Limits...
Topical KGF treatment as a therapeutic strategy for vaginal atrophy in a model of ovariectomized mice.

Ceccarelli S, D'Amici S, Vescarelli E, Coluccio P, Matricardi P, di Gioia C, Benedetti Panici P, Romano F, Frati L, Angeloni A, Marchese C - J. Cell. Mol. Med. (2014)

Bottom Line: One of the most frequent complaints for post-menopausal women is vaginal atrophy, because of reduction in circulating oestrogens.We demonstrated that KGF is able to induce proliferation of vaginal mucosa, and we gained insight on its mechanism of action by highlighting its contribution to switch ERα signalling towards non-genomic pathway.RM2012A000404) has been applied for this study.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.

Show MeSH

Related in: MedlinePlus

Effect of KGF and E2 on human vaginal mucosa cells (HVMs) proliferation. Immunofluorescence analysis with polyclonal antibodies against Ki67 was performed on: (A) HVMs treated with KGF (0.5, 10 or 20 ng/ml; 2.6 × 10−11, 1.1 × 10−10 and 1.1 × 10−9 M, respectively) or E2 (0.5, 10 or 20 ng/ml; 1.8 × 10−9, 7.3 × 10−9 and 7.3 × 10−8 M, respectively) for 24 hrs; (B) HVMs derived from biopsies of pre- or post-menopausal women treated with KGF (20 ng/ml) or E2 (20 ng/ml); (C) HVMs treated with KGF (20 ng/ml), E2 (20 ng/ml) or a combination of them for 24 hrs; (D) HVMs pre-treated with tamoxifen (Tam, 100 nM) for 30 min. and then treated with KGF (20 ng/ml), E2 (20 ng/ml), Tam, KGF plus Tam or E2 plus Tam for 24 hrs, which were also stained with 1% crystal violet (E). The percentage of Ki67-positive cells was determined by counting the number of Ki67-positive nuclei versus total number of nuclei in 10 different areas randomly taken from three different experiments. Error bars represent standard deviations. *P < 0.05; **P < 0.01; ***P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4196664&req=5

fig01: Effect of KGF and E2 on human vaginal mucosa cells (HVMs) proliferation. Immunofluorescence analysis with polyclonal antibodies against Ki67 was performed on: (A) HVMs treated with KGF (0.5, 10 or 20 ng/ml; 2.6 × 10−11, 1.1 × 10−10 and 1.1 × 10−9 M, respectively) or E2 (0.5, 10 or 20 ng/ml; 1.8 × 10−9, 7.3 × 10−9 and 7.3 × 10−8 M, respectively) for 24 hrs; (B) HVMs derived from biopsies of pre- or post-menopausal women treated with KGF (20 ng/ml) or E2 (20 ng/ml); (C) HVMs treated with KGF (20 ng/ml), E2 (20 ng/ml) or a combination of them for 24 hrs; (D) HVMs pre-treated with tamoxifen (Tam, 100 nM) for 30 min. and then treated with KGF (20 ng/ml), E2 (20 ng/ml), Tam, KGF plus Tam or E2 plus Tam for 24 hrs, which were also stained with 1% crystal violet (E). The percentage of Ki67-positive cells was determined by counting the number of Ki67-positive nuclei versus total number of nuclei in 10 different areas randomly taken from three different experiments. Error bars represent standard deviations. *P < 0.05; **P < 0.01; ***P < 0.001.

Mentions: We first performed KGF and E2 treatments in vitro on HVMs obtained from biopsies of the vaginal mucosa from n = 3 healthy donors [25], to compare their effect on cell proliferation. Cell proliferation was determined by counting cells positive for Ki67 antigen, which identifies cycling cells, and reported in graph as percentage of positive cells (Fig. 1A). Our results show a dose-dependent increase in HVMs proliferation following treatment with both KGF and E2, as compared to untreated cells. To exclude an involvement of patients' age in the extent of proliferative response to KGF or E2 treatments, we performed the proliferation assays separately on HVMs obtained from n = 10 donors in pre-menopausal age and n = 10 in post-menopausal age. As shown in Figure 1B, despite a difference in the basal proliferation levels of untreated cells, as expected, KGF and E2 exerted their proliferative potential in both pre- and post-menopause-derived cells. These results indicate that KGF is able to stimulate the proliferation of vaginal epithelium in vitro as well as estradiol.


Topical KGF treatment as a therapeutic strategy for vaginal atrophy in a model of ovariectomized mice.

Ceccarelli S, D'Amici S, Vescarelli E, Coluccio P, Matricardi P, di Gioia C, Benedetti Panici P, Romano F, Frati L, Angeloni A, Marchese C - J. Cell. Mol. Med. (2014)

Effect of KGF and E2 on human vaginal mucosa cells (HVMs) proliferation. Immunofluorescence analysis with polyclonal antibodies against Ki67 was performed on: (A) HVMs treated with KGF (0.5, 10 or 20 ng/ml; 2.6 × 10−11, 1.1 × 10−10 and 1.1 × 10−9 M, respectively) or E2 (0.5, 10 or 20 ng/ml; 1.8 × 10−9, 7.3 × 10−9 and 7.3 × 10−8 M, respectively) for 24 hrs; (B) HVMs derived from biopsies of pre- or post-menopausal women treated with KGF (20 ng/ml) or E2 (20 ng/ml); (C) HVMs treated with KGF (20 ng/ml), E2 (20 ng/ml) or a combination of them for 24 hrs; (D) HVMs pre-treated with tamoxifen (Tam, 100 nM) for 30 min. and then treated with KGF (20 ng/ml), E2 (20 ng/ml), Tam, KGF plus Tam or E2 plus Tam for 24 hrs, which were also stained with 1% crystal violet (E). The percentage of Ki67-positive cells was determined by counting the number of Ki67-positive nuclei versus total number of nuclei in 10 different areas randomly taken from three different experiments. Error bars represent standard deviations. *P < 0.05; **P < 0.01; ***P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4196664&req=5

fig01: Effect of KGF and E2 on human vaginal mucosa cells (HVMs) proliferation. Immunofluorescence analysis with polyclonal antibodies against Ki67 was performed on: (A) HVMs treated with KGF (0.5, 10 or 20 ng/ml; 2.6 × 10−11, 1.1 × 10−10 and 1.1 × 10−9 M, respectively) or E2 (0.5, 10 or 20 ng/ml; 1.8 × 10−9, 7.3 × 10−9 and 7.3 × 10−8 M, respectively) for 24 hrs; (B) HVMs derived from biopsies of pre- or post-menopausal women treated with KGF (20 ng/ml) or E2 (20 ng/ml); (C) HVMs treated with KGF (20 ng/ml), E2 (20 ng/ml) or a combination of them for 24 hrs; (D) HVMs pre-treated with tamoxifen (Tam, 100 nM) for 30 min. and then treated with KGF (20 ng/ml), E2 (20 ng/ml), Tam, KGF plus Tam or E2 plus Tam for 24 hrs, which were also stained with 1% crystal violet (E). The percentage of Ki67-positive cells was determined by counting the number of Ki67-positive nuclei versus total number of nuclei in 10 different areas randomly taken from three different experiments. Error bars represent standard deviations. *P < 0.05; **P < 0.01; ***P < 0.001.
Mentions: We first performed KGF and E2 treatments in vitro on HVMs obtained from biopsies of the vaginal mucosa from n = 3 healthy donors [25], to compare their effect on cell proliferation. Cell proliferation was determined by counting cells positive for Ki67 antigen, which identifies cycling cells, and reported in graph as percentage of positive cells (Fig. 1A). Our results show a dose-dependent increase in HVMs proliferation following treatment with both KGF and E2, as compared to untreated cells. To exclude an involvement of patients' age in the extent of proliferative response to KGF or E2 treatments, we performed the proliferation assays separately on HVMs obtained from n = 10 donors in pre-menopausal age and n = 10 in post-menopausal age. As shown in Figure 1B, despite a difference in the basal proliferation levels of untreated cells, as expected, KGF and E2 exerted their proliferative potential in both pre- and post-menopause-derived cells. These results indicate that KGF is able to stimulate the proliferation of vaginal epithelium in vitro as well as estradiol.

Bottom Line: One of the most frequent complaints for post-menopausal women is vaginal atrophy, because of reduction in circulating oestrogens.We demonstrated that KGF is able to induce proliferation of vaginal mucosa, and we gained insight on its mechanism of action by highlighting its contribution to switch ERα signalling towards non-genomic pathway.RM2012A000404) has been applied for this study.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.

Show MeSH
Related in: MedlinePlus