Expression of epithelial growth factors and of apoptosis-regulating proteins, and presence of CD57+ cells in the development of inflammatory periapical lesions

Abstract The mechanisms that stimulate the proliferation of epithelial cells in inflammatory periapical lesions are not completely understood and the literature suggests that changes in the balance between apoptosis and immunity regulation appear to influence this process. Objective: To evaluate the expression of the epidermal growth factor (EGF), its receptor (EGFR) and of the keratinocyte growth factor (KGF), the presence of CD57+ cells, the epithelial cell proliferation index, and the expression of the Bcl-2 protein in inflammatory periapical lesions (IPL) at different stages of development. Methodology: Our sample was composed of 52 IPLs (22 periapical granulomas - PG - and 30 periapical cysts - PC), divided into three groups: PGs, small PCs, and large PCs. Specimens were processed for histopathologic and immunohistochemical analyses. Sections were evaluated according to the amount of positive staining for each antibody. Results: We found no significant differences among the groups regarding Bcl-2 (p=0.328) and Ki-67 (p>0.05) expression or the presence of CD57+ cells (p=0.748). EGF (p=0.0001) and KGF (p=0.0001) expression was more frequent in PCs than in PGs, and CD57+ cells were more frequent in IPLs with intense inflammatory infiltrates (p=0.0001). We found no significant differences in KGF (p=0.423), Bcl-2 (p=0.943), and EGF (p=0.53) expression in relation to inflammatory infiltrates or to the type of PC epithelial lining, but observed greater KGF expression (p=0.0001) in initial PCs. EGFR expression was similar among the groups (p>0.05). Conclusion: More frequent EGF and KGF expression in PCs and the greater presence of CD57+ cells in lesions with intense inflammatory infiltrates suggest that these factors influence IPL development. The greater KGF expression in initial PCs suggests its importance for the initial stages of PC formation.

Treatment of radiation-induced intestinal mucosal injury with keratinocyte growth factor 2 mutant in mice / 军事医学

Objective To investigate the therapeutic effect of KGF-2 mutants on radiation-induced intestinal mucosal injury.Methods Specific pathogen free (SPF) female C57BL/6J mice were randomly divided into 4 groups, which were treated with/without KGF2 mutant and/or bone marrow cell transplantation.All the 4 groups were radiated with one-time whole bodyγ-ray exposure of 12 Gy.Also one untreated group was included as control.The therapeutic effect of recombi-nant human KGF-2 mutants on radiation-induced intestinal mucosal injury was analyzed by the survival rate at the 30th day, the pathological change and the apoptosis as well as autophagy status of small intestine at 72 hours after exposure.Results After irradiation, all the mice in the untreated group died within 9 days while the mice treated with KGF-2 mutant combined with bone marrow cell transplantation showed a 70%survival rate at the 30th day.We also found that intestinal mucosa of the mice in this group had a more intact structure, a higher level of autophagy and a lower level of apoptosis via HE staining and immunohistochemistry.Conclusion KGF-2 mutant has a significant therapeutic effect on radiation-induced intestinal mucosal injury.

Effects of Keratinocyte Growth Factor 2(KGF-2) on Keratinocyte Growth, Migration and on Excisional Wound Healing / 生物化学与生物物理进展

Keratinocyte growth factor 2 (KGF-2) is a member of the FGF family that is mainly synthesized by mesenchymal cells and acta predominantly on epithelial cells in a paracrine manner. It is known to play an important role in fetal limb and lung development; skin wound healing and prostatic epithelial cell growth. The KGF-2 coding sequence were isolated from human kidney cDNA library, revealing that the Kgf-2 gene is also expressed in the kidney apparatus. Purified from prokaryotic E. coli cells, the effects of the recombinant KGF-2 protein in cultured keratinocyte were analyzed by using MTT assay and in situ TUNEL assay. Interestingly, results revealed that KGF-2 promoted keratinocyte cell growth by stimulating cell proliferation and attenuating cell apoptosis. These findings supported a few evidences that KGF-2 could contribute to alveolar epithelial cells against apoptosis. Cell migration assays for the first time revealed that KGF-2 could stimulate keratinocyte cell migration in vitro. In addition, in the pilot animal test, recombinant KGF-2 incorporated within the hydrogel dressing exhibited significantly stimulatory effect on cutaneous wound healing. These combined effects implicate a potential therapeutic application of human recombinant KGF-2 in the future.

Effects of Keratinocyte Growth Factor (KGF), Epidermal Growth Factor (EGF), and Extracellular Calcium on the Growth of Cultured Psoriatic Keratinocytes / 대한피부과학회지

We investigated the effects of KGF and EGF, together with extracellular calcium, on the growth of cultured psoriatic keratinocytes, in order to establish a new chemically defined medium for culturing psoriatic keratinocytes using a modified form of MCDB 153 media. We compared the cell growth pattern under various culture conditions, including growth factors (KGF or EGF), and extracellular calcium concentrations, attachment and/or matrix factors (type I collagen coating or 3T3 fibroblast layering), culture durations, and types of cultured cells such as normal human epidermal keratinocytes (NHEK) and psoriatic keratinocytes. In order to achieve the above objective, semiquantitative RT-PCR for K16 mRNA, direct immunofluorescence with K8.12 as the markers of regenerating keratinocytes, and microscopic observation for cell colony formation were performed. The results are summarized as follows: 1. Psoriatic keratinocytes were grown optimally at 0.15 mM calcium, irrespective of growth factors or even in free control. They also grew well at 20 nM KGF. 2. KGF and/or EGF played an active role in cell growth, especially in the 5 days' culture. The growth stimulatory effect of EGF was suppressed by 0.5 mM calcium, but the effect of KGF was sustained at this calcium concentration. Furthermore, KGF exhibited a cell Survival effect of 18 days on type I collagen coating, and also in 12 days' culture on 3T3 fibroblast layering. 3. Cultured psoriatic keratinocytes were more vulnerable to extracellular calcium than NHEK with regard to optimal calcium concentration; they grew best at 0.15 mM, which was much lower than 1.5 mM in NHEK. 4. 3T3 fibroblasts exerted a favorable effect on cell growth and survival, especially in 12 days' culture, which may have been due to the paracrine effect of endogenous KGF from the 3T3 feeder cells, and cell reattachment/pile-up properties. To improve the culture method for psoriatic keratinocytes, the study suggests we should consider the optimal extracellular calcium concentration, introduce feeder cell layering to increase cell reattachment/pile-up, and supplement the mesenchymal paracrine growth factors such as KGF to exert a favorable effect on cell growth and survival.

The Effects of Keratinocyte Growth Factor and Its Receptors Antisense Oligonucleotide on Prolifera- tion of HaCat Cells / 中华皮肤科杂志

Objective To investigate the effects of keratinocyte growth factor (KGF) and KGF receptor (KGFR) antisense oligonucleotide (ASODN) on cell cycle and apoptosis of HaCat cells. Methods HaCaT cell, an immortalized keratinocyte cell strain, was cultured in vitro. Flow cytometry was used to measure the cell cycle and apoptosis mediated by KGF and ASODN. Results The rates of S phase and apoptosis in the group treated with KGF increased significantly than those in the control group (both P

The Effect of Amniotic Membrane Transplantation on Corneal Epithelial Cell Proliferation

PURPOSE: to investigate the effect of amniotic membrane transplantation (AMT) on corneal epithelial healing. METHODS: A 4-mm epithelial debridement was made in central rabbit cornea. Then, human amniotic membrane was transplanted (AMT group) or a contact lens was applied (contact lens group). The contralateral eyes were unwounded as controls. After surgery 5-bromo-2'-deoxyuridine (BrdU) was injected via ear vein. Each corneal tissue including the limbus was obtained on postoperative 12, 24 and 48 hours. Proliferating cell nuclear antigen (PCNA) in limbal and peripheral corneal epithelium and keratinocyte growth factor (KGF) in limbal stroma were immunolocalized by immunohistochemical method. Incorporation of BrdU in limbal and peripheral corneal epithelium was visualized by indirect immunofluorescent method. RESULTS: AMT group significantly accelerated the expression of PCNA and BrdU at limbal and peripheral corneal epithelial cells. The expression of PCNA and BrdU showed a peak at 24hr in both groups and increased in limbal epithelial cells more than peripheral corneal epithelial cell in AMT group. The expression of KGF on limbal keratocyte increased in AMT group more than contact lens group and coincided wiht the expression pattern of PCNA and BrdU. The number of keratocyte in significantly decreased in contanct lens group compared wiht AMT group. CONCLUSION: AMT enhanced corneal epithelial wound healing in vivo by stimulating limbal epithelial proliferation which is indirectly mediated in part by upregulating the expression of KGF, which is a potent epithelial mitogen secreted by limbal keratocytes.

A study on expression of KGF mRNA in human NSCLC and its role in the development of lung cancer / 中国癌症杂志

Purpose:To investigate expression of keratinocyte growth factor(KGF) messenger ribonuclunc acid in human NSCLC and to study the role of KGF in the development of NSCLC. Methods:The expression of KGF mRNA in 50 cases with NSCLC was detected by in situ hybridization (ISH). Results:On ISH slides, positive KGF mRNA was mainly shown as fusiform stain in plasma of fibroblast and blood vessel smooth muscle cell in mesenchymal of NSCLC, also, some parenchyma cell plasma was stained. The positive rate of KGF mRNA in tumor(86%) is statistically higher than that in normal tissue(24%)(P