Psychological Distress and Negative Emotions in Post-COVID Infection: A Comparative Study of the COVID and NO-COVID Young Patients.

According to the literature, mental health assumed urgent relevance, and several scholars are debating on the enduring of the neurological and psychiatric symptoms in post COVID patients. Our study aimed to investigate the emotional dimensions in young population to the COVID exposition: primary endpoint was to detect the psychological distress up to 3 months in post-COVID-19. A comparative study was conducted among young adults in Italy. We also assessed dysphoria, depression, anxiety, stress symptoms, pessimism, and positive personality traits. The participants were 140 Italian young aged 18-30 years (mean = 22.1, SD ± 2.65; 65.0% female). The sample was distinguished in two groups: COVID and NO-COVID groups. The results revealed that young who have been exposed to COVID-19 infection evidenced emotional vulnerability by higher psychological distress (depression, anxiety, stress), dysphoria signs (irritability, discontent, interpersonal resentment, and feelings of renunciation/surrender) then No COVID-19 infection young. Furthermore, COVID patients showed higher negative emotions about the expected life, uncertain for future, and loss of motivation (characterized no desires) than NO-COVID infection. In conclusion, the vulnerability of young exposed to COVID infection even in mild severity should be considered as emerging unmet need of mental health recovering: urgent health policy actions to boost the psychological, biological and social strategic pillar for young generation.

Early Diagnosis of Melanoma and Breast Cancer in Women: Influence of Body Image Perception.

The diagnosis of melanoma and breast cancer may impact many aspects of life with significant reductions in emotional functioning and quality of life. The aim of the study was to analyze the emotional traits of female patients with oncological in early-stage diagnosis, investigating predictors for psychological distress and analyzing body image perception. An observational study was conducted, A sample of 84 female cancer patients (age range 30-55 years) with melanoma (n = 42) and breast cancer diagnosis (n = 42). The examined emotional variables were psychological distress; depression, stress, and anxiety; metacognitions; and body self-perception. Findings showed higher psychological distress in breast cancer than in melanoma patients (p = 0.00), which was related to lower positive self-perception of body image (p = 0.03). Furthermore, psychological distress was negatively correlated with consequences of clinical treatment on body image, and low well-being affected the social interaction and well-being with own body. There was no significant difference between cancer staging and timing from diagnosis. Prevention and therapeutic psychological protocols might be adapted and tailored to the unmet needs of the patients in medical treatments to promote and enhance the Quality of Life in survivorship.

Effects of P4 Antagonist RU486 on VEGF and Its Receptors' Signaling during the In Vivo Transition from the Preovulatory to Periovulatory Phase of Ovarian Follicles.

The development of an adequate blood vessel network is crucial for the accomplishment of ovarian follicle growth and ovulation, which is necessary to support the proliferative and endocrine functions of the follicular cells. Although the Vascular Endothelial Growth Factor (VEGF) through gonadotropins guides ovarian angiogenesis, the role exerted by the switch on of Progesterone (P4) during the periovulatory phase remains to be clarified. The present research aimed to investigate in vivo VEGF-mediated mechanisms by inducing the development of periovulatory follicles using a pharmacologically validated synchronization treatment carried out in presence or absence of P4 receptor antagonist RU486. Spatio-temporal expression profiles of VEGF, FLT1, and FLK1 receptors and the two major MAPK/ERKs and PI3K/AKT downstream pathways were analyzed on granulosa and on theca compartment. For the first time, the results demonstrated that in vivo administration of P4 antagonist RU486 inhibits follicular VEGF receptors' signaling mainly acting on the theca layer by downregulating the activation of ERKs and AKTs. Under the effect of RU486, periovulatory follicles' microarchitecture did not move towards the periovulatory stage. The present evidence provides new insights on P4 in vivo biological effects in driving vascular and tissue remodeling during the periovulatory phase.

Mediator Effect of Affinity for E-Learning on Mental Health: Buffering Strategy for the Resilience of University Students.

The pandemic affected the quality of life and wellness of the population, changing living habits through restriction measures. This study aimed to analyze the psychological impact of the fear of the COVID-19 pandemic and the adoption of e-learning for university students. The study was articulated in two research applications: the first application was a rapid review on the psychological effects of the pandemic on the emotional dimension of undergraduate students; the second application was an observational study on the effect of e-learning adoption in the pandemic emergency. In the first step, we performed a systematic search of MEDLINE through PubMed and the Web of Science [Science Citation Index Expanded (SCI-EXPANDED); Social Sciences Citation Index (SSCI); Emerging Sources Citation Index (ESCI)] of all scientific literature published from May 2020 to February 2021. The reviewed articles suggest the impact of the pandemic and lockdown measures on university students due to several mental symptoms, including anxiety, stress, depression, event-specific distress, and a decrease in psychological well-being. Psychological symptoms were related to the experience of several stressors, such as the risk for a reduction of academic perspectives, massive e-learning adoption, economic issues, social restrictions, and implications for daily life related to the COVID-19 outbreak. The second scientific application was conducted to evaluate the affinity for e-learning on a sample composed of Italian undergraduates exposed to massive e-learning adoption. The results evidence the positive influence of e-learning in academic programs for the wellbeing of undergraduates. The mediator effect of the affinity of youth for e-learning can be considered to have had a buffering effect for professional advancement and for the mental health of university students in a public health emergency.

Therapeutic potential of hAECs for early Achilles tendon defect repair through regeneration.

Cell-based therapy holds great promise for tendon disorders, a widespread debilitating musculoskeletal condition. Even if the cell line remains to be defined, preliminary evidences have proven that amniotic-derived cells possess in vitro and in vivo a great tenogenic potential. This study investigated the efficacy of transplanted human amniotic epithelial cells (hAECs) by testing their early regenerative properties and mechanisms involved on a validated ovine Achilles tendon partial defect performed on 29 animals. The injured tendons treated with hAECs recovered rapidly, in 28 days, structural and biomechanical properties undertaking a programmed tissue regeneration, differently from the spontaneous healing tissues. hAECs remained viable within the host tendons establishing with the endogenous progenitor cells an active dialogue. Through the secretion of modulatory factors, hAECs inhibited the inflammatory cells infiltration, activated the M2 macrophage subpopulation early recruitment, and accelerated blood vessel as well as extracellular matrix remodelling. In parallel, some in situ differentiated hAECs displayed a tenocytelike phenotype. Both paracrine and direct hAECs stimulatory effects were confirmed analysing their genome profile before and after transplantation. The 49 human up-regulated transcripts recorded in transplanted hAECs belonged to tendon lineage differentiation (epithelial-mesenchymal transition, connective specific matrix components, and skeleton or muscle system development-related transcripts), as well as the in situ activation of paracrine signalling involved in inflammatory and immunomodulatory response. Altogether, these evidences support the hypothesis that hAECs are a practicable and efficient strategy for the acute treatment of tendinopathy, reinforcing the idea of a concrete use of amniotic epithelial cells towards the clinical practice.

Amniotic epithelial stem cell biocompatibility for electrospun poly(lactide-co-glycolide), poly(ε-caprolactone), poly(lactic acid) scaffolds.

Three biodegradable thermoplastic polymers, poly(ε-caprolactone) (PCL), poly(l-lactide-co-d,l-lactide) (PLA) and poly(l-lactide-co-glycolide) (PLGA), have been used to produce nonwovens scaffolds with uniform micrometer fibres. Scaffolds' physical and morphological characterization was performed by X-ray diffraction, Scanning Electron Microscopy and Contact-Angle test. Morphological investigations revealed that all produced fibres were randomly orientated with interconnected pores ranging between 5 and 12µm in diameter. An average fibre diameter of 1.5, 0.75 and 1.2µm was found for PCL, PLA and PLGA, respectively. Moreover, experiments were designed to verify whether the fabricated electrospun substrates were biocompatible for ovine amniotic epithelial stem cells (oAECs) under in vitro conditions. Cell adhesion, survival, spatial organization on fibres, proliferation index, and DNA quantification after 48h culture, showed an enhanced adhesion and proliferation, especially for PLGA scaffolds. The favourable interaction between oAECs and the fibrous scaffolds was attributed to the greatly improved porosity and pore size distribution of the electrospun scaffolds. In addition, AECs can be considered ideal for tissue engineering especially when using biocompatible and opportunely produced scaffolds.

Thermal balneotherapy in Antsirabe-Madagascar: water analysis and its applications in an African context.

The Orthopaedic Rehabilitation Centre of Madagascar can be found in the city of Antsirabe. The health care facility is equipped with 2 thermal sources of bicarbonate water used for post-operative rehabilitation and for the ailment of chronic degenerative illnesses. The aim of this study, which derives from the cooperation between the School of Specialization in Thermal Therapy of the University of Parma and Antsirabe Orthopaedic Hospital, is to analyze the real properties of thermal waters utilized in this centre and to do an overview of its possible applications in this particular African context.

M1 and M2 macrophage recruitment during tendon regeneration induced by amniotic epithelial cell allotransplantation in ovine.

Recently, we have demonstrated that ovine amniotic epithelial cells (oAECs) allotransplanted into experimentally induced tendon lesions are able to stimulate tissue regeneration also by reducing leukocyte infiltration. Amongst leukocytes, macrophages (Mφ) M1 and M2 phenotype cells are known to mediate inflammatory and repairing processes, respectively. In this research it was investigated if, during tendon regeneration induced by AECs allotransplantation, M1Mφ and M2Mφ phenotype cells are recruited and differently distributed within the lesion site. Ovine AECs treated and untreated (Ctr) tendons were explanted at 7, 14, and 28 days and tissue microarchitecture was analyzed together with the distribution and quantification of leukocytes (CD45 positive), Mφ (CD68 pan positive), and M1Mφ (CD86, and IL12b) and M2Mφ (CD206, YM1 and IL10) phenotype related markers. In oAEC transplanted tendons CD45 and CD68 positive cells were always reduced in the lesion site. At day 14, oAEC treated tendons began to recover their microarchitecture, contextually a reduction of M1Mφ markers, mainly distributed close to oAECs, and an increase of M2Mφ markers was evidenced. CD206 positive cells were distributed near the regenerating areas. At day 28 oAECs treated tendons acquired a healthy-like structure with a reduction of M2Mφ. Differently, Ctr tendons maintained a disorganized morphology throughout the experimental time and constantly showed high values of M1Mφ markers. These findings indicate that M2Mφ recruitment could be correlated to tendon regeneration induced by oAECs allotransplantation. Moreover, these results demonstrate oAECs immunomodulatory role also in vivo and support novel insights into their allogeneic use underlying the resolution of tendon fibrosis.

Integrated treatment of the severely injured patient with coexistent comorbidities. A case report and literature review.

Major trauma represents a paradigmatic clinical condition, needing multidisciplinary and structured approach. When a patient is affected by significant comorbidities such as diabetes mellitus and other conditions that compromise the immune competence the risk of infections in the presence of exposed fractures is remarkably high. Although usually managed by multidisciplinary teams, these patients often have unfavorable functional outcomes. We describe here the case of a young diabetic and immunosuppressed man, treated for a destroying injury of the left lower limb, with more than 60 centimeters of multiple fractures exposition. The prompt and fast integration of several different competences led to favorable functional outcome, without any severe local or systemic complication. Therefore, in severely injured patients, especially when affected by significant comorbidities, a quick and highly integrated treatment, provided by a polyspecialistic team, is crucial for reducing unfavorable outcomes and improving the recovery rate.

Niccolò Paganini: the hands of a genius.

Niccolò Paganini was a man stricken by many diseases and according to Myron Schonenfield and Renzo Mantero, he may have also suffered from Marfan Syndrome. The following paper is based on actual physical examinations reports made by his contemporary physicians, documented in personal letters In addition, the current investigation presents a comparison between a post-mortem mold of Paganini's right hand and several paintings of the Maestro drawn by Lyser, his personal portraitist, and the images of the hand of a well-known musician affected by Marfan Syndrome which was treated by the authors. (www.actabiomedica.it).

Costs optimization in anaesthesia.

OBJECTIVES: The aim of this study is to analyze the direct cost of different anaesthetic techniques used within the Author's hospital setting and compare with costs reported in the literature. METHODS: Mean cost of drugs and devices used in our local Department of Anaesthesia was considered in the present study. All drugs were supplied by the in-house Pharmacy Service of Parma's General Hospital. All calculation have been made using an hypothetical ASA1 patient weighting 70 kg. The quality of consumption and cost of inhalation anaesthesia with sevoflurane or desflurane at different fresh gas flow were analyzed, and the cost of total venous anaesthesia (TIVA) using propofol and remifentanil with balanced anaesthesia were also analyzed. In addition, direct costs of general, spinal and sciatic-femoral nerve block anaesthesia used for common plastic surgery procedures were assessed. RESULT: The results of our study show that the cost of inhalational anaesthesia decreases using fresh gas flow below 1L, and the use of desflurane is more expensive. In our Hospital, the cost of TIVA is more or less equivalent to the costs of balanced anaesthesia with sevoflurane in surgical procedure lasting more than five hours. The direct cost was lower for the spinal anaesthesia compared with general anaesthesia and sciatic- femoral nerve block for some surgical procedures. (www.actabiomedica.it).

Cellular and molecular maturation in fetal and adult ovine calcaneal tendons.

Processes of development during fetal life profoundly transform tendons from a plastic tissue into a highly differentiated structure, characterised by a very low ability to regenerate after injury in adulthood. Sheep tendon is frequently used as a translational model to investigate cell-based regenerative approaches. However, in contrast to other species, analytical and comparative baseline studies on the normal developmental maturation of sheep tendons from fetal through to adult life are not currently available. Thus, a detailed morphological and biochemical study was designed to characterise tissue maturation during mid- (2 months of pregnancy: 14 cm of length) and late fetal (4 months: 40 cm of length) life, through to adulthood. The results confirm that ovine tendon morphology undergoes profound transformations during this period. Endotenon was more developed in fetal tendons than in adult tissues, and its cell phenotype changed through tendon maturation. Indeed, groups of large rounded cells laying on smaller and more compacted ones expressing osteocalcin, vascular endothelial growth factor (VEGF) and nerve growth factor (NGF) were identified exclusively in fetal mid-stage tissues, and not in late fetal or adult tendons. VEGF, NGF as well as blood vessels and nerve fibers showed decreased expression during tendon development. Moreover, the endotenon of mid- and late fetuses contained identifiable cells that expressed several pluripotent stem cell markers [Telomerase Reverse Transcriptase (TERT), SRY Determining Region Y Box-2 (SOX2), Nanog Homeobox (NANOG) and Octamer Binding Transcription Factor-4A (OCT-4A)]. These cells were not identifiable in adult specimens. Ovine tendon development was also accompanied by morphological modifications to cell nuclei, and a progressive decrease in cellularity, proliferation index and expression of connexins 43 and 32. Tendon maturation was similarly characterised by modulation of several other gene expression profiles, including Collagen type I, Collagen type III, Scleraxis B, Tenomodulin, Trombospondin 4 and Osteocalcin. These gene profiles underwent a dramatic reduction in adult tissues. Transforming growth factor-ß~1 expression (involved in collagen synthesis) underwent a similar decrease. In conclusion, these morphological studies carried out on sheep tendons at different stages of development and aging offer normal structural and molecular baseline data to allow accurate evaluation of data from subsequent interventional studies investigating tendon healing and regeneration in ovine experimental models.

In Vivo Behavior of a Custom-Made 3D Synthetic Bone Substitute in Sinus Augmentation Procedures in Sheep.

In this study, the in vivo behavior of a custom-made three-dimensional (3D) synthetic bone substitute was evaluated when used as scaffold for sinus augmentation procedures in an animal model. The scaffold was a calcium phosphate ceramic fabricated by the direct rapid prototyping technique, dispense-plotting. The geometrical and chemical properties of the scaffold were first analyzed through light and electron scanning microscopes, helium picnometer, and semi-quantitative X-ray diffraction measurements. Then, 6 sheep underwent monolateral sinus augmentation with the fabricated scaffolds. The animals were euthanized after healing periods of 45 and 90 days, and block sections including the grafted area were obtained. Bone samples were subjected to micro computerized tomography, morphological and histomorphometric analyses. A complete integration of the scaffold was reported, with abundant deposition of newly formed bone tissue within the biomaterial pores. Moreover, initial foci of bone remodeling were mainly localized at the periphery of the implanted area after 45 days, while continuous bridges of mature lamellar bone were recorded in 90-day specimens. This evidence supports the hypothesis that bone regeneration proceeds from the periphery to the center of the sinus cavity. These results showed how a technique allowing control of porosity, pore design, and external shape of a ceramic bone substitute may be valuable for producing synthetic bone grafts with good clinical performances.

Gestational stage affects amniotic epithelial cells phenotype, methylation status, immunomodulatory and stemness properties.

Stem cells isolated from amniotic epithelium (AECs) have shown great potential in cell-based regenerative therapies. Because of their fetal origin, these cells exhibit elevated proliferation rates and plasticity, as well as, immune tolerance and anti-inflammatory properties. These inherent attitudes make AECs well-suited for both allogenic and xenogenic cellular transplants in animal models. Since in human only at term amnion is easily obtainable after childbirth, limited information are so far available concerning the phenotypic and functional difference between AECs isolated from early and late amnia. To this regard, the sheep animal model offers an undoubted advantage in allowing the easy collection of both types of AECs in large quantity. The aim of this study was to determine the effect of gestational age on ovine AECs (oAECs) phenotype, immunomodulatory properties, global DNA methylation status and pluripotent differentiation ability towards mesodermic and ectodermic lineages. The immunomodulatory property of oAECs in inhibiting lymphocyte proliferation was mainly unaffected by gestational age. Conversely, gestation considerably affected the expression of surface markers, as well the expression and localization of pluripotency markers. In detail, with progression of gestation the mRNA expression of NANOG and SOX2 markers was reduced, while the ones of TERT and OCT4A was unaltered; but at the end of gestation NANOG, SOX2 and TERT proteins mainly localized outside the nuclear compartment. Regarding the differentiation ability, LPL (adipogenic-specific gene) mRNA content significantly increased in oAECs isolated from early amnia, while OCN (osteogenic-specific gene) and NEFM (neurogenic-specific gene) mRNA content significantly increased in oAECs isolated from late amnia, suggesting that gestational stage affected cell plasticity. Finally, the degree of global DNA methylation increased with gestational age. All these results indicate that gestational age is a key factor capable of influencing morphological and functional properties of oAECs, and thus probably affecting the outcome of cell transplantation therapies.

Effect of antiprogesterone RU486 on VEGF expression and blood vessel remodeling on ovarian follicles before ovulation.

BACKGROUND: The success of ovarian follicle growth and ovulation is strictly related to the development of an adequate blood vessel network required to sustain the proliferative and endocrine functions of the follicular cells. Even if the Vascular Endothelial Growth Factor (VEGF) drives angiogenesis before ovulation, the local role exerted by Progesterone (P4) remains to be clarified, in particular when its concentration rapidly increases before ovulation. AIM: This in vivo study was designed to clarify the effect promoted by a P4 receptor antagonist, RU486, on VEGF expression and follicular angiogenesis before ovulation, in particular, during the transition from pre to periovulatory follicles induced by human Chorionic Gonadotropins (hCG) administration. MATERIAL AND METHODS: Preovulatory follicle growth and ovulation were pharmacologically induced in prepubertal gilts by combining equine Chorionic Gonadotropins (eCG) and hCG used in the presence or absence of RU486. The effects on VEGF expression were analyzed using biochemical and immunohistochemical studies, either on granulosa or on theca layers of follicles isolated few hours before ovulation. This angiogenic factor was also correlated to follicular morphology and to blood vessels architecture. RESULTS AND CONCLUSIONS: VEGF production, blood vessel network and follicle remodeling were impaired by RU486 treatment, even if the cause-effect correlation remains to be clarified. The P4 antagonist strongly down-regulated theca VEGF expression, thus, preventing most of the angiogenic follicle response induced by hCG. RU486-treated follicles displayed a reduced vascular area, a lower rate of endothelial cell proliferation and a reduced recruitment of perivascular mural cells. These data provide important insights on the biological role of RU486 and, indirectly, on steroid hormones during periovulatory follicular phase. In addition, an in vivo model is proposed to evaluate how periovulatory follicular angiogenesis may affect the functionality of the corpus luteum (CL) and the success of pregnancy.

Return to run after partial amputation of the ankle: clinical assessment and instrumental evaluation.

BACKGROUND AND AIM OF THE WORK: the traumatic amputation or partial amputation of a portion of the lower limb is one of the most serious and not so rare road accident and job injury. There are few cases reported of replantation of the lower extremities rather than amputation surgery. This work describes a case of partial amputation of the right ankle. METHODS: the emergency treatment consisted of rigorous lavage and debridement, reduction, stabilization of the ankle and restoreation of the neurovascular and soft tissues lesions. Because of cutaneous necrosis another surgical treatment of reverse rotation flap was needed. The good outcome has been documented with foot pressure analysis, stabilometric evaluation and gait and jump analysis based on measurements of acceleration. RESULTS: after seven months of personalized rehabilitation program the patient walks without devices, has recovered functional of range of movement and had no neurological deficit or subjective problems. Currently the subject has returned to his job and runs without pain. CONCLUSIONS: emergency foot salvage treatment was possible thanks to a close collaboration among different physicians with specialized skills, good emergency management and an adequate infrastructure.

Synthetic bone substitute engineered with amniotic epithelial cells enhances bone regeneration after maxillary sinus augmentation.

BACKGROUND: Evidence has been provided that a cell-based therapy combined with the use of bioactive materials may significantly improve bone regeneration prior to dental implant, although the identification of an ideal source of progenitor/stem cells remains to be determined. AIM: In the present research, the bone regenerative property of an emerging source of progenitor cells, the amniotic epithelial cells (AEC), loaded on a calcium-phosphate synthetic bone substitute, made by direct rapid prototyping (rPT) technique, was evaluated in an animal study. MATERIAL AND METHODS: Two blocks of synthetic bone substitute (∼0.14 cm(3)), alone or engineered with 1×10(6) ovine AEC (oAEC), were grafted bilaterally into maxillary sinuses of six adult sheep, an animal model chosen for its high translational value in dentistry. The sheep were then randomly divided into two groups and sacrificed at 45 and 90 days post implantation (p.i.). Tissue regeneration was evaluated in the sinus explants by micro-computer tomography (micro-CT), morphological, morphometric and biochemical analyses. RESULTS AND CONCLUSIONS: The obtained data suggest that scaffold integration and bone deposition are positively influenced by allotransplantated oAEC. Sinus explants derived from sheep grafted with oAEC engineered scaffolds displayed a reduced fibrotic reaction, a limited inflammatory response and an accelerated process of angiogenesis. In addition, the presence of oAEC significantly stimulated osteogenesis either by enhancing bone deposition or making more extent the foci of bone nucleation. Besides the modulatory role played by oAEC in the crucial events successfully guiding tissue regeneration (angiogenesis, vascular endothelial growth factor expression and inflammation), data provided herein show that oAEC were also able to directly participate in the process of bone deposition, as suggested by the presence of oAEC entrapped within the newly deposited osteoid matrix and by their ability to switch-on the expression of a specific bone-related protein (osteocalcin, OCN) when transplanted into host tissues.

H3K9 trimethylation precedes DNA methylation during sheep oogenesis: HDAC1, SUV39H1, G9a, HP1, and Dnmts are involved in these epigenetic events.

The oocyte, to become a fully mature gamete, has to acquire a correct pattern of DNA methylation on its genome; this epigenetic event represents the major point of the molecular mechanisms that occur during postnatal oogenesis. It is known that an intimate link exists between DNA methylation and histone posttranslational modifications, such as trimethylation of lysine 9 on histone 3 (H3K9me3), that is essential in the silencing of gene transcription. What remains unclear is the precise sequence of these two epigenetic events and the protein expression of the enzymes that catalyze this epigenetic maturation during oogenesis. To identify the key molecules involved in global DNA methylation and H3K9me3, a biological network-based computational model was realized. Then, the spatiotemporal distribution of the proteins, identified from the biological network, was assessed during postnatal oogenesis. The results obtained suggest the existence of a sequential cascade of events in which H3K9me3 is the primary step followed by DNA methylation. These two epigenetic marks are realized due to the recruitment of the HDAC1, SUV39H1, G9a, HP1, and Dnmt3a, which were always localized in the nuclei of the oocytes and were dependent on chromatin configuration. These results involving DNA methylation and H3K9me3 are crucial in defining the oocyte developmental competence.

Role of amniotic fluid mesenchymal cells engineered on MgHA/collagen-based scaffold allotransplanted on an experimental animal study of sinus augmentation.

OBJECTIVES: The present research has been performed to evaluate whether a commercial magnesium-enriched hydroxyapatite (MgHA)/collagen-based scaffold engineered with ovine amniotic fluid mesenchymal cells (oAFMC) could improve bone regeneration process in vivo. MATERIALS AND METHODS: Bilateral sinus augmentation was performed on eight adult sheep in order to compare the tissue regeneration process at 45 and 90 days after implantation of the oAFMC-engineered scaffold (Test Group) or of the scaffold alone (Ctr Group). The process of tissue remodeling was analyzed through histological, immunohistochemical, and morphometric analyses by calculating the proliferation index (PI) of oAFMC loaded on the scaffold, the total vascular area (VA), and vascular endothelial growth factor (VEGF) expression levels within the grafted area. RESULTS: MgHA/collagen-based scaffold showed high biocompatibility preserving the survival of oAFMC for 90 days in grafted sinuses. The use of oAFMC increased bone deposition and stimulated a more rapid angiogenic reaction, thus probably supporting the higher cell PI recorded in cell-treated sinuses. A significantly higher VEGF expression (Test vs. Ctr Group; p = 0.0004) and a larger total VA (p = 0.0006) were detected in the Test Group at 45 days after surgery. The PI was significantly higher (p = 0.027) at 45 days and became significantly lower at 90 days (p = 0.0007) in the Test Group sinuses, while the PI recorded in the Ctr Group continued to increase resulting to a significantly higher PI at day 90 (CTR day 45 vs. CTR day 90; p = 0.022). CONCLUSIONS: The osteoinductive effect of a biomimetic commercial scaffold may be significantly improved by the presence of oAFMC. CLINICAL RELEVANCE: The amniotic fluid mesenchymal cell (AFMC) may represent a novel, largely and easily accessible source of mesenchymal stem cells to develop cell-based therapy for maxillofacial surgery.

Blood vessel remodeling in pig ovarian follicles during the periovulatory period: an immunohistochemistry and SEM-corrosion casting study.

BACKGROUND: The present research aims to describe the process of vascular readjustment occurring in pig ovary during the periovulatory phase (from LH surge to ovulation) that drives the transformation of the follicle, a limited blood supplied structure, into the corpus luteum, a highly vascularised endocrine gland required to maintain high levels of progesterone in pregnancy. The swine model was chosen because it is characterized by a long periovulatory window (about 40-44 hrs-similar to human) that permits to recover follicles at a precise endocrinological timing. METHODS: By validated hormonal protocol (eCG+hCG), able to mimic the physiologic gonadotropin stimulation, preovulatory follicles (PreOFs, 60 h-eCG), follicles in the middle (early periovulatory follicles, EPerOFs, 18 h-hCG) or late (LPerOFs, 36 h-hCG) periovulatory phase were isolated from prepubertal gilts. To understand the angiogenic process, morphological/morphometrical analyses were performed by combining immunohistochemistry (IHC) and SEM of vascular corrosion casts (VCC) techniques. RESULTS: PreOFs showed a vascular plexus with proliferating endothelial cells (EPI). This plexus was characterized by a dense inner capillary network, with angiogenic figures, connected to the outer network by anastomotic vessels (arterioles and venules of the middle network). EPerOFs decreased their EPI, blood vessel extension in the outer network, and evidenced a reduced compactness of blood vessels. In LPerOFs, a rapid neovascularization was associated to an intensive tissue remodeling: the follicle acquired an undulated aspect presenting arterioles/venules near the basal membrane, increased vascular extension by EPI, sprouting and non-sprouting angiogenesis.The analysis of vascular geometric relations and branching angles evidenced similar values at all stages. CONCLUSION: These data allow us to hypothesize that EPerOFs are in a quiescent status. LPerOFs represent the "metamorphic" follicles that rapidly turn-on angiogenesis to sustain a successful corpus luteum formation. Particularly, it is interesting to underlie that the non-sprouting angiogenesis, typical of structures in rapid neovascularization, occurred only in the LPerOFs. Moreover, vascular geometric relations showed as blood vessel remodeling occurs with the "maximum output and the minimum energetic expense".This knowledge will allow to better understand the mechanisms regulating the reproductive success and to clarify the complex physiological angiogenic process in adult tissues.