Growth Factor-β1 In Fibroblasts Research Articles

The association of infant urinary adrenal steroids with the risk of childhood asthma development

BackgroundAdrenal steroids play important roles in early-life development. However, our understanding of the effects of perinatal adrenal steroids on the development of childhood asthma is incomplete. ObjectiveTo evaluate the associations between early-life adrenal steroid levels and childhood asthma. MethodsParticipants included the Infant Susceptibility to Pulmonary Infections and Asthma following Respiratory Syncytial Virus Exposure birth cohort children with untargeted urinary metabolomics data measured during early infancy (n = 264) and nasal immune mediator data measured concurrently at age 2 to 6 months (n = 76). A total of 11 adrenal steroid compounds were identified using untargeted metabolomics and 6 asthma-relevant nasal immune mediators from multiplex assays were a priori selected. Current asthma at ages 5 and 6 years was ascertained using validated questionnaires. Associations were tested using logistic and linear regression with confounders adjustment. ResultsPregnenetriol disulfate (adjusted odds ratio [aOR] = 0.20, 95% CI = 0.06-0.68) and 3a,21-dihydroxy-5b-pregnane-11,20-dione-21-glucuronide (aOR = 0.34, 95% CI = 0.14-0.75) were inversely associated with childhood asthma at 5 and 6 years after multiple testing adjustment. There was a significant interaction effect of pregnanediol-3-glucuronide by biological sex assigned at birth (aOR = 0.11, 95% CI = 0.02-0.51, for those with female sex) on childhood asthma. Pregnenetriol disulfate was inversely associated with granulocyte-macrophage colony-stimulating factor (β = −0.45, q-value = 0.05). 3a,21-dihydroxy-5b-pregnane-11,20-dione 21-glucuronide was inversely associated with interleukin [IL]-4 (β = −0.29, q-value = 0.02), IL-5 (β = −0.35, q-value = 0.006), IL-13 (β = −0.26, q-value = 0.02), granulocyte-macrophage colony-stimulating factor (β = −0.35, q-value = 0.006), and fibroblast growth factor-β (β = −0.24, q-value = 0.01) after multiple testing adjustment. ConclusionThe inverse association between adrenal steroids downstream of progesterone and 17-hydroxypregnenolone and the odds of childhood asthma and nasopharyngeal type 2 immune biomarkers suggest that increased early-life adrenal steroids may suppress type 2 inflammation and protect against the development of childhood asthma.

Experimental conditions and protein markers for redifferentiation of human coronary artery smooth muscle cells.

A phenotype switch from contractile type to proliferative type of arterial smooth muscle cells is known as dedifferentiation, but to the best of our knowledge, little is known about redifferentiation of coronary artery smooth muscle cells. The purpose of the present study was to determine in vitro culture conditions for inducing redifferentiation of coronary artery smooth muscle cells. In addition, the present study aimed to determine protein markers for detection of redifferentiated arterial smooth muscle cells. Human coronary artery smooth muscle cells (HCASMCs) were cultured in the presence or absence of growth factors, including epidermal growth factor, fibroblast growth factor-B and insulin. Protein expression and migration activity of HCASMCs were evaluated using western blotting and migration assay, respectively. In HCASMCs 5 days after 100% confluency, expression levels of α-smooth muscle actin (α-SMA), calponin, caldesmon and SM22α were significantly increased, while expression levels of proliferation cell nuclear antigen (PCNA) and S100A4 and migration activity were significantly decreased, compared with the corresponding levels just after reaching 100% confluency, indicating that redifferentiation occurred. Redifferentiation was also induced in a low-density culture of HCASMCs in the medium without growth factors. When the culture medium for confluent cells was replaced daily with fresh medium, the expression levels of α-SMA, caldesmon, SM22α, PCNA and S100A4 and migration activity were not significantly different but the calponin expression was significantly increased compared with the levels in dedifferentiated cells just after reaching 100% confluency. Thus, redifferentiation was induced in HCASMCs by deprivation of growth factors from culture medium. The results suggested that α-SMA, caldesmon and SM22α, but not calponin, are markers of redifferentiation of HCASMCs.

Expression of fibroblast growth factor-β and transforming growth factor-β in mauli banana stem (Musa Acuminate) extract gel - treated traumatic ulcer

Purpose: To investigate the effect of mauli banana (Musa acuminate) stem topical gel extract application on the expression of transforming growth factor - β (TGF - β) and fibroblast growth factor-β (FGF-β) during the healing process in traumatic oral ulcers.
 Methods: The work represented a true experimental study incorporating a post test - only control group design. Four groups of male Wistar rats (Rattus norvegivus) (n = 20) with traumatic oral ulcers were given mauli banana stem extract gel of varying concentrations the negative control group: 0 %; treatment group I: 25 %; treatment group II: 37.5 %; and treatment group III: 50 %. The animals were subsequently sacrificed prior to conducting biopsy on day 5. Immunohistochemical staining was performed in order to analyze the degree of FGF-β and TGF–β expressions.
 Results: TGF–β was strongly expressed in treatment group II (16.80 ± 1.30). TGF-β expression was significantly different, except between treatment groups II and III (Table 2). FGF-β was strongly expressed in treatment group II (15.60 ± 3.97). There was significant difference in FGF-β expression between all the groups with the exception of treatment groups I and III.
 Conclusion: Topical application of mauli banana stem extract gel (37.5 % concentration) stimulates FGF-β and TGF-β expression on day 5 of traumatic oral ulcer healing process. Thus, the extract gel has potentials for clinical application for the therapy of traumatic oral ulcers

The effect of halofugınone and pirfenidone on wound healing in experimental glaucoma filtration surgery

To investigate the effects of halofuginone and pirfenidone on wound healing in experimental glaucoma filtration surgery (GFS). Animal experimentation. A total of 42 male New Zealand albino rabbits were separated into 6 equal groups. A limbal-based trabeculectomy was performed on 5 groups, and Group I (control group) underwent no surgery and received no postoperative medication. For Group II (sham group), 1 drop 0.9% NaCl was instilled qid for 14 days. For Group III, 1% topical corticosteroid (prednisolone acetate) was instilled 1 drop qid for 14 days. For Group IV, 0.4mg/mL mitomycin-C (MMC) was applied intraoperatively to the region of the scleral flap. For Group V, 0.5% pirfenidone was instilled 1 drop qid for 14 days postoperatively. For Group VI, a sponge soaked in 10ng/mL halofuginone was applied to the surgical region for 3 mins. In addition, 1% topical corticosteroid was instilled ×1 drop qid for 14 days postoperatively for Groups IV, V and VI. After 14 days, sections prepared from the bleb regions of the enucleated eyes were evaluated histopathologically and immunohistochemically. Statistical analyses of the study were performed with Kruskal-Wallis variance analysis and the Mann-Whitney U test. With regard to fibroblasts, suppression of the number of mononuclear cells and immunohistochemical staining intensity of transforming growth factor-b (TGF-β), fibroblast growth factor-b (FGF-β) and platelet derived growth factor (PDGF), the corticosteroid, MMC, pirfenidone and halofuginone groups were seen to exhibit more effect than the sham group (P<0.05). Compared to the pirfenidone and fuginone groups, inhibition of fibroblast and monocyte proliferation was determined to be lower in the MMC group (P<0.05). The intensity of TGF-β and FGF-β staining was seen to be lower in these two treatment groups than in the MMC group (P<0.05). Halofuginone and pirfenidone may be used as effective alternative agents in delaying wound healing in glaucoma filtration surgery.

Differentiation of Mesenchymal Stem Cells from Humans and Animals into Insulin-producing Cells: An Overview In Vitro Induction Forms.

The use of stem cells in cell therapies has shown promising results in the treatment of several diseases, including diabetes mellitus, in both humans and animals. Mesenchymal stem cells (MSCs) can be isolated from various locations, including bone marrow, adipose tissues, synovia, muscles, dental pulp, umbilical cords, and the placenta. In vitro, by manipulating the composition of the culture medium or transfection, MSCs can differentiate into several cell lineages, including insulin-producing cells (IPCs). Unlike osteogenic, chondrogenic, and adipogenic differentiation, for which the culture medium and time are similar between studies, studies involving the induction of MSC differentiation in IPCs differ greatly. This divergence is usually evident in relation to the differentiation technique used, the composition of the culture medium, the cultivation time, which can vary from a few hours to several months, and the number of steps to complete differentiation. However, although there is no "gold standard" differentiation medium composition, most prominent studies mention the use of nicotinamide, exedin-4, ß-mercaptoethanol, fibroblast growth factor b (FGFb), and glucose in the culture medium to promote the differentiation of MSCs into IPCs. Therefore, the purpose of this review is to investigate the stages of MSC differentiation into IPCs both in vivo and in vitro, as well as address differentiation techniques and molecular actions and mechanisms by which some substances, such as nicotinamide, exedin-4, ß-mercaptoethanol, FGFb, and glucose, participate in the differentiation process.

Abstract 1423: The addition of bevacizumab to chemoimmunotherapy prolongs progression-free survival in patients with chronic lymphocytic leukemia (CLL) through modulation of the microenvironment

Abstract Introduction. First-line chemoimmunotherapy (CIT) has significantly increased the complete remission (CR) rate for patients with CLL; however, relapse is a common event and strategies aimed at eradicating disease more effectively are needed. Vascular Epithelial Growth Factor (VEGF) plays a crucial role in the cross talk between CLL B cells and their microenvironment. In addition low baseline VEGF levels predict a better response to CIT. Testing the combination of an anti-VEGF agent with CIT was therefore assessed by us in upfront CLL patients. Methods. Here we report the results of a phase II open-label randomized trial comparing the combination of bevacizumab, an anti-VEGF monoclonal antibody, with pentostatin, cyclophosphamide and rituximab (PCR-B) to PCR alone as front-line therapy in previously untreated patients with CLL. VEGF, b-fibroblast growth factor (FGF), thrombospondin (TSP)-1, chemokine ligand (CCL)-3 and CCL-4 plasma levels were measured at baseline and at time of response assessment. Results. Between 01/2009 and 01/2013, 62 patients were accrued in the study, 32 in the arm A (PCR-B) and 30 in arm B (PCR alone). A higher rate of grade 3-4 cardiovascular toxicity was observed with the use of PCR-B (34% vs 0%, p&amp;lt;0.001): this included 7 cases of hypertension, 2 cases of congestive heart failure, 1 myocarditis, and 1 case of torsade de point. A higher CR rate (50% vs 33%, p = 0.21) and a significantly longer median progression-free (p = 0.04) and treatment-free (p = 0.05) survival were achieved with the use of PCR-B. No difference in chemokine kinetics was observed between the 2 arms, except for VEGF, which significantly increased after PCR-B. In the PCR-B arm, chemokine kinetics for CCL-3 (median drop 5.0 vs. median drop 31.8, respectively; p = 0.02) and CCL-4 (median drop 23.8 vs. median drop 120.9, respectively; p = 0.01) were significantly associated with achievement of CR. No significant correlations were found between cytokine plasma levels and responses for patients treated in the PCR only arm. Conclusions. The addition of bevacizumab to CIT is safe and effective, although associated with higher cardiovascular toxicity, in particular hypertension. The addition of bevacizumab produces higher CR rates, translating into improved progression-free survival, and is associated with decreased plasma levels of CCL-3 and CCL-4. Citation Format: PAOLO STRATI, TAIT D. SHANAFELT, BETSY LAPLANT, ADAM PETTINGER, CONNIE LESNICK, DANIEL NIKCEVICH, TIMOTHY CALL, WEI DING, CURTIS HANSON, NEIL KAY. The addition of bevacizumab to chemoimmunotherapy prolongs progression-free survival in patients with chronic lymphocytic leukemia (CLL) through modulation of the microenvironment. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1423.

Mesenchymal Cell Associated Fibrosis in Experimental mplW515L Mouse Model of Myelofibrosis

Background: Bone marrow fibrosis caused by excessive collagen deposition represents a significant indicator of poor prognosis in myeloproliferative neoplasms (MPNs). Genetic driver mutations have been identified as JAK2, CALR and mpl, but the precise mechanism and cell type leading to fibrosis remains unanswered. TGFβ production by mutant hematopoietic cells is the primary factor responsible for fibrosis initiation. Although effective JAK2 inhibitor therapy has been adopted to manage disease symptoms and decrease abnormal hematopoietic cells, this pharmacological inhibitor does not reverse fibrosis. Our previous observations in patients with Large granular lymphocyte (LGL) leukemia indicate that mesenchymal-derived fibroblasts, distinguished by the capacity to undergo trilineage differentiation into osteoblasts, adipocytes and chondrocytes, are responsible for the fibrosis observed in these patients. In this study, we report the identification of a similar fibrosis-initiating mesenchymal cell population in a mouse model of MPN induced by transplantation of gfp-mplW515L retrovirus-expressing hematopoietic cells and indicate a mechanism to reverse the fibrotic phenotype.Methods: Primary mesenchymal stromal cells (MSCs) were isolated from mice with MPN caused by the transplantation of gfp-mplW515L -transduced bone marrow cells (MPN mice) and from control mice that received transplants with gfp-wild-type(wt)-mpl-expressing bone marrow. MSCs were harvested 17 days post-transplant when WBC counts exceeded at least >20,000 cells/μl. Fibrillar collagen I (detected with trichrome stain in bone marrow biopsies) and collagen III (detected with reticulin stain in bone marrow biopsies) were identified as the major proteins produced in the bone marrow and by cultured MSCs in MPN mice consistent with pathologic examination. The amount of collagen was quantified in MSCs grown under reduced oxygen in undifferentiating culture conditions. To determine pluripotency, cells were grown with differentiation-specific culture media and stained with Alizarin Red S (to detect osteoblasts) and Alcian blue (chondrocytes). Collagen production was determined by qRT-PCR and polychromatic immunofluorescence staining of three-dimensional cultures and TGFβ was measured by qRT-PCR and ELISA.Results: To isolate purified populations of MSCs, gfp-positive hematopoietic cells were removed and MSCs were maintained in long-term culture in the absence of malignant cells. MSCs were isolated in micro-cultures from gfp-wt-mpl (ie, control) and mplW515L transplanted mice. Using gfp-wt-mpl-derived MSCs, we first showed that the addition of TGFβ induced collagen I and III expression as expected from previous studies. Original undifferentiated MSCs derived from control and MPN mice grew continuously in undifferentiated culture conditions with a similar life expectancy, and both populations were capable of further differentiation into osteoblasts and chondrocytes. Collagen I (p<0.0001) and III (p=0.0008) mRNA and fiber formation were increased in the MSCs from mplW515L -transplanted mice compared to control. Without exogenous cytokines or HSC supernatant, the MSCs from mutant-mpl-mice maintained greater than 2-fold increase in collagen suggesting that intrinsic reprogramming occurred in the MSC population independent of tumor cells that are normally considered the source of TGFβ and other fibrosis-initiating cytokines. TGFβ mRNA (0.9878±0.129, control vs 1.023±0.319, MPN p=ns) and total TGFβ secreted protein detected by ELISA (165.0±22.7pg/ml, control vs 144.1±21.8pg/ml, MPN p=ns) was similar in control and MPN-derived MSCs indicating that excessive TGFβ production does not maintain the fibrotic phenotype in culture. We have shown previously that collagen from fibrotic MSCs in LGL leukemia is inhibited by culturing the cells with fibroblast growth factor-b (FGFb, FGF-2). Similar to LGL leukemia-dervied MSCs, the addition of FGFb stimulated proliferation and restored normal levels of collagen I and III in the MSCs from MPN mice.Conclusions: This study indicates that expanded MPN-derived MSCs can be re-programmed to produce normal amounts of collagen, which may reverse fibrosis. This study has implications for the development of MSC-based cell therapies in MPN. DisclosuresNo relevant conflicts of interest to declare.

Abstract 19213: Epigenetically Reprogrammed Mesenchymal Stem Cells Improve Myocardial Remodeling and Cardiac Function After Myocardial Infarction by Modulating Post-infarct Inflammation and Neoangiogenesis

Background: The efficacy of transplantation of default mesenchymal stem cells (MSCs) is controversial because of their limited plasticity and decreased function in elderly patients with myocardial infarction (MI). We investigated whether epigenetically reprogrammed-MSCs ameliorate myocardial remodeling in a mouse model of MI. Methods: Bone marrow-derived MSCs were sequentially treated with 5 mM/L of valproic acid and 100 nM/L of 5-azacytidine for 48 hours. Quantitative RT-PCR was performed to evaluate the effect of epigenetic modifiers on the gene expression of MSCs. 2 x 105 default MSCs, 2 x 105 epigenetically reprogrammed-MSCs, or phosphate-buffered saline were injected into peri-infarct zone immediately after ligating proximal portion of left anterior descending artery. On days 28 after MI, in vivo cardiac magnetic resonance (CMR) and harvest of heart tissue was sequentially performed. Results: Epigenetic modification of MSCs induced gene expression of anti-inflammatory markers such as transforming growth factor-β, indoleamine 2,3-dioxygenase. Fibroblast growth factor-β increased 37.5% and myocyte-specific enhancer factor 2C increased 36.8% at the mRNA level. Transplantation of modified-MSCs showed improved ejection fraction on CMR. In histopathologic analysis, infarct size was significantly decreased in modified-MSCs transplanted mice (p=0.002). When assessing capillary density of peri-infarct zone, larger number of CD31+ staining vascular structures was observed in modified-MSCs transplanted mice. Immunofluorescence stain showed marked increase of CD4+CD25+Foxp3+ regulatory T cells as well as CD68+MR+ M2 macrophage and decrease of CD68+iNOs+ M1 macrophage in modified-MSCs transplanted mice. Conclusions: Transplantation of epigenetically reprogrammed-MSCs significantly improves cardiac function by modulating post-infarct inflammation and neoangiogenesis in a preclinical model of AMI.

The Suppression of Wound Healing Response with Sirolimus and Sunitinib Following Experimental Trabeculectomy in a Rabbit Model

Purpose: To investigate the effects of sirolimus and sunitinib on wound healing in experimental glaucoma filtering surgery (GFS).Material and Methods: Thirty-five male New Zealand pigmented rabbits were randomly assigned to five groups, each including seven rabbits: The rabbits in the control group were not operated on and did not receive any treatment. The rabbits in the sham group underwent trabeculectomy and had one drop of saline instilled four times a day for 14 days. The rabbits in the mitomycin-C (MMC) group underwent trabeculectomy, and a sponge soaked in 0.4 mg/mL MMC was applied intraoperatively to the scleral surgical site for three minutes. The rabbits in the sirolimus group underwent trabeculectomy and 30 ng/mL sirolimus-soaked sponge was applied intraoperatively to the scleral surgical site for three minutes. Sunitinib 0.5 mg/mL four drops in a day were applied in the sunitinib group for 14 days after surgery. On day 14 of the experiment, eyes were enucleated and histologically and immunohistochemically analyzed. Statistical analyses of the study were performed with Kruskal–Wallis variance analysis and Mann–Whitney U test.Results: The mean fibroblast and MNC numbers and the mean immunostaining intensities of transforming growth factor-β (TGF-β), fibroblast growth factor-β (FGF-β) and platelet derived growth factor (PDGF) in the MMC, sirolimus and sunitinib groups were statistically significantly lower than those of the sham group (p < 0.01). The mean fibroblast and MNC numbers and the mean immunostaining intensities of TGF-β, FGF-β and PDGF in the MMC, sirolimus and sunitinib groups were similar (p > 0.05).Conclusions: Our study suggests that the applications of sirolimus and sunitinib effectively suppress the subconjunctival scarring after experimental GFS.

Layer-by-layer assembled cell instructive nanocoatings containing platelet lysate

Great efforts have been made to introduce growth factors (GFs) onto 2D/3D constructs in order to control cell behavior. Platelet lysate (PL) presents itself as a cost-effective source of multiple GFs and other proteins. The instruction given by a construct-PL combination will depend on how its instructive cues are presented to the cells. The content, stability and conformation of the GFs affect their instruction. Strategies for a controlled incorporation of PL are needed. Herein, PL was incorporated into nanocoatings by layer-by-layer assembling with polysaccharides presenting different sulfation degrees (SD) and charges. Heparin and several marine polysaccharides were tested to evaluate their PL and GF incorporation capability. The consequent effects of those multilayers on human adipose derived stem cells (hASCs) were assessed in short-term cultures. Both nature of the polysaccharide and SD were important properties that influenced the adsorption of PL, vascular endothelial growth factor (VEGF), fibroblast growth factor b (FGFb) and platelet derived growth factor (PDGF). The sulfated polysaccharides-PL multilayers showed to be efficient in the promotion of morphological changes, serum-free adhesion and proliferation of high passage hASCs (P > 5). These biomimetic multilayers promise to be versatile platforms to fabricate instructive devices allowing a tunable incorporation of PL.

Impact of δ-Tocotrienol on Inflammatory Biomarkers and Oxidative Stress in Hypercholesterolemic Subjects

Background: Tocotrienols have hypocholesterolemic, anti-inflammatory, and anti-cancer properties. Clinical studies using tocotrienol-rich fraction (TRF) from palm oil yielded inconsistent results with regards to its efficacy due to presence of tocopherols in TRF mixture. Objectives: The impact of tocopherol-free δ-tocotrienol on inflammatory and oxidative stress biomarkers, plasma cytokines/proteins, their gene expression, and microRNAs was studied in hypercholesterolemic subjects. Design: Hypercholesterolemic (n=31; serum cholesterol >5.2 mmol/L) subjects were enrolled in the study. All hypercholesterolemic subjects were given increasing doses of δ-tocotrienol (125, 250, 500, 750 mg/d) plus AHA Step-1 diet for 4 weeks each during a 30 week study period. Serum nitric oxide (NO), C-reactive protein (CRP), malondialdehyde (MDA), δ-glutamyl-transferase (δ-GT), total antioxidant status (TAS), cytokines/proteins, cDNA, and microRNAs were determined. Results: All concentrations of δ-tocotrienol reduced serum levels of NO, CRP, MDA, δ-GT. The most effective dose (250 mg/d) decreased serum NO (40%), CRP (40%), MDA (34%), δ-GT (22%) significantly (P<0.001), while TAS levels increased 22% (P<0.001). The 500 mg/d and 750 mg/d doses were less effective in improving oxidative stress compared to the 250 mg/d dose. Inflammatory plasma cytokines (resistin, IL-1α, IL-12, IFN-γ) were reduced 15-17% (P<0.05-0.01), while cardiac angiogenic fibroblast growth factor-b (FGF-b) and platelet-derived growth factor (PDGF) were decreased by 11% and 14% (P<0.05-0.01), respectively, with 250 mg/d δ-tocotrienol treatment. Similar results were obtained for cytokine gene expression. Several plasma miRNAs (miRNA-16-1, miRNA-125a, miRNA-133, miRNA-155, miRNA-223, miRNA-372, miRNA-10b, miRNA-18a, miRNA-214) associated with cardiovascular disease and cancer were modulated by δ-tocotrienol treatment. Conclusions: In a dose-dependent study of 125-750 mg/d, δ-tocotrienol maximally reduced inflammation and oxidative stress parameters with a 250 mg/d dose in hypercholesterolemic subjects, and may be a potential therapeutic alternative natural product for the maintenance of health during aging process.

Risk factors and glycation end products in patients with different forms of coronary heart disease and type 2 diabetes mellitus

To determine the levels of growth factors and glycation end products in patients with different forms of coronary heart disease (CHD) and type 2 diabetes mellitus (T2DM). A total of 134 patients with CHD and T2DM, including 38 patients with non-ST-elevation acute coronary syndrome (ACS), were examined. The arterial and venous serum levels of basic fibroblast growth factor-β (FGF-β), transforming growth factor-β (TGF-β), placental growth factor (PlGF), advanced glycation end products (AGEs) and their receptors (RAGE) were estimated in all the patients. A direct correlation was found between the degree of arterial stenosis and the level of growth factors and AGEs in the patients with T2DM; there was also a direct correlation of the examined factors with lipid metabolic parameters. There was a significant two-fold increase in FGF-β, PlGF, and RAGE levels in the patients with ACS. Hyperglycemia was found to negatively affect the progression of atherosclerotic changes in the vessel wall and on that of fibrotic processes.

Topical infliximab for the suppression of wound healing following experimental glaucoma filtration surgery.

BackgroundThe purpose of this work was to look into the effects of infliximab on wound healing in experimental glaucoma filtration surgery and to compare the antifibrotic effects of this agent to that of mitomycin-C (MMC).MethodsTwenty-eight male New Zealand White rabbits were randomly assigned to four groups, each including seven rabbits: control group, sham group, MMC group, and infliximab group. The rabbits in the control group were not operated on and did not receive any treatment. The rabbits in the sham group underwent trabeculectomy and had one drop of saline instilled four times a day for 14 days. The rabbits in the MMC treatment group underwent trabeculectomy, and a sponge soaked in 0.4 mg/mL MMC was applied intraoperatively to the scleral surgical site for three minutes. The rabbits in the infliximab treatment group underwent trabeculectomy and one drop of 10 mg/mL infliximab was instilled four times a day for 14 days after surgery. On day 14 of the experiment, the operated and control eyes were enucleated and histologically and immunohistochemically analyzed.ResultsThe mean fibroblast and mononuclear cell (MNC) numbers and the mean immunostaining intensities of transforming growth factor-β (TGF-β), fibroblast growth factor-β (FGF-β), and platelet-derived growth factor (PDGF) in the sham group were higher than those of the control group (P<0.01). The mean fibroblast and MNC numbers and the mean immunostaining intensities of TGF-β, FGF-β, and PDGF in the MMC and infliximab groups were statistically significantly lower than those of the sham group (P<0.01). The mean fibroblast and MNC numbers and the mean TGF-β, FGF-β, and PDGF immunostaining intensities of the MMC and infliximab groups were similar (P>0.05).ConclusionOur study suggests that topical infliximab effectively suppresses the subconjunctival wound healing response after experimental glaucoma filtration surgery, reducing the MNC and fibroblast numbers and immunostaining intensities of TGF-β, FGF-β, and PDGF.

HGF, sIL-6R and TGF-β1 Play a Significant Role in the Progression of Multiple Myeloma.

Background. In the last few years, it has been widely reported that proinflammatory and angiogenic cytokines are important for the development and progression of multiple myeloma (MM).Objectives. To further validate and acquire more insight into this view we decided to check whether plasma levels of certain cytokines and their soluble receptors differ between MM patients and healthy subjects.Patients and Methods. The study was conducted in 76 MM patients aged 22 to 77 years (60±10 years) and 35 healthy controls aged 20 to 63 years (33±10 years). Plasma levels of interleukin-6 (IL-6), b-fibroblast growth factor (b-FGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1), as well as soluble receptors for IL-6 (sIL-6R) and VEGF (sVEGF-R2) were measured using enzyme-linked immunosorbent assay (ELISA).Results. Significantly higher plasma levels of IL-6 (13.65±42.61 vs. 1.04±1.12 pg/ml, p=0.006), HGF (2174±2714 vs. 648±130 pg/ml, p<0.001), b-FGF (7.92±10.78 vs. 2.54±5.38 pg/ml, p<0.001) and sIL-6R (37.1±14.2 vs. 25.3±6.4 ng/ml, p=0.003) were observed in MM patients vs. healthy controls, respectively. Plasma sVEGF-R2 was significantly lower in MM patients than in controls (7518±2119 vs. 8725±1281 pg/ml, respectively; p<0.001). We observed an inverse correlation between length of treatment and the level of sIL-6R, and TGF-β1 in plasma.Conclusions. Plasma levels of HGF, b-FGF, IL-6 and sIL-6R in MM patients were higher ​​when compared to the control group. Antineoplastic therapy leads to a time-dependent decrease in plasma levels of sIL-6R, and TGF-β1 in MM patients. Blood plasma level of HGF is an optimal measure to differentiate patients in whom disease is progressing versus patients who respond to therapy.

Rat bone marrow mesenchymal stem cells improve regeneration of thin endometrium in rat

To investigate whether bone marrow mesenchymal stem cell (BMSC) treatment could improve the regeneration of endometrium and improve the endometrial receptivity in an experimental model of thin endometrium. Randomized, control trial, animal research. National key laboratory. Sprague-Dawley rats. Bone marrow mesenchymal stem cell transplantation by tail vein IV injection. Endometrial thickness, the expression of mark proteins for endometrial cell, and endometrial receptivity. The endometrium was significantly thicker and the expression of cytokeratin, vimentin, integrin αγβ3, and leukemia inhibitor factor were significantly stronger compared with the control group. Some proinflammatory cytokines, such as tumor necrosis factor-α (TNF-α) messenger RNA (mRNA) and interleukin-1β mRNA, were significantly down-regulated, and anti-inflammatory cytokines, such as fibroblast growth factor-β (bFGF) mRNA and interleukin-6 mRNA, were significantly up-regulated in the experimental group compared with the control group. The BMSCs have beneficial effect on thin endometrium, and may play a role through migration and immunomodulatory of BMSCs.

Effects of PDGF-BB and b-FGF on the production of cytokines, hyaluronic acid and the proliferation of orbital fibroblasts in thyroid ophthalmopathy

The effects of Platelet-derived growth factor-BB(PDGF-BB) and Basic-fibroblast growth factor (b-FGF) on the proliferation and production of hyaluronic acid and, cytokine productions in cultured human orbital fibroblasts were evaluated. Orbital fibroblasts were cultured (5 from eyes with thyroid-associated ophthalmopathy and, 5 from normal eyes) and classified into 3 groups-: (1) the Unstimulated group, (2) the Platelet-derived growth factor-BB stimulation group and, (3) the b-Fibroblast growth factor stimulation group. The hyaluronic acid concentration of each group was measured by ELISA. The cytokine production (IL-1β, IL-2, IL-6, IL-8 and RANTES) of each group was measured in the cultured medium. The proliferation of orbital fibroblasts was measured in each group using a thymidine analog EdU, which can be incorporated into DNA during active DNA synthesis. In conclusion, PDGF-BB and b-FGF induced orbital fibroblasts increased production of hyaluronic acid in thyroid-associated ophthalmopathy. In addition to the induction of orbital fibroblast proliferation, PDGF-BB increased the production of IL-8. Further studies on the inhibitory factors of orbital fibroblasts with a larger sample size are warranted to aid in the development of fundamental treatments for thyroid-associated ophthalmopathy.

Expression of adhesion-related cytokines in the uterine fluid after transcervical resection of adhesion

To study the concentration of adhesion-related cytokines, transforming growth factor (TGF) β(1), platelet-derived growth factor (PDGF) BB, b-fibroblast growth factor (bFGF) in intrauterine fluid and their roles in the intrauterine-adhesion reformation. From August 2009 to February 2010, 18 patients with moderate or severe intrauterine adhesions underwent transcervical resection of intrauterine adhesion as study group, in the mean time, 20 patients with incomplete septate uterus underwent transcervical resection of septum as control group. The charge of each patient's intrauterine fluid was collected at the time of 3, 6, 9, 12, 24, 48 and 72 hours, respectively, then the concentration of the three cytokines was measured by enzyme-linked immunosorbent assay (ELISA). The concentration of the expression of TGF-β(1) in the intrauterine fluid at 3, 6, 9, 12 hours after operation in study group was (3.6 ± 0.9), (10.4 ± 1.1), (7.6 ± 1.2), (7.2 ± 1.3) ng/ml, respectively; PDGF-BB was (2.6 ± 0.6), (3.5 ± 0.5), (5.4 ± 1.0), (5.7 ± 0.8) ng/ml, respectively; bFGF was (16.9 ± 1.3), (95.8 ± 17.8), (330.9 ± 70.5), (1303.3 ± 117.4) ng/ml, respectively. The expression of TGF-β(1) in control group was (3.0 ± 0.6), (7.5 ± 0.6), (5.4 ± 0.6), (4.6 ± 0.8) ng/ml, respectively; PDGF-BB was (2.5 ± 0.4), (2.6 ± 0.5), (4.7 ± 0.6), (4.4 ± 0.4) ng/ml, respectively; bFGF was (19.1 ± 2.4), (82.9 ± 21.8), (249.0 ± 54.2), (775.6 ± 178.8) ng/ml, respectively. The concentration of TGF-β(1), PDGF-BB and bFGF reached the peak during the time of 3 - 24 hours, 6 - 48 hours, 6 - 72 hours after surgery, respectively. The concentration of TGF-β(1), PDGF-BB and bFGF in study group were significantly higher than that in control group (P < 0.05). Concentration of TGF-β(1), PDGF-BB and bFGF was positively associated with area of transcervical resection of intrauterine adhesion, those cytokines might be involved in reformatin of intrauterine-adhesion.

The Key Role of Insulin-Like Growth Factor I in Limbal Stem Cell Differentiation and the Corneal Wound-Healing Process

Limbal stem cells (LSC), which reside in the basal layer of the limbus, are thought to be responsible for corneal epithelial healing after injury. When the cornea is damaged, LSC start to proliferate, differentiate, and migrate to the site of injury. To characterize the signaling molecules ensuring communication between the cornea and LSC, we established a mouse model of mechanical corneal damage. The central cornea or limbal tissue was excised at different time intervals after injury, and the expression of genes in the explants was determined. It was observed that a number of genes for growth and differentiation factors were significantly upregulated in the cornea rapidly after injury. The ability of these factors to regulate the differentiation and proliferation of limbal cells was tested. It was found that the insulin-like growth factor-I (IGF-I), which is rapidly overexpressed after injury, enhances the expression of IGF receptor in limbal cells and induces the differentiation of LSC into cells expressing the corneal cell marker, cytokeratin K12, without any effect on limbal cell proliferation. In contrast, the epidermal growth factor (EGF) and fibroblast growth factor-β (FGF-β), which are also produced by the damaged corneal epithelium, supported limbal cell proliferation without any effect on their differentiation. Other factors did not affect limbal cell differentiation or proliferation. Thus, IGF-I was identified as the main factor stimulating the expression of IGF receptors in limbal cells and inducing the differentiation of LSC into cells expressing corneal epithelial cell markers. The proliferation of these cells was supported by EGF and FGF.

Modulation of Postoperative Scarring with Tacrolimus and Octreotide in Experimental Glaucoma Filtration Surgery

Purpose: The purposes of this study were to investigate the effects of topically administrated Tacrolimus and Octreotide on modulation of postoperative scarring in experimental glaucoma filtration surgery and to compare the antifibrotic properties of these agents with mitomycin-C (MMC).Material and methods: A total of 28 New Zealand rabbits weighing 2.5–3 kg were randomly divided into a surgical control (SC) group and three experimental groups. Standard filtration surgeries were performed on the right eyes of all the rabbits. The rabbits in the SC group received only vehicle after the surgeries, whereas the rabbits in the three experimental groups were treated either with 0.4 mg/mL MMC during the surgery (MMC group) or with 0.3 mg/mL Tacrolimus drop four times a day (TT group) or with 10 µg/mL Octreotide drop three times a day (OT group) for 14 days. The animals were killed on day 14, eyes were enucleated and histologically and immunohistochemically analyzed.Results: In SC group mean fibroblast, mononuclear cell number and fibroblast growth factor-β (FGF-β), transforming growth factor-β (TGF-β) immunostaining intensity was higher than all treatment groups. In OT group mean fibroblast number was lesser than MMC (p < 0.01) and TT (p < 0.05) group. In TT group mean fibroblast number was lesser than MMC group (p < 0.05). Mean mononuclear cell number was similar between MMC, OT and TT groups (p > 0.05). In MMC, OT and TT groups mean TGF-β and FGF-β immunostaining intensity was similar (p > 0.05).Conclusions: Topically administration of Tacrolimus and Octreotide effectively reduced the subconjuntival scarring response 2 weeks after experimental glaucoma filtration surgery.

Evidence for distinct histologic profile of nasal polyps with and without eosinophilia

To evaluate the histology, RNA, and protein signatures of nasal polyps (NPs) in order to demonstrate specific subtypes of disease and differentiate "idiopathic" NPs based on tissue eosinophilia. Prospective laboratory-based study. NP tissue was obtained from patients referred to the University of Virginia Health System for sinus surgery. Histology analyses included hematoxylin-eosin, Gomori's trichrome, toluidine blue, and chloroacetate staining. RNA and protein were extracted from tissue and cytokine transcript or protein concentrations determined. Idiopathic NPs can be divided into distinct subsets characterized by absence (NE) and presence (E) of prominent eosinophilia. The validity of this distinction is supported by the demonstration that NE polyps are further distinguished by glandular hypertrophy, dense collagen deposition, and mononuclear cellular infiltrate. In contrast, E-NP display edema, rare glandularity, and minimal collagen deposition except within the basement membrane. Total mast cell numbers were reduced in E-NP, whereas connective tissue mast cells were increased in NE-NP. Consistent with the distinctive pattern of increased fibrosis, NE-NP displayed increased transforming growth factor-β and vascular endothelial growth factor transcripts. Similarly, NE-NPs had higher concentrations of transforming growth factor-β, fibroblast growth factor-β, and platelet-derived growth factor protein. Idiopathic NPs can be distinguished by NE and E and are supported by the observations that these display distinct histologic, gene, and protein expression patterns. The findings suggest that as unique diseases, idiopathic NPs will require distinct therapeutic interventions.