The effect of mesenchymal stem cells and sildenafil on flap viability in perforator-based flaps for ischemia/reperfusion injury: An experimental study.

PURPOSE: In this study, we evaluated the individual and combined effects of mesenchymal stem cells (MSCs) and sildenafil citrate (SC) on the viability of pedicled perforator flaps in which ischemia/reperfusion injury developed after induction of primary ischemia. MATERIALS AND METHODS: Seven Sprague-Dawley rats were used as donors of cells. Rectangular flaps (7 × 7 cm2 ) were created featuring the right second epigastric musculocutaneous perforator in 63 male Sprague-Dawley rats. The animals were randomly divided into two experimental groups (based on the ischemia time of 4 or 8 hours) and a control group. Each of the experimental group was further divided into four subgroups with no treatment, subcutaneous administration of MSCs after termination of ischemia, intraperitoneal administration of SC after termination of ischemia, and combined MSCs and SC treatments at the end of the period of ischemia (n = 7 for each subgroup). A sham group with no-ischemia to flap was used as the control (n = 7). On day 7, viable areas on the flaps were calculated from photographs. The levels of the antioxidative enzymes, such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX), were analyzed in tissue samples obtained from the most distal regions of the flap prior to ischemia and on day 7 after induction of ischemia. RESULTS: No difference was detected between the no-ischemia group and 4-hours SC-treated subgroup, 4-hours combined MSC and SC treated subgroup, 8-hours MSC-treated subgroup, or 8-hours SC-treated subgroup (P > 0.05). In 4-hours ischemia group, the viable flap area of combined MSC and SC treated subgroup was significantly greater than that of the ischemia subgroup (17.17 ± 12.56 cm2 vs. 7.24 ± 7.17 cm2 ; P = 0.015). However, in 8-hours ischemia group, the viable flap area of MSC- treated subgroup was significantly greater than that of the ischemia subgroup (2.69 ± 3.71 cm2 vs. 14.52 ± 8.57 cm2 ; P = 0.004). There were no significant differences in SOD, CAT, and GPX levels detected between no-ischemia group and any of the treated subgroups in 4- and 8-hours ischemia groups (P > 0.05). However, SOD, CAT, and GPX levels in the no-ischemia group were lower than that in 4-hours ischemia control subgroup or 8-hours ischemia control subgroup (P < 0.05). CONCLUSION: In this rat pedicled perforator based abdominal flap, we found that after primary ischemia, application of MSCs and SC, either individually or in combined form, significantly enhanced antioxidant enzyme levels compared with those in the control group, and provided protection against ischemia/reperfusion injury. The two treatments acted synergistically to protect against damage after 4 hours of ischemia, but either treatment alone more effectively enhanced viable flap area after 8 hours of ischemia, although some flap damage was apparent. © 2015 Wiley Periodicals, Inc. Microsurgery 36:402-409, 2016.

Keratinocyte growth factor-2 and autologous serum potentiate the regenerative effect of mesenchymal stem cells in cornea damage in rats.

AIM: To investigate the healing process after severe corneal epithelial damage in rats treated with mesenchymal stem cells (MSCs) cultured with or without keratinocyte growth factor (KGF-2) and autologous serum (AS) on amniotic membrane (AM). Many patients are blind and devastated by severe ocular surface diseases due to limbal stem cell deficiency. Bone marrow-derived MSCs are potential sources for cell-based tissue engineering to repair or replace the corneal tissue, having the potential to differentiate to epithelial cells. METHODS: The study included 5 groups each including 10 female "Sprague Dawley" rats in addition to 20 male rats used as bone marrow donors. Group I rats received AM+MSCs, Group II rats AM+MSCs cultured with KGF-2, Group III rats AM+MSCs cultured with KGF-2+AS, Group IV rats only AM and Group V rats, none. AS was derived from blood drawn from male rats and bone marrow was obtained from the femur and tibia bones of the same animals. Therapeutic effect was evaluated with clinical, histopathological and immunohistochemical assessment. MSC engraftment was demonstrated via detection of donor genotype (Y+) in the recipient tissue (X) with polymerase chain reaction. RESULTS: Corneal healing was significantly better in Groups I-III rats treated with MSC transplantation compared to Group IV and Group V rats with supportive treatment only. The best results were obtained in Group III rats with 90% transparency, 70% lack of neovascularization, and 100% epithelium damage limited to less than 1/4 of cornea. CONCLUSION: We suggest that culture of MSCs with KGF-2 and AS on AM is effective in corneal repair in case of irreversible damage to limbal stem cells.

Disruption of HDX gene in premature ovarian failure.

We present a case of a 19-year-old phenotypically normal girl with premature ovarian failure. Cytogenetic analysis using G banding and fluorescence in situ hybridization (FISH) from cultured peripheral blood lymphocytes of the patient and the family revealed a de novo X;15 translocation and the imbalance to be 46,X,t(X;15)(Xpter → Xq21::15q11 → 15qter;15pter → 15q11::Xq21 → Xqter). ish (CEPX+, wep15+, ISNRPN+, PML+, D15S10+, wcp15-, SNRRN-, PML-)[20]. The X chromosome inactivation (XCI) assay revealed a completely skewed XCI pattern in which selective pressure favors an active maternal allele. The Affymetrix 2.7 M cytogenetics whole-Genome array confirmed the chromosomal imbalance and identified disruption of the HDX gene at Xq21, the translocation breakpoint.

Two males with SRY-positive 46,XX testicular disorder of sex development.

The 46,XX testicular disorder of sex development (46,XX testicular DSD) is a rare phenotype associated with disorder of the sex chromosomes. We describe the clinical, molecular, and cytogenetic findings of a 16- and a 30-year-old male patient with sex-determining region Y (SRY)-positive 46,XX testicular DSD. Chromosomal analysis revealed 46,XX karyotype. Fluorescence in situ hybridization (FISH) showed the SRY region translocated to the short arm of the X chromosome. The presence of the SRY gene was also confirmed by polymerase chain reaction (PCR). The X chromosome inactivation (XCI) assay showed that both patients have a random pattern of X chromosome inactivation. This report compares the symptoms and features of the SRY-positive 46,XX testicular DSD patients.

A 19-year follow-up of a patient with type 3 ectrodactyly-ectodermal dysplasia-clefting syndrome who developed non-Hodgkin lymphoma.

The ectrodactyly-ectodermal dysplasia-clefting (EEC) syndrome is characterized by ectrodactyly, ectodermal dysplasia, and clefting. The development of a malignancy with EEC syndrome is very rare. Here we present follow-up on a Turkish boy with EEC syndrome type 3 who developed malignant lymphoma with high expression of p63. He had chronic renal failure due to recurrent urinary infections caused by ureterovesical reflux. Cervical, diffuse, large, B-cell non-Hodgkin lymphoma with high expression of p63 was diagnosed, and the patient died at 19 years of age. The transcription factor p63 is a key regulator of ectodermal, orofacial, and limb development. Mutations in the p63 gene can cause syndromes of ectodermal dysplasia, ectrodactyly, and orofacial clefting. Malignant lymphoma is a very rare complication of EEC syndrome. We suggest that p63 gene mutation analysis should be performed in every EEC syndrome patient with the possibility of developing malignant tumors.

An epileptic case with mosaic ring chromosome 6 and 6q terminal deletion.

Ring chromosomes are rare chromosome disorders that arise usually de novo. Children with ring chromosome 6 have a wide range of intellectual functioning and congenital anomalies. We report an epileptic case of a 10-year-old boy to be mild psychomotor retardation and dysmorphic traits including microcephaly, brachycephaly, flat occiput, large and apparently low set ears, and bilateral syndactyly between his second and third fingers with mosaic ring chromosome 6 and 6q terminal deletion. Peripheral chromosome and fluorescent in situ hybridisation (FISH) analysis of the patient showed mos46,XY,r(6)(p24;q26),del(6)(q27) [30]/46,XY,del(6)(q27) [20] de novo. We presented the patient in the light of literature because the mosaic ring 6 and 6q terminal deletion was different caryotypically from other mosaic ring 6 patients.

Germ cell tumor showing partial trisomy 1 in a gonadectomized intersex child with monosomy X and double Y mosaicism.

High incidence of germ cell tumors arising from dysgenetic gonads in patients with sexual chromosome abnormalities has been described, especially in patients with a Y chromosome bearing cell line. Here we report a 14-year-old patient with ambiguous genitalia. Constitutional karyotype showed 45,X/46,X,derY [?t(Yp;Yq)] mosaicism. The patient developed an abdominally located mixed malignant germ cell tumor 5 years after the removal of the dysgenetic gonads. Tumor karyotype showed partial trisomy 1q, a derivative 8q, and a hyperdiploidy with +X, +7, +12, +15, +19, +21, and an unidentified marker.