Platelet Rich Plasma in the Treatment of Frontal Fibrosing Alopecia

Frontal Fibrosing Alopecia (FFA) represents a form of scaring alopecia more frequent in postmenopausal women that presents with frontal hairline recession. It is typically classified as a variant of lichen planopilaris. Treatment of FFA can be challenging with poor long-term outcomes. Platelet-Rich Plasma (PRP) consists of an autologous concentration of platelets in a small volume of plasma. Activated platelets secrete cytokines and growth factors and thus may have a potential role in the treatment of inflammatory scarring alopecia such as FFA. A 68-year-old female with multi-resistant FFA was treated with lesional PRP injections every 4 weeks for 16 weeks. Baseline LPPAI score and phototrichograms targeting a representative area of disease activity were compared at baseline and at 16 weeks. After 16 weeks, no significant change in follicular units or follicular density from baseline to week 16 was noted. Only a minimal improvement in inflammatory activity observed clinically and through the Lichen Planopilaris Activity Index was observed. The discordance between the follicular density count and observed inflammatory activity suggests a longer treatment and observational period is needed. Additionally, the frequency of PRP injections potentially may also need to be increased. Given the limited efficacy of current therapies for FFA, PRP injections may be an option in patients with refractory disease, as an adjunct to systemic therapy. Additional investigation is needed to optimize frequency of PRP injections in FFA and to better assess its true anti-inflammatory effect.

A New Bioreactor to Promote Follicular Growth With or Without Activin A

Background: The aim of the present study was to evaluate the effect of Activin A on the activation of in vitro folliculogenesis of human ovarian tissues with or without our new three-dimensional structure (3D-structure). Methods: Five fresh ovarian human tissues were cultured in vitro in 4 groups with 100ng/mL Activin A or without Activine A and within or without a 3D-structure for 20 or 22 days of in vitro culture. Follicular density and quality were evaluated, and follicular diameters were measured. Estradiol secretion was quantified. Proliferation and apoptosis through immunostaining were performed.Results: The proportion of primordial follicles was significantly reduced, and the proportion of primary and secondary follicles was significantly increased in all four groups (p<0.001). Antral cavities were observed in the four culture groups. Activin A supplementation did not significantly affect the follicular density, follicular quality, follicular growth, or estradiol secretion (p>0.05). The 3D-structure increased the density of primary follicles and decreased the estradiol secretion (p<0.001). Tissular proliferation was significantly lower in the 3D-structure group compared to the non-3D-structure group (p=0.008). Regarding tissular apoptosis, it was significantly higher in the Activin group compared to the non-Activin group (0.006). Conclusion: The presence of Activin A did not seem to play a key role in in vitro folliculogenesis activation. However, the results may indicate that the 3D-structure could be more physiological and could prevent a pejorative in vitro folliculogenesis flare-up.

When to Cryopreserve Ovarian Tissue: Effects of Chemotherapy on the Ovarian Reserve by Studying Follicular Density, Tissue Damage and Follicular Apoptosis

BackgroundPatients scheduled to receive chemotherapy should be counseled on fertility preservation. Known gonadotoxic chemotherapies such as alkylating agents have a high risk of altering ovarian reserve. In some cases, the urgency of treatment requires the use of chemotherapy before fertility preservation, which will be carried out at a later stage. Most often the ovarian tissue is cryopreserved. The aim of our study is to investigate the impact of chemotherapies on follicular density, tissue damage and apoptosis of reserve follicles.ResultsWe included 140 patients: 63 patients, mean age 18.8 years, were included in the group no chemotherapy (group A) and 77 patients, mean age 17.1 years, in the group presence of chemotherapy before ovarian conservation (group B). None of the patients had had pelvic radiotherapy prior to ovarian cryopreservation. The histological parameters studied were: follicular density, the presence of cortical fibrosis and the presence of vessel abnormalities. We selected 12 patients from group A and 15 patients from group B, comparable in age and pathology, for whom we evaluated follicle apoptosis by immunostaining cleaved caspase 3. We demonstrated an inverse relationship between follicular density and age (p<0.0001), as well as a lack of effect of chemotherapy on follicular density (p=0.87). There was no difference in other histological parameters. On the other hand, we showed an impact of chemotherapies, especially alkylating agents, on the apoptosis of ovarian follicles (p<0.0001).ConclusionOur study is the largest cohort reported to date. This work underlines that conservation of ovarian tissue after chemotherapy remains possible.

Human Ovarian Cryopreservation: Vitrification Versus Slow Freezing From Histology To Gene Expression.

Background: Cryopreservation of ovarian tissue is one of the strategies offered to girls and women needing gonadotoxic treatment, as a means of preserving their fertility. There are two methods of ovarian tissue cryopreservation: slow freezing, the reference method, and vitrification, an alternative method. The aim of the present study was to evaluate which of the two is the best method for human ovarian tissue cryopreservation. Each ovary was divided into 3 groups: Fresh, Slow freezing and Vitrification. In each group a histological study to evaluate follicular density and quality; and an evaluation of 6 gene expression (CYP11A, STAR, GDF9, ZP3, CDK2, CDKN1A) were performed. Results: We observed no significant difference in follicular density within these 3 groups. Slow freezing altered the pool of primordial follicles compared to the Fresh tissue (31.8% vs 55.9%, p = 0.046, respectively). The expression of genes involved in steroidogenesis varied after cryopreservation compared to the fresh group; CYP11A was under-expressed in both freezing groups compared to the fresh group and significantly under-expressed in the slow freezing group (p = 0.01), STAR was over-expressed in the slow freezing group and significantly under-expressed in the vitrification group (p = 0.01). Regarding the expression of genes involved in cell cycle regulation, CDKN1A was significantly under-expressed in both freezing groups (slow freezing: p = 0.0008; vitrification: p = 0.03) compared to the fresh group. Conclusion: Vitrification had no effect on the histological quality of the follicles at any stage of development compared to Fresh tissue. There was no significant difference in gene expression between the two techniques.

Ovarian Tissue Reservation and Risk of Reimplantation in a Young Girl with Acute Lymphocytic Leukemia after 6-Month Chemotherapy: A Case Report

In the fertility preservation programs, ovarian cryopreservation is usually offered when the risk of premature ovarian failure is high (>30–50%) while the risk of ovarian metastasis is low. According to the guidelines, it must be done before the patient receives chemotherapy. A 22-year-old girl with acute lymphocytic leukemia was a candidate for ovarian cryopreservation after 6 months of chemotherapy. Despite chemotherapy, the anti-Mullerian hormone survey was within normal range. Ovarian tissue cryopreservation was done. In the histology survey, follicular density was 7.48. This case shows that only having a history of chemotherapy does not exclude the patient from the fertility preservation program. Regarding referring the patients for fertility preservation, cumulative factors such as a history of gonadotoxic treatment, age, and treatment protocol should be considered. In addition, the case was negative for assessing of CD45 marker. New data may challenge previous strict criteria, and extend the indications of this effective method in preserving fertility among cancer patients.

Effects of low-dose X-ray medical diagnostics on female gonads: Insights from large animal oocytes and human ovaries as complementary models

Diagnostic imaging has significantly grown over the last thirty years as indispensable support for diagnostic, prognostic, therapeutic and monitoring procedures of human diseases. This study explored the effects of low-dose X-ray medical diagnostics exposure on female fertility. To aim this, cumulus-oocyte complexes (COCs) recovered from the ovaries of juvenile sheep and human ovaries were used as complementary models for in vitro studies. In the sheep model, the effects of low-dose X-rays on oocyte viability and developmental competence were evaluated. In human ovaries originated from two age group (21–25 and 33–36 years old) subjects with gender dysphoria, X-rays effects on tissue morphology, follicular density and expression of apoptosis-related (NOXA, PUMA, Bcl2, Bak, γH2AX) and cell cycle-related genes (p21 and ki67) were investigated. It was noted that in sheep, the minimum dose of 10 mGy did not influence most of examined parameters at oocyte and embryo levels, whereas 50 and 100 mGy X-ray exposure reduced oocyte bioenergetic/oxidative activity but without any visible effects on oocyte and embryo development. In addition, blastocyst bioenergetic/oxidative status was reduced with all used doses. Overall data on human ovaries showed that low-dose X-rays, similarly as in sheep, did not alter any of examined parameters. However, in women belonging to the 33–36 year group, significantly reduced follicular density was observed after exposure to 50 and 100 mGy, and increased NOXA and Bax expression after exposure at 50 mGy. In conclusion, used low-doses of X-ray exposure, which resemble doses used in medical diagnostics, produce weak damaging effects on female fertility with increased susceptibility in advanced age.

Adaptation of a trap door technique for the recovery of ovarian cortical biopsies from Cebus apella (capuchin monkey)

SummaryThere is a paucity of efficient cryopreservation protocols for primordial follicles enclosed in the ovarian tissue from non-human primates (NHP), in special New World primates. Our objective was to establish an optimal procedure for the recovery of ovarian biopsies from capuchin monkeys. To this end, we adapted a trap door biopsy method. Follicular density and quality of the biopsies were evaluated and ultrasound analysis was performed before and continuously after surgery to assess ovarian structure. Ovarian tissue biopsies recovered by the trap door technique allowed the successful harvesting of primordial follicles from capuchin monkeys, and no complication was recorded. The female cycle was not affected by surgery and no adherence was found thereafter. In conclusion, the adaptation of a trap door biopsy method is a safe procedure and allows recovery of healthy primordial follicles.