Search

Background: The healing of diabetic wounds is poor due to a collagen deposition disorder. Matrix metalloproteinase-9 (MMP-9) is closely related to collagen deposition in the process of tissue repair. Many studies have demonstrated that extracellular vesicles derived from adipose-derived stem cells (ADSC-EVs) promote diabetic wound healing by enhancing collagen deposition. Objective: In this study, we explored whether ADSC-EVs could downregulate the expression of MMP-9 in diabetic wounds and promote wound healing by improving collagen deposition. The potential effects of ADSC-EVs on MMP-9 and diabetic wound healing were tested both in vitro and in vivo. Methods: We first evaluated the effect of ADSC-EVs on the proliferation and MMP-9 secretion of HaCaT cells treated with advanced glycation end product-bovine serum albumin (AGE-BSA) using CCK-8, western blot and MMP-9 enzyme-linked immunosorbent assay(ELISA). Next, the effects of ADSC-EVs on healing, re-epithelialisation, collagen deposition, and MMP-9 concentration in diabetic wound fluids were evaluated in an immunodeficient mouse model via MMP-9 ELISA and haematoxylin and eosin, Masson’s trichrome, and immunofluorescence staining for MMP-9. Results: In vitro, ADSC-EVs promoted the proliferation and MMP-9 secretion of HaCaT cells. In vivo, ADSC-EVs accelerated diabetic wound healing by improving re-epithelialisation and collagen deposition and by inhibiting the expression of MMP-9. Conclusion: ADSC-EVs possess the potential of healing of diabetic wounds in a mouse model by inhibiting downregulating MMP-9 and improving collagen deposition. Thus, ADSC-EVs are a promising candidate for the treatment of diabetic wounds.

Purpose A "dead bag syndrome" has been recently described, in which the capsular bag appears to be clear many years after surgery, becoming diaphanous and floppy, and unable to support the intraocular lens (IOL) within it. The aim of this study was to describe findings of this syndrome. Setting John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. Design Case series with clinicopathological correlation. Methods Out of 10 cases suspected to be from dead bag syndrome, 8 IOLs/7 capsular bags were removed due to subluxation/dislocation. The seven capsular bags available for analysis were fixed in formalin and submitted to histopathological examination (hematoxylin/eosin and Masson's trichrome stains). The associated explanted IOLs in 5 cases were also examined microscopically. Results Histopathologic examination of the seven capsular bags showed capsular thinning and/or splitting. Lens epithelial cells (LECs) were completely absent on two specimens while the other five specimens had rare LECs on the inner surface of the capsule. Explanted IOLs were three-piece silicone lenses or single-piece hydrophobic acrylic lenses. One IOL optic showed a small amount of granular pigment deposition, but the optics of the other four lenses were unremarkable. Conclusions In this syndrome, there appears to be an absence of secondary proliferation of LECs, as well as fibrotic changes. The capsule shows some signs of degradation, such as thinning and/or splitting. Weakness of zonular attachments appears to be an associated finding, with subsequent in-the-bag IOL dislocation. Further studies are necessary to ascertain the etiology of this condition.

Aim: The authors aimed to quantitatively assess the variation with age of three of the main components of the aortic wall, namely elastic fibers (FE) and collagen fibers [FC(COL)], and smooth muscle cells (SMCs). Materials and Methods: Four aortic cross sections (base, cross, thoracic, abdominal) were collected during autopsy from 90 cases of all ages, processed using the classical histopathological (HP) technique (formalin fixation and paraffin embedding) and stained with Orcein and Goldner’s trichrome. The obtained histological slides were transformed into virtual slides. Quantitative measurements of the three components were made on identical regions of interest (ROIs) selected from two-paired slides stained with the above-mentioned techniques using custom-made software, developed in MATLAB (MathWorks, USA). Results and Discussions: FE revealed an obvious decreasing general trend with age, present at all four levels of investigation. Smooth muscle fibers (FM) density showed almost no variation with age regardless of the level at which the measurement was made. FC(COL) density had an obvious increasing trend with age, expressed in all four aortic regions FE densities and FM densities were higher in men than in women, while FC(COL) densities were higher in women than in men in three of the aortic regions excepting, in all cases, the cross region. Conclusions: There were differences between men and women concerning the composition of aortic wall cellular and extracellular compartments. FE and FC(COL) dominated the age-related remodeling process of the aortic wall. The process evolved in the same way in all regions of the aorta. Studies need to be continued to define more clearly this complex process of vascular wall remodeling with aging.

The primo vascular system (PVS) is observed in different parts of the body under different physiological and disease conditions. Previously, the PVS was not observed in the vagina. The vaginal samples of this study were collected from the female genitalia of healthy New Zealand white rabbits from the animal house, Faculty of Medicine, Assiut University. The vaginal samples were fixed in Bouin's solution. The sections were stained with hematoxylin and eosin and Crossmon's trichrome. Additionally, the sections were immunohistochemically stained with neuron-specific enolase (NSE) and vascular endothelial growth factor (VEGF). A primo node was observed on the lymph vessel of the vagina and has several characteristics that resemble those of the previously discovered primo nodes. The primo node in this study was surrounded by mesothelial cells that provide positive immunoreactivity to NSE and VEGF. Sinuses of different sizes, floating cells, telocyte-like cell, and primo microcells were observed as the main constituents of the primo node. Additionally, migratory cells were detected, which passed from the primo node to the enclosing lymph vessel.

The purpose of this study was to determine the efficacy of highly concentrated tricalcium phosphate (TCP) biocomposite screws on local bone formation in a rabbit model of osteoporosis induced by bilateral ovariohysterectomy (OHE). Fourteen 24-week-old female New Zealand rabbits (weight, 3-3.5 kg) were divided into two groups: 1) OHE and biodegradable poly(lactic-co-glycolic acid) (PLGA) without s-TCP plate or screw insertion (OHE/BSR) group; and 2) OHE and biocomposite PLGA with highly concentrated s-TCP plate and screw insertion (OHE/HCCP). Both groups underwent bilateral OHE and had two different types of screws and plates inserted at the proximal tibia. Bilateral tibiae were extracted at 25 weeks post-OHE. The extracted tibiae were scanned with ex-vivo micro-computed tomography (micro-CT). Parameters including bone mineral density (BMD), trabecular bone volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp) were evaluated after staining the tibial samples with hematoxylin and eosin (H&E) and Masson's trichrome. We then performed pathological assessments. Micro-CT images revealed improved new bone formation near the implant in the OHE/HCCP group with higher values of BMD, BV/TV, and Tb.N but lower values of Tb.Th and Tb.Sp than the OHE/BSR group. Analyses of H&E and Masson's trichrome staining showed better new bone formation around the implant in the OHE/HCCP group than in the OHE/BSR group. The use of highly concentrated TCP biocomposite screw and plate might improve local bone formation and facilitate osteoconductivity in an osteoporotic rabbit model. This article is protected by copyright. All rights reserved.

Background Arteriolosclerotic ulcers of Martorell are histologically characterized by hyaline arteriolosclerosis resulting in concentric occlusion of the arteriolar lumina. Although several authors have previously reported on hyaline changes in hypertensive arteriolopathies, so far, little information is available on the molecular composition of hyaline wall depositions. Objectives This study aimed at the molecular characterization of hyaline arteriolar deposits in patients with hypertensive arteriolopathy using confocal Raman spectroscopy. Methods Samples of patients diagnosed with arteriolosclerotic ulcers of Martorell were analysed using confocal Raman spectroscopy. The findings were correlated with histological analyses. Skin samples from healthy, non-hypertensive patients served as controls. Results Confocal Raman spectroscopy analysis revealed that subendothelial hyaline deposits in arteriolosclerotic ulcers are mainly composed of collagen and phospholipids, in particular phosphatidylcholine. The presence of collagen within hyaline deposits was confirmed by Masson's Trichrome and Picrosirius Red staining. Additionally, the presence of collagen could also be shown for hypertensive nephrosclerosis. Actin was markedly decreased in hyalinized compared to control vessels, corresponding to the loss of smooth muscle cells in the process of hyalinization. This was confirmed by immunofluorescence staining for α-smooth muscle actin and desmin. Conclusion The present findings suggest that arteriolar hyaline deposits in hypertensive arteriolopathy are mainly composed of collagen and phospholipids, in particular phosphatidylcholine. Together with the concurrent absence of actin, these findings suggest that potentially critical disease mechanisms involve pressure-induced vascular smooth muscle cell apoptosis with subsequent deposition of collagen.

Objective Dientamoeba fragilis (D. fragilis) is a flagellated protozoan with an amoeba-like morphology, located in the gastrointestinal tract. The hypothesis was that the parasite was transported by Enterobius vermicularis (E. vermicularis) eggs. This study aimed to determine the association of D. fragilis and E. vermicularis with the genotypes of the identified strain of D. fragilis. Results of trichrome staining were compared with those of polymerase chain reaction (PCR), which is widely used in the diagnosis of D. fragilis. Methods A total of 391 samples were obtained. The stool and cellophane slide samples were sent together to the Parasitology Department Laboratory, Faculty of Medicine, Aydin Adnan Menderes University, between 1 October 2017 and 1 October 2018. Stool samples of all patients with E. vermicularis (n=74) and without E. vermicularis (n=74) infection were used. All samples were examined for the presence of D. fragilis by trichrome staining and PCR. The 18S ribosomal RNA region of D. fragilis isolates was sequenced. Demographic characteristics and clinical findings of the patients were evaluated. Results D. fragilis was detected in 42 (28.37%) of 148 samples; 28 (66.6%) of them were detected in patients with E. vermicularis infection. The coexistence of two parasites was significant (p 0.05). Conclusion D. fragilis is frequently associated with E. vermicularis, so the presence of D. fragilis should be also considered in affected patients. The use of high-sensitivity molecular methods such as PCR is important in preventing false results. Amac Dientamoeba fragilis (D. fragilis), amip benzeri morfolojiye sahip, gastrointestinal yerlesimli, kamcili bir protozoondur. Parazitin Enterobius vermicularis (E. vermicularis) yumurtalariyla tasindigi hipotezi kabul gormektedir. Calismamizda D. fragilis ve E. vermicularis birlikteligini incelemek, bulunan D. fragilis’lerin genotiplerini belirlemek ve D. fragilis tanisinda yaygin olarak kullanilan trikrom boyama ile polimeraz zincir reaksiyon (PZR) yontemlerini karsilastirmak amaclanmistir. Yontemler Calismamizda Aydin Adnan Menderes Universitesi Tip Fakultesi, Parazitoloji Laboratuvari’na 1 Ekim 2017-1 Ekim 2018 tarihleri arasinda diski ve selofan lam ornegi birlikte gonderilmis toplam 391 olgu ornegi incelenmistir. Selofanli lam orneklerinde E. vermicularis saptanan tum gonullu olgularin (74 olgu) diski ornegi ile E. vermicularis negatif 74 olgunun diski ornegi calisilmistir. Tum diskilar trikrom boyama ve PZR yontemleri ile D. fragilis varligi acisindan incelenmistir. Saptanan D. fragilis izolatlarinin 18S ribozomal RNA bolgesi sekanslanmistir. Olgularin demografik ozellikleri ve klinigi degerlendirilmistir. Bulgular Toplam 148 olgunun 42’sinde (%28,37) D. fragilis saptanmistir. D. fragilis pozitif olan 42 olgunun %66,6’sini E. vermicularis pozitif olgular olusturmus ve iki parazitin birlikteligi anlamli bulunmustur (p 0,05). Sonuc Arastirmamizda D. fragilis’in siklikla E. vermicularis ile birliktelik gosterdigi ve bu olgularda D. fragilis varligina ayrica dikkat edilmesi gerektigi vurgulanmistir. Yanlis sonuclari engellemede, yuksek duyarliliga sahip PZR gibi yontemlerin onemi bir kez daha gorulmustur.

The current study investigated the morphological, histochemical, and immunohistochemical characteristics of alarm cells and their precursors in ruby-red-fin shark (rainbow shark), Epalzeorhynchos frenatum (Teleostei: Cyprinidae). Precursor alarm cells were shown to be small, cuboidal, pyramidal, or round in shape, with eosinophilic cytoplasm, resting on the basement membrane of the epidermis. The cells later elongated to become columnar in shape. Subsequently, they enlarged and became large oval-shaped cells. They then underwent shrinkage and vacuolation. The superficial alarm cells were collapsed. Alarm cells were found to have an affinity for different histochemical stains, including bromophenol blue, iron hematoxylin, Sudan black, Mallory triple trichrome, Crossman's trichrome, Safranin O, and Weigert's stains, as well as lipase and alkaline phosphatase. Endocrine properties of the alarm cells were identified by silver staining and synaptophysin immunostaining. Alarm cells exhibited stemness activities and had a strong immunoaffinity for CD117. They had a proteolytic function, as identified by lysosome-specific staining with acridine orange and strong immunoaffinity for matrix metalloproteinase (MMP-9). They also exhibited proliferatively, reflected by immunological staining by proliferating cell nuclear antigen. In conclusion, alarm cells are unique epidermal cells with multiple functions. They play immunological, and endocrine, roles. They also retain stemness and proliferative properties.

The His bundle is a part of the specialized electrical conduction system that provides a connection between the atrial and ventricular myocardial compartments in both normal and abnormal hearts. The aim of this study was to perform a morphometric analysis of His bundle characteristics of in humans, dogs, horses and pigs and compare them in these studied species. Histological sections of 5 μm thickness were obtained and stained with hematoxylin–eosin and Masson's trichrome; the desmin and periodic acid–Schiff methods were also used for precise identification of cells. The His bundle was found to be longer in horses (2.85 ± 1.02 mm) and pigs (1.77 ± 0.9 mm) than in dogs (1.53 ± 0.8 mm) or humans, in which it was shortest (1.06 ± 0.6 mm). The area and diameters in His bundle cells, were significantly larger in pigs and horses than in humans (p

Background: Gingivectomy is a periodontal surgical procedure that eliminates excess gingival tissues by excision, usually to remove a gingival pocket which leads to scars and recontours the gingival to its proper anatomical form. Wound healing is a process to restore the missing functions and structures of a tissue due to injuries or inflammations. Robusta coffee bean contains many bioactive compounds that can accelerate the wound healing process. The study aimed to determinate the effectiveness of robusta coffee bean extract gel with concentration 40%,50%,60% on collagen fibers density in post-gingivectomy wound healing Method: 36 Wistar rats were divided into 9 groups: control, gingivectomy without therapy, gingivectomy +40% robusta coffee bean extract gel, gingivectomy +50% robusta coffee bean extract gel, and gingivectomy+60% robusta coffee bean extract gel. The last 4 groups were divided again into day 3 and 7 euthanasia. The results were processed by Trichrome Mallory to visualized the collagen fibers and then measured by ImageJ software. Result: Robusta coffee beans extract gel therapy with concentration 40%, 50% and 60% had higher average collagen fibers density compared to group without therapy. Conclusion: Robusta coffee bean extract gel increased the density of collagen fibers in post-gingivectomy wound healing

To identify and describe histological and immunohistochemical criteria that may differentiate between 4 skin and lymph node lesions associated with Mycobacterium (M.) bovis and M. microti in a diagnostic 5 pathology setting. 6Materials and Methods 7Archived skin and lymph node biopsies of tuberculous lesions were stained with haematoxylin and 8 eosin, Ziehl-Neelsen and Masson’s Trichrome. Immunohistochemistry was performed to detect 9 expression of calprotectin, CD3 and Pax5. Samples were scored for histological parameters (i.e. 10 granulomas with central necrosis vs small granulomas without central necrosis, percentage necrosis, 11 and/or multinucleated giant cells), number of acid-fast bacilli (bacterial index) and lesion percentage of 12 fibrosis and positive immunohistochemical staining. 13Results 14Twenty-two samples were examined (M. bovis n = 11, M. microti n = 11). When controlling for age, 15 gender and tissue, feline M. bovis-associated lesions more often featured large multi-layered 16 granulomas with central necrosis. Conversely, this presentation was infrequent in feline M. microti-associated lesions, where small granulomas without central necrosis predominated. The presence of an outer fibrous capsule was variable in both groups, as was the bacterial index. There were no differences in intralesional expression of immunohistochemical markers.Clinical SignificanceDifferences in the histological appearance of skin and lymph node lesions may help to infer feline infection with either M. bovis or M. microti at an earlier stage when investigating these cases, informing clinicians of the potential zoonotic risk. Importantly, cases of tuberculosis can present with numerous acid-fast bacilli. This implies that a high bacterial index does not infer infection with non-zoonotic non-tuberculous mycobacteria.

Background Microsporidia are a large family of obligate intracellular protozoa; these medically important species are recognized as opportunistic agents in intestinal complications in HIV+/AIDS patients. Methods The current cross-sectional study was designed and conducted from October 2018 to June 2019 to determine intestinal microsporidia in HIV+/AIDS patients by trichrome/Zeihl-Neelsen staining and SYBR Green-based real-time PCR. Results Out of 80 HIV+/AIDS patients, 23.75% (n=19) and 12.5% (n=10) were identified by molecular and microscopic methods, respectively. The predominant species in patients was Encephalitozoon (94%), which was found by quantitative real-time PCR and its high resolution melting tool. Conclusion As far as we know, this is the first report from the Alborz region. The prevalence of intestinal microsporidiosis in this area in HIV+/AIDS patients was higher than both the global and national average. In addition to the need for further studies to prove protozoan pathogenicity in the aforementioned group, preventive measures should be considered.

Background: Diabetes Mellitus (DM) is a worldwide common disease and it is considered a serious public health disorder .The reproductive function is highly sensitive to changes in metabolic status, with defective reproductive capacity. Aim of Work: To demonstrate the histological changes in the prostate of rats with STZ induced diabetes type and to examine to what extent insulin could reverse such changes. Materials and methods: Twenty six male albino rats were divided into three groups. Group I: control; Group II: Diabetic was exposed to induction of diabetes by a single intraperitoneal injection of Streptozotocin(STZ) 60 mg/kg in 0.1 M citrate buffer; Group III: Diabetic then received insulin (3IU/day S.C( (for 4 weeks). The rats of each group were sacrificed after 4 weeks. The prostates were processed for electron microscopic sections and other sections stained with H&E, Masson's trichrome, immunohistochemical staining of α smooth muscle actin, Vimentin and PCNA and statistical analysis were applied. Results: Diabetes resulted in deterioration of normal structure of the prostatic acini and some prostatic acini showed cell proliferation with increase Masson's trichrome staining and increase immunoreactivity for α smooth muscle actin, Vimentin and PCNA. Electron microscopic examination reavealed dilatation of the cisternae of the Golgi complex, endoplasmic reticulum, and some epithelial cells showed proliferation and irregular contour of their nuclei. Insulin administration mostly reversed these changes. Conclusion: insulin administration greatly improved the histological architecture of the prostate gland.

Aseptic osteonecrosis of the hip (AON) is a rare, but well-known pathology in rheumatology and orthopedic surgery that is a necrosis of the articular cartilage secondary to a necrosis of the subchondral bone. The microscopic aspect is well known, but the microCT aspect has never been reported or correlated with histopathological findings. The objective of this study was to improve the knowledge of the pathophysiology of AON using histochemistry and microCT. One hundred and sixty femoral heads with stage 3 or 4 AON were analyzed: one half of the head was sent for microCT analysis after impregnation with phosphotungstic acid (PTA) and the other half was used for histological analysis without decalcification. The microCT analysis provides relevant information on the cracked articular cartilage and the relationship with the necrotic subchondral trabecular bone well illustrated on three videos. In histology, Goldner's trichrome showed that the articular cartilage remains well preserved for a long time. In addition, toluidine blue staining reveals a modeling process, i.e. the apposition of new bone without prior resorption by osteoclasts. Rhodamine B staining (fluorescence analysis) reveals that the osteonecrotic trabeculae and subchondral bone were devoid of osteocytes. Areas of peri-necrotic osteosclerosis are due to direct bone formation on the surface of pre-existing necrotic trabeculae.

PURPOSE To investigate the reflectivity of the structural optical coherence tomography images of the inner choroid as it relates to potential structural composition. METHODS The reflectivity of slab images 29 to 49 µm and 45 to 65 µm below the retinal pigment epithelium, obtained with the Zeiss Plex Elite 9000, was evaluated. The mean and SD of a group of subjects with no ocular disorders were determined. Binarization of the images was performed and a threshold level established at the mean plus one SD for each slab depth. The proportion of area binarized was evaluated with generalized estimating equations. Representative histologic images obtained from autopsy donors were stained with Masson's trichrome, a staining method helpful in evaluating collagen and ground substance of tissue. RESULTS There were 67 eyes of 38 subjects with a mean age of 44.5 (range 22-82) years. Using generalized estimating equations, age was found to be a significant predictor for the proportion of binarized pixels in both the 29 to 49 µm (P = 0.034) and the 45 to 65 µm (P < 0.001) slabs. The histologic specimens illustrated the loss of ground substance with increasing compaction of collagen fibers in the choroidal stroma with advancing age. CONCLUSION The reflectivity from the inner choroid is not uniform and changes with age. As suggested by the histologic specimens, we propose the optical coherence tomography reflectance from the inner choroid is related, in part, to the packing density of collagen fibers present there.

Colpodella spp. are free-living flagellates closely related to the apicomplexans. Human infections by Colpodella sp. have been reported. A biflagellated trophozoite and cyst stage comprise the known life cycle stages of Colpodella sp. However, the process of encystation and excystation within the life cycle is unclear. Life cycle stages initiating human infections are unknown. We performed a detailed investigation of the life cycle of Colpodella sp. (ATCC 50594) in culture using Sam-Yellowe's trichrome stains and differential interference contrast (DIC) for light microscopy and fluorescence microscopy of Congo red-stained cells and investigated ultrastructure using transmission electron microscopy (TEM). We report previously undocumented stages of Colpodella sp. Asymmetric and asynchronous division was detected inside cysts by trichrome staining and by TEM. Odd-numbered juveniles and cysts containing more than four juvenile trophozoites were identified. Live imaging of active cultures captured the excystation and egress of juvenile trophozoites and confirmed the presence of multinucleate cysts. The ultrastructure of the multinucleate cyst is reminiscent of apicomplexan schizonts. Insights gained from the life cycle stages observed in culture allowed the construction of the life cycle of Colpodella sp. Knowledge of the life cycle will aid biochemical and molecular characterization of Colpodella sp. and help identify stages in human infections.

Background Intimal calcification typically develops in advanced atherosclerosis, and microcalcification may promote plaque progression and instability. Conversely, intraplaque hemorrhage and erythrocyte extravasation may stimulate osteoblastic differentiation and intralesional calcium phosphate deposition. The presence of erythrocytes and their main cellular components (membranes, hemoglobin, and iron) and colocalization with calcification has never been systematically studied. Methods and Results We examined three types of diseased vascular tissue specimens, namely, degenerative aortic valve stenosis (n = 46), atherosclerotic carotid artery plaques (n = 9), and abdominal aortic aneurysms (n = 14). Biomaterial was obtained from symptomatic patients undergoing elective aortic valve replacement, carotid artery endatherectomy, or aortic aneurysm repair, respectively. Serial sections were stained using Masson–Goldner trichrome, Alizarin red S, and Perl's iron stain to visualize erythrocytes, extracelluar matrix and osteoid, calcium phosphate deposition, or the presence of iron and hemosiderin, respectively. Immunohistochemistry was employed to detect erythrocyte membranes (CD235a), hemoglobin or the hemoglobin scavenger receptor (CD163), endothelial cells (CD31), myofibroblasts (SMA), mesenchymal cells (osteopontin), or osteoblasts (periostin). Our analyses revealed a varying degree of intraplaque hemorrhage and that the majority of extravasated erythrocytes were lysed. Osteoid and calcifications also were frequently present, and erythrocyte membranes were significantly more prevalent in areas with calcification. Areas with extravasated erythrocytes frequently contained CD163-positive cells, although calcification also occurred in areas without CD163 immunosignals. Conclusion Our findings underline the presence of extravasated erythrocytes and their membranes in different types of vascular lesions, and their association with areas of calcification suggests an active role of erythrocytes in vascular disease processes.

The olfactory organs of two African lungfishes,Protopterus amphibiusandP. dolloi, were investigated using a stereo microscope and a compound light microscope and were described anatomically, histologically, and histochemically. Like other lungfishes, these species present the following general features: i) elongated olfactory chamber (OC), ii) anterior nostril at the ventral tip of the upper lip, iii) posterior nostril on the palate of the oral cavity, iv) lamellae with multiple cell types such as olfactory receptor neurons, supporting cells, basal cells, lymphatic cells, and mucous cells (MC), and vi) vomero-like epithelial crypt (VEC) made of glandular epithelium (GE) and crypt sensory epithelium. Some of these features exhibit differences between species: MCs are abundant in both the lamellar and inner walls of the OC inP. amphibiusbut occur only in lamellae inP. dolloi. On the other hand, some between feature differences are consistent across species: the GE of bothP. amphibiusandP. dolloiis strongly positive for Alcian blue (pH 2.5)-periodic acid Schiff (deep violet coloration), and positive with hematoxylin and eosin and with Masson’s trichrome (reddish-brown staining), unlike the MCs of the two species which stain dark red with both Alcian blue (pH 2.5)-periodic acid Schiff and Masson’s trichrome but respond faintly to hematoxylin and eosin. The differing abundance of MCs in the two lungfishes might reflect different degrees in aerial exposure of the olfactory organ, while the neutral and acid mucopolysaccharide-containing VEC, as indicated by staining properties of the MCs, is evolutionary evidence thatP. amphibiusandP. dolloiare the closest living relatives to tetrapods, at least in the order Dipnoi.

Venous invasion (VI) is a powerful prognostic factor in colorectal cancer (CRC) that is widely underreported. The ability of elastin stains to improve VI detection is now recognized in several international CRC pathology protocols. However, concerns related to the cost and time required to perform and evaluate these stains in addition to routine hematoxylin and eosin (HE) stains remains a barrier to their wider use. We therefore sought to determine whether an elastin trichrome (ET) stain could be used as a "stand-alone" stain in CRC resections, by comparing the sensitivity, accuracy, and reproducibility of detection of CAP-mandated prognostic factors using ET and HE stains. Representative HE- and ET-stained slides from 50 CRC resections, including a representative mix of stages and prognostic factors, were used to generate 2 study sets. Each case was represented by HE slides in 1 study set and by corresponding ET slides from the same blocks in the other study set. Ten observers (3 academic gastrointestinal [GI] pathologists, 4 community pathologists, 3 fellows) evaluated each study set for CAP-mandated prognostic factors. ET outperformed HE in the assessment of VI with respect to detection rates (50% vs. 28.6%; P0.0001), accuracy (82% vs. 59%, P0.0001), and reproducibility (k=0.554 vs. 0.394). No significant differences between ET and HE were observed for other features evaluated. In a poststudy survey, most observers considered the ease and speed of assessment at least equivalent for ET and HE for most prognostic factors, and felt that ET would be feasible as a stand-alone stain in practice. If validated by others, our findings support the use of ET, rather than HE, as the primary stain for the evaluation of CRC resections.

The principal function of the ventricular conduction system is rapid electrical activation of the ventricles. The aim of this study is to conduct a morphometric study to pinpoint the morphological parameters that define cardiac conduction cells, allowing us to distinguish them from other cells. Five male horse hearts and five male dog hearts were used in the study. The hearts were fixed in a 5% formaldehyde solution. Histological sections of 5 μm thickness were acquired and stained with hematoxylin-eosin and Masson's trichrome and cardiac conduction cells and their junctions were identified by desmin, connexin 40 and a PAS method. We found statistically significant differences in cardiac conduction fibers density and thickness, which was much higher in horses than in dogs (p = 0.000 for both values). By comparing the measured parameters of the cells in both species, we determined that cardiac conduction cells area and diameters were greater in horses than in dogs (p = 0.000 for all values). In dogs there are more junctions (30.8%) than in horses (26.1%), a statistically significant difference (p = 0.041). Our findings regarding the cardiac conduction fibers distribution in the animal species studied becomes new knowledge that contributes to the morphological study of this component of the cardiac conduction system and also makes it possible to locate exactly the site with the highest density of cardiac conduction fibers as a contribution to the cardiological study of these structures that lead to the prevention of ventricular arrhythmias and the identification of their treatment site.

The study has aimed to investigate and determine the anatomical position, shape, size, and histological features of the ductus venosus, and its role as a shunt in the fetal circulatory system in domestic ruminants. The research was conducted on 19 bovine, 11 sheep and 5 goat fetuses, aborted at the late stage of pregnancy or deceased just after delivery. The general anatomy of the ductus venosus was investigated by in-situ dissection of the corrosive cast obtained by injection of 25% solution of Vinylite mass through the umbilical vein. For histological examination, the fetal tissue samples were stained with Hematoxylin and Eosin, Masson’s trichrome, Verhoeff-Van Gieson and Gomoriꞌs silver stain. The results showed that ruminant fetal ductus venosus is a curved, trumpet-shaped vessel, situated in the central part of the liver, above the porta hepatis. Its ventral part is constricted in the form of an isthmus, having a prominent lip-like thickening at the junction with the portal sinus. Histological examination showed the dominant presence of collagen and elastic fibers in its tunica media, with thin bands of smooth muscle fibers oriented in a longitudinal and circular direction indicating ability for vasoconstriction and vasodilatation.

Introduction: Tacrolimus is a macrolide calcineurin inhibitor which has been shown to decrease acute and chronic rejection, improved renal function over the longterm post-transplant. Tacrolimus is considered the drug of choice in immunosuppressive therapy for chronic diseases irresponsive to corticosteroids, autoimmune diseases and organ transplantation system. Aim: The aim of the investigation was to study the effect of the immunosuppressive drug Tacrolimus on the lingual mucous membrane and salivary glands of treated rats through hematoxylin and eosin stained sections to detect any microscopic changes in the structure of the mucous membrane and salivary glands of the tongues & Masson’s trichrome stained sections for detection of the integrity of collagen. Material and Methods: Thirty healthy adult male albino rats with body weight ranging from 150 to 180 gm were used in this investigation. The animals were divided into two groups 15 animals each. Group 1: served as control, group 2: animals were treated with the immunosuppressive drug tacrolimus dissolved in distilled water in a daily oral dose of 0.5 mg/kg body weight. At the end of the experiment the rats of different groups were euthanized by cervical dislocation. The tongues were dissected out, fixed, processed, embedded in paraffin, sectioned and stained with hematoxylin and eosin for histological evaluation & Masson’s trichrome stain for demonstration of collagen fibers. Results: The light microscopic examination of group I showed the normal histological features of both surface epithelium and underlying lamina propria, while group II Treated with tacrolimus revealed atrophic and degenerative changes that involved the surface epithelium and lamina propria of both the dorsal and ventral surfaces of the tongue as well as the lingual salivary glands. Conclusion: Tacrolimus administration showed structural alterations in the lingual mucous membrane and the lingual salivary glands which will certainly affect the taste sensation.

Purpose Low free testosterone (T) level in men is independently associated with presence and severity of Non-Alcoholic Steatohepatitis (NASH). The histological and molecular effects of oral testosterone prodrug LPCN 1144 treatment on hepatic fibrosis and NASH features are unknown. A metabolic syndrome-induced NASH model in rabbits consuming high fat diet (HFD) has been previously used to assess treatment effects of injectable T on hepatic fibrosis and NASH features. Here we present results on LPCN 1144 in this HFD-induced, NASH preclinical model. Methods Male rabbits were randomly assigned to five groups: regular diet (RD), HFD, HFD + 1144 vehicle (HFD + Veh), HFD + 1144 (1144), and HFD + 1144 + α-tocopherol (1144 + ALPHA). Rabbits were sacrificed after 12 weeks for liver histological, biochemical and genetic analyses. Histological scores were obtained through Giemsa (inflammation), Masson’s trichrome (steatosis and ballooning), and Picrosirius Red (fibrosis) staining. Results Compared to RD, HFD and HFD + Veh significantly worsened NASH features and hepatic fibrosis. Considering HFD and HFD + Veh arms, histological and biomarker features were not significantly different. Both 1144 and 1144 + ALPHA arms improved mean histological scores of NASH as compared to HFD arm. Importantly, percentage of fibrosis was improved in both 1144 (p p = 0.05) treatment arms vs. HFD. Both treatment arms also reduced HFD-induced inflammation and fibrosis mRNA markers. Furthermore, 1144 treatments significantly improved HFD-induced metabolic dysfunctions. Conclusions Histological and biomarker analyses demonstrate that LPCN 1144 improved HFD-induced hepatic fibrosis and NASH biochemical, biomolecular and histochemical features. These preclinical findings support a therapeutic potential of LPCN 1144 in the treatment of NASH and of hepatic fibrosis.

The present study evaluated the role of dietary folate modulations in the development of hepatocellular carcinoma (HCC) in a rat model. Male Wistar rats were given diethylnitrosamine (DEN) carcinogen for a period of 18 weeks in addition to different folate modulations. Biochemical parameters were assayed and liver tissues were examined using various histopathological stains viz. Hematoxylin and eosin (H&E), Masson’s trichrome, Immunohistochemistry (IHC) staining for arginase-1 and α-smooth muscle actin (SMA). Serum folate and hepatic folate stores were decreased and increased in folate deficiency (FD) and folate oversupplemented (FO) group respectively. Analysis of serum liver function tests revealed deranged liver functioning in all the groups. H&E staining of rat liver demonstrated vague nodularity from 2nd to 8th week, fibrosis from 10th to 15th week, cirrhosis and HCC from 16th to 18th week. Combining the observations of H&E with IHC for arginase-1, 14 (50%), 11 (39.3%) and 17 (58.6%) rats showed HCC positivity in FN (folate normal), FD and FO diets respectively. IHC for α-SMA depicted increased staining with progression of the disease from fibrosis to cirrhosis in all the dietary groups. Collectively, findings of all the histopathological stains, revealed increase in the number of cirrhotic cases and decrease in the number of HCC cases in FD group, indicating delayed progression of HCC with FD. Moreover, FO led to more number of HCC and reduction in the number of cirrhotic cases, signifying early progression of HCC.

Objective: To evaluate the biological effects of coral scaffold with or without adipose derived stem cells (ADSCs) on healing of bony defects in albino rats. Methods: Fifty four male albino rats weighing approximately 250-300 gm were selected and randomly divided into 3 groups. Each rat received one bone defect on the left side of mandible near the diastema. The defects were either left empty (group A) n=18, treated with coral only (group B) n=18 or received a combination of coral and adipose derived stem cells (group C) n=18 which were harvested from the rats peri renal region. Bone healing was evaluated using heamatoxylin and eosin and trichrome stain, then the results were subjected to digital image analysis followed by one- way ANOVA statistical analysis. Results: Bony defects received the combination of coral scaffold and stem cells show better healing in respect to the quantity and the quality of the newly formed bone. Conclusion: ADSCs loaded onto coral granule scaffold have ameliorating effect on the repair of the mandibular defects in rat model.

Introduction: The goal of this study was to histopathologically evaluate the appearance of degrading MIRAgel scleral buckles so that they can be reliably distinguished by ophthalmic pathologists from other foreign materials. Methods: Retrospective chart review and histopathologic study with special stains, including Alcian blue, periodic acid-Schiff, Masson’s trichrome, and Perls’ Prussian blue for iron, of 4 cases between 2017 and 2019. Results: Hydrolyzed MIRAgel scleral buckles from 4 patients had a consistent histopathologic appearance. They had a honeycomb structure with the walls of the lattice ranging from distinct to poorly defined. The walls of the lattice were positive for Alcian blue, while the contents of each cell contained periodic acid-Schiff-positive material. Other special stains were not as valuable in highlighting the material. Discussion: Although the capsules of MIRAgel scleral buckles have been well studied, the material itself has not been. While clinical history and radiographic appearance are often diagnostic of a hydrolyzed MIRAgel scleral buckle, there are instances of preoperative ambiguity where histopathologic confirmation can be useful. Conclusions: MIRAgel scleral buckles have a distinct histopathologic appearance that can be readily distinguished from that of other foreign materials.

Summary Objective This novel study sought to untangle the association between hydration state and the estrous cycle in the vocal folds, since the voice is reported to negatively change in speakers during the estrous cycle and with dehydration. We hypothesized that there would be alterations in vocal fold tissue morphology depending on hydration state and that these changes would vary with the estrous cycle. Study Design Prospective, in vivo study design. Methods Female Sprague Dawley rats (n = 30) were used in this study. Sixteen rats were systemically dehydrated to an average of 10% reduction in body weight by withholding water (range of body weight loss: 8%–13%). Fourteen rats were assigned to euhydrated, control condition. Estrous stage of female Sprague Dawley rats (n = 30) was determined via cytological evaluation of vaginal smears. Following euthanization, larynges were prepared for histological staining with hematoxylin and eosin, Masson's trichrome and alcian blue (pH 2.5). To quantify hyaluronan, alcian blue staining was completed pre- and posthyaluronidase incubation. The change in staining percent was quantified with image analysis algorithms and reported as the hyaluronan quantity. Relative collagen distribution (index of dehydration), hyaluronan quantity, and tissue morphology were the outcome measures. Results Systemic dehydration was associated with changes in hyaluronan quantity in the rat vocal fold lamina propria. Dehydration did not significantly affect the collagen distribution nor the tissue morphology. Estrous stage alone does not impact the quantity of vocal fold hyaluronan, alter tissue morphology, or change collagen distribution. Conclusion Decreases in hyaluronan quantity in the lamina propria of the rat vocal fold may play a role in tissue fluid balance during systemic dehydration. Future studies will expand this work to investigate additional components of the vocal fold extracellular matrix to fully elucidate the impact of hydration state on the vocal fold.

The evaluation of the morphology and organization of collagen fibers is critical in understanding wound healing and tissue remodeling after a thermal injury of the skin. However, histological analysis conducted by pathologists is often labor-intensive and limited to qualitative evaluations and scoring within a narrow field of view. In this study, we propose a convolutional neural network (CNN) model to classify Masson’s trichrome (MT)-stained histology images of burn-induced scar tissue and to characterize the microstructures of normal tissue and scar tissue in a quantitative manner. The scar tissue is created on in vivo rodent models and prepared for MT-stained histology slides after wound healing. A CNN model is developed, trained, and tested with various sizes of the histology images for classification and characterization. The proposed model classifies both normal tissue (i.e., without burn, as the control) and scar tissue at various scales with over 97% accuracy. The features acquired from the proposed CNN model visually characterizes the density and directional variance of the collagen fibers distributed in the dermal layers from whole histology images. The proposed deep learning technique can provide an objective and reliable method to rapidly assess and quantify wound repair and remodeling.

The application of deep learning for automated segmentation (delineation of boundaries) of histologic primitives (structures) from whole slide images can facilitate the establishment of novel protocols for kidney biopsy assessment. Here, we developed and validated deep learning networks for the segmentation of histologic structures on kidney biopsies and nephrectomies. For development, we examined 125 biopsies for Minimal Change Disease collected across 29 NEPTUNE enrolling centers along with 459 whole slide images stained with Hematoxylin & Eosin (125), Periodic Acid Schiff (125), Silver (102), and Trichrome (107) divided into training, validation and testing sets (ratio 6:1:3). Histologic structures were manually segmented (30048 total annotations) by five nephropathologists. Twenty deep learning models were trained with optimal digital magnification across the structures and stains. Periodic Acid Schiff-stained whole slide images yielded the best concordance between pathologists and deep learning segmentation across all structures (F-scores: 0.93 for glomerular tufts, 0.94 for glomerular tuft plus Bowman's capsule, 0.91 for proximal tubules, 0.93 for distal tubular segments, 0.81 for peritubular capillaries, and 0.85 for arteries and afferent arterioles). Optimal digital magnifications were 5X for glomerular tuft/tuft plus Bowman's capsule, 10X for proximal/distal tubule, arteries and afferent arterioles, and 40X for peritubular capillaries. Silver stained whole slide images yielded the worst deep learning performance. Thus, this largest study to date adapted deep learning for the segmentation of kidney histologic structures across multiple stains and pathology laboratories. All data used for training and testing and a detailed online tutorial will be publicly available.

Objective: This study aimed at examining whether topical treatment of colon anastomosis with hyaluronic acid can influence the healing of colonic anastomosis in rats treated with systemic corticosteroid. Methods: Three groups of Wistar rats weighing 252 ± 7g were used, with six rats each, all submitted to anastomosis of the proximal colon: Group 1 - control rats, without treatment. Group 2 - rats treated with subcutaneous (s.c) corticosteroid, and topical application of 0.9% saline solution over the anastomosis. Group 3 - rats treated with corticosteroid s.c. and topical application of 0.4% hyaluronic acid (10 mg/ml) on colonic anastomosis. On the 7th postoperative day, under anesthesia and laparotomy, the anastomosis site was subjected to the determination of rupture pressure. Then, samples containing the anastomosis site were resected and fixed in 10% buffered formaldehyde and embedded in paraffin. Masson H-E and trichrome staining. Histometry evaluated the infiltration of inflammatory cells at the anastomosis site, using a numerical scale from 0 to 4. Continuous variables were assessed using the Tukey test. Differences considered significant with p0.05). In the analysis of intraluminal pressure of colonic anastomosis, there was a statistically significant difference when comparing groups 1 (286.8±9.9 mmHg) and 2 (155±6.0 mmHg), 1 (286.8±9,9 mmHg) and 3 (199.1±7.7) and 2 (155±6.0 mmHg) and 3 (199.1±7.7), with p

Introduction: Vitiligo: An autoimmune disorder of pigmentation characterised by loss of functional melanocytes and melanin in epidermis. The disease has incidence of 0.5%–2% with no ethnic/sex predilection. Aim: Morphological dermoscopic patterns in vitiligo and its correlation with stability. Setting & Desing: An observational study conducted on patients attending C. R. Gardi Hospital, Ujjain during January’19-December’19. Materials and Methods: Dermoscopic examination was done after taking consent in 50 clinically diagnosed cases of vitiligo including stable vitiligo and unstable vitiligo. Statistical Analysis Used : Fisher’s Exact Test Sig. Results: On dermoscopy, patterns like perifollicular pigmentation (44%), perilesional/marginal hyperpigmentation (50%), intra/perilesional erythema with telangiectasia (86%), perifollicular depigmentation (64%), trichrome pattern (50%), leukotrichia (46%), starburst appearance (24%), comet tail appearance (48%), micro-koebner’s phenomenon (42%), tapioca sago appearance (28%) were observed. Perilesional/marginal hyperpigmentation was observed more frequently in stable disease. Perifollicular depigmentation, trichrome pattern, comet tail appearance, micro koebner’s phenomenon and tapioca sago appearance are the findings significant in unstable disease. Perifollicular pigmentation and intra/perilesional erythema with telangiectasia though are findings of stable disease, were observed in both stable and unstable disease in similar numbers. Leukotrichia and starburst appearance were present in both stable and unstable disease despite being the findings of unstable disease. Conclusion: Dermoscopy serves as an auxiliary tool in confirming the diagnosis and assessing stability of the disease. Dermoscopy allows appreciation of subtle features invisible to naked eye. This is the first ever study to find out the statistically significant findings in stable and unstable vitiligo. We conclude that dermoscopic st

Background: Periodontitis affects approximately 20%–50% of the global population and is caused by gram-negative bacteria, such as Porphyromonas gingivalis (P. gingivalis). Host modulation therapy (HMT) is part of a periodontal therapy that is used as an adjunct to conventional periodontal treatment to reduce tissue damage. Lemuru fish oil containing EPA and DHA can reduce the formation of MMPs and will further increase the number of fibroblasts there by stimulating collagen formation. Purpose: To determine the effect of lemuru fish oil gel on the collagen density and width of the periodontal tissue induced by P. gingivalis and the correlation between these parameters. Methods: Thirty male Wistar rats were divided into five groups. Induction of P. gingivalis was carried out first, then lemuru fish oil gel was applied to the gingival sulcus for 14 days, according to collagen scores in histological preparations using Masson's trichrome (MT). The width of the periodontal ligament was measured with an image raster program in µm. The d ata were analysed using statistics to test hypotheses using SPSS version 24. Results: Significant differences in the results of the c ollagen density were observed between groups K- and K+ and groups K+ and P2. Meanwhile, no significant difference was observed between groups K- and P2, P3, P2 and P3 and K+ and P1 . The mean values of the periodontal ligament widths were K- (299.61 ± 51.82µm), K+ (425.85 ± 61.54µm), P1 (346.93 ± 33.53µm), P2 (370.15 ± 49.42µm) and P3 (379.6 ± 49.26). Conclusion: Lemuru fish oil can affect the width of the ligament and the collagen density with an optimal concentration of 20%. The correlation between the collagen density and the periodontal ligament width was negative and not significant.

This study assessed intestinal protozoa with stress on Blastocystis among patients with CKD andthose on hemodialysis (HD) in Nephrology Unit, Medical Research Institute, Alexandria.The patients were divided into three groups. GI: 50 patients on HD for at least one year, GII: 50 CKDpatients not on HD, and GIII: 50 healthy controls. Fresh stool samples were collected in labeled plasticboxes and examined as direct smears after staining with trichrome, Modified Ziehl-Neelsen (MZN)stains and modified trichrome (MT) stains. Besides, stool samples were cultured for Blastocystis hominisand Copro-antigen (ICT) was used to detect E. histolytica /dispar, G. lamblia and C. parvum.The results showed that the parasitic infections were detected in 79% of the patients; 40% in HD and39% in CKD patients. The positive samples were (47%) detected by direct wet mount followed byiodine smear (43%) and then trichrome stain (42%). Microsporidiosis was detected in 55 (55%) HDand CKD patients using the MT stain. ICT agreed with microscopic examination in diagnosis ofE.histolytica/ dispar and G. lamblia and showed perfect agreement with MZN in C. parvum diagnosis.

The aim of this study was evaluating the effects of vacuum on microstructure and biocompatibility of bovine decellularized pericardium. So the bovine pericardia were decellularized and then the vacuum was applied for two periods of time; 90 and 180 min. DNA, glucose amino glycan, collagen and elastin content assay, scanning electron microscopy (SEM) examination, hematoxylin and eosin (H&E) and Masson's trichrome stainings performed to evaluate microstructure of tissues. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test, subcutaneous implantation, and tensile test were used to assay biocompatibility and mechanical properties of decellularized tissues. The results showed that applying vacuum reduced residual DNA significantly. Vacuum after 180 min reduced more residual DNA. There were no significant differences in the content of glucose amino glycan (GAG), collagen, and elastin between the vacuumed and control groups. SEM examination was revealed that vacuum for 180 min increased pore size and porosity more than 90 min and control groups. H&E and Masson's trichrome stainings revealed extracellular matrix preservation after decellularization in all groups. Cell viability was increased in vacuumed samples significantly after 72 h in vaccumed samples. H&E staining and tensile test after implantation of tissues were showed less inflammation in the vacuum applied tissues and increased durability. The vacuum increased DNA removal, pore size, porosity, and biocompatibility in vitro and in vivo and durability of bovine decellularized pericardium in vivo. Considering the important role of time, more studies should be performed to optimize time, intensity, and method of application of vacuum in decellularization of different tissues as well as bovine pericardium.

Background and Aim: Jatropha gaumeri Greenm. is commonly used to treat mouth blisters and skin rashes, its latex has analgesic and anti-inflammatory activity on buccal ulcer. This study aimed to demonstrate the wound healing activity of a cream formulation of Jatropha gaumeri Greenm. latex in a murine model, provide a histological assessment of its scarring effects, and identify the family of phytochemicals involved in these effects. Materials and Methods: Latex was obtained from the cut stalk leaves and young stems of J. gaumeri and stored in sterile tubes with protection from light. Chloroform, ethyl acetate, and aqueous fractions of the latex were obtained. Fifty male Balb/c mice aged 10-12 weeks were divided into 10 groups of five mice: Group 1 corresponded to healthy mice with wounds; Group 2 corresponded to mice with wounds and treated with A-Derma®; and from Group 3 to group 10 corresponded to mice treated with a different latex fraction. A circular skin wound of about 1 cm was made on the paravertebral region of each mouse under anesthetized and aseptic conditions. The wounds were topically treated every 24 h with the respective extracts for 22 days, after which skin tissue specimens were obtained and stained with hematoxylin-eosin and Masson's trichrome. The efficiency of healing was measured by quantifying the tensile strength of the scars. The phytochemicals in the latex were elucidated using thin chromatography. Results: The aqueous latex fraction produced the best wound healing activity and was superior to the positive control. Reepithelialization at the histological level resulted in tissue that resembled healthy skin in terms of the appearance of collagen, the regeneration of hair follicles, and cellularity of the dermis, which showed organized epithelialization. A wound healing efficacy of 97% was observed, and it seems that alkaloids were the phytochemicals mostly likely responsible for these effects. Conclusion: J. gaumeri latex exhibited wound healing activity, possibly mediated by phytochemicals such as alkaloids in the aqueous fraction.

Objectives Many studies of neonates have shown that renal pelvis ectasia is more common in boys. The aim of this study was to determine whether there are structural differences in the renal pelvis between male and female fetuses in the second trimester of gestation. Material and methods We studied 34 renal pelvises obtained from 34 human fetuses (17 males and 17 females), ranging in age from 13 to 23 weeks postconception. The renal pelvis tissue was stained with Masson’s trichrome to quantify connective and smooth muscle cells (SMC). The tissue also was fixed for scanning electron microscopy (SEM) in a modified Karnovsky solution. The images were captured with an Olympus BX51 microscope and Olympus DP70 camera. The stereological analysis was done with the Image-Pro and ImageJ programs, using a grid to determine volumetric densities (Vv). Means were statistically compared using simple linear correlation and the Mann-Whitney test (p Results Quantitative analysis indicated differences (p = 0.0275) in Vv of connective tissue in male renal pelvises (mean = 55.3%) compared to female ones (mean = 51.46%). Quantitative analysis indicated a significant difference (p = 0.0002) in SMC in male renal pelvises (mean = 12.57%) compared to female ones (mean = 16.22%). When we compared the SMC at different ages, we did not find any correlation in male (r2 = 0.2657, p = 0.3027) or female fetuses (r2 = 0.3798, p = 0.1326). When we compared the connective tissue at different ages, we did not find any correlation in female fetuses (r2 = 0.3798, p = 0.2870), but we did observe a positive correlation between the connective tissue and age in male fetuses (r2 = 0.8308, p Conclusion The renal pelvis presents significant structural differences between male and female fetuses. The renal pelvis in males had less SMC and presented a positive correlation of connective tissue with age and the renal pelvis in female had less connective tissue without correlation with the age. Level of evidence III

This study was conducted to evaluate the bone regeneration effects of collagen/strontium-doped Bioglass (BG-Sr) scaffolds. The present study comprised four parts including designing and fabricating scaffolds, characterization tests such as biodegradation, water uptake, porosity, FTIR (Fourier transform infrared spectroscopy), and SEM (Scanning electron microscope), in vitro tests such as MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)), alkaline phosphatase, calcium assay, Alizarin Red staining, and real-time PCR (Polymerase chain reaction), and ultimately in vivo and histopathological tests such as evaluating bone regeneration, mineralization with Hematoxylin-Eosin, Masson's trichrome and Alizarin Red staining, and histomorphometric studies 4, 8 and 12 weeks after implanting the scaffolds in the skull and femur of 36 rabbits. Significant reductions were observed in degradability, water absorption, and porosity in the BG-Sr samples. In vitro results showed that the synthetic scaffolds were less toxic, stimulated, and induced cell growth. The ALP (Alkaline phosphatase) activity was higher on day 14 than on day 7, and the highest calcium content was observed in the BG-Sr samples on day 14. In line with the SEM and MTT results, the DAPI (Diamidino-2-phenylindole) results confirmed cell proliferation. Moreover, osteocalcin and ALP gene expression were high in the BG-Sr group. The results of the histopathological tests showed high levels of bone formation in the BG-Sr scaffolds. As a scaffold in regenerative medicine, the BG-Sr composite can be an effective product for bone regeneration.