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1.
Int Wound J ; 2020 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-33314661

RESUMO

We aimed to explore the mechanism of circular RNAs (circRNAs) and provide potential biomarkers for molecular therapy of diabetic foot ulcers (DFU). Gene expression profile of GSE114248, including five normal samples and five DFU samples, was downloaded from GEO database. Differentially expressed circRNAs (DEcircRNAs) between two groups were identified. Then, DEcircRNA-miRNA and miRNA-mRNA interaction was revealed, followed by the circRNA-miRNA-mRNA network construction. Moreover, functional and pathway analysis were performed based on mRNAs, followed by the DM-related pathway exploration. Specific binding sites for key circRNAs and associated miRNAs were under investigation. Finally, RT-qPCR was used to verify the candidate the relative expression level of circRNA between normal tissues and DFU. Totally, 65 DEcircRNAs were revealed between two groups, followed by 113 circRNA-miRNA-mRNA interactions explored. The mRNAs in these interactions were mainly assembled in functions like cell proliferation and pathways. Moreover, a total of 11 DM-related pathways were revealed. Finally, circRNA-miRNA specific binding-site analysis revealed two key circRNAs, for example, circRNA_072697 and circRNA_405463, corresponding to their miRNAs. These two circRNAs were novel biomarkers for DFU. circRNA_072697 acted as a sponge of miR-3150a-3p in the progression of DFU via regulating KRAS. MAPK signaling pathway might contribute to the development of DFU.

2.
Int J Low Extrem Wounds ; : 1534734620944512, 2020 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-32734789

RESUMO

The purpose of this study was to investigate the clinical efficacy of compound polymyxin B ointment for treating chronic refractory wounds. A retrospective analysis was performed on 111 patients who underwent chronic refractory wound treatment. Patients were divided into 2 groups, with 45 patients included in the experimental group (compound polymyxin B group) and 66 patients included in the control group (silver sulfadiazine group). After thorough debridement in both groups, either compound polymyxin B ointment or silver sulfadiazine cream was evenly applied to the patient's wound and covered with sterile gauze. In both groups, dressing changes were dependent on the wound's condition and secretions. Using the Bates-Jensen Wound Assessment Tool (BWAT), patients in both groups were scored, after which wound healing, infection, and healing time were compared. There was no significant difference in BWAT scores between the 2 groups on the 7th or 14th day; however, on the 21st day, the BWAT score in the experimental group was significantly lower than that of the control group. The difference was statistically significant (P < .05). There was no significant difference in the BWAT-I scores between the 2 groups on the seventh day. The healing time in the experimental group was significantly shorter than that of the control group, and the difference was statistically significant (P < .05). For the treatment of chronic refractory wounds, thorough debridement followed by compound polymyxin B ointment topical application can reduce and control wound infection effectively and accelerate the process of wound repair.

3.
Int Wound J ; 17(5): 1428-1438, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32515909

RESUMO

Negative pressure wound therapy (NPWT) has been widely used in various lesions. This study aimed to explore the biological effects of negative pressure on the polymorphonuclear neutrophils (PMNs), macrophages, and epidermal keratinocyte cells involved in wound healing. PMNs differentiated from HL-60, macrophages were derived from THP-1 monocytes, and keratinocytes were cultured in vitro, and they were treated with 0, -0.03 mp, and -0.05 mp, respectively. Cell ultrastructure; viability; apoptosis; and protein factors such as tumour necrosis factor-α (TNF-α), interferon-γ (IFN-γ), epidermal growth factor (EGF), epidermal growth factor receptor (EGFR), interleukin-17 (IL-17), and cell division cycle 42 (Cdc42) were determined by transmission electron microscopy (TEM), CCK8, flow cytometry (FCM), ELISA, and simple Western assays, respectively. After negative pressure stimulation, the cell ultrastructure of PMNs and macrophages cells was presented with a marked increase of lysosomes and a relative decrease of mitochondria. In addition, the cell viability was enhanced in PMNs and macrophages in a pressure-dependent manner and apoptosis ratios were significantly reduced in PMNs and macrophages. In addition, under -0.05 negative pressure, IFN-γ and IL-17 were significantly increased in PMNs or macrophages. Moreover, increased EGF and EGFR and Cdc42 levels in keratinocytes induced by the -0.05 mpa were detected, indicating that the migration chemotaxis of keratinocyte cells was enhanced. Negative pressure might promote cell proliferation, accelerate inflammatory responses, and promote epithelialisation during wound healing by increasing IFN-γ, IL-17, Cdc42, EGF, and EGFR in PMNs, macrophages, or keratinocytes under different negative pressures.

4.
Wound Repair Regen ; 28(4): 532-538, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32386345

RESUMO

The vascular causes of lower-extremity ulcers cannot be neglected because they can directly affect treatment methods. No detailed epidemiological statistics have described vascular etiological diagnosis in China. This study aimed to explore the prevalence of clinical vascular etiological examination of lower-extremity ulcers and improve the diagnosis and treatment effectiveness of lower-extremity ulcers. Data were collected from the WoundCareLog database, which includes 2413 cases of lower-extremity ulcers from 478 hospitals nationwide. Data analysis revealed that 1698 (70.4%) lower-extremity blood flow examinations (including physical examination [PE] and assistant examinations [AE]) were performed, of which 61.7% were PE, 10.4% were AE only, and 27.9% were the combined PE and AE[PAE]. The proportion of nonexaminations was higher in the nondiabetic group than in the diabetic group (χ2 = 34.5; P < .01). The positive rates of vascular etiological examination in the diabetic and nondiabetic groups were 69.7% and 70.7%, respectively. Among the four economic regions of China, there were statistically significant differences in the use of the different examination methods. The examination of vascular diseases in lower-extremity ulcers in China has not been fully popularized and requires improvement; there was no statistically significant difference between examination rates by doctors and nurses, which is mainly based on PE. However, PE has certain rates of misdiagnosis and missed diagnosis. The false-positive and false-negative rates were 25.7% and 57.6%, respectively. The use of an AE can compensate for this deficiency by making diagnosis more precise, while the quantitative diagnostic criteria allow disease diagnosis to transcend geographical and operator differences and maximize uniformity. The vascular B-ultrasound examination is more suitable for the medical environment in China because of its mature technology, high hospital penetration rate, and low cost.

5.
Chin J Traumatol ; 22(5): 296-299, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31481277

RESUMO

The incidence of chronic wounds has been increasing over the past 20 years. However, the standardized diagnosis and treatment practice of chronic refractory wounds have not been established. In addition, the properties of the wound are characterized by morphology and thus correct description of the wound in medical history collection plays a vital role, which directly affects the definitive diagnosis. To develop more accurate format of clinical history record which can correctly reflect a patient's course and treatment progress, and to standardize the medical history record of chronic refractory wounds, at the national or regional level, we designed the WoundCareLog APP. It acts as a recording and communication tool for wound healing specialists at all levels of medical institutions in China. The WoundCareLog APP is fully compatible to meet the criteria and requirements of conventional medical records by embedding 9 modules. In addition, the demands for morphological description of wounds in wound healing diagnosis and treatment have been fulfilled by enroll of digital imaging technology to overcome the inadequacies of traditional medical history records.


Assuntos
Aplicativos Móveis , Cicatrização , Ferimentos e Lesões/diagnóstico , China , Doença Crônica , Humanos , Ferimentos e Lesões/patologia , Ferimentos e Lesões/fisiopatologia , Ferimentos e Lesões/terapia
6.
Mol Biol Rep ; 46(1): 67-76, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30374768

RESUMO

Scar formation and wound non-healing often occur during wound repair after skin injury, which are still unresolved. Clinic indicated that the structure played an important role in the wound repair. Our previous research showed that the wound over-healed (scar formation) when the integrity and continuity of dermal tissues was destroyed by injury. Other evidences showed that wound healing was impaired in diabetes because the underlying alternation in their skin tissues occurred caused by advanced glycation end products (AGES) aggregation. In order to explore the changes of the structure of skin at nanoscale, the small angle X-ray scattering (SAXS), compared with transmission electron microscopy (TEM), was applied to observe the skin in different pathological status. The results showed that there were some regular patterns in the structure of dermal tissue. The patterns were changed by different pathological status, which would result in wound healing disorder. These will be beneficial for clarifying the pathological mechanisms of wound healing.


Assuntos
Pele/patologia , Cicatrização/fisiologia , Difração de Raios X/métodos , Adulto , China , Cicatriz/patologia , Derme/patologia , Feminino , Humanos , Masculino , Microscopia Eletrônica de Transmissão/métodos , Pessoa de Meia-Idade , Espalhamento a Baixo Ângulo , Pele/metabolismo , Raios X
7.
J Diabetes Res ; 2017: 1428537, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29119117

RESUMO

Advanced glycosylation end products (AGEs) accumulate in diabetic wounds. Interactions between AGEs and their receptor (RAGE) leads to dermatologic problems in diabetes. Macrophage, which plays important roles in wound healing, highly expresses RAGE. Therefore, we investigated whether RAGE-expressing macrophages might be responsible for impaired wound healing on diabetes. We used anti-RAGE antibody applied topically on diabetic wounds. After confirming that wound healing was improved in anti-RAGE antibody group compared with normal mice, our results showed that macrophages appeared insufficient in the early stage and fading away slowly in the later proliferative phase compared with the control group, which was ameliorated in anti-RAGE antibody-applied wounds. Blocking AGE-RAGE signaling also increased neutrophils phagocytized by macrophages and promoted the phenotypic switch of macrophages from proinflammatory to prohealing activities. In vitro, phagocytosis of THP-1 (M0) and lipopolysaccharide- (LPS-) induced (M1) macrophages was impaired by treatment with AGEs, while IL-4- and IL-13-induced (M2) macrophages was not. Finally, AGEs increased the proinflammatory response of M1 macrophages, while inhibiting the polarization and anti-inflammatory functions of M2 macrophages. In conclusion, inhibition of AGE-RAGE signaling improved functional disorders of macrophages in the early inflammatory phase, which promoted the healing of wounds in diabetic mice.


Assuntos
Anticorpos/farmacologia , Diabetes Mellitus Experimental/metabolismo , Produtos Finais de Glicação Avançada/metabolismo , Macrófagos/efeitos dos fármacos , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Transdução de Sinais/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Animais , Macrófagos/metabolismo , Camundongos , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Receptor para Produtos Finais de Glicação Avançada/imunologia
8.
Int J Low Extrem Wounds ; 16(2): 94-103, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28682730

RESUMO

The aim of this study was to evaluate the changes of proliferation, apoptosis, homeostasis, and differentiation of human adipose-derived stem cells (hASCs) in the simulated diabetic microenvironment and discuss the potential of the mesenchymal stem cell in the treatment of chronic diabetic wound. We simulated diabetic microenvironment with glycation end products (AGEs) in vitro and studied the changes of hASCs in proliferation and apoptosis. We found that AGEs inhibited the proliferation and lead to hASCs apoptosis, and the endothelial cell directed differentiation was also inhibited. AGEs upregulated growth-related oncogene and monocyte chemoattractant protein-1 and downregulated urokinase-type plasminogen activator receptor, which may inhibit the proliferation and transference of endothelial cells. The simulated diabetic microenvironment affects the proliferation, apoptosis, and homeostasis of hASCs, the endothelial cell migration, and the synthesis of collagen protein, leading to delayed wound healing.


Assuntos
Colágeno , Pé Diabético , Células Endoteliais/fisiologia , Células-Tronco Mesenquimais/fisiologia , Nicho de Células-Tronco/fisiologia , Cicatrização/efeitos dos fármacos , Adipócitos/metabolismo , Apoptose/fisiologia , Diferenciação Celular , Movimento Celular , Proliferação de Células , Colágeno/biossíntese , Colágeno/metabolismo , Pé Diabético/metabolismo , Pé Diabético/patologia , Pé Diabético/terapia , Produtos Finais de Glicação Avançada , Homeostase/fisiologia , Humanos , Transplante de Células-Tronco Mesenquimais , Modelos Biológicos
9.
Chin J Traumatol ; 20(4): 202-206, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28526612

RESUMO

PURPOSE: To further study the mechanism of epithelization on the fascia side of the flap after surgical incision and the treatment of the negative pressure therapy. METHODS: With the patients' informed consent, parts of tissue samples were obtained from a 51-year-old diabetic patient who was suffering lower extremity ulcers. The samples were processed with hematoxylin and eosin (HE) staining and Masson trichrome staining. The keratin 19, keratin 15 and carcino-embryonic antigen (CEA) were immunohistochemically detected. RESULTS: The results of HE staining showed that the specimen was divided into two regions, newborn area and original epithelial area. There were more inflammatory cells infiltrating in the dermis in the newborn epithelial area, compared with the original epithelial area. Cells in newborn epithelial area were more active and many dinuclear and polynuclear cells were observed in newborn epithelial area. But there were more cuticular layers and obvious rete pegs in original epithelial area. In addition, the cells with keratin 19 and CEA positive were found around hair follicle, while keratin 15 was negative. Masson trichrome staining showed that there was a lot of de novo collagen in newborn epithelial area. CONCLUSION: Epidermal cells on the fascia side of the flap could be derived from the stem cells. Negative pressure wound therapy would attract not only cells but also other elements such as growth factors, cytokines, some nutrients and extracellular matrix. With the formation of the appropriate microenvironment after debridement, the migrated cells can grow, differentiate and spread, eventually leading to the epithelization on the fascia side of the flap in diabetic foot.


Assuntos
Desbridamento/métodos , Pé Diabético/terapia , Tratamento de Ferimentos com Pressão Negativa , Pé Diabético/patologia , Humanos , Imuno-Histoquímica , Queratina-15/análise , Queratina-19/análise , Pessoa de Meia-Idade
10.
Arch Med Sci ; 12(1): 179-87, 2016 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-26925135

RESUMO

INTRODUCTION: The aim of the study was to explore the effect of aminoguanidine cream on the skin tissue microenvironment in diabetic rats. MATERIAL AND METHODS: A total of 51 healthy male Sprague Dawley (SD) rats were randomly divided into three groups: the diabetes group (n = 18), the aminoguanidine group (n = 18) and the control group (n = 15). Rats in the diabetes group and aminoguanidine group were injected with 65 mg/kg streptozotocin to induce the diabetes model, and in the control group with citrate buffer. After successful induction of diabetes, the back hair of all rats was stripped by barium sulfide, and the aminoguanidine group was treated with aminoguanidine cream using disinfected cotton swabs twice every day for 40 days, while the diabetes and control groups were treated with the cream matrix. The pathological changes of skin were observed by HE staining, while the content of inflammatory cytokines (TNF-α, IL-8, ICAM and IL-1α) and the antioxidant indexes (T-AOC, GSH-PX, MPO MDA H2O2) were examined using commercial kits. RESULTS: After 40 days of treatment, the diabetes group manifested tissue lesions, whereas the aminoguanidine group seemed normal. Compared with the diabetes group, the content of inflammatory cytokines TNF-α, IL-8, ICAM and IL-1α was dramatically lower in the aminoguanidine group. T-AOC in all groups underwent dramatic changes and returned to normal finally. The activities of GSH-PX and MPO and content of H2O2 in the diabetes group were all higher than those in the aminoguanidine group. CONCLUSIONS: Aminoguanidine may have a good systemic effect on alleviating the pathological changes of skin tissue in diabetic rats, which may be attributed to the regulation of GSH-PX, TNF-α, IL-8, ICAM and IL-1α.

11.
J Burn Care Res ; 37(2): e115-24, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-25407384

RESUMO

Inflammation, initiated by polymorphonuclear neutrophil (PMNs) infiltration, is the first step in wound healing. The aim of this study is to investigate the function of neutrophils in a diabetes-impaired wound healing model and to explore the underlying mechanisms leading to neutrophil dysfunction. Superficial second-degree burns were created in the streptozotocin (STZ)-induced diabetic rat model, and the changes in the levels of advanced glycation end products (AGE), receptor of AGE (RAGE), inflammatory cytokines and oxidative markers, as well as cell apoptosis were determined. The effects of AGE on isolated PMNs were also determined in vitro. We found that deposition of AGE in diabetic rat skin activated the neutrophils before injury. However, the dense inflammatory band failed to form in the diabetic rats after injury. Compared with the controls, enhanced expression of RAGE and accelerated cell apoptosis were observed in the burned skin of diabetic rats. The altered expression pattern of inflammatory cytokines (tumor necrosis factor-alpha and interleukin-8) and oxidative markers (glutathione peroxidase, myeloperoxidase, hydrogen peroxide, and malondialdehyde) between burned skin of diabetic and control rats revealed delayed neutrophil chemotaxis and respiratory burst. Furthermore, the results in vitro showed that exposure to AGE inhibited the viability of PMNs, promoted RAGE production and cell apoptosis, and prevented the migration of PMNs, consistent with the findings in vivo. Besides, AGE-treated neutrophils showed increased secretion of inflammatory cytokines and increased oxidative stress. Combined, our results suggest that an interaction between AGE and its receptors inhibits neutrophil viability and function in the diabetic rat burn model.


Assuntos
Queimaduras/fisiopatologia , Diabetes Mellitus Experimental/fisiopatologia , Infiltração de Neutrófilos , Cicatrização/fisiologia , Animais , Apoptose , Biomarcadores/metabolismo , Movimento Celular , Quimiotaxia , Citocinas/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Produtos Finais de Glicação Avançada/metabolismo , Inflamação/fisiopatologia , Estresse Oxidativo , Ratos , Explosão Respiratória , Coloração e Rotulagem
12.
Zhonghua Shao Shang Za Zhi ; 30(2): 109-15, 2014 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-24989654

RESUMO

OBJECTIVE: To study the infiltration of macrophages and their phenotype in the healing process of full-thickness wound in rat. METHODS: Thirty healthy SD rats were divided into control group (n = 6) and injury group (n = 24) according to the random number table. Two round full-thickness skin defects (11 mm diameter) were created on both sides of dorsal spine of rats in injury group with surgical scissors and homemade trephine. After injury, wound area was measured immediately. The wounds were disinfected with iodophor every day. Rats in control group received anesthesia and hair removal only. On post injury day (PID) 1, 3, 7, and 13, respectively, 6 rats of injury group were sacrificed after the measurement of wound area (wound healing rate was calculated). Wound samples were obtained by excision down to healthy fascia along wound edge. Histological study was done with HE staining. The expression of CD68 (the surface marker of macrophage) in the wound tissue was observed with immunohistochemical staining. The double positive expressions of induced nitric oxide synthase (iNOS) plus CD68 (type I macrophage) and arginase 1 (Arg-1) plus CD68 (type II macrophage) were observed with immunofluorescence staining. The levels of interferon-γ (IFN-γ), TNF-α, IL-4, IL-13, IL-10, and IL-12 in wound tissue were assayed by double-antibody sandwich ELISA, and the ratio of IL-10/IL-12 was calculated. Full-thickness skin tissues (11 mm diameter) in rats of control group were excised at the same site as rats in injury group, and the histological observation and cytokines assay were performed as well. Data were processed with one-way analysis of variance or LSD- t test. RESULTS: Wound area of rats in injury group was gradually reduced after injury, and the overall difference of the wound healing rate on each PID was statistically significant (F = 358.55, P < 0.01). No abnormal appearance of skin tissue was observed in rats of control group. In injury group, inflammatory cell infiltration was obvious in wound tissue on PID 1 and 3; vascular structure and fresh collagen were observed in wound tissue on PID 7 and 13. Numbers of CD68 positive cells in skin tissue of rats in control group and wound tissue of rats in injury group on PID 1, 3, 7, and 13 were respectively (2.7 ± 1.5), (31.8 ± 3.5), (40.8 ± 4.7), (20.8 ± 2.8), (3.2 ± 2.4) per 200 times visual field (F = 180.55, P < 0.01). Compared with that in control group, the number of CD68 positive cells of rats in injury group was increased on PID 1, 3, and 7 (with t values respectively 18.81, 18.79, 14.05, P values below 0.01). No double positive expression of iNOS plus CD68 or Arg-1 plus CD68 was observed in normal tissue of rats in control group. In injury group, proportions of iNOS plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively (12.2 ± 2.8)%, (16.5 ± 2.9)%, (4.2 ± 2.3)%, (0.7 ± 0.8)% (F = 72.50, P < 0.01); proportions of Arg-1 plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively 0, (8.2 ± 1.9)%, (21.5 ± 3.4)%, (4.7 ± 2.0)% (F = 120.93, P < 0.01). In injury group, proportion of iNOS plus CD68 double positive cells on PID 3 was significantly higher than that on other PID (with t values respectively 2.65, 8.17, 12.95, P values below 0.05); proportion of Arg-1 plus CD68 double positive cells on PID 7 was higher than that on other PID (with t values respectively 15.27, 8.25, 10.38, P values below 0.01). Compared with that of Arg-1 plus CD68 double positive cells, proportion of iNOS plus CD68 double positive cells was higher on PID 1 and 3 (with t values respectively 10.71 and 5.88, P values below 0.01) and lower on PID 7 and 13 (with t values respectively 10.24 and 4.60, P values below 0.01). The overall differences of IFN-γ, TNF-α, IL-4, IL-13, and IL-10/IL-12 ratio in skin tissue of rats in control group and wound tissue of rats in injury group on every PID were statistically significant (with F values from 14.08 to 631.03, P values below 0.01). Compared with those in control group, levels of IFN-γ, TNF-α, IL-4, and IL-13 in wound tissue of rats in injury group were significantly higher on every PID (with t values from 4.58 to 9.17, P values below 0.05), while IL-10/IL-12 ratio was significantly higher on PID 1, 3, and 7 (with t values respectively 27.70, 30.51, 9.49, P values below 0.05) . In injury group, IFN-γ level on PID 1 [(61 ± 5) pg/mL] and IL-10/IL-12 ratio on PID 3 (1.647 ± 0.098) were significantly higher than those of control group and those on other PID in injury group [with IFN-γ level respectively (32 ± 4), (54 ± 6), (46 ± 7), (47 ± 4) pg/mL and IL-10/IL-12 ratio respectively 0.328 ± 0.045, 0.960 ± 0.034, 0.530 ± 0.028, 0.289 ± 0.040, with t values respectively from 3.19 to 8.20 and from 16.59 to 31.84, P values below 0.05]. CONCLUSIONS: Macrophage infiltration increases in the healing process of full-thickness wound in rat with different phenotypes, among which type I macrophage appears in the inflammatory stage, and type II macrophage predominates in the proliferative stage.


Assuntos
Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Macrófagos , Pele/lesões , Cicatrização/genética , Animais , Antígenos CD/genética , Antígenos de Diferenciação Mielomonocítica/genética , Colágeno , Ensaio de Imunoadsorção Enzimática , Interferon gama , Interleucina-10 , Interleucina-12 , Interleucina-13 , Interleucina-4 , Masculino , Fenótipo , Ratos , Fator de Necrose Tumoral alfa/sangue
13.
Front Biosci (Landmark Ed) ; 19: 727-33, 2014 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-24389216

RESUMO

Extracellular matrix is one of the key environmental factors influencing cell survival and provides signals for cell morphological change, migration, proliferation and differentiation. However, the mechanism through which denatured collagen modulates the biological properties of fibroblasts, is unclear. We investigated the regulation of human fibroblast differentiation in vitro grown in collagen gels with different properties. The break modulus of collagen with denatured collagen and half-load normal collagen was reduced compared with that of normal collagen gel. Fibroblasts cultured in denatured collagen gels showed increased expression of matrix metalloproteinase9 ( MMP-9), tissue inhibitor of metalloproteinase 2 (TIMP2), α-smooth muscle actin (α-SMA), osteoblast cadherin, phosphorylated Myosin phosphatase target subunit1 (p-MYPT1), connective tissue growth factor, type I collagen, type III collagen, α-smooth muscle actin messenger RNA, RhoA, rho-associated protein kinase, and transforming growth factor ß receptors 1 and 2 compared with that in cells cultured in normal collagen gel. But there was no significant difference regarding expression level between denatured collagen gel and half-load normal collagen gel .These findings suggest that the change in break modulus caused by decreasing normal collagen concentration may be the key factor inducing fibroblast differentiation.


Assuntos
Diferenciação Celular , Colágeno/metabolismo , Fibroblastos/citologia , Sequência de Bases , Células Cultivadas , Primers do DNA , Humanos , Reação em Cadeia da Polimerase em Tempo Real
14.
Int J Low Extrem Wounds ; 11(3): 224-30, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23008344

RESUMO

Dermatological problems in diabetes might play an important role in the spontaneous ulcers and impaired wound healing that are seen in diabetic patients. Investigation of the cause of diabetic skin disorders is critical for identifying effective treatment. The abdominal full-thickness skin tissues of 33 patients (14 nondiabetic and 19 diabetic) were analyzed. The cell viability and malondialdehyde (MDA) production of fibroblasts were measured after advanced glycosylation end product (AGE)-bovine serum albumin (BSA) exposure. Cutaneous histological observation showed reduced thickness of the diabetic abdominal dermis with morphological characteristics of obscured multilayer epithelium and shortened, thinned, and disorganized collagen fibrils with focal chronic inflammatory cell infiltration when compared with controls of the same age. Accumulation of AGEs in diabetic skin was prominent. Less hydroxyproline, higher myeloperoxidase activity, and increased MDA content were detected in diabetic skin. In vitro, the time- and dose-dependent inhibitory effects of AGE-BSA on fibroblast viability as well as the fact that AGE-BSA could promote MDA production of fibroblasts were shown. It is shown that the accumulation of AGEs in diabetic skin tissue induces an oxidative damage of fibroblasts and acts as an important contributor to the thinner diabetic abdominal dermis. The authors believe that diabetic cutaneous properties at baseline may increase the susceptibility to injury, and diabetic wounds possess atypical origin in the repair process.


Assuntos
Complicações do Diabetes/patologia , Produtos Finais de Glicação Avançada/metabolismo , Dermatopatias/metabolismo , Dermatopatias/patologia , Abdome/anatomia & histologia , Abdome/patologia , Estudos de Casos e Controles , Complicações do Diabetes/metabolismo , Feminino , Fibroblastos , Humanos , Masculino , Malondialdeído/metabolismo , Pessoa de Meia-Idade , Soroalbumina Bovina/análise , Pele/anatomia & histologia , Pele/metabolismo , Pele/patologia , Dermatopatias/etiologia , Fatores de Tempo , Cicatrização
15.
Zhonghua Shao Shang Za Zhi ; 28(1): 32-5, 2012 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-22490537

RESUMO

OBJECTIVE: To investigate the accumulation of advanced glycation end products (AGE) and the inflammatory response of skin and wound in diabetic patients, and to analyze their relationship in vitro. METHODS: Histological staining and immunohistochemical staining was respectively performed on skin and wound tissue specimens collected from 10 patients with Type II diabetes mellitus (diabetes group) and 12 non-diabetic patients with skin injury (control group) to observe the arrangement of collagen and the distribution of inflammatory cells, and to determine the expression levels of AGE and its receptor (RAGE). Malondialdehyde (MDA) levels in skin and wound tissue homogenates were assayed by enzyme-linked immunosorbent assay. In vitro, human neutrophils were isolated and treated with RPMI-1640 culture medium or that containing AGE-human serum albumin in the concentration of 0.315, 0.625, 1.250 mg/mL, and they were identified as normal control (NC) group, low concentration (L) group, moderate concentration (M) group, and high concentration (H) group. Cell viability in each group was determined by MTT colorimetric assay, and the reactive oxygen species (ROS) in cell was measured with 2', 7'-dichlorofluorescein-diacetate. Data were processed with t test. RESULTS: Compared with those of skin in control group, collagens of skin tissues in diabetes group atrophied and disorderly arranged. Inflammatory cells in wounds in diabetes group were dispersed, in which collagens arranged loosely and irregularly, as compared with those of wounds in control group. Expression levels of AGE and RAGE of skin in diabetes group were higher than those in control group. In diabetes and control groups, especially in diabetes group, the numbers of RAGE-positive cells in wound tissue were more than those in skin tissue. Large amount of inflammatory cells with positive expression of RAGE were observed in diabetes group. MDA level of skin and wound tissue in diabetes group was respectively (6.3 ± 1.0), (7.1 ± 2.4) nmol per milligram protein, which were obviously higher than those in control group [(2.9 ± 1.0), (3.6 ± 1.4) nmol per milligram protein, with t value respectively 8.017, 4.349, P < 0.05 or P < 0.01]. Cell viability and ROS levels in neutrophils were increased in L, M, and H groups [(59 ± 8)%, (77 ± 5)%, (67 ± 6)% and 1.67 ± 0.14, 2.13 ± 0.17, 3.48 ± 0.48] as compared with those in NC group [(34 ± 5)% and 0.58 ± 0.06, with t value respectively 7.195, 14.890, 11.130 and 20.195, 24.905, 16.864, P < 0.05 or P < 0.01]. CONCLUSIONS: Abnormal oxidative stress in diabetic skin leads to an atypical origin of wound repair. AGE-RAGE effect is a critical mediator for oxidative stress in diabetic wound tissue during wound healing.


Assuntos
Diabetes Mellitus Tipo 2 , Produtos Finais de Glicação Avançada/metabolismo , Estresse Oxidativo , Receptores Imunológicos/metabolismo , Cicatrização , Idoso , Estudos de Casos e Controles , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Espécies Reativas de Oxigênio/metabolismo , Receptor para Produtos Finais de Glicação Avançada , Albumina Sérica/metabolismo , Albumina Sérica Humana , Pele/metabolismo , Pele/patologia
16.
Burns ; 37(6): 1015-22, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21641116

RESUMO

OBJECTIVE: This study observed the degree of neo-vascularisation and differential expression of angiogenesis growth factors and their receptor in deep partial-thickness scald wound with diabetes mellitus. METHODS: Sixty Sprague-Dawley rats were randomised into a control group and an STZ-induced diabetic group, inflicted with partial-thickness scalding of 20% total body surface area (20% TBSA) on the back. Wound specimens were harvested immediately after scald and on 1, 3, 7, 10, 14 and 21 post-scald days (PSDs) to observe histological changes, and wound healing rates were calculated. The degree of neo-vascularisation in wound (labelled with blue microsphere) and the quantity of vascular endothelial cells (labelled with red CD31) were also measured by double-labelling immunofluorescence. Angiopoietin-1 (Ang-1), Angiopoietin-2 (Ang-2), Tie-2, vascular endothelial growth factor (VEGF) and Flt-1 messenger RNA (mRNA) were analysed by real-time RT-PCR. Ang-1, Ang-2 and VEGF protein expressions were measured by Western blotting. RESULTS: Wound healing was markedly impaired in diabetic rats. The diabetic rats show inhibited vascularity in the wound edge at every time point (the quantitation of vascularity was 60.0±3.0 in the control group and 12.0±1.4 in the diabetic group, p<0.01 on day 7). Although neo-vascularisation in the number of endothelial cells was not significantly different compared with the normal group, part of new vascular endothelial cells did not form the vascular function. After injury, expression of Ang-2 mRNA and protein were increased in both groups, and the normal group showed decreases on day 7, 14 and 21, whereas the diabetic group showed significant increases. Although the expression VEGF and its receptors before injury was higher than the normal group, the level at 1, 3 and 7 days after injury was significantly lower than that 14 days, and that at 21 days after injury was significantly higher than the normal group. CONCLUSION: Vascular endothelial cells can proliferate actively in the diabetic wound with deep partial-thickness burns, but it is still poor in blood supply due to lack of functional capillaries. The mechanism may be related to sustained abnormal high expression of Ang-2 and down-regulated VEGF.


Assuntos
Queimaduras/fisiopatologia , Diabetes Mellitus Experimental/fisiopatologia , Neovascularização Patológica/fisiopatologia , Proteínas Angiogênicas/metabolismo , Animais , Biomarcadores/metabolismo , Western Blotting , Queimaduras/metabolismo , Queimaduras/patologia , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Masculino , Neovascularização Patológica/patologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Reação em Cadeia da Polimerase , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Cicatrização/fisiologia
17.
Zhonghua Shao Shang Za Zhi ; 25(4): 275-80, 2009 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-19951546

RESUMO

OBJECTIVE: To investigate the effect of Thymosin and growth hormone(GH) on inflammatory response in burn rats or burn rats with sepsis. METHODS: Sixty-four SD rats were randomly divided into normal control group (NC, without treatment), sepsis group (S, with injection of LPS), sepsis + Thymosin group (ST, with successive injection of Thymosin and LPS), sepsis + GH group [SGH, with successive injection of recombinant human GH (rhGH) and LPS], burn group, burn + sepsis group (BS, with injection of LPS after burn), burn + sepsis + Thymosin group (BST, with successive injection of Thymosin and LPS after burn), burn + sepsis + GH (BSGH, with successive injection of rhGH and LPS after burn), with 8 rats in each group. Specimens of spleen tissues were harvested to determine HLA-DR in lymphocyte and evaluate inflammatory cell infiltration (score). Specimens of peripheral blood were collected to determine Toll-like receptor 4 (TLR4) level in monocyte and serum level of TNF-alpha, IL-4, IL-6, IL-10. RESULTS: Compared with those in NC group, serum level of IL-10 in S group decreased obviously, while other indices increased obviously (P < 0.01). The levels of HLA-DR and TLR4 and serum level of TNF-alpha were similar between SGH and ST groups (P > 0.05). Compared with those in SGH group [(2.87 +/- 0.04) score, and IL-6 (0.0083 +/- 0.0018) microg/mg, IL-4 (0.0102 +/- 0.0021) microg/mg, IL-10 (0.0310 +/- 0.0027) microg/mg, respectively], degree of inflammatory cell infiltration (1.50 +/- 0.76) score and serum levels of IL-6, IL-4, IL-10 of rats in ST group decreased obviously (0.0064 +/- 0.0012, 0.0058 +/- 0.0024, 0.0230 +/- 0.0021 microg/mg, respectively, P < 0.01). The levels of HLA-DR, TLR4 and inflammatory cell infiltration degree of spleen in B group were respectively higher than those in NC group and lower than those in BS group. Compared with those in NC group, serum levels of TNF-alpha, IL-6 in B group increased significantly, while IL-4, IL-10 showed an opposite tendency. There was no obvious difference between BST and BSGH groups in serum levels of HLA-DR and IL-6 (P > 0.05). Compared with those in BST group, inflammatory cell infiltration degree in spleen and the levels of TLR, TNF-alpha obviously decreased (P < 0.01), while IL-4 and IL-10 levels increased in BSGH group (P < 0.01). CONCLUSIONS: Inhibitive effects between Thymosin and GH on extensive inflammatory reaction were similar with or without trauma, and GH has better effect as compared with Thymosin when with trauma.


Assuntos
Queimaduras/imunologia , Hormônio do Crescimento Humano/farmacologia , Inflamação/imunologia , Sepse/imunologia , Timosina/farmacologia , Animais , Anti-Inflamatórios/farmacologia , Masculino , Ratos , Ratos Sprague-Dawley
18.
Zhonghua Shao Shang Za Zhi ; 24(1): 9-12, 2008 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-18512551

RESUMO

OBJECTIVE: To investigate the effects of advanced glycation end products (AGE) on the biological behavior of neutrophils in vitro, to look for the relationship between accumulation of AGE and abnormal inflammation in wound healing in diabetic mellitus patients. METHODS: Neutrophils were isolated from SD rats and incubated in vitro. The cells were divided into four groups according to different concentrations of AGE in cell suspension: control group (C, with treatment of RPMI - 1640), A group (with treatment of 0.315 mg/mL AGE + RPMI - 1640), B group (with treatment of 0.625 mg/mL AGE + RPMI - 1640), D group (with treatment of 1.250 mg/mL AGE + RPMI - 1640). Activity of neutrophils were determined by MTT colorimetric assay. Selectin-L mRNA expressions were analyzed by reversible transcription polymerase chain reaction (RT -PCR) technique. The levels of reactive oxygen species (ROS) in neutrophils were measured with DCFH-DA method. The protein concentration of neutrophil elastase (NE) was assayed by ELISA. RESULTS: The activity of neutrophils were obviously increased in A, B, and D groups when compared with that in C group [(0.170 +/- 0.040) in C group, (0.320 +/- 0.030) in A group, (0.380 +/- 0.020) in B group, (0.290 +/- 0.010) in D group, P <0. 05]. The expression of Selectin-L mRNA in A, B, D groups were significantly higher than that in C group (0.95 +/- 0.08, 1.36 +/- 0.27, 0.50 +/- 0.26.vs.0.36 +/- 0.26, P < 0.05. respectively). The ROS levels in A, B, D groups was markedly higher than that in C group (1.64 +/- 0.20, 2.16 +/- 0.26, 3.26 +/- 0.75. vs. 0.72 +/- 0.15, P <0.05, respectively). The levels of NE in A, B, D groups were significantly increased when compared with that in C group(1.98 +/- 0.43, 2.50 +/- 0.43, 2.01 +/- 0.18 vs 0.91 +/- 0. 21, P <0.05, respectively). CONCLUSION: AGE can enhance the activity of neutrophil, with change in cellular biological behaviors, which may be one of main reasons for abnormal inflammation in wounds of diabetes mellitus patients.


Assuntos
Produtos Finais de Glicação Avançada/farmacologia , Ativação de Neutrófilo , Neutrófilos/metabolismo , Animais , Células Cultivadas , Produtos Finais de Glicação Avançada/metabolismo , Selectina L/metabolismo , Elastase de Leucócito/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo
19.
Zhonghua Shao Shang Za Zhi ; 23(1): 6-12, 2007 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-17605245

RESUMO

Dermal defection and the degree of its loss determine the natural process of wound healing, which is the key reason leading to excess scar hyperplasia. The function of tri-dimensional structure in dermis acts as a template to regulate the properties of reparative cells. The template structure induces the reparative cells to grow into the structure which changes the skin mechanic status on wound area. Also, the component of extracellular matrix can affect behaviours of fibroblasts negatively or positively, for the reason that the structure of dermal tissue has a permissive effect on the dermal components in regulating behaviours of reparative cells. Therefore, the behaviors of cells depend on the structure of the template. The suitable tri-dimensional structure of dermis facilitates normal cell cycling. The more the structure of dermis closed to its physiological status, the better the biological behaviors of cells act. Moreover, the integrity as well as the continuity of dermal tissue is the prerequisite for serving as a template. The damage to the integrity and the continuity of dermal tissue may be one of the key reasons to lead abnormal tissue repair and scar formation. Thus, we hypothesize that the loss of dermal template may be one of the mechanism of abnormal scar formation and propose the theory of extracellular matrix framework deficiency or destruction.


Assuntos
Cicatriz/patologia , Derme/patologia , Cicatrização , Epiderme/patologia , Humanos
20.
Wound Repair Regen ; 12(6): 607-12, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15555051

RESUMO

Functional damage to microvascular endothelial cells by hyperglycemia is thought to be one of the critical risk factors in the impaired wound healing seen with diabetes mellitus. It is also thought that free radical stress plays a significant role in this endothelial cell dysfunction. In the present study, the effect of a free radical scavenger, MCI 186, on the endothelial cell dysfunction of cultured cells induced by high-glucose conditions was studied. Human dermal microvascular endothelial cells were cultured with high-glucose medium (50 mM) with or without MCI-186 (10 microM) for 7 days. Fifty mM mannitol was used as an osmotic control in this study. After this treatment, cell proliferation, activation of mitogen-activated protein kinase (MAPK), the level of apoptosis, and caspase-3 activation induced by removal of growth factors or tumor necrosis factor-alpha treatment were studied. High-glucose conditions significantly decreased cell proliferation and increased apoptosis levels with the activation of caspase-3 induced by growth factor removal. The high-glucose condition significantly activated MAPK. MCI-186 treatment improved cellular proliferation and reduced apoptosis and caspase-3 activation induced by high-glucose conditions. MCI-186 also inhibited the activation of MAPK. On the other hand, MCI-186 did not alter the level of apoptosis and caspase-3 activation induced by TNF-alpha treatment. In conclusion, we suggest that MCI-186 may be beneficial for improving the endothelial cell dysfunction induced by hyperglycemia.


Assuntos
Antipirina/análogos & derivados , Antipirina/farmacologia , Células Endoteliais/efeitos dos fármacos , Depuradores de Radicais Livres/farmacologia , Hiperglicemia/fisiopatologia , Pele/irrigação sanguínea , Apoptose , Western Blotting , Caspase 3 , Caspases/metabolismo , Proliferação de Células , Células Cultivadas , Edaravone , Humanos , Microcirculação , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
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