying mechanisms. Our results suggested that severe burns could induce acute kidney injury, which could be partially reversed by melatonin. Melatonin attenuated oxidative stress, inflammation and apoptosis accompanied by the increased expression of SIRT1. The protective effects of melatonin were abrogated by the inhibition of SIRT1. In conclusion, we demonstrate that melatonin improves severe burn-induced AKI via the activation of SIRT1 signaling.

Asunto(s)

Lesión Renal Aguda/prevención & control , Quemaduras/complicaciones , Melatonina/farmacología , Sirtuina 1/metabolismo , Acetilación/efectos de los fármacos , Lesión Renal Aguda/etiología , Animales , Apoptosis/efectos de los fármacos , Citocinas/metabolismo , Riñón/efectos de los fármacos , Riñón/patología , Masculino , Proteínas de Neoplasias/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Proteínas de Transporte Nucleocitoplasmático/metabolismo , Estrés Oxidativo/efectos de los fármacos , Sustancias Protectoras/farmacología , Ratas Sprague-Dawley , Proteína p53 Supresora de Tumor/metabolismo

RESUMEN

Keloid, a skin benign tumor, is characterized by overgrowth of fibroblasts and the excessive deposition of extracellular matrix in wounded skin. Peroxisome proliferator-activated receptor-γ (PPAR-γ) agonist was recently evaluated to inhibit fibrosis. This study explored the underlying mechanisms. Fibroblasts isolated from 25 keloid patients (KFs) and fibroblasts isolated from healthy controls (NSFBs) were also subjected to treatment with PPAR-γ agonist troglitazone and antagonist GW9662 or for transfection with miR-92 mimics or inhibitor, Axl siRNA, and miR-92b or Axl promoter constructs, as well as being subjected to qRT-PCR, ELISA, Western blot, protein array, luciferase, and ChIP assays. The data demonstrated that TGF-ß1 and Axl proteins were significantly elevated in samples from keloid patients, while troglitazone treatment significantly reduced levels of TGF-ß1 and Axl mRNA and proteins in KFs. Moreover, knockdown of Axl expression reduced expression of TGF-ß1 and its pathway genes (such as α-SMA and Snail). PPAR-γ regulation of Axl expression was through transcriptional activation of miR-92b. miR-92b expression downregulated Axl expression at both mRNA and protein levels, whereas GW9662 completely reversed the inhibitory effects of miR-92b mimics on Axl expression. Gene ontology analysis of miR-92b targeting genes showed that TGF-ß and Axl were both potential targets of miR-92b, as confirmed by luciferase assay. These findings demonstrated that PPAR-γ-induced miR-92b expression inhibited Axl expression and in turn reduced expression of TGF-ß1 and the downstream genes in KFs, suggesting that targeting of this novel gene pathway may be useful for therapeutic control of fibrosis or keloid.

RESUMEN

A keloid is a benign skin tumor formed by an overgrowth of granulation tissue in affected patients. Peroxisome proliferator-activated receptor-γ (PPAR-γ) agonists were reported to be able to regulate extracellular matrix production in human dermal fibroblasts. This study explored the underlying molecular mechanism of PPAR-γ agonist troglitazone treatment for fibroblasts obtained from keloid patients. The data revealed that troglitazone treatment of keloid fibroblasts (KFs) downregulated the expression of early growth response-1 (Egr1) and collagen-1 (Col1). Level of Egr1 were closely associated with KF-induced fibrosis. The miRNA profiling data revealed that miR-543 was transcriptionally activated after troglitazone treatment. Bioinformatic analysis and experimental data showed that miR-543 was able to target Egr1. ELISA data confirmed that Col1 protein in the supernatant were modulated by the feedback regulatory axis of PPAR-γ agonist-induced miR-543 to inhibit Egr1 expression, whereas PPAR-γ antagonist treatment abolished such effect on Col1 suppression in KFs. This study demonstrated that the PPAR-γ agonist-mediated miR-543 and Egr1 signaling plays an important role in the suppression of collagen synthesis in KFs. Future in vivo studies are needed to confirm these in vitro data.

RESUMEN

Recent microRNA expression profiling studies have documented an up-regulation of miR-146a in several angiogenesis models. However, the underlying molecular mechanism of miR-146a in the angiogenic activity of endothelial cells has not been clearly elucidated. The present study was aimed to evaluate whether miR-146a promotes angiogenesis in HUVECs by increasing FGFBP1 expression via directly targeting CREB3L1. miR-146a was over expressed in HUVECs via lentiviral-miR-146a. Expression profiling analysis found miR-146a over expression resulted in up-regulation of angiogenesis and cytokine activity associated genes including FGF2. Further a combination of bioinformatics and experimental analyses demonstrated the CREB3L1 as a bona fide functional target of miR-146a during angiogenesis. Moreover, CREB3L1 inhibited luciferase expression from FGFBP1 promoter containing only CRE elements. Furthermore, CREB3L1 inhibited FGFBP1 expression by binding to two CRE-like sites located at approximately -1780-1777 and -868-865 bp relative to the FGFBP1 transcription start site. Additionally, ectopic expression of CREB3L1 decreased miR-146a-induced FGF2 secretion. These findings indicate that the miR-146a-CREB3L1-FGFBP1 signaling axis plays an important role in the regulation of angiogenesis in HUVECs and provides a potential therapeutic target for anti-angiogenic therapeutics.

Asunto(s)

Proteínas Portadoras/metabolismo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Células Endoteliales de la Vena Umbilical Humana/fisiología , Péptidos y Proteínas de Señalización Intercelular/metabolismo , MicroARNs/metabolismo , Neovascularización Fisiológica , Proteínas del Tejido Nervioso/metabolismo , Transducción de Señal , Regulación hacia Arriba , Citocinas/metabolismo , Humanos

RESUMEN

Compelling evidence from animal models and clinical studies suggest that transplantation of olfactory ensheathing cells (OECs), specialized glia in the olfactory system, combined with specific training may be therapeutically useful in the central nervous system (CNS) injuries and neurodegenerative diseases. The unique function of OECs could mainly attribute to both production of cell adhesion molecules and secretion of growth factors in OECs, which support neuron survival and neurite outgrowth. However, little is known about whether engulfment of neuronal degenerative debris by OECs also equally contributes to neuronal survival and neurite outgrowth. Furthermore, the molecular mechanisms responsible for neuronal degenerative corpses' removal remain elusive. Here, we used an in vitro model of primary culture of spinal cord neurons to investigate the effect of engulfment of degenerative neuron debris by OECs on neuronal survival and neurite outgrowth and the possible molecular mechanisms. Our results showed that OECs can engulf an amount of degenerated neuron debris, and this phagocytosis can make a substantial contribution to neuron growth, as demonstrated by increased number of neurons with longer neurite length and richer neurite branches when compared with the combination of neuron debris and OEC conditioned medium (OECCM). Moreover, p38 mitogen-activated protein kinase (p38MAPK) signaling pathway may mediate the OEC engulfment of debris because the p38MAPK-specific inhibitor, SB203580, can abrogate all the positive effects of OECs, including clearance of degenerated neuron debris and generation of bioactive molecules, indicating that p38MAPK is required for the process of phagocytosis of the neuron debris. In addition, the OEC phagocytic activity had no influence on its generation of bioactive molecules. Therefore, these findings provide new insight into further investigations on the OEC role in the repair of traumatic CNS injury and neurodegenerative diseases.

Asunto(s)

Citofagocitosis/fisiología , Neuritas/enzimología , Bulbo Olfatorio/citología , Bulbo Olfatorio/enzimología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Animales , Supervivencia Celular/fisiología , Activación Enzimática/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Neuronas/enzimología

RESUMEN

OBJECTIVE: To explore the methods of systemic treatment of defects of skin and soft tissue on the knees after severe trauma or burn. METHODS: Twenty patients with defects of skin and soft tissue on the knees after severe trauma or burn hospitalized in our center from January 2009 to December 2011. The injury areas on the knees ranged from 5 cm×4 cm to 30 cm×20 cm. The wounds were treated with radical debridement, vacuum sealing drainage, and douche through dripping to control infection in early stage. Then they were covered with transplantation of skin grafts plus flap or only with flap. Totally 8 local flaps (including 6 local rotation or transposition flaps and 2 saphenous artery flaps) and 12 free flaps (including 8 anterolateral thigh flaps and 4 latissimus dorsi musculocutaneous flaps) were used. The flap size ranged from 6 cm×5 cm to 32 cm×22 cm. The rehabilitation training of the knee joints was carried out in the early stage after wound healing. RESULTS: All free skin grafts and flaps used in 15 patients survived. Thirteen of them were primarily healed, while some small parts of skin grafts of the other two patients were in poor condition because of infection, and they healed after another session of skin transplantation. Infection occurred under the free flap in one of the 5 patients transplanted with flaps only, which was healed after continuous douche through dripping and another surgical debridement following wet dressing. The knee joints were in good function during the follow-up period of 1 - 3 years. CONCLUSIONS: The systemic therapy of radical debridement, vacuum sealing drainage technique, douche through dripping, transplantation of large autologous grafts and flaps, and the early rehabilitation training are effective and reliable in repairing defects of skin and soft tissue at the knee region after severe injuries.

Asunto(s)

Quemaduras/complicaciones , Traumatismos de la Rodilla/cirugía , Traumatismos de los Tejidos Blandos/cirugía , Colgajos Quirúrgicos , Adolescente , Adulto , Niño , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Trasplante de Piel , Adulto Joven

RESUMEN

The generation of induced neuronal cells from human bone marrow stromal stem cells (hBMSCs) provides new avenues for basic research and potential transplantation therapies for nerve injury and neurological disorders. However, clinical application must seriously consider the risk of tumor formation by hBMSCs, neural differentiation efficiency and biofunctions resembling neurons. Here, we co-cultured hBMSCs exposed to retinoic acid (RA) with human olfactory ensheathing cells (hOECs) to stimulate its differentiation into neural cells, and found that hBMSCs following 1 and 2 weeks of stimulation promptly lost their immunophenotypical profiles, and gradually acquired neural cell characteristics, as shown by a remarkable up-regulation of expression of neural-specific markers (Tuj-1, GFAP and Galc) and down-regulation of typical hBMSCs markers (CD44 and CD90), as well as a rapid morphological change. Concomitantly, in addition to a drastic decrease in the number of BrdU incorporated cells, there was a more elevated synapse formation (a hallmark for functional neurons) in the differentiated hBMSCs. Compared with OECs alone, this specific combination of RA and hOECs was significantly potentiated neuronal differentiation of hBMSCs. The results suggest that RA can enhance and orchestrate hOECs to neural differentiation of hBMSCs. Therefore, these findings may provide an alternative strategy for the repair of traumatic nerve injury and neurological diseases with application of the optimal combination of RA and OECs for neuronal differentiation of hBMSCs.

Asunto(s)

Células Madre Pluripotentes Inducidas/efectos de los fármacos , Neurogénesis/efectos de los fármacos , Bulbo Olfatorio/citología , Mucosa Olfatoria/citología , Células del Estroma/efectos de los fármacos , Tretinoina/farmacología , Antígenos de Diferenciación/análisis , Astrocitos/citología , Linaje de la Célula , Transdiferenciación Celular/efectos de los fármacos , Células Cultivadas/citología , Células Cultivadas/efectos de los fármacos , Técnicas de Cocultivo , Galactosilceramidasa/biosíntesis , Galactosilceramidasa/genética , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Proteína Ácida Fibrilar de la Glía/biosíntesis , Proteína Ácida Fibrilar de la Glía/genética , Humanos , Células Madre Pluripotentes Inducidas/citología , Proteínas del Tejido Nervioso/biosíntesis , Proteínas del Tejido Nervioso/genética , Cresta Neural/citología , Neuronas/citología , Oligodendroglía/citología , Células del Estroma/citología , Tubulina (Proteína)/biosíntesis , Tubulina (Proteína)/genética

RESUMEN

OBJECTIVE: To investigate the clinical application of free superficial iliac circumflex artery skin flaps, as well as the management of donor site defects. METHODS: 17 free superficial iliac circumflex artery skin flaps were applied for the traumatic defects or deformities on face, neck, foot, hand, ankle and lower leg, respectively. The donor site defects were closed directly or covered by paraumbilical island flaps. RESULTS: The 17 flap size ranged from 5 cm x 3 cm to 19 cm x 14 cm. 16 flaps survived completely except 1 flap with partial necrosis, which was closed by free skin graft. The donor site defects were closed directly in 10 cases, and covered by paraumbilical island flaps in 7 flaps without no flap necrosis. The abdomen had a good appearance. CONCLUSIONS: Good appearance can be achieved with free superficial iliac circumflex artery skin flaps for the defects on face, neck, foot, hand, ankle and lower leg. Paraumbilical island flap can be used for the donor site defects.

Asunto(s)

Colgajos Tisulares Libres/trasplante , Sitio Donante de Trasplante/cirugía , Heridas y Lesiones/cirugía , Arterias , Pie , Colgajos Tisulares Libres/irrigación sanguínea , Humanos , Procedimientos de Cirugía Plástica , Piel , Trasplante de Piel

Asunto(s)

Etnicidad/genética , Frecuencia de los Genes , Genética de Población , Repeticiones de Microsatélite , China , Dermatoglifia del ADN , Sitios Genéticos , Heterocigoto , Humanos , Reacción en Cadena de la Polimerasa Multiplex

RESUMEN

Killer cell immunoglobulin-like receptors (KIRs) are expressed on natural killer cells and as such regulate their response against infection and malignancy. KIR genes are variable in gene content and type, which results in different KIR haplotypes, and can be used to discriminate individuals and populations from different regions or ethnic groups. In the present study, we represent the first report on the KIR gene frequency and content diversities of 14 KIR genes (KIR2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 3DL1, 3DL2, 3DL3, 3DS1) and 2 pseudogenes (KIR3DP1 and 2DP1) in the Chinese Mongolian population. The 16 detected KIR genes were all observed. All the individuals were typed positive for the four framework genes KIR3DL3, 3DL2, 2DL4 and the pseudogene KIR3DP1, as well as for the pseudogene KIR2DP1. The observed carrier gene frequencies (OF) of the other KIR genes ranged from 16% at the KIR2DL2 locus to 93% at the KIR3DL1 locus. Over all, 48 different gene profiles were found in the study population and the most commonly observed KIR gene profile with a frequency of 14% consisted of KIR2DL4, 3DL2, 3DL3, 2DP1, 3DP1, 2DL1, 2DL3 and 3DL1 which belongs to the AA genotype. Principal component analysis (PCA) and the dendrogram illustrated the genetic distances between our study population and previously published populations from other ethnic groups or regions. The results of the present study show that the KIR gene family is highly polymorphic and can be a valuable tool for enriching the Chinese ethnical gene information resources, for anthropological studies, as well as for KIR gene related disease research.

Asunto(s)

Pueblo Asiatico/etnología , Pueblo Asiatico/genética , Frecuencia de los Genes , Variación Genética , Receptores KIR/genética , Etnicidad/genética , Genotipo , Haplotipos , Humanos , Mongolia/etnología , Seudogenes , Receptores KIR2DL4/genética , Receptores KIR3DL1/genética , Receptores KIR3DL2/genética

RESUMEN

OBJECTIVE: To study the safety and effects of free composite tissue flaps in repairing devastating wounds in early stage. METHODS: One hundred and twenty-three patients with 128 devastating wounds hospitalized in our burns center from 2005 to 2009 were repaired with free flaps or composite tissue flaps. Flap types used included 58 latissimus dorsi muscular flaps, 32 anterolateral thigh flaps, 21 circumflex scapular flaps, 6 dorsalis pedis composite flaps, 3 big toe nail skin flaps, 3 forearm flaps, and 1 lateral thoracic flap. One wound was repaired with lateral lower leg flap with fibula, and 3 wounds with free latissimus dorsi muscular flap plus skin graft. RESULTS: Vascular crisis was observed in 10 transplanted flaps 1 to 5 days after operation; 6 flaps with this complication were saved after emergency surgical exploration. Total survival rate of transplanted flaps and composite tissue flaps was 95.3% (122/128). All patients were followed up for 3 months to 4 years; satisfactory appearance and restoration of partial function were found in all of them. CONCLUSIONS: Free composite tissue transplantation reduces amputation rate, achieves primary reconstruction of function with good appearance, shortens length of hospital stay, and reduces surgical operation time, complications, and treatment cost. It is a good approach in the repair of massive devastating soft tissue injury.

Asunto(s)

Quemaduras/cirugía , Colgajos Tisulares Libres , Traumatismos de los Tejidos Blandos/cirugía , Adolescente , Adulto , Anciano , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Procedimientos de Cirugía Plástica , Trasplante de Piel , Cicatrización de Heridas , Adulto Joven

RESUMEN

OBJECTIVE: To investigate the feasibility of debridement at early stage by combining lavation and drainage under continuous negative pressure in the limb salvage surgery of patients suffering from diabetic foot complicated by plantar abscess. METHODS: Ten patients suffering from diabetic foot (10 feet affected) combined with plantar abscess were involved. After being debrided, wounds of patients were lavaged and drained for 7 - 14 ds under continuous negative pressure through a detained stomach tube. Suction under negative pressure continued for 3 - 5 ds after lavation, and then drainage tube was removed. Stitches on wound were removed about 3 weeks after surgery. The condition of the wounds was observed. RESULTS: After above-mentioned treatment, local inflammatory response of patients was obviously ameliorated, and the wounds healed with satisfactory preservation of function and shape of the feet. CONCLUSIONS: Debridement combined with local lavation and drainage under continuous negative pressure is satisfactory for the treatment of diabetic foot complicated by plantar abscess, and it can preserve the length of the affected limb.

Asunto(s)

Absceso/cirugía , Infecciones Bacterianas/cirugía , Desbridamiento , Pie Diabético/cirugía , Úlcera del Pie/cirugía , Terapia de Presión Negativa para Heridas , Absceso/complicaciones , Anciano , Infecciones Bacterianas/complicaciones , Pie Diabético/complicaciones , Estudios de Factibilidad , Femenino , Úlcera del Pie/complicaciones , Humanos , Masculino , Persona de Mediana Edad

RESUMEN

OBJECTIVE: To reproduce a model of heat injured KC in vitro and explore its apoptosis rate of KC due to heat injury at different temperature. METHODS: Human KCs were cultured in vitro, and they were incubated at 37, 41, 43, 45, 48, and 51 degrees C respectively for 10 mins in water bath. Trypan blue staining and Hoechst 33258 fluorescence staining were used respectively to determine necrosis and apoptosis of KC. Rates of apoptosis and necrosis of KC were analyzed quantitatively by flow cytometer. The proliferation activity of KC after heat injury was detected by MTT test. RESULTS: The results of trypan blue staining, Hoechst 33258 fluorescence staining, and flow cytometer demonstrated that number of apoptotic and necrotic KC increased gradually along with a rise of water bath temperature. The rates of apoptosis and necrosis of KC were respectively (12.3 +/- 3.2)% and (14.1 +/- 1.6)% at 45 degrees C, (27.7 +/- 5.1)% and (58.0 +/- 4.2)% at 48 degrees C. Rate of KC necrosis reached up to (83.0 +/- 5.3)% at 51 degrees C. Inhibition of KC growth reached a stationary phase when the injurious temperature reached 45 degrees C as observed with MTT test. CONCLUSIONS: Heat injury can induce apoptosis and growth inhibition of KC in vitro. Incubating KC at 45 degrees C for 10 mins is a good condition to reproduce a model of heat injured KC in vitro. This model may be used to study the biological character and apoptosis of KC after burn injury.

Asunto(s)

Apoptosis , Supervivencia Celular , Calor , Queratinocitos/citología , Quemaduras , Proliferación Celular , Células Cultivadas , Citometría de Flujo , Humanos

RESUMEN

OBJECTIVE: To explore the methods of repair of massive deep skin and soft tissue injuries. METHODS: Fifty-six patients with deep skin and soft tissue injuries were hospitalized from July 2006 to January 2008. Among them, 23 cases were caused by burn, 17 cases by electric injury, 7 cases by hot crush injury, 6 cases by avulsion injury, and 3 cases due to other reasons (including traffic accident, crush injury, soft tissue infection respectively). Sixty-five skin flaps were raised to repair and reconstruct the injured tissues, including 21 local flaps, 18 distant pedicled skin flaps, and 26 free skin flaps. The area of skin flaps ranged from 1.5 cm x 1.0 cm to 39.0 cm x 23.0 cm. RESULTS: Sixty skin flaps survived completely, partial necrosis occurred in 3 flaps, and complete necrosis in 2 flaps. There was no obvious difference in average survival rate among local skin flaps (95.2%), distant pedicled skin flaps (88.8%), and free skin flaps (92.3%, P > 0.05). CONCLUSIONS: Skin flap transposition can be still considered as the major effective method in repair of massive deep skin and soft tissue injury. On the premises of high survival rate, free skin flap transposition can be considered as the first choice.

Asunto(s)

Quemaduras/cirugía , Piel/lesiones , Traumatismos de los Tejidos Blandos/cirugía , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Procedimientos de Cirugía Plástica/métodos , Trasplante de Piel/métodos , Colgajos Quirúrgicos , Adulto Joven

RESUMEN

OBJECTIVE: To investigate the optimal operation method for the management of various chronic wounds in legs and feet. METHODS: Fifty-one chronic wounds were evaluated according to infection, inflammatory response, and distribution in different areas of the leg and foot. Preoperative treatment was given accordingly, then transposition of skin flap, skin grafting, or amputation was performed. The healing rate after single session operation and average hospitalization were statistically analyzed. RESULTS: The wound healing rate after single session operation was 86. 3% , the average hospital stay was (17. 8 +/- 2. 1) days, and the appearance and function of the leg and foot after operation was satisfactory. CONCLUSION: The appropriate preoperative treatment and operation method conforming to the wound location and evaluation are of vital importance in the management of chronic wounds in the leg and foot. Operation is one of the most effective ways to repair chronic wounds in the leg and foot, and it can shorten the wound healing process and restore the function.

Asunto(s)

Úlcera del Pie/patología , Úlcera del Pie/cirugía , Úlcera de la Pierna/patología , Úlcera de la Pierna/cirugía , Adulto , Enfermedad Crónica , Humanos , Longevidad , Masculino , Colgajos Quirúrgicos , Cicatrización de Heridas

RESUMEN

OBJECTIVE: Injecting the EPC into the corresponding skin flap to study EPC biological characteristics and its effect on neovascularization in ischemia skin flap. METHODS: CD133 + cells were enriched from human umbilical cord blood by immunomagnetic sorting, and cultured with EGM - 2MV media. After labeled with PKH26 (fluorescent cell linker), the EPC were injected into the over-length flap models made on athymic mice. Observing the EPCs trace and their participating in the flap vascularization using a fluorescent microscope. The potential of EPC neovascularization in ischemic tissue of skin flap was evaluated through measuring the necrotic area and vessel diameter and quantity in the skin flap. RESULTS: The skin flap necrosis area of EPC group is significantly smaller than that of control (P < 0.05), the dermal and hypodermal blood perfusion of EPC group is significantly more than that of control (P < 0.05). Immunohistological and label fluorescent analyses showed vWF antigen-positive cells and labeled cells constructing blood vessels of flap. CONCLUSIONS: Our data support the EPC may contribute to angiogenesis, speed up ischemic tissue vascularization.

Asunto(s)

Trasplante de Células Madre de Sangre del Cordón Umbilical , Supervivencia de Injerto , Células Madre/citología , Colgajos Quirúrgicos , Animales , Células Cultivadas , Femenino , Sangre Fetal , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos

RESUMEN

OBJECTIVE: To explore the endothelial progenitor cell markers and biological characteristics of human CD133 umbilical cord blood cells( EPC). METHODS: CD133+ cells were enriched from human umbilical cord blood by immunomagnetic sorting, and cultured with EGM-2MV medium containing epidermal growth factor, vascular endothelial growth factor and fibroblast growth factor 2. The percentage of CD133+ cells in cord blood monocytes, the growth curve and growth characteristics of primary EPCs were measured by flow cytometry and immunochemistry method. Weibel-Palade body was observed with transmission electron microscope. The mixture of EPCs and human stomach cancer cell line GC7901 were injected into athymic mice to observe the tumor growth and vascularization. RESULTS: The percentage of CD133+ cells in cord blood monocytes was 0.91%, and after sorting, the percentage of CD133+ cells was raised to 85.52%. The cultured cells showed a typical spindle-shaped morphology in 3 post-culture days (PCD) and areas of clusters of cobblestone-like cells in 10 PCD. The number of EPC increased from 7 PCD on, peaked on 17 PCD. Obvious amplification and clone-like growth on 7 PCD were observed by light microscope. Typical Weibel-Palade body was observed in the cells under transmission electron microscope. Tumor forming experiment in athymic mice showed that the tumor size of EPC group was larger than that of control with smaller necrosis area and more and larger blood vessels. Immuno-fluorescent staining showed many human vWF antigen-positive endothelial cells being involved in the tumor vascularization. CONCLUSION: Immunomagnetic sorting can efficiently enrich EPC from human umbilical cord blood. Our data support that the EPC may contribute to angiogenesis, speed up vascularization of ischemic tissue.

Asunto(s)

Células Endoteliales/citología , Sangre Fetal/citología , Monocitos/citología , Células Madre/citología , Antígeno AC133 , Animales , Antígenos CD , Antígenos CD34 , Técnicas de Cultivo de Célula , Diferenciación Celular , Línea Celular Tumoral , Separación Celular , Células Cultivadas , Femenino , Glicoproteínas , Humanos , Ratones , Ratones Endogámicos BALB C , Neovascularización Patológica , Péptidos

RESUMEN

OBJECTIVE: To explore the preliminary methods of in vitro isolation, culture and identification of sebocytes and eccrine sweat gland cells from human fetal skin. METHODS: Human fetal skin was digested with dispase or type II collagenase, and then by micro - sieving to isolate human sebaceous gland and eccrine sweat gland cells. DMEM/F12 (1: 1) was used as the basic culture medium, supplemented with fetal bovine serum, recombinant human epidermal growth factor, L-glutamine, Hydrocortisone, choleratoxin, penicillin and streptomycin as the medium for sebocytes, or fetal bovine serum, recombinant human epidermal growth factor, triiodothyronine, hydrocortisone, insulin, transferrin, sodium selenite to the medium for eccrine sweat gland duct cells. Primary cultures and subcultures were incubated at 37 degrees C in humidified atmosphere of 5% CO2/95% oxygen. Cell morphology was observed by inverted phase contrast microscopy, and the cultured cells were identified with cell clone efficiency determination. The cultured sebocytes were identified with oil red staining and CK4.62, Epithelia Membrane Antigen (EMA) immunohistochemistry staining. The cultured eccrine sweat gland duct cells were identified with CK7, CK19 immunohistochemistry staining. RESULTS: The isolated sebocytes and eccrine sweat gland cells from human fetal skin could grow by adhering to the wall and proliferate in vitro. The cell clone efficiency of human fetal sebocytes was 2.7%, which was obviously lower than that of human fetal keratinocytes (8.0%, P < 0.01). There was no obvious difference in the cell clone efficiency between human fetal eccrine sweat gland cells (7.3%) and human fetal keratinocytes (7.7%, P > 0.05) . The results of oil red staining indicated that a small quantity of lipid droplets in sebocytes, and immunohistochemistry staining of CK4.62, EMA were positive in subculture sebocytes. The immunohistochemistry staining of CK7, CK19 was positive in subculture eccrine sweat gland duct cells. CONCLUSION: In vitro cultured human fetal sebocytes and eccrine sweat gland duct cells displayed the markers and biological characteristics of epithelial lineage, but human fetal sebocytes proliferated more

Asunto(s)

Técnicas de Cultivo de Célula , Glándulas Ecrinas/citología , Glándulas Sebáceas/citología , Vernix Caseosa/citología , Feto/citología , Humanos , Piel/citología