Immunohistochemical and gene expression analysis of autologous platelet rich fibrin for distal limb wound defects healing in donkeys (Equus asinus).

2020 ◽  
Vol 21 (1) ◽  
pp. 46-55
Author(s):  
Mohamed Albahrawy ◽  
Khaled Abouelnasr ◽  
Mohamed Hamed ◽  
Mohamed EL-Adl ◽  
Esam Mosbah ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Expression Patterns ◽  
Growth Factor Receptor ◽  
Immunohistochemical Analysis ◽  
Tissue Growth ◽  
Control Group ◽  
Distal Limb ◽  
Platelet Rich Fibrin ◽  
Significant Difference

Objective: To evaluate the effect of platelet-rich fibrin (PRF) in the promotion of distal limb wound defects healing in donkeys. Design: A randomized experimental design Animals: Twelve clinically healthy male donkeys, weighing, 130–230 kg and aged 4 –5 years were allocated into three groups(4 animals/each) and undergo a 6cm2 (2cm X 3cm) 2 wound defects on the dorsolateral surface of right metacarpal and metatarsal regions for each donkey. Control (group A): the wound defects were left for spontaneous healing. In groups B and C, the wound defects were treated with either one application of PRF (B) or with three consecutive applications of PRF (a week interval) (C). Wound defects healing were evaluated clinically, histologically and immunohistochemically, in addition to gene expression patterns of angiogenic and myofibroblastic genes vascular endothelial growth factor (VEGF-A), collagen type 3 α1 (COL3α1), and fibroblast growth factor 7 (FGF-7) and tissue growth factor β1 (TGFβ1) were performed. Results: The healing percentage of single and three PRF applications was significantly higher (P <0.05) (84.6%, and 93.7% respectively) than in control one (66.7%). The number of days needed for complete wound healing was considerably shorter in repeated PRF treated wound defects (63.2±2.8) compared with single PRF and untreated wound defects (71.6±3 and 86.3±3, respectively). Semi-quantitative evaluation of histological sections at 15 and 45 days post-operative showed a significant difference (P<0.05) in epithelization, PMNL, fibroblasts, tissue macrophages, neo-angiogenesis and new collagen scores in both PRF groups compared to control one. Qualitative analysis of immunohistochemical views of the wound defects showed a significant immunostaining difference against EGFR, VEGF, and TGFβ stain between both PRF treated groups and control one. Immunohistochemical analysis of cells stained for epidermal growth factor receptor (EGFR), VEGF, and TGFβ at 15 and 45 days after interference was higher in both PRF treated groups compared to control one, but three PRF application showed the highest rates. The relative expression of FGF-7, TGFβ1, VEGF-A, and COL3α1 genes was higher in both PRF groups compared to control one, but the triple PRF group revealed the highest expression. Conclusion and clinical relevance: Application of PRF could improve the healing of distal limb wound defects in donkeys.

The effect of bevacizumab on targeting of anti-epidermal growth factor receptor and anti-insulin-like growth factor 1 receptor antibodies.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 3035-3035
Author(s):  
Sandra Heskamp ◽  
Otto C. Boerman ◽  
Janneke D.M. Molkenboer-Kuenen ◽  
Wim J.G. Oyen ◽  
Winette T.A. Van Der Graaf ◽  
...  
Keyword(s):  
Growth Factor ◽  
Ex Vivo ◽  
Growth Factor Receptor ◽  
Progression Free Survival ◽  
Immunohistochemical Analysis ◽  
Control Group ◽  
Vascular Density ◽  
Tumor Vascularity ◽  
Bevacizumab Treatment ◽  
Ex Vivo Biodistribution

3035 Background: Bevacizumab and cetuximab are approved antibodies for treatment of patients with metastasized colorectal cancer. However, the combination of bevacizumab and cetuximab does not improve progression free survival (Tol et al. NEJM 2009). This may be explained by the disruption of tumor vascularity by bevacizumab, thereby reducing targeting of other antibodies to the tumor. The aim of this study was to determine the effect of bevacizumab on the targeting of anti-EGFR and IGF-1R antibodies in tumors with SPECT/CT imaging. Methods: Mice with subcutaneous EGFR and IGF-1R-expressing SUM149 xenografts were injected intraperitoneally with a single dose of bevacizumab (10 mg/kg). After four days, mice received an intravenous injection of 17 MBq 111In-labeled cetuximab, an anti-EGFR antibody, or R1507, an anti-IGF-1R antibody. A control group was injected with labeled hLL2 anti-CD22, an irrelevant IgG . Three days after injection, SPECT/CT images were acquired and mice were dissected for ex vivo biodistribution. Tumors were analyzed immunohistochemically to determine vascular density (CD34), EGFR and IGF-1R expression. Results: SPECT imaging revealed that bevacizumab treatment reduced targeting of anti-EGFR and anti-IGF-1R antibodies by 42% and 35%, respectively. Ex vivo biodistribution showed that uptake of 111In-cetuximab in untreated tumors was 35.2 ± 1.6 %ID/g, compared with 19.7 ± 5.3 %ID/g for bevacizumab treated tumors (p = 0.009). A similar effect was observed for 111In-R1507 (control: 26.7 ± 2.8 %ID/g, bevacizumab:18.9 ± 2.8 %ID/g, p =0.009). No significant differences in tumor uptake were observed in mice that received the irrelevant IgG. Immunohistochemical analysis showed that vascular density decreased with 40%, while EGFR and IGF-1R expression was unaltered. Conclusions: Bevacizumab treatment can significantly reduce targeting of other antibodies to tumors. This underlines the importance of timing and sequencing of bevacizumab therapy in combination with other antibodies.

212 EXPRESSION OF FIBROBLAST GROWTH FACTOR 18 (FGF18) AND COGNATE RECEPTORS (FGFR3C AND FGFR4) DURING LUTEAL DEVELOPMENT AND INDUCED LUTEOLYSIS IN CATTLE

2010 ◽  
Vol 22 (1) ◽  
pp. 264
Author(s):  
D. M. Guerra ◽  
A. C. S. Castilho ◽  
M.F. Machado ◽  
B. Berisha ◽  
D. Schams ◽  
...  
Keyword(s):  
Growth Factor ◽  
Mrna Expression ◽  
Blood Vessels ◽  
Fibroblast Growth Factor ◽  
Fibroblast Growth Factor Receptor ◽  
Expression Patterns ◽  
Growth Factor Receptor ◽  
Immunohistochemical Analysis ◽  
Fibroblast Growth ◽  
Santa Cruz

Fibroblast growth factor receptor 2 (FGFR2) has been shown to induce luteinization in granulosa cells, luteal angiogenesis, and luteal growth. Alternative splicing of 4 genes give rise to 7 subtypes of fibroblast growth factor receptors (FGFR) with varying affinity for different fibroblast growth factors (FGF). Fibroblast growth factor receptor 2 and FGF18 efficiently activate FGFR3C and FGFR4 and may act in cooperation in tissues expressing these receptors. We aimed to determine mRNA expression patterns for FGF18, FGFR3C, and FGFR4 during bovine luteal development and following induced luteolysis. In addition, we assessed FGF18 localization in the bovine CL. Bovine CL were obtained from abattoir ovaries and classed into 4 stages of development: stage 1 =corpus hemorragicum; stage 2 = developing CL; stage 3 = mature or early functional luteolysis CL; and stage 4 = structural luteolysis. To assess FGF18 and FGFR mRNA expression during induced luteolysis, adult cows (Bos taurus Holstein-Friesians) were injected with the PGF2 analogue cloprostenol (500 mg i.m. Intervet, Unterschleissheim, Germany) during the mid-luteal phase of the cycle (Days 8-12). Corpus luteum were collected by transvaginal ovariectomy at 0, 0.5, 2, 4, 12, 24, 48, and 64 hr (n = 5/time point) after PGF2 injection. Tissue samples were submitted to total RNA extraction. Expression of FGF18, FGFR3C, and 4 mRNA during the bovine CL lifespan and induced luteolysis were measured by real-time RT-PCR with oligo-dT in the RT and bovine-specific primers in the PCR. Expression of cyclophilin was used as internal control. The effect of developmental stage and time post-PGF2 on gene expression was tested by ANOVA, followed by Tukey- Kramer HSD test. Immunohistochemical analysis was performed with a commercial human antibody (anti-FGF18; Santa Cruz Biotechnology, Santa Cruz, CA, USA). Fibroblast growth factor 18, FGFR3C, and FGFR4 mRNA was detected in all 4 developmental stages; FGF18 mRNA abundance was higher in stage 3 (2.89 ± 0.05; mean ± SEM) compared with stages 1 (0.3 ± 0.27), 2 (0.56 ± 1.27), and 4 (0.99 ± 0.32). Fibroblast growth factor 18 and FGFR4 mRNA expression did not significantly change during induced luteolysis. Fibroblast growth factor receptor 3C mRNA abundance peaked 4 h after PGF2 injection and significantly decreased at 24 h post-treatment in comparison with peak levels. Immunohistochemical analysis revealed the presence of FGF18 in small and large luteal cells and in blood vessels. In conclusion, the mRNA expression patterns of FGF18 and its receptors suggest their participation in the control of luteal differentiation, particularly during functional luteolysis. The localization of FGF18 protein to blood vessels suggests it may play a role in the control of angiogenesis in the bovine CL. Supported by CAPES/FAPESP.

scholarly journals Protective effect of trimetazidine in radiation-induced cardiac fibrosis in mice

2020 ◽  
Vol 61 (5) ◽  
pp. 657-665
Author(s):  
Jinmeng Zhang ◽  
Xinjia He ◽  
Xinya Bai ◽  
Yang Sun ◽  
Peng Jiang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Molecular Mechanism ◽  
Cardiac Fibrosis ◽  
Tissue Growth ◽  
Control Group ◽  
High Dose ◽  
X Rays ◽  
Radiation Induced ◽  
Tgf Β1

Abstract Radiation-induced heart damage is a serious side effect caused by radiotherapy, especially during the treatment of cancer near the chest. Trimetazidine is effective at reducing inflammation in the heart, but how it affects radiation-induced cardiac fibrosis (RICF) is unknown. To investigate the potential effect and molecular mechanism, we designed this project with a C57BL6 male mouse model supposing trimetazidine could inhibit RICF in mice. During the experiment, mice were randomly divided into six groups including a control group (Con), radiation-damaged model group (Mod) and four experimental groups receiving low-dose (10 mg/kg/day) or high-dose (20 mg/kg/day) trimetazidine before or after radiation treatment. Apart from the control group, all mice chests were exposed to 6 MV X-rays at a single dose of 20 Gy to induce RICF, and tissue analysis was done at 8 weeks after irradiation. Fibroblast or interstitial tissues and cardiac fibrosis-like characteristics were determined using haematoxylin and eosin and Masson staining, which can be used to assess myocardial fibrosis. Immunohistochemical analysis and RT-PCR were used to determine gene expression and study the molecular mechanism. As a result, this study suggests that trimetazidine inhibits RICF by reducing gene expression related to myocyte apoptosis and fibrosis formation, i.e. connective tissue growth factor (CTGF), transforming growth factor (TGF)-β1, smad2 and smad3. In conclusion, by regulating the CTGF/TGF-β1/Smad pathway, trimetazidine could be a prospective drug for clinical treatment of RICF.

scholarly journals Dopamine adjusts the circadian gene expression of Per2 and Per3 in human dermal fibroblasts from ADHD patients

2021 ◽  
Author(s):  
Frank Faltraco ◽  
Denise Palm ◽  
Adriana Uzoni ◽  
Lena Borchert ◽  
Frederick Simon ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dermal Fibroblast ◽  
Sleep Onset ◽  
Dermal Fibroblasts ◽  
Control Group ◽  
Adhd Group ◽  
Healthy Controls ◽  
Circadian Gene ◽  
Significant Difference ◽  
Circadian Preference

AbstractA link between dopamine levels, circadian gene expression, and attention deficit hyperactivity disorder (ADHD) has already been demonstrated. The aim of this study was to investigate the extent of these relationships by measuring circadian gene expression in primary human-derived dermal fibroblast cultures (HDF) after dopamine exposure. We analyzed circadian preference, behavioral circadian and sleep parameters as well as the circadian gene expression in a cohort of healthy controls and participants with ADHD. Circadian preference was evaluated with German Morningness-Eveningness-Questionnaire (D-MEQ) and rhythms of sleep/wake behavior were assessed via actigraphy. After ex vivo exposure to different dopamine concentrations in human dermal fibroblast (HDF) cultures, the rhythmicity of circadian gene expression (Clock, Bmal1, Per1-3, Cry1) was analyzed via qRT-PCR. We found no statistical significant effect in the actigraphy of both groups (healthy controls, ADHD group) for mid-sleep on weekend days, mid-sleep on weekdays, social jetlag, wake after sleep onset, and total number of wake bouts. D-MEQ scores indicated that healthy controls had no evening preference, whereas subjects with ADHD displayed both definitive and moderate evening preferences. Dopamine has no effect on Per3 expression in healthy controls, but produces a significant difference in the ADHD group at ZT24 and ZT28. In the ADHD group, incubation with dopamine, either 1 µM or 10 µM, resulted in an adjustment of Per3 expression to control levels. A similar effect also was found in the expression of Per2. Statistical significant differences in the expression of Per2 (ZT4) in the control group compared to the ADHD group were found, following incubation with dopamine. The present study illustrates that dopamine impacts on circadian function. The results lead to the suggestion that dopamine may improve the sleep quality as well as ADHD symptoms by adjustment of the circadian gene expression, especially for Per2 and Per3.

Long-term treadmill training ameliorates endothelium-dependent vasorelaxation mediated by insulin and insulin-like growth factor-1 in hypertension

2015 ◽  
Vol 119 (6) ◽  
pp. 663-669 ◽  
Author(s):  
Yi-Yuan Lin ◽  
Shin-Da Lee ◽  
Chia-Ting Su ◽  
Tsung-Lin Cheng ◽  
Ai-Lun Yang
Keyword(s):  
Growth Factor ◽  
Insulin Receptor ◽  
Treadmill Training ◽  
Control Group ◽  
Insulin Like Growth Factor ◽  
Hypertensive Rats ◽  
Protein Levels ◽  
Significant Difference ◽  
Nos Inhibitors

Dysfunction of insulin and insulin-like growth factor-1 (IGF-1) is associated with the pathophysiology of hypertension. The influence of long-term exercise on vascular dysfunction caused by hypertension remains unclear. We investigated whether long-term treadmill training improved insulin- and IGF-1-mediated vasorelaxation in hypertensive rats. Eight-week-old male spontaneously hypertensive rats (SHR) were randomly divided into sedentary and exercise (SHR-EX) groups. The SHR-EX group was trained on a treadmill for 60 min/day, 5 days/wk, for 8 wk. Wistar-Kyoto rats (WKY) were used as the normal control group. After training, aortic insulin- and IGF-1-mediated vasorelaxation was evaluated in organ baths. Additionally, the roles of phosphatidylinositol 3-kinase (PI3K), nitric oxide synthase (NOS), and aortic protein expression were examined in the three groups. Compared with sedentary SHR and WKY groups, insulin- and IGF-1-mediated vasorelaxation was significantly enhanced to a nearly normal level in the SHR-EX group. After endothelial denudation, blunted and comparable vasorelaxation was found among the three groups. Pretreatment with selective PI3K and NOS inhibitors attenuated insulin- and IGF-1-mediated vasorelaxation, and no significant difference was found among the three groups after the pretreatment. The aortic protein levels of the insulin receptor (IR), IGF-1 receptor (IGF-1R), insulin receptor substrate-1 (IRS-1), and endothelial NOS (eNOS) were also significantly increased in the SHR-EX group compared with the other two groups. These results suggested that treadmill training elicited the amelioration of endothelium-dependent insulin/IGF-1-mediated vasorelaxation partly via the increased activation of PI3K and NOS, as well as the enhancement of protein levels of IR, IGF-1R, IRS-1, and eNOS, in hypertension.

scholarly journals Insulin-like Growth Factor Receptor 1 (IGF1R) Gene Copy Number Is Associated With Survival in Operable Non–Small-Cell Lung Cancer: A Comparison Between IGF1R Fluorescent In Situ Hybridization, Protein Expression, and mRNA Expression

2010 ◽  
Vol 28 (13) ◽  
pp. 2174-2180 ◽  
Author(s):  
Rafal Dziadziuszko ◽  
Daniel T. Merrick ◽  
Samir E. Witta ◽  
Adelita D. Mendoza ◽  
Barbara Szostakiewicz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Protein Expression ◽  
Squamous Cell ◽  
Copy Number ◽  
Gene Copy Number ◽  
Growth Factor Receptor ◽  
Gene Copy ◽  
Cell Versus

PurposeThe purpose of this study was to characterize insulin-like growth factor-1 receptor (IGF1R) protein expression, mRNA expression, and gene copy number in surgically resected non–small-cell lung cancers (NSCLC) in relation to epidermal growth factor receptor (EGFR) protein expression, patient characteristics, and prognosis.Patients and MethodsOne hundred eighty-nine patients with NSCLC who underwent curative pulmonary resection were studied (median follow-up, 5.3 years). IGF1R protein expression was evaluated by immunohistochemistry (IHC) with two anti-IGF1R antibodies (n = 179). EGFR protein expression was assessed with PharmDx kit. IGF1R gene expression was evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR) from 114 corresponding fresh-frozen samples. IGF1R gene copy number was assessed by fluorescent in situ hybridization using customized probes (n = 181).ResultsIGF1R IHC score was higher in squamous cell carcinomas versus other histologies (P < .001) and associated with stage (P = .03) but not survival (P = .46). IGF1R and EGFR protein expression showed significant correlation (r = 0.30; P < .001). IGF1R gene expression by qRT-PCR was higher in squamous cell versus other histologies (P = .006) and did not associate with other clinical features nor survival (P = .73). Employing criteria previously established for EGFR copy number, patients with IGF1R amplification/high polysomy (n = 48; 27%) had 3-year survival of 58%, patients with low polysomy (n = 87; 48%) had 3-year survival of 47% and patients with trisomy/disomy (n = 46; 25%) had 3-year survival of 35%, respectively (P = .024). Prognostic value of high IGF1R gene copy number was confirmed in multivariate analysis.ConclusionIGF1R protein expression is higher in squamous cell versus other histologies and correlates with EGFR expression. IGF1R protein and gene expression does not associate with survival, whereas high IGF1R gene copy number harbors positive prognostic value.

Sign in / Sign up

Export Citation Format

Share Document