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1.
Tumour Biol ; 36(6): 4167-73, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25592378

RESUMEN

Sex-determining region Y-related high mobility group box 4 (SOX4) has been proven to serve as a critical role in cancer development and progression. However, little is known about the pathological role of SOX4 in breast cancer patients. The purpose of this study is to measure the expression of SOX4 in breast cancer patients and to explore the clinical significance of SOX4. Using RT-PCR and Western blot, messenger RNA (mRNA) and protein expression of SOX4 were measured in breast cancer tissues and adjacent normal mammary tissues. The relationship of SOX4 expression with clinical characteristics of 148 breast cancer patients was analyzed by immunohistochemistry. In the present study, our results indicated that SOX4 mRNA and protein were highly expressed in breast cancer tissues compared with adjacent normal mammary tissues and positively correlated with clinical stage (I-II vs. III-IV; P = 0.008), T classification (T1-T2 vs. T3-T4; P = 0.013), N classification (N0-N1 vs. N2-N3; P < 0.001), M classification (M0 vs. M1; P = 0.001), estrogen receptor (negative vs. positive; P = 0.029), progesterone receptor (negative vs. positive; P = 0.004), and histological grade (G1 vs. G2-G3; P = 0.033) in breast cancer patients. Furthermore, we also found that SOX4 protein overexpression was an unfavorable prognostic factor in breast cancer patients (P < 0.001), regardless of clinical stage, tumor size, lymph node metastasis, and distant metastasis. Finally, high SOX4 expression was an independent poor prognostic factor for pancreatic patients through multivariate analysis (P = 0.033). In conclusion, SOX4 overexpression serves as an unfavorable prognostic biomarker in breast cancer patients.


Asunto(s)
Biomarcadores de Tumor/biosíntesis , Neoplasias de la Mama/genética , Pronóstico , Factores de Transcripción SOXC/biosíntesis , Adulto , Anciano , Biomarcadores de Tumor/genética , Neoplasias de la Mama/patología , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Estimación de Kaplan-Meier , Metástasis Linfática , Persona de Mediana Edad , Factores de Transcripción SOXC/genética
2.
J Mol Med (Berl) ; 102(10): 1255-1266, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39158595

RESUMEN

Crouzon syndrome (CS), a syndromic craniosynostosis, is a craniofacial developmental deformity caused by mutations in fibroblast growth factor receptor 2 (FGFR2). Previous CS mouse models constructed using traditional gene editing techniques faced issues such as low targeting efficiency, extended lineage cycles, and inconsistent and unstable phenotypes. In this study, a CRISPR/Cas9-mediated strategy was employed to induce a functional augmentation of the Fgfr2 point mutation in mice. Various techniques, including bone staining, micro-CT, histological methods, and behavioral experiments, were employed to systematically examine and corroborate phenotypic disparities between mutant mice (Fgfr2C361Y/+) and their wild-type littermates. Confirmed via PCR-Sanger sequencing, we successfully induced the p.Cys361Tyr missense mutation in the Fgfr2 IIIc isoform of the extracellular domain (corresponding to the p.Cys342Tyr mutation in humans) based on Fgfr2-215 transcript (ENSMUST00000122054.8). Fgfr2C361Y/+ mice exhibited characteristics consistent with the phenotypic features associated with CS, including skull-vault craniosynostosis, skull deformity, shallow orbits accompanied by exophthalmos, midface hypoplasia with malocclusion, and shortened skull base, notably without any apparent limb defects. Furthermore, mutant mice displayed behavioral abnormalities encompassing deficits in learning and memory, social interaction, and motor dysfunction, without anxiety-related disorders. Histopathological examination of the hippocampal region revealed structural abnormalities, suggesting possible brain development impairment secondary to craniosynostosis. In conclusion, we constructed a novel gene-edited Fgfr2C361Y/+ mice strain based on CRISPR/Cas9, which displayed skull and behavioral abnormalities, serving as a new model for studying genetic molecular mechanisms and exploring treatments for CS. KEY MESSAGES: CRISPR/Cas9 crafted a Crouzon model by enhancing Fgfr2-C361Y in mice. Fgfr2C361Y/+ mice replicate CS phenotypes-craniosynostosis and midface anomalies. Mutant mice show diverse behavioral abnormalities, impacting learning and memory. Fgfr2C361Y/+ mice offer a novel model for cranial suture studies and therapeutic exploration.


Asunto(s)
Sistemas CRISPR-Cas , Disostosis Craneofacial , Modelos Animales de Enfermedad , Receptor Tipo 2 de Factor de Crecimiento de Fibroblastos , Cráneo , Animales , Receptor Tipo 2 de Factor de Crecimiento de Fibroblastos/genética , Disostosis Craneofacial/genética , Ratones , Cráneo/anomalías , Cráneo/patología , Fenotipo , Conducta Animal , Edición Génica , Masculino , Femenino
3.
Technol Health Care ; 31(4): 1333-1342, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36847032

RESUMEN

BACKGROUND: There are two minimally invasive ways of treating cholecystolithiasis combined with choledocholithiasis, but there remains some controversy regarding which technique is better, since they both have advantages and disadvantages. The one-step method involves laparoscopic cholecystectomy, laparoscopic common bile duct exploration, and primary close (LC + LCBDE + PC), while the two-step procedure consists of endoscopic retrograde cholangiopancreatography, endoscopic sphincterotomy, and laparoscopic cholecystectomy (ERCP + EST + LC). OBJECTIVE: This multicenter retrospective study aimed to analyze and compare the effects of the two techniques. METHODS: The data of patients who underwent either one-step LCBDE + LC + PC or two-step ERCP + EST + LC treatment for gallstones in the gallbladder and bile duct at the Shanghai Tenth People's Hospital, Shanghai Tongren Hospital, and Taizhou Fourth People's Hospital between January 1, 2015 and December 31, 2019 were collected, and the preoperative indicators of the two groups were compared. RESULTS: The surgical success rate of the one-step laparoscopic group was 96.23% (664/690), the transit abdominal opening rate was 2.03% (14/690), and there were 21 cases of postoperative bile leakage. The success rate of the two-step endolaparoscopic surgery was 78.95% (225/285), the transit opening rate was 2.46% (7/285), and there were 43 postoperative cases of pancreatitis and five of cholangitis. Postoperative cholangitis, pancreatitis, postoperative stone recurrence, postoperative hospitalization, and treatment costs were significantly lower (P< 0.05) in the one-step laparoscopic group than in the two-step endolaparoscopic group. However, the amount of intraoperative bleeding, the postoperative extraction time of the abdominal drainage tube, and the incidence of bile leakage were higher (P< 0.05) in the one-step laparoscopic group than in the two-step endolaparoscopic group. CONCLUSION: The two methods of treating choledocholithiasis combined with choledocholithiasis that were analyzed in this study were safe and effective, and each method had its own advantages.


Asunto(s)
Colangitis , Coledocolitiasis , Laparoscopía , Humanos , China , Colangitis/complicaciones , Colangitis/cirugía , Coledocolitiasis/cirugía , Coledocolitiasis/complicaciones , Conducto Colédoco/cirugía , Tiempo de Internación , Estudios Retrospectivos
4.
Am J Transl Res ; 13(10): 11255-11270, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34786056

RESUMEN

BACKGROUND: FGFR2 (fibroblast growth factor receptor 2) mutations are implicated in the etiopathogenesis of syndromic craniosynostosis, and C278F- or C342Y-FGFR2 mutations can lead to Crouzon syndrome. The dura mater exerts crucial effects in the regulation of cranial suture development. However, the underlying mechanisms of these biological processes are rarely studied. This research explored and analyzed the biological function of FGFR2 overexpressed by dura cells on cranial osteoblasts. METHODS: Dura cells and cranial osteoblasts from C57BL/6 mice aged 6 days were obtained and cultured respectively. Lentivirus-FGFR2 constructs were engineered with C278F- and C342Y-FGFR2 mutations. The dura cells were infected with the constructs and co-cultured with osteoblasts in a trans-well system. Four experimental groups were established, namely the Oste group, the Oste+Dura-vector group, the Oste+Dura-C278F group, and the Oste+Dura-C342Y group. FACS, CCK8, and EdU assays were used to evaluate the osteoblast proliferation levels. Western blot and RT-qPCR were used to measure the expressions of the factors related to proliferation, differentiation, and apoptosis. Furthermore, the expression levels of the key factors in the Hippo/YAP-PI3K-AKT proliferation pathway were measured and analyzed. Finally, rescue experiments were performed with an RNA interfering assay. RESULTS: The proliferation and differentiation levels of the osteoblasts in the Oste+Dura-C278F and Oste+Dura-C342Y groups were significantly up-regulated, but the apoptosis levels in the four groups were not significantly different. The YAP, TEADs1-4, p-PI3K, and p-AKT1 expressions in the mutant FGFR2 groups were higher than the corresponding expressions in the control groups, and the results of the rescue experiments showed a reverse expression tendency, which further confirmed the effects of the FGFR2 mutations in the dura cells on the proliferation of the osteoblasts and the underlying possible mechanisms. CONCLUSION: Our studies suggest that the Crouzon mutations (C278F- and C342Y-) of FGFR2 in dura cells can enhance osteoblast proliferation and differentiation and might influence the pathogenesis of craniosynostosis by affecting the Hippo/YAP-PI3K-AKT proliferation signaling pathway.

5.
Inflammation ; 44(1): 358-370, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33044666

RESUMEN

The inflammatory response involving interleukin-1ß (IL-1ß) has been thought to play an important role in the development of late-phase sepsis. However, in this study, we wanted to explore the possibility of using IL-1ß to improve the prognosis of sepsis by triggering local differentiation of bone marrow cells (BMCs) into regulatory dendritic cells (DCs) in vivo, thereby reversing the immune paralysis in late-phase sepsis. Sepsis mouse models were induced by cecal ligation and puncture (CLP) and lethal Escherichia coli O18 infection. Mice were injected intraperitoneally with IL-1ß after CLP and after the lethal infection. Septic BMCs and liver immune cells were isolated at 0, 3, 6, 9, and 14 days post-CLP. BMCs and liver cells isolated from septic mice treated with IL-1ß were adoptively transferred into CLP mice. GFP+-C57BL/6 parabiosis models were established. Serum IL-1ß levels were determined by enzyme-linked immunosorbent assay (ELISA) kit, and the number, ratio, and phenotype of immune cells were observed by flow cytometry. IL-1ß treatment improved the survival of sepsis and increased the numbers of BMCs and liver immune cells in septic mice. Moreover, IL-1ß stimulation increased the number and the percentage of CD11c-CD45RBhigh DCs in septic BM and liver. Adoptive transfer of septic BMCs, liver immune cells, and CD11c-CD45RBhigh DCs treated with IL-1ß into CLP mice attenuated sepsis. IL-1ß triggered the redistribution of CD11c-CD45RBhigh DCs as well as BMCs in parabiosis models. IL-1ß protects against sepsis by stimulating local proliferation and differentiation of BMCs into CD11c-CD45RBhigh DCs at immune organs and non-immune organs.


Asunto(s)
Modelos Animales de Enfermedad , Interleucina-1beta/uso terapéutico , Sepsis/prevención & control , Animales , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/metabolismo , Movimiento Celular/efectos de los fármacos , Movimiento Celular/fisiología , Relación Dosis-Respuesta a Droga , Interleucina-1beta/farmacología , Masculino , Ratones , Ratones Endogámicos C57BL , Sepsis/inducido químicamente , Sepsis/metabolismo
6.
RSC Adv ; 9(15): 8239-8245, 2019 Mar 12.
Artículo en Inglés | MEDLINE | ID: mdl-35518665

RESUMEN

Porous organic polymers have attracted significant attention owing to their large specific surface area, excellent chemical and thermal stability, and controllable skeletons. phenanthroline-based microporous organic polymer (Phen-MOP) has been synthesized via a cost-effective method based on the Scholl reaction. The Phen-MOP polymer exhibits high surface area and good stability. Owing to the phenanthroline skeleton embedding into the microporous polymer framework, the Phen-MOP can serve as a platform to support a transition metal catalyst. After being post-modified with palladium acetate, the synthesized Phen-Pd-MOP framework can serve as a highly efficient heterogeneous catalyst for the Suzuki-Miyaura coupling reaction and the Heck coupling reaction. Moreover, the Phen-Pd-MOP catalyst could be reused at least 10-12 times without any significant loss of the catalytic activity.

7.
Huan Jing Ke Xue ; 40(9): 4230-4237, 2019 Sep 08.
Artículo en Zh | MEDLINE | ID: mdl-31854889

RESUMEN

Nitrous oxide (N2O) is an important greenhouse gas and has a great impact on global warming. Affected by human activities, rivers, and reservoirs have become active sites for N2O production and emission. In Three Gorges Reservoir and the downstream Yangtze River, the concentrations and fluxes of dissolved N2O were investigated in September-October 2009 and October 2016 to identify the factors controlling the distribution of N2O in the reservoir and the effect of damming and reservoir operation on N2O emission from the Yangtze River. The dissolved N2O concentration of the reservoir ranged from 9.74 to 16.36 nmol·L-1 with an average of (12.49±1.75) nmol·L-1 in the surface water (about 0 m) and ranged from 9.99 to 14.00 nmol·L-1 with an average of (11.21±0.91) nmol·L-1 in the bottom water (ranging from 6 m to 103 m). The dissolved N2O in the water column of Three Gorges Reservoir varied little, and no significant difference was noted between the overall N2O concentration in the reservoir and that in the downstream river. A positive correlation was noted between N2O and nitrate (NO3-) (P<0.01), although negative correlations were found between N2O and ammonium (NH4+) (P<0.05) and nitrite (NO2-) (P<0.01) in Three Gorges Reservoir. The surface N2O in the Three Gorges Reservoir was overall saturated, with saturation ranging from 122% to 170% in 2009 and from 114% to 187% in 2016. The mean emission rate of N2O from the surface water was (4.6±2.4) µmol·(m2·d)-1 in 2016 and (16.6±4.9) µmol·(m2·d)-1 in October 2009. The Three Gorges Reservoir is a non-negligible source of atmospheric N2O. No obvious difference was noted between upstream and downstream N2O concentrations, which implies that no degassing emission occurs when water passes through turbines and spillways.

8.
Sci Total Environ ; 666: 1209-1219, 2019 May 20.
Artículo en Inglés | MEDLINE | ID: mdl-30970486

RESUMEN

Rivers and reservoirs are affected by human activities and are sources of the greenhouse gas nitrous oxide (N2O). Concentrations of N2O in the middle and lower reaches of the Yellow River and Xiaolangdi Reservoir, China, were measured in June and December 2017. Fluxes were estimated by boundary layer method to explore their controlling factors, especially the impact of damming and reservoir operation. In the middle and lower reaches of the Yellow River, N2O concentrations in surface waters were 26.65 ±â€¯14.67 nmol L-1 in summer and 21.16 ±â€¯5.35 nmol L-1 in winter. In comparison, the concentrations of N2O in the reservoir were 32.94 ±â€¯17.32 nmol L-1 in summer and 23.73 ±â€¯5.60 nmol L-1 in winter. The longitudinal distribution of N2O along the river exhibited different patterns with surface N2O decreasing downstream towards the dam in summer but increasing in winter. Vertical profiles of N2O concentrations in the reservoir showed an increase with depth in summer but were almost vertically uniform in winter. In winter, N2O that had accumulated in the bottom water in summer was transported to the surface by vertical mixing and released into the atmosphere. Dissolved oxygen (DO), water temperature, and in situ biological production were the main factors affecting the distribution of N2O. The mean emissions rates of N2O from the surface waters were 13.7 ±â€¯8.8 µmol m-2 d-1 in summer and 13.2 ±â€¯7.6 µmol m-2 d-1 in winter. Approximately 1.31 × 106 mol N2O was released from the reservoir surface in 2017, which represents 0.12% of the annual N2O emissions from global reservoirs. The construction of dams increased N2O emission from the lower reaches of the river by 4.53 × 105 mol and 1.22 × 105 mol due to the discharge of the bottom water and the water and sediment regulation, respectively. This study demonstrates that the construction of dams and reservoir operation practices have made the Xiaolangdi Reservoir a key area for N2O emissions.

9.
Int J Ophthalmol ; 11(12): 1922-1925, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-30588423

RESUMEN

AIM: To evaluate surgical outcomes of modified Z-epicanthoplasty with blepharoplasty that we previously reported from the patient's perspective using patient-reported outcome measures (PROMs) and patient satisfaction scores. METHODS: A total of patients (n=180) who underwent the surgery between January 2013 and June 2016 were randomly selected. Standardized patient satisfaction forms (total score, 40) and validated PROMs questionnaires (total score, 12) were sent to patients for completion. PROMs assesses the severity of scarring, pain and asymmetry, as well as functional and appearance issues. RESULTS: All patients were female, ranging from 18 to 35 years old (mean=24). The response rate was 73.3% (n=132). The majority of patients reported good or excellent outcomes based on PROM analysis. Patients reported minimum or non-visible scarring at both the double eyelid surgical scar (85.6%) and the inner canthus (80.3%). Issues concerning function and appearance were minimal as 80.3% reported satisfaction with both domains. Notably, the majority of patients reported either a high or very high satisfaction rate to yield a mean score of 104 out of 120 (P<0.05). CONCLUSION: Integration of our modified Z-epicanthoplasty with blepharoplasty produces good outcomes based on PROM results, which shows a positive linear relationship with patient satisfaction scores.

10.
Chin Med J (Engl) ; 130(6): 691-697, 2017 Mar 20.
Artículo en Inglés | MEDLINE | ID: mdl-28303852

RESUMEN

BACKGROUND: Retroperitoneal fibrosis (RPF) and lymphoma presenting as retroperitoneal mass may closely resemble each other and misdiagnosis may occur. This study investigated the differential imaging features of RPF and lymphoma which presented as a retroperitoneal soft tissue using multidetector-row computed tomography (MDCT). METHODS: The 42 consecutive patients were included in this retrospective review, including 19 RPF patients (45.2%; including 13 males and 6 females; mean age: 56.7 ± 6.2 years) and 23 patients with lymphoma (54.8%; including 14 males and 9 females; mean age: 57.4 ± 12.3 years). An array of qualitative computed tomography (CT) features of lesions in 42 consecutive patients with newly diagnosed untreated RPF and lymphoma were retrospectively analyzed. The quantitative size of the lesion at the para-aortic region and attenuation in the precontrast, arterial, and portal phases were calculated in regions of interest and compared between the patients with newly diagnosed untreated RPF and with lymphoma. Receiver operating characteristic curve analysis was used to assess the potential diagnostic value of each quantitative parameter. Inter-reader concordance was also calculated. RESULTS: Mean ages between patients with RPF and lymphoma were not significantly different (56.7 ± 6.2 years vs. 57.4 ± 12.3 years P = 0.595). Compared to those in patients with lymphoma, homogeneous enhancement (65.2% vs. 94.7%, P = 0.027) and pelvic extension (52.2% vs. 89.5%, P = 0.017) were significantly more common while the involvement of additional nodes (78.3% vs. 5.3%, P < 0.001), suprarenal extension (60.9% vs. 15.8%, P = 0.004), and aortic displacement (43.5% vs. 5.3%, P = 0.006) were significantly less common in patients with RPF. Lesion size at the para-aorta was significantly greater in patients with lymphoma, compared with RPF patients (3.9 ± 1.2 cm vs. 1.8 ± 0.6 cm; P < 0.001). The attenuation values in three phases were not significantly different between patients with RPF and lymphoma. Inter-reader concordance for subjective features ranged from very good to excellent (range: 85.7-100.0%). CONCLUSIONS: This study showed that MDCT can help differentiate between untreated RPF and lymphoma on the basis of qualitative CT features and lesion sizes. Differentiating RPF from lymphoma on the basis of attenuation values in the precontrast, arterial, and portal phases was difficult to accomplish.


Asunto(s)
Linfoma/diagnóstico , Tomografía Computarizada Multidetector/métodos , Fibrosis Retroperitoneal/diagnóstico , Anciano , Diagnóstico Diferencial , Femenino , Humanos , Masculino , Persona de Mediana Edad , Curva ROC , Estudios Retrospectivos
11.
Huan Jing Ke Xue ; 35(1): 157-62, 2014 Jan.
Artículo en Zh | MEDLINE | ID: mdl-24720199

RESUMEN

Different phosphorus compositions (TP, IP and OP) were determined in surface sediments of the Yellow Sea (YS) and the East China Sea (ECS),sampled from two cruises between March and June, 2011. The results showed that, contents of TP were from 10.50 micromol x g(-1) to 24.10 micromol x g(-1), IP ranged from 7.14 micromol x g(-1) to 17.10 micromol x g(-1) was the major phosphorus speciation accounting for more than 70% in TP. The percent of IP in TP at most stations was between 50% and 90% , only four stations which lied in the East China Sea with the percent was over 90%. The dominant factors affecting the phosphorus concentration and distribution in surface sediments were anthropogenic activities, sources of input materials, grain size, depositional environments and hydrological conditions. TP burial flux (TPBF) that appeared regional differences was affected by sedimentation rates, porosity and oxygen in bottom water, but TP content and sedimentation rates were crucial factors for TPBF.


Asunto(s)
Monitoreo del Ambiente , Sedimentos Geológicos/química , Fósforo/análisis , China , Océanos y Mares , Agua de Mar/química
12.
Oncol Lett ; 7(6): 1936-1940, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24932263

RESUMEN

Detection of metastasis of hepatocellular carcinoma (HCC) is crucial for early diagnosis. Epithelial-mesenchymal transition (EMT) is a common event in the metastasis of tumor cells. Slug and Snail are homologous proteins, which play an important role in EMT. The present study aimed to investigate whether Slug and Snail overexpression is associated with the invasiveness of HCC in vitro and in vivo. Invasion, colony formation and wound healing assays, as well as flow cytometry analysis, were performed to examine the invasiveness and proliferation capabilities of HepG2 cells following transfection with cNDA or the siRNA of Slug or Snail. The effects of Slug on HCC in vivo were examined using a xenograft model. Slug upregulation increased the percentage of cluster of differentiation (CD)133+ cells among HepG2 cells, and induced cell invasion and proliferation; whereas Snail upregulation did not affect the cells in vitro. The Slug overexpression group exhibited the highest rate of tumor growth compared with the Snail overexpression and control groups in vivo. These findings demonstrated that Slug increases the percentage of CD133+ cells, promotes the clonigenicity of HCC cells and induces a stronger stemness in Slug-overexpressing cells. These changes activate dormant developmental pathways in invading tumor cells. Thus, Slug may serve as a novel target for HCC prognosis and therapy.

13.
Artículo en Inglés | MEDLINE | ID: mdl-22668427

RESUMEN

OBJECTIVE: We investigated the effectiveness and safety of topical application of 5 mg allicin adhesive tablets in the treatment of minor recurrent aphthous ulcerations (MiRAU). MATERIAL AND METHODS: A randomized, double-blinded, placebo-controlled, clinical trial was performed. Tablets containing 5 mg allicin or vehicle only were consecutively applied 4 times per day for 5 days. The size and pain level of ulcers were measured and recorded on days 1, 2, 4, and 6. RESULTS: A total of 96 subjects with MiRAU fulfilled the study. Allicin adhesive tablets significantly reduced ulcer size (P < .005, P < .003, P < .001 for days 2, 4, and 6, respectively) and alleviated ulcer pain score (P < .03, P < .001, P < .05 for days 2, 4, and 6, respectively) compared with vehicle tablets. Minor and major adverse reactions were not observed. CONCLUSIONS: Allicin adhesive tablets were effective in reducing ulcer size and alleviating ulcer pain of the patients in the treatment of MiRAU without significant side effects.


Asunto(s)
Adhesivos/uso terapéutico , Estomatitis Aftosa/tratamiento farmacológico , Ácidos Sulfínicos/uso terapéutico , Adolescente , Adulto , Distribución de Chi-Cuadrado , Disulfuros , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estadísticas no Paramétricas , Comprimidos , Resultado del Tratamiento , Adulto Joven
14.
Zhonghua Shao Shang Za Zhi ; 28(5): 329-35, 2012 Oct.
Artículo en Zh | MEDLINE | ID: mdl-23290756

RESUMEN

OBJECTIVE: To observe the changes in the structure and cytocompatibility of porcine acellular dermal matrix, which was prepared with dermal reticular layer, treated with matrix metalloproteinase 7 (MMP-7), genipin, and vacuum freeze-drying. METHODS: Fifty-four pieces of porcine dermal reticular layer, prepared with lateral abdominal skin were obtained from healthy large Yorkshire pig with mechanical method under sanitary condition, each 10.0 mm×5.0 mm in size and 0.5 - 0.6 mm in thickness. They were divided into normal control group (A(1), without treatment, n = 6), decellularization group (B, decellularized, n = 12), decellularization + MMP-7 group (C, treated with MMP-7 after decellularization, n = 12), decellularization + MMP-7 + genipin group (D, treated with MMP-7 and genipin after decellularization, n = 12), and decellularization + MMP-7 + genipin + vacuum freeze-drying group (E, treated with MMP-7, genipin, and vacuum freeze-drying after decellularization, n = 12) according to the random number table. Meanwhile, 6 pieces of human acellular dermal matrix, with the same size and thickness as listed above, were taken as control group (A(2), without treatment) in the cytocompatibility tests. HE staining and scanning electron microscope were used to detect the cell number and the change in tissue structure in dermal scaffold in groups A(1) and B-E. Immunohistochemical staining was used to determine residual vimentin, laminin and collagen IV in groups A(1), B, and C. Cytotoxicity tests were employed to test the cytotoxicity of the leaching solutions of groups B-E. Human fibroblasts were seeded on the surface of dermal scaffold in groups A(2) and B-E. The proliferation of fibroblasts were determined on post culture day (PCD) 3, 7, and 14, and the content of IL-6 and IL-8 in the supernatant were determined on PCD 3 and 7 with enzyme-linked immunosorbent assay. Data were processed with two-way analysis of variance and LSD- t test. RESULTS: Granular structure with hair follicle in pale yellow color was observed in group A(1). Small amount of hair, epithelial root sheath, nuclei, cell debris-like structure, vimentin, laminin, and collagen IV were observed in group B but not in group C, D, or E, which had been treated with MMP-7. The toughness of dermal scaffold was stronger in groups D, E than in groups B and C as observed in gross condition observation. The collagen fibers of dermal scaffold in groups C-E maintained their structural integrity with similar arrange as that of group A(1). The interspaces among collagen fibers in groups C-E were all increased, while those of groups C and D were similar but larger than that in group B; the interspace in group E was the largest. Groups B-E scored level 0 or 1 in the cytotoxicity test. Fibroblasts could proliferate on the surface of dermal scaffold in groups A(2) and B-E. Furthermore, with the extension of culture time, fibroblasts gradually became to be stratified to form multiple layers, and they proliferated toward the dermis. High density of fibroblasts was observed on the surface in groups D and E and in the deep layer in groups A(2) and C. On PCD 7, the contents of IL-6 [(132 ± 14), (104 ± 9), (122 ± 14), (120 ± 12), (128 ± 17) pg/mL] and IL-8 [(135 ± 18), (102 ± 17), (127 ± 18), (134 ± 23), (141 ± 24) pg/mL] in the supernatant in groups A(2) and B-E were significantly higher than those on PCD 3 [(55 ± 13), (34 ± 8), (48 ± 8), (50 ± 13), (49 ± 12) pg/mL] and [(93 ± 19), (63 ± 11), (82 ± 15), (82 ± 16), (89 ± 16) pg/mL], with F values respectively 98.869, 184.038, 125.531, 93.237, 87.265 and 15.694, 23.451, 22.801, 19.607, 18.808, P values below 0.05. The differences among groups A(2) and B-E in the levels of IL-6 and IL-8 at each time point were statistically significant (with F values respectively 2.809, 3.301 and 3.757, 3.266, P values below 0.05). The differences among groups A(2), C, D, and E in amount of IL-6 and IL-8 at each time point were not statistically significant (with t values respectively 0.058 - 1.905 and 0.034 - 1.295, P values above 0.05), but they were all higher than those in group B (with t values respectively 3.707 - 5.612 and 2.785 - 4.079, P values below 0.05). CONCLUSIONS: The low immunogenic porcine dermal scaffold treated with MMP-7, genipin, and vacuum freeze-drying after decellularization, has good cytocompatibility. The growth of only a few fibroblasts in the dermal scaffold may be correlated with genipin, which increases tissue toughness.


Asunto(s)
Dermis/trasplante , Histocompatibilidad , Andamios del Tejido , Dermis Acelular , Animales , Células Cultivadas , Humanos , Trasplante de Piel , Porcinos , Cicatrización de Heridas
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