Gross and histological findings in the canine placenta and amnion at term: What's normal, abnormal or pathological?

The canine placenta is an underexamined organ. Placental abnormalities can affect foetus development and may be responsible for a low weight of the infant at birth; however, knowledge on their clinical significance in the canine species is limited. We aimed to describe macroscopic and microscopic findings in the canine placenta and amnion at term in clinically uncomplicated pregnancies and to evaluate their relationship with birth weight of healthy puppies. During natural delivery or C-section, the birth weight of 82 puppies was recorded, 72 placentas and 66 amnions were recovered. The foetal and maternal surfaces of the placental girdle, marginal haematoma and amnion were evaluated. Each gross finding was recorded, morphometrically assessed and sampled for histological diagnosis. Furthermore, specimens of placenta and amnion were collected from representative areas and microscopic deviations from normal structure were evaluated in haematoxylin and eosin sections. Gross examination revealed 'abnormalities' in the 75.4% of the collected placentas. Necrosis was the gross change most commonly observed in the placental girdle (72.5%). Congestion (17.4%) and clotted blood/fibrinoid material (2.9%) were also observed. No gross changes of either the marginal haematoma or the amnion were recorded. Histologically, placental girdle showed necrosis (62.3%), mineralization (52.2%), congestion (36.2%) and neutrophilic infiltration (27.5%). Marginal haematoma exhibited mineralization (11.6%) and neutrophils (29%), while necrotic foci were rarely observed (4.3%). In the amnion, the most frequent alteration observed was hypertrophy of the epithelium (35.9%) followed by oedema (31.2%), mineralized foci (28.1%), fibrosis (23.4%), congestion (15.6%) and more rarely neutrophils (12.5%). Puppies' birth weight was not statistically affected by either gross or histological abnormalities. Our study revealed that macroscopic and microscopic 'abnormalities' of the placenta and amnion may be common in uncomplicated pregnancies at term; however, no implications on puppies' birth weight were observed. Deviations from 'normal' morphology of canine foetal adnexa warrant further investigation to assess their clinical implications if present.

Effect of umbilical cord length on early fetal biomechanics.

The umbilical cord suspends the fetus within the amniotic cavity, where fetal dynamics is one of its many functions. Hence, the umbilical cord is a viable index in determining fetal activity. Fetal movements result in mechanical loads that are fundamental for fetal growth. At present, mechanical environment during early human fetal development is still largely unknown. To determine early fetal movement dynamics at given physiological (0.060 m) and pathological umbilical cord lengths (0.030 m, 0.020 m, 0.017 m and 0.014 m) a 2D computational model was created to simulate dynamic movement conditions. Main findings of this computational model revealed the shortest umbilical cord length (0.014 m) with a 6(10-6)N, twitch force amplitude had a two-fold increase on linear velocity (0.12 m/s) in comparison with other lengths (0.05m/s). Moreover, umbilical cord length effect presented an increasing exponential tension on the fetus body wall from longest to shortest, from 0 N in the control length to 0.05 N for the shortest umbilical cord. Last, tension was always present over a period of time for the shortest cord (0.03 N to 0.08 N). Collectively, for all variables evaluated the shortest umbilical cord (0.014 m) presented remarkable differences with other lengths in particular with the second shortest umbilical cord (0.017 m), suggesting a 0.003 m difference represents a greater biomechanical effect. In conclusion, this computational model brings new insights required by clinicians, where the magnitude of these loads could be associated with different pathologies found in the clinic.

Perinatal outcomes in dichorionic diamniotic twins with multifetal pregnancy reduction versus expectant management: A systematic review and meta-analysis.

BACKGROUND: Published findings on perinatal outcomes of multifetal pregnancy reduction (MPR) of dichorionic diamniotic (DCDA) twin pregnancy to singleton are controversial. We performed a meta-analysis to appraise the effects of MPR of DCDA twin pregnancy versus expectant management on perinatal outcomes. METHODS: Four electronic databases were searched from their inception to June 15, 2019, to identify publications that appraised MPR before 15 weeks of gestation. Studies reporting perinatal outcomes of both MPR of DCDA twin pregnancy to singleton and expectant management were considered. The relative risks (RRs) and mean differences with 95% confidence intervals (CIs) were pooled using a random-effects model. RESULTS: Six studies involving 7398 participants showed that MPR of DCDA twin pregnancy to singleton was associated with a lower risk of preterm birth (5 studies with 7297 participants; RR: 0.30, 95% CI: 0.22-0.40; P < .001) and higher birth weight (4 studies with 5763 participants; mean differences: 548.10 g, 95% CI: 424.04-672.15; P < .001) than expectant management; there was no difference in the occurrence of miscarriages (5 studies with 7355 participants; RR: 1.57, 95% CI: 0.90-2.75; P = .11). Sensitivity analysis showed that all the results were stable and reliable, with the omission of 2 studies with serious risk of bias. CONCLUSION: Compared to expectant management, MPR of DCDA twin pregnancy to singleton prevents preterm birth and low birth weight, without increasing the risk of miscarriages. Regarding perinatal morbidity related to preterm birth, MPR can be reserved as a remediation measure to improve the perinatal outcomes of DCDA twin pregnancies.

Super-Thick Amniotic Membrane Graft for Ocular Surface Reconstruction.

PURPOSE: The purpose of this study was to evaluate super-thick amniotic membrane grafts (ST-AMGs) for ocular surface reconstruction. DESIGN: Retrospective, interventional case series. METHODS: This was a single-center study of clinical practice that included select patients with typically large ocular surface abnormalities that required reconstruction. The intervention studied was surgical insertion of a ST-AMG for reconstruction or repair of the ocular surface. Main outcome measures included intraoperative handling, graft position at 1 week post implantation, graft dissolution at 3 weeks, epithelialization of the ocular surface and symblepharon. RESULTS: Eleven ST-AMGs were implanted after resection with cryotherapy: 5 conjunctival melanoma, 4 squamous cell carcinoma, 1 sebaceous carcinoma, and 1 atypical pterygium. In addition, 1 was implanted for scleral necrosis. ST-AMGs were up to nine times thicker than standard amniotic grafts and were therefore amenable to both running and interrupted 7-0 Vicryl sutures without cheese-wiring. All cases had a well-positioned ST-AMG at 1 week and 75% (n = 9) had partial graft dissolution at 3 weeks. Complete epithelialization without wound dehiscence was noted in all cases. However, secondary (after additional tumor treatment) symblepharon formed in 16.7% (n = 2). In all cases, the mean visual acuity and intraocular pressures remained unchanged during conjunctival reconstruction and subsequent secondary treatments. Post epithelialization adjuvant topical chemotherapy was given to extend treatment margins and treat presumed occult disease in 50% (n = 6). At mean follow-up of 25.5 months (median 10, range 3-90), 10 cases (83.3%) showed complete local tumor control, 1 showed revascularization of the scleral melt, and 1 required orbital exenteration. CONCLUSION: ST-AMGs were easy to suture and relatively persistent. Epithelialization of the ocular surface without primary symblepharon formation was noted. ST-AMGs should be considered an alternative for ocular surface reconstruction.

Similarities between induced membrane and amniotic membrane: Novelty for bone repair.

Previous clinical studies have shown the efficacy of a two-stage surgical procedure - the induced membrane (IM) technique - for reconstruction of large bone defects or bone non-union. The first stage involves radical debridement and insertion of a cement spacer into the bone defect. The second stage, performed weeks to months later, consists of removing the spacer while leaving the foreign body membrane induced by the cement in place, and then filling the cavity with bone autograft. The IM has been shown to (1) act as a protective physical barrier by preventing bone autograft resorption and (2) act as a bioreactor by promoting healing through revascularisation and growth factor secretion, and by concentrating mesenchymal stem cells (MSC) with osteogenic properties. New solutions to reduce this surgical procedure to a single step are being explored, for example by using an IM-like bioactive and protective barrier inserted into the bone defect at the same time as bone graft.

Liver diseases: what is known so far about the therapy with human amniotic membrane?

Liver, the largest intern organ of the human body, is responsible for several vital tasks such as digestive and excretory functions, as well as for nutrients storage and metabolic functions, synthesis of new molecules and purification of toxic chemicals. Cirrhosis, fibrosis and hepatocellular carcinoma are the most prevalent liver diseases. Despite all the studies performed so far, treatment options for these diseases are very limited. For this reason, it is urgent to find effective therapies for these pathologies. Several studies have been performed during the last decade about the possible application of human amniotic membrane in hepatic diseases therapy. Promising results about human amniotic membrane or its derived cells, in vitro and in vivo, applications in fibrosis, cirrhosis and hepatocellular carcinoma were already published. Since it is an attractive study area, it is becoming a dynamic scientific subject. However, the action mechanisms of human amniotic membrane and its derived cells in hepatic diseases therapy must be precisely known in order that this promising therapy could be clinically used.

Outcome of monochorionic-biamniotic twins conceived by assisted reproduction: a population-based study.

Objecive: To evaluate the perinatal outcome of monochorionic-diamniotic twins conceived by assisted reproduction technology (ART). METHODS: We compared data from a national population-based perinatal registry on perinatal outcomes of monochorionic-diamniotic twins conceived by ART with their dichorionic counterparts and with spontaneous monochorionic-diamniotic twins. We compared maternal characteristics, pregnancy complications and perinatal outcomes. RESULTS: Monochorionic-diamniotic twin pregnancies (n=45) comprised 5.5% of all ART twins, and 9.3% of all monochorionic twins in this dataset. ART does not appear to increase the already high risk of monochorionicity compared to spontaneous conception, with the latter having an increased incidence for birth weight <1500 g [odds ratio (OD) 2.6, 95% confidence interval (CI) 1.2-5.4]. CONCLUSION: Our results disagree with hospital-based data and suggest that monochorionic-diamniotic twins following ART are not at increased risk of adverse perinatal outcomes compared to spontaneous monochorionic-diamniotic twins as well as to dichorionc twins conceived by ART.

Perinatal outcome of dichorionic-triamniotic as compared to trichorionic triplets.

OBJECTIVE: To evaluate if the perinatal outcomes of dichorionic-triamniotic (DC) triplets are significantly different than that of trichorionic (TC) triplets. STUDY DESIGN: Comparison of maternal and neonatal data of 44 DC to 46 TC triplets, using univariate analysis. RESULTS: DC triplets were significantly more common after spontaneous conception but all other maternal characteristics as well complications and cesarean section rates were similar. Both groups had similar incidence of birth at <32 and <28 weeks as well as similar incidence of very low and extremely low birth weight. There was similar incidence of neonatal morbidity except for twin-twin transfusion syndrome (13.6%) in the DC group. The stillbirth rate was 45/1000 and 29/1000, the early neonatal mortality rates were 63/1000 and 45/1000, and the perinatal mortality rate was 106/1000 and 72/1000 for DC and TC triplets, respectively (all not significantly different). CONCLUSIONS: Our data indicate that DC twins are not significantly disadvantaged compared to TC triplets and the similar outcomes might be reassuring for those who consider continuing their DC triplet pregnancy.

The clinical applications of human amnion in plastic surgery.

Since the early 1900s, human amnion has been applied to a wide variety of clinical scenarios including burns, chronic ulcers, dural defects, intra-abdominal adhesions, peritoneal reconstruction, genital reconstruction, hip arthroplasty, tendon repair, nerve repair, microvascular reconstruction, corneal repair, intra-oral reconstruction and reconstruction of the nasal lining and tympanic membrane. Amnion epithelial and mesenchymal cells have been shown to contain a variety of regulatory mediators that result in the promotion of cellular proliferation, differentiation and epithelialisation and the inhibition of fibrosis, immune rejection, inflammation and bacterial invasion. The full repertoire of biological factors that these cells synthesise, store and release and the mechanisms by which these factors exert their beneficial effects are only now being fully appreciated. Although many commercially available biological and synthetic alternatives to amnion exist, ethical, religious, and financial constraints may limit the widespread utilisation of these products. Amnion is widely available, economical and is easy to manipulate, process and store. Although many clinical applications are of historical interest only, amnion offers an alternative source of multi-potent or pluripotent stem cells and therefore may yet have a great deal to offer the plastic surgery and regenerative medicine community. It is the purpose of this article to review the clinical applications of human amnion relevant to plastic surgery.

Vaginal delivery through annular placenta -- case report.

Annular placenta is an extremely rare morphological type of human placenta. It is commonly related to placental vessel abnormalities frequently causing antenatal and postnatal hemorrhage and operative delivery. Gravida 4 para 1 had an uneventful course of pregnancy and normal vaginal delivery followed by moderate postpartum hemorrhage. Hemorrhage was found to be local in origin but the placenta was annular in shape and the newborn was delivered through one of the openings. Annular placenta was not recognized before delivery. Its implantation site was in the lower uterine segment but high enough to allow the passage of the fetus through its annular defect and vaginal birth. To our knowledge, this is a first report of annular placenta ending in normal vaginal delivery.

Correlation between cord insertion type and chorionic villus vascularization of the co-twin in diamniotic-monochorionic twin pregnancies.

BACKGROUND: Recent evidence suggests that cord insertion type of one twin correlates with chorionic plate vascularization of the monochorionic co-twin. Specifically, for twins with paracentral cords, chorionic plate vascularization is significantly greater when the co-twin has a velamentous, rather than paracentral cord insertion. AIMS: To determine whether this correlation between cord insertion type and vascularization of the co-twin also extends to the deeper chorionic villus tree. STUDY DESIGN: Morphometric analysis of chorionic villus vascularization in CD31-immunostained sections of a retrospective cohort of gestational age-matched third trimester monochorionic placentas with discordant paracentral/velamentous (PC/V) or concordant paracentral/paracentral (PC/PC) cord insertions. OUTCOME MEASURES: Vascular numerical density (number of vascular profiles per unit villus stromal area) of intermediate villi (>80 µm diameter) and terminal villi (<80 µm). RESULTS: For twins with paracentral cord insertion, the vascular numerical density of intermediate villi was significantly higher for twins in a discordant PC/V relationship than for those in a concordant PC/PC relationship (P<0.05), thus replicating previous findings in superficial chorionic vessels. For terminal villi, in contrast, the vascular numerical density of twins with paracentral cords in a PC/V combination was significantly lower than of those in a PC/PC combination, and similar to that of their co-twins with velamentous cord insertion. CONCLUSIONS: Early placental angiogenesis in monochorionic twin gestations may be influenced by implantation and cord localization of the co-twin. The regulation of terminal villus angiogenesis appears to be dissociated from more proximal villus angiogenesis and independent of cord insertion of the co-twin.

Ceratoplastia lamelar em cães usando membrana amniótica equina: Estudo clínico e morfológico / Lamelar keratoplasty in dogs using equine amniotic membrane: Clinical and morphological study

A membrana amniótica (MA) consolidou-se no tratamento de afecções na superfície ocular. O objetivo deste estudo foi avaliar a viabilidade do implante de MA equina em ceratoplastia lamelar de cães. As membranas amnióticas foram preservadas em glicerina (98%). A eficácia do implante foi acompanhada por avaliação clínica, tempo de cicatrização, resposta inflamatória e reconstrução da arquitetura da córnea. Foram selecionados 12 cães, que foram divididos em quatro grupos de três animais. Em cada animal, foi realizada ceratotomia lamelar com 5 mm de diâmetro, seguida de aplicação do implante de MA. Após cirurgia, os animais foram avaliados em diferentes tempos: 2, 7, 21 e 40 dias. Duran- te o período de observação, os exames oftalmológicos foram realizados com intervalo de 48 h e, após a última avaliação, os animais foram submetidos à eutanásia. Os olhos foram enucleados, fixados e corados com hematoxilina-eosina (HE), ácido periódico de Schiff (PAS) e picrossirius. Os implantes foram completamente epitelizados em cerca de 10 dias após a cirurgia. Os neovasos apresentaram involução progressiva a partir de 21 dias e não foram detectados ao final de 40 dias pós-cirurgia, restando apenas uma nébula no local da lesão. À microscopia óptica, observou-se resposta inflama- tória moderada, presença de epitélio pavimentoso estratificado aos sete dias e epitelização completa aos 21 dias. Aos 40 dias, a membrana basal do epitélio apresentou-se reconstituída. Assim, concluímos que a membrana amniótica equina é viável como implante em córnea de cão, sendo incorporada ao estroma e resultando em restabelecimento parcial da transparência.

Amniotic membranes have been successfully used in the treatment of superficial corneal diseases. The purpose of this study was to assess the feasibility of use of equine amniotic membrane (AM) in lamellar keratoplasty of dogs. Amniotic membranes were preserved in glycerol (98%). Implant efficacy was assessed by clinical evaluation, time of healing, in-flammatory response, and the corneal architectural configuration. Twelve dogs were divided into four groups of three animals. In each animal, a 5-mm diameter lamellar keratectomy was performed, and followed by implantation of the AM. The animals were evaluated at the 2th, 7th, 21th, and 40th days after surgery. During this period, ophthalmological exams were performed at 48-h intervals, and the animals were euthanized after the last evaluation. The eyes were enu-cleated, included, fixed, and stained with hematoxylin-eosin (HE), periodic acid-Schiff (PAS), and picrosirius. Epitheli-zation of the implants was completed in about ten days after surgery. New vessels progressively diminished from 21 days on, were not detected on day 40 after surgery, and only a nebula was observed in the implant area. Optical microscopy revealed a mild inflammatory response, presence of squamous epithelium on day seven, and total epithelization on day 21 after surgery. On day 40, the basal epithelial membrane was shown to be reformed. Therefore, we conclude that use of equine amniotic membranes is feasible as implant for dog cornea since it is incorporated to the corneal stroma, and a relative transparency can be obtained.

Dichorionic pregnancy: delayed interval delivery with endoloop ligation.

We describe two cases of delayed delivery in dichorionic, diamniotic pregnancies, where we used an Endoloop ligature to clamp the umbilical cord with excellent maternal and fetal long-term outcome.

Prediction of abortion using three-dimensional ultrasound volumetry of the gestational sac and the amniotic sac in threatened abortion.

PURPOSE: To determine whether gestational sac volume (GSV) or amniotic sac volume (ASV) and/or the difference between them can predict abortion in women with first-trimester threatened abortion. METHODS: Ninety patients between 6 and 12 weeks of gestation presenting with vaginal bleeding were studied. Seventy-six delivered after 24 weeks of gestation (group A) and 14 aborted before 20 weeks of gestation (group B). All patients had a singleton viable pregnancy demonstrated by transvaginal ultrasound. Gestational sac and amniotic sac volumes were measured in all the patients using three-dimensional transvaginal ultrasound with Virtual Organ Computer-aided Analysis software, and the gestational sac volume - amniotic sac volume (GSV - ASV) was calculated. RESULTS: The groups did not differ in terms of age, parity, number of previous abortions, or term deliveries. The GSV (group A: mean 32.0 ± 27.7 cm(3) ; group B: 26.7 ± 29.1 cm(3) ) and the ASV (group A: 21.1 ± 25.5 cm(3) ; group B: 20.6 ± 26.0 cm(3) ) were not statistically different, while the GSV - ASV was significantly smaller in group B (aborting before week 20) (group A: 10.9 ± 10.9 cm(3) ; group B: 6.1 ± 8.6 cm(3) ; p < 0.05). Using receiver operator curves, the area under the curve for predicting normal pregnancy outcome of the GSV - ASV measurement was 0.654. When the GSV - ASV was 1.8 cm(3) or less, abortion was predicted with 84% sensitivity and 43% specificity. CONCLUSIONS: The measurement of the GSV and the ASV are not good predictors of abortion in patients with first-trimester vaginal bleeding, whereas the use of the GSV - ASV may be helpful in predicting the outcome of pregnancy.

Prospective risk of stillbirth: monochorionic diamniotic twins vs. dichorionic twins.

AIMS: To determine reliably the risk of stillbirth among twin pregnancies. METHODS: A data analysis of 3241 and 6581 women with monochorionic diamniotic (MD) and dichorionic (DD) twins, respectively, who gave birth at ≥22 weeks of gestation. RESULTS: The prospective risk (per 1000 women) of stillbirth, 25.3 and 11.5 at gestational week 22, decreased to <10.0 at gestational weeks 31 and 28 among the MD and DD twins, respectively. After single intrauterine fetal death, the co-twin died in utero or within 7 days of life more frequently among MD twins than among DD twins [42.7% (35/82) vs. 2.6% (2/76); relative risk, 16.2; 95% confidence interval, 4.0-65.1]. CONCLUSION: Women with MD twins were 2.2-fold more likely to experience stillbirth than women with DD twins. The prospective risks of stillbirth were <1.0% for both groups of women at ≥32 weeks of gestation.

The effects of preservation procedures on amniotic membrane's ability to serve as a substrate for cultivation of endothelial cells.

Amniotic membrane (AM) has been used as a scaffold for the ex vivo expansion of different types of cells and a cell delivery matrix in regenerative medicine. Since the preservation procedures can influence the AM properties for experimental and clinical purposes, this study was established to investigate the feasibility of using the AM after different preservation methods to serve as substrates for endothelial cell expansion ex vivo. The effects of cryopreservation and lyophilization were evaluated on mechanical and histological characteristics of the AM, and the results were compared with the fresh AM. The ECM components of the basement membrane were well conserved in all groups. Although lyophilization resulted in more histological changes and lower level of physical variables including thickness, F(max), elongation at break and suture retention than the fresh and cryopreserved AM, endothelial cells grown on the lyophilized AM were better attached to the basement membrane. Cytotoxicity assay by MTT showed that the lyophilized AM is a compatible substrate for endothelial cells cultivation. The findings of this study suggest that the lyophilized AM is a suitable matrix for cultivation of endothelial cells due to this fact that lyophilization led to exposure of basement membrane of the AM.

The impact of compositional topography of amniotic membrane scaffold on tissue morphogenesis of salivary gland.

Amniotic membrane (AM) has been widely used in the reconstruction of oral epithelial defects. However, whether it is also effective in facilitating tissue formation of salivary gland, an appendix of oral epithelia, has never been explored. To investigate the effects and the underlying mechanism of AM on salivary gland morphogenesis, murine fetal submandibular gland (SMG) explants were cultured on different preparations of AM scaffolds. It was found that, on AM stromal scaffold, SMG demonstrated well-developed branching morphogenesis. Nonetheless, on AM epithelial scaffold, SMG epithelial cell converted to a spindle-shape, lost intercellular connection, changed cytoskeletal organization, and exhibited scattering behaviors. Meanwhile, the integrity of SMG basement membrane was dismantled as well. However, when acellular AM epithelial scaffold was used, cultured SMG demonstrated organized morphology, indicating that AM epithelial component provided specific surface features for SMG morphogenesis. To further investigate AM scaffold morphogenetic effect, it was found hepatocyte growth factor (HGF), an epithelial scattering factor, was expressed abundantly in cultivated AM epithelia. After blocking HGF function of AM, cultured SMG regained branching activity, reorganized cell adhesion and subcellular organization, and reproduced basement membranes. Therefore, AM-derived bioactive factor profoundly influences cell behaviors and structure formation of SMG. Together, this study showed that compositional topography of AM scaffold is important in affecting SMG morphogenesis. By understanding the effects of AM scaffold on SMG morphogenesis, it provides important information for rationally designing and fabricating AM scaffold for salivary gland regeneration.

The amniotic membrane as a source of stem cells.

Cellular therapy has emerged as a new potential tool for curing a wide range of degenerative diseases and tissue necrosis. Embryonic stem cells possess potential for differentiation into a wide range of cell lineages, but the ethical issues associated with establishment of this human cell line have to be resolved prior to any use. The bone marrow (BM) is the usual source of adult stem cells for hematopoietic stem cell transplants and cellular therapy, but the BM harvest is a surgical procedure that requires general anesthesia or sedation, and there seems to be a reduction of the proliferative potential and differentiation capacity of the marrow mesenchymal stem cells in older donors. For these reasons there is an increasing interest in other sources of stem cells from adult and fetal tissues. The amniotic membrane (AM) or amnion is a tissue of particular interest because its cells possess characteristics of stem cells with multipotent differentiation ability, and because of low immunogenicity and easy procurement from the placenta, which is a discarded tissue after parturition, thus avoiding the current controversies associated with the use of human embryonic stem cells. Therefore, amniotic membrane has been proposed as a good candidate to be used in cellular therapy and regenerative medicine.