Healing of particulate allografts mixed with platelet concentrates in ridge preservation and sinus lift: a prospective histomorphometric study.

The objective of the present study was to evaluate the bone quality of sinus and alveolar grafts following filling with particulate allogenous bone (DFDBA 300-500µm) and platelet concentrate (platelet-rich fibrin, PRF). A prospective interventional clinical study was carried out. A total of 40 bone cores, 2mm in diameter, were taken from 21 patients: 22 from grafted alveoli, 7 from grafted sinus sites, and 11 from native bone used as a control. Fixed, paraffin-embedded samples were subjected to histological staining with hematoxylin-eosin and Masson's trichrome. Bone maturity of the samples was evaluated by two independent operators using histomorphometric analysis. There existed a greater proportion of lamellar neoformed bone than woven neoformed bone as the healing time increased. Moreover, there was also an increasing proportion of newly formed bone in the grafted sockets as a function of healing time (average: 41.22% ≤ 5 months, 55.89% Ëƒ 5 months). Resorption of DFDBA particles also appears to be correlated with healing time in the grafted socket (average: 15.43 ≤ 5 months, 13.72% Ëƒ 5 months). In conclusion, performing sinus lift and alveolar socket preservation techniques using DFDBA and PRF results in high quality, mature bone tissue according to histological criteria.

Are birds pseudoteeth and denticulations related to touch papilla? An investigation in parrot, goose, and chicken.

We first studied the morphology and the development of goose denticulations, which develop mainly by a ripple process, and the touch papillae of the bill tip organ, which appears through an evagination process at the end of the beak. During their development, we observed the specific expression of PAX9, PITX2, and BMP4, while SHH was expressed mainly in the basal layer of the epithelium in a non-specific manner. Adult goose denticulations are associated with numerous columns. The goose denticulations and columns were filled with numerous Herbst and Grandry corpuscles, as well the touch papillae of the bill tip organ. Histological analysis of adult parrot pseudoteeth revealed that the osseous pseudoteeth were extended by similar columns filled with Herbst and Grandry corpuscles. We also examined adult and embryonic chicken beaks. During ontogeny, we observed a process of rostral evagination with folding associated with discrete ripples in the anterior part of the beak rudiment, in which PAX9, PITX2, and BMP4 are expressed. In the corresponding adult areas, there were numerous sensory corpuscles with rostral columns, which were similar to the features observed in goose. These observations support the hypothesis that pseudoteeth and denticulations constitute sensory organs, and that the touch papillae exhibit some similarities with pseudoteeth.

Effects of HSP90 inhibition on primordial germ cells migration: A study in the gonad of the chick embryo.

INTRODUCTION: Primordial Germ Cells (PGCs) differentiate into spermatozoa or oocytes. They appear early during embryonic development before migrating to the gonadal ridges. Because of their long migration, PGCs have been proposed as a valuable model to study long distance cell migration. Some species also present a vascular phase in the migration of the germline and could therefore be compared to metastatic migration. HSP90 is a heat shock protein involved in the stabilization of several client-proteins, including oncoproteins. HSP90 inhibition has been proved to decrease PGCs migration in mouse and zebrafish. MATERIAL AND METHODS: We investigated the effect of geldanamycin on PGCs migration in a species with a vascular phase, the chicken. Geldanamycin was injected in the egg at 48h of incubation, PGC's were detected in blood using of blood smears, and in the embryo by immunohistochemistry using anti-HSP90 antibody. RESULTS: The effects of the treatment were similar to those observed in mouse and zebrafish. We show the presence of ectopic germs cells in the vasculature and in the dorsal mesentery, and some deformities of the gonads. CONCLUSION: Inhibition of HSP90 decreases the migration of PGCs and proposed the migration of PGCs in the chick embryo as an interesting model to study metastatic invasion.

Is Vasa such a highly specific marker for primordial germ cells? A comparison of VASA and HSP90 proteins expression in young chicken embryos.

INTRODUCTION: Primordial germ cells (PGCs) have been studied since the 19th century with several different methods. The earliest works were based on the morphological criteria of these cells associated or not with a particular staining. Different markers have been proposed in immunohistochemistry among which we can quote the Stage-specific embryonic antigene-1 (SSEA-1), the embryonic mouse antigen-1 (EMA-1) or the heat shock protein 90. Unfortunately, none of them are germline specific. The VASA protein is considered as one of the most reliable marker for PGCs by some authors with its expression being considered to limited to the germ cells. However, other studies have reported its expression in somatic cells. Here, we described the expression of the heat shock protein, HSP90, and the VASA protein in the early chick embryo. MATERIAL AND METHODS: Embryos from stages Hamburger-Hamilton (HH) 19, 21 and 28 were collected. Embryos were dissected and fixed in Serra's medium. Sections were placed on slides for PAS staining and for double immunohistochemistry with HSP90 and VASA. RESULTS: VASA and HSP90 expression have been observed in germ cells but as well in other cell lineages with a spatio-temporal gradient in respect to the characteristics of development of each organ. The conclusion is that VASA expression is not limited to the germ line in chick embryo.

A reviewed chronology of primordial germ cells migration in the chick embryo.

INTRODUCTION: Primordial Germ Cells (PGCs) are present in all sexually reproducing animals. They differentiate into spermatozoa or oocytes and are therefore responsible for the transmission of genetic and epigenetic information across generations. In birds, PGCs are first observed in the center of the blastodisc at stage Eyal-Giladi X. With the formation of the primitive streak, germ cells are translocated anteriorly to the germinal crescent. At stage Hamburger- Hamilton 10-12, they enter the vasculature before migrating through the dorsal mesentery towards the genital ridges. MATERIAL AND METHODS: Embryos from stages Hamburger-Hamilton (HH) 16 to 22 were collected. Blood samples were taken from the dorsal aorta and from the heart in order to perform blood smears and PAS staining. Embryos were dissected and fixed in Serra's medium. Sections were placed on slides for PAS staining. A sample of each embryo was collected for DNA extraction and PCR in order to determine the sex of the embryos. RESULTS: PGCs were observed in blood circulation until stage HH 20 on blood smears and until stage HH 19 on histological sections. The first PGCs arrived in the genital ridges were observed from stage HH 17. A few germ cells were still migrating in the dorsal mesentery at stage HH 22. The aim of this study was to review the chronology of the migration of PGCs in chick embryos.

Expression of Wnt and beta-catenin in mouse tooth germs and chick facial primordia: a comparison / Expresión de Wnt y beta-catenina en gérmenes de dientes de ratón y primordios faciales de polluelos: una comparación

In order to compare Wnt/ beta-catenin expression in mouse and chick facial primordia during development, we performed an immunohistochemical analysis of both protein expressions in E12 to E17 mouse embryos and in E3 and E10 chick embryos. During odontogenesis, from bud to bell stage, both proteins exhibit similar fixation patterns, with epithelial and mesenchymal immunoreactivity, consistant with literature data. Double labelling demonstrates that the same cells express both antigens, even in undifferentiated mesenchyme. The enamel knot, and the ameloblastic and odontoblastic layers are stained at the same manner. In the chick, Wnt and beta-catenin are diffusely present on craniofacial mesenchyme. In both species, premuscular blastemata express Wnt and b-catenin, but Wnt is specifically expressed on the perichondrium and ossification centers, suggesting a role independent from beta-catenin pathway.

Geldanamycin administration reduces the amount of primordial germ cells in the mouse embryo.

INTRODUCTION: Heat shock proteins (HSPs) are expressed or overexpressed in response to exposure to stress. They act as molecular chaperones, ensuring the correct folding of numerous client proteins. HSP90 is one of the most conserved HSPs. Its role extends beyond stress tolerance. HSP90 also contributes to development, differenciation, apoptosis and oncogenesis. Numerous tumors are associated with an overexpression of HSP90 and this expression can be used to evaluate its metastatic capacity. Primordial germ cells (PGCs) exhibit HSP90 expression under normal conditions. PGCs arise early in development and migrate by a combination of passive and active movements towards the gonads. The aim of this work was to study the impact of an inhibition of HSP90 on the migration of the PGCs. Geldanamycin, a well established HSP90 inhibitor with potent antitumor properties was used to achieve this inhibition. MATERIEL AND METHODS: 5mg of Geldanamycin were administered to E8 pregnant mice. E17 embryos were removed and fixed for staining and Immunohistochemistry with anti-HSP90 and anti-VASA antibodies. RESULTS: Geldanamycin-treated mouse embryos exhibited less VASA-immunopositive cells compared to the non-treated ones. These results suggest that geldanamycin administration at the time of PGCs migration reduces the number of PGCs in the gonads. HSP90 and VASA stainings were identical. We therefore expressed the idea that HSP90 could be used as a reliable marker for PGCs.

[Immunohistochemical study of Dlx1 and Msx1 expression during cephalic development of Dumbo and Wistar rats. Correlation with morphological data]. / Étude immuno-histochimique de l'expression des protéines Dlx1 et Msx1 durant la céphalogenèse des rats Wistar et Dumbo Corrélation avec les données morphologiques.

The Dumbo rat is characterized by a short snout, low ears and relative hypoplasia of maxillar and zygomatic bones. It corresponds to an autosomal recessive genotype. Previous study demonstrated a global deficit of Dlx1 and Msx1 genes expression in comparison to Wistar embryos as considered as control animals. We performed a histological study of cephalic development of Dumbo rats compared to Wistar embryos and an immunohistochemical analysis of Dlx1 and Msx1 protein expression during cephalogenesis. Our data indicate that the pattern of expression of both genes is similar in both strains, but that quantitative differences in gene expression can be the result of delayed organogenesis in Dumbo rat in comparison to Wistar. Some data about gene expressions are discussed at the light of the postulated function of Dlx1 and Msx1 in cephalic development.

[Geldanamycin administration reduces the number of HSP86-positive germ cells in the mouse embryo: preliminary results]. / L'administration de geldanamycine réduit le nombre de gonocytes exprimant l'HSP86 chez l'embryon de souris: premiers résultats.

5 mg of Geldanamycin, an inhibitor of stress protein HSP86 which express on mammalian germ cells, were administered to E8 pregnant mice. E17 embryos were removed, and a quantitative analysis of HSP90-immunoreactive cells in the gonad was performed, in comparison to control embryos. First, we observed that the number of germ cells is lower in male than in female embryos, as well in control and experimental embryos. External features of experimental and control embryos did not display any difference. Embryos exposed to geldanamycin exhibit a significant decrease of immunoreactive germ cells. In two embryos, we observed a group of ectopic immunoreactive cells in the pelvic area. We conclude that geldanamycin inhibits germ cells migration, and suggest that this inhibition can lead to ectopic germ cell populations, similar to teratomas.

Correlation of Hsp110 expression with caspase-3 and -9 during apoptosis induced by in vivo embryonic exposition to retinoic acid or irradiation in early mouse craniofacial development.

OBJECTIVE: To analyze the expression and role of three proteins (HSP110, caspase-3 and caspase-9) during craniofacial development. DESIGN: Seven pregnant C57Bl/6J mice received, by force-feeding at gestation day 9 (E9), 80 mg/kg of all-trans retinoic acid mixed to sesame oil. Seven pregnant NMRI mice received two grays irradiation at the same gestation day. Control mice of both strains (seven mice for each strain) were not submitted to any treatment. Embryos were obtained at various stages after exposition (3, 6, 12 and 24 h), fixed, dehydrated and embedded. Coronal sections (5 microm) were made. Slide staining occurred alternatively using anti-Hsp110, anti-caspase-3 and anti-caspase-9 immunohistochemistry. RESULTS: Expression of HSP110, caspase-3 and caspase-9 was found in cells of well-known locations of programmed cell death. After retinoic acid exposure, expressions were increased especially in neural crest cells of mandibular and hyoid arches. Quantification of positive cells shows that caspase-9 and Hsp110 were expressed before caspase-3. After irradiation, the expression of the three proteins quickly increased with a maximum 3 h after irradiation. For all three models of apoptosis (physiological, retinoic-induced and irradiation-induced) HSP110 positive cells were more numerous than caspase-3 positive cells. Caspase-3 positive cells were more numerous than caspase-9 positive cells especially in mesectodermal irradiation-induced apoptotic cells. CONCLUSION: The findings show a potential function of HSP110 in apoptosis during embryo development. Caspase-3-expressing cells are more numerous than cells expressing caspase-9, especially irradiation-induced apoptotic neural crest cells. This suggests that other caspases, still to be identified, may activate caspase-3 in this model.