Elastin Staining Research Articles

Urethrovaginal Septum: A Histological Study on a Cadaver and Its Clinical Significance.

The academic literature concerning the tissue bridging the anterior vaginal wall and the posterior urethral wall, the urethrovaginal septum, is lacking and inconsistent, not only when compared to non-reproductive anatomy but also when compared to male reproductive anatomy. This knowledge gap must be addressed, given the implication of this tissue in numerous female reproductive pathologies and functions, including pelvic organ prolapse, intercourse and orgasm, urethral strictures, vaginal cancers, and stress urinary incontinence. This study seeks to characterize the histological composition of the urethrovaginal septum, including type I and III collagen proportions, elastin content, neurovascular distribution, and smooth muscle arrangement. Specimens were resected bilaterally from the bisected pelvic region of a postmenopausal female cadaver, spanning antero-posteriorly from the lumen of the urethra to the lumen of the vagina. The specimens were divided into medial and lateral portions, sectioned, and stained with hematoxylin and eosin, trichrome, reticulin, and elastin stains. Images were obtained through virtual microscopy. Type I collagen was confirmed as the major connective tissue component. Reticular fibers were mainly limited to blood and lymphatic vessel walls, as well as the perineurium and epineurium of nerve fibers. The elastic fiber content was minimal, appearing mainly in the lamina propria and vascular walls and interspersed with type I collagen in the connective tissue matrix of the septum. Smooth muscle bundles were mainly observed in the muscularis layers of both walls, but to a greater extent in the anterior vagina. These findings largely confirm those of the few published histological studies of this tissue while contributing to the current state of knowledge regarding the distribution of elastin and reticulin. Further research in premenopausal females and increased awareness of the microanatomy of the urethrovaginal septum is advised.

An Unusual Skin Finding in a Rare Case of Goltz Syndrome

Abstract Introduction/Objective Goltz Syndrome (Focal Dermal Hypoplasia) is a rare X linked dominant genetic disorder with various dermatologic, musculoskeletal and neurologic manifestations, secondary to a mutation in the PORCN gene. Skin findings include telangiectasias, raspberry-like papillomas (angiofibroma like lesions), nodular fat herniation, xerosis and photosensitivity. Lesions are bilateral, and asymmetrically distribututed along the lines of Blasco. The adipocytic lesions can be diagnosed via a skin biopsy showing adipose tissue in the superficial epidermis and dermis or via molecular techniques showing a mutation in the PORCN gene. Here we report a case of Goltz Syndrome with unique dermal findings. Methods/Case Report A male infant presented at birth with mild diffuse xerosis, large hypopigmented and depigmented atrophic patches and plaques and multiple pink yellow red exophyitic soft papules distributed asymetrically across the body along the lines of Blascho. Additionally the patient had clinodactly, widely spaced nipples, extra pads of fat in the toes and philtrum and fingernails with longitudal ridges and lateral nail plate deviation. A diagnosis of Goltz syndrome was made via molecular analysis that showed mosiacism at C.502G>A (p.Gly168Arg) in the PORCN gene. The patient had a very superficial 2-3 mm erosion at the intersection between the scrotum and perineum. At 18 months, this had grown to finger like projections and eventually grew into multiple tan red exophytic warty papillomatous telangactatic like lesions measuring 3.0 x 0.8 cm that were painful and bled easily. A biopsy of this lesion showed focal mild epidermal hyperplasia and elongation of the rete ridges with wide spread diffuse vascular proliferation with multiple ectactic vessels along with mild dermal and perivascular inflammatory infiltrates. Also noted was infiltration of mature adipose tissue in the superficial epidermis and dermis. We preformed elastin and trichrome stains to identify if this lesion was reactive in nature or de novo. Our results showed retained elastin staining with a negative trichrome eluding to a denovo process. Additionally GLUT1 was negative, ruling out an infantile hemangioma. The above findings in this lesion do not represent any previously described dermatologic entity. Results (if a Case Study enter NA) NA Conclusion This is a rare and unique dermatologic finding of a case of Goltz sydrome, which itself is a rare disorder. In our literature review, this is the only case of Goltz Syndrome thusfar to have presented with this unique skin lesion.

Stereologic consequences of iatrogenic collapse: The morphology of adenocarcinoma in situ overlaps with invasive patterns. Proposal for a necessary modified classification of pulmonary adenocarcinomas

Recognizing non-invasive growth patterns is necessary for correct diagnosis, invasive size determination and pT-stage in resected non-small cell lung carcinoma. Due to iatrogenic collapse after resection, the distinction between adenocarcinoma in-situ (AIS) and invasive adenocarcinoma may be difficult. The aim of this study is to investigate the complex morphology of non-mucinous non-invasive patterns of AIS in resection specimen with iatrogenic collapse, and to relate this to follow-up.The effects of iatrogenic collapse on the morphology of collapsed AIS were simulated in a mathematical model. Three dimensional related criteria applied in a modified classification, using also cytokeratin 7 and elastin as additional stains, in two independent retrospective cohorts of primary pulmonary adenocarcinomas ≤3 cm resection specimen with available follow-up information.The model demonstrated that infolding of alveolar walls occurs during iatrogenic collapse and lead to a significant increase in tumor cell heights in maximal collapse areas, compared to less collapsed areas. The morphology of infolded AIS overlaps with patterns described as papillary and acinar adenocarcinoma according to the WHO classification, necessitating an adaptation.The modified classification incorporates recognition of iatrogenic and biologic collapse, tangential cutting effect true invasion and surrogate markers of invasion i.e. grey zone, covering a multilayering falling short of micropapillary, cribriform and solid alveolar filling growth. The use of elastin and CK7 staining aids in the morphologic recognition of iatrogenic collapsed AIS and the distinction from invasive adenocarcinoma. Out of a total of 70 resection specimens 1 case was originally classified as AIS and 9 were reclassified as iatrogenic collapsed AIS. Patients with collapsed AIS showed a 100 % recurrence-free survival after a mean follow-up time of 69.5 months.With the current WHO classification, AIS is overdiagnosed as invasive adenocarcinoma due to infolding. The modified classification facilitates the diagnosis of AIS.

Investigating the role of various extracellular matrix in androgenetic alopecia: Insights from immunostaining and quantitative analysis – A pilot study

Abstract Androgenetic alopecia (AGA) is the most common hair loss, characterized by miniaturization of hair follicles, shortened anagen, and prolonged telogen phases. Clinically, distinct patterns of hair loss distribution are evident between males and females. The pathogenesis of AGA is considered a mutual interplay of genetic, hormonal, and environmental factors. Mechanical properties of the scalp, for example, skin stiffness, have also been proposed as an important factor. To date, no comprehensive study has been conducted to demonstrate the density and distribution of various extracellular matrix (ECM) components at different dermal depths in AGA. To address this knowledge gap, we conducted immunohistochemical staining in patients with pattern hair loss to analyze the distribution of ECM components. Our findings reveal distinct distribution patterns of collagen I, collagen III, and elastin fibers within different dermal layers in male and female AGA cases. Notably, the intensity of elastin staining was significantly increased in all dermal layers in the male AGA skin, compared to the normal scalp skin. In summary, our study highlights the significance of ECM components in both male and female AGA.

Assessing venous invasion in stage II colon cancer: optimal elastin stains and survival analysis

AimsVenous invasion (VI) in colorectal carcinoma influences treatment strategies, especially in early stages. Despite elastin staining effectiveness in detecting VI, guidelines for its routine application, including the optimal number of...

Morphologic Features of Invasion in Lung Adenocarcinoma: Diagnostic Pitfalls.

Reproducibility of pulmonary invasive adenocarcinoma diagnosis is poor when applying the World Health Organization (WHO) classification. In this article, we aimed first to explain by 3-dimensional morphology why simple pattern recognition induces pitfalls for the assessment of invasion as applied in the current WHO classification of pulmonary adenocarcinomas. The underlying iatrogenic-induced morphologic alterations in collapsed adenocarcinoma in situ overlap with criteria for invasive adenocarcinoma. Pitfalls in seemingly acinar and papillary carcinoma are addressed with additional cytokeratin 7 and elastin stains. In addition, we provide more stringent criteria for a better reproducible and likely generalizable classification.

Delineation of the healthy rabbit tongue by immunohistochemistry – A technical note

In the oral cavity the tongue is an important muscular organ that supports the swallowing of food and liquids. It is responsible for the sense of taste, based on the many different taste buds it contains. Research in the field of tongue diseases demands for suitable preclinical models. The healthy rabbit tongue may therefore serve as baseline and reference for the pathological situation. With this consideration, we covered the fixation and histological stainings as well as the immunohistochemical labelling of the healthy rabbit tongue. In this technical note, initial choice of the fixative is discussed, with a comparison of formalin fixation and subsequent paraffin embedding versus cryopreservation. Moreover, we delineate the effect of an antigen retrieval step for formalin fixation by several examples. Finally, we provide ECM markers collagen I, collagen III, fibronectin, α-SMA and elastin staining as well as ki67 for proliferative status and PAR-2 protein expression as a marker for inflammatory status and nociception in tongue sections, mainly from the tongue body. Technically, we found superiority of paraffin sections for collagen I, collagen III, fibronectin, ki67 and α-SMA labelling, for selected detections systems. As for ECM components, the lamina propria was very rich in collagen and fibronectin, while the muscular body of the tongue showed only collagen and fibronectin positive areas between the muscle fibers. Moreover, α-SMA was clearly expressed in the walls of arteries and veins. The inflammatory marker PAR-2 on the other hand was prominently expressed in the salivary glands and to some extent in the walls of the vessels. Particular PAR-2 expression was found in the excretory ducts of the tongue. This technical note has the aim to provide baseline images that can be used to compare the pathological state of the diseased rabbit tongue as well as for inter-species comparison, such as mouse or rat tongue. Finally, it can be used for the comparison with the human situation.

The Usefulness of Elastin Staining to Detect Vascular Invasion in Cancer.

Tumor prognosis hinges on accurate cancer staging, a pivotal process influenced by the identification of lymphovascular invasion (LVI), i.e., blood vessel and lymphatic vessel invasion. Protocols by the College of American Pathologists (CAP) and the World Health Organization (WHO) have been established to assess LVI in various tumor types, including, but not limited to, breast cancer, colorectal cancer (CRC), pancreatic exocrine tumors, and thyroid carcinomas. The CAP refers to blood vessel invasion as "angioinvasion" (vascular invasion) to differentiate it from lymphatic vessel invasion (lymphatic invasion). For clarity, the latter terms will be used throughout this review. The presence of lymphatic and/or vascular invasion has emerged as a pivotal prognostic factor; therefore, its accurate identification is crucial not only for staging but also for providing the patient with an honest understanding of his/her prognosis. Given the prognostic importance of the correct identification of LVI, specific staining techniques are employed to distinguish lymphatic vessel invasion from angioinvasion and to differentiate true LVI from artifact. These encompass hematoxylin and eosin (H&E) staining, elastic staining, Factor VIII staining, Ulex europaeus I agglutinin staining, CD31, CD34, D2-40, ERG, and D2-40 (podoplanin) immunohistochemical (IHC) stains among others. Based on a review of numerous publications regarding the efficacy of various methods for LVI detection, elastin staining demonstrated superior accuracy and prognostic value, allowing for more targeted treatment strategies. The clinical significance of accurately detecting LVI cannot be overstated, as it is strongly linked to higher cancer-related mortality and an increased risk of tumor recurrence. This review aims to examine the existing literature on the use of elastin stains in the detection of vascular invasion among different types of tumors and its prognostic value.

An approach to reduce Descemet's membrane scrolling: Relevance to Descemet's membrane endothelial keratoplasty (DMEK).

We aimed to determine whether Descemet's membrane (DM) scrolling occurs primarily along the vertical or horizontal axis and establish whether oval trephination along the axis of least scrolling can reduce the grade of the scroll. The longest limbus-to-limbus axis on 28 sclerocorneal discs was taken as the horizontal axis. The horizontal (n = 7) or (right angles to it) vertical (n = 6) axis was marked on DM before peeling it off. The direction and grade of scrolling was observed. Narrow strips (3-4 mm wide) were then cut along the two axes (n = 4 each) and the scrolling pattern was observed. Ellipses (7 × 9 mm) of DM were punched along the two axes (n = 6 each) and the scrolls graded. Immunofluorescent staining for elastin on horizontal and vertical tissue sections from three DM samples was performed. The intensity and thickness of elastin staining were measured. Twenty-four (85.72%) DM samples showed scrolling along the horizontal axis, none showed scrolling along the vertical axis, and four (14.28%) samples showed a spiral scroll, regardless of which axis was marked (grade 3.7 and 3.6). Vertically oval discs showed significantly reduced scrolling (grade 1.2) compared to horizontally oval discs (grade 3.5). Narrow strips of DM showed a similar scrolling pattern. Immunohistology showed no difference in any of the parameters examined along the two axes or from the center to the periphery. DM scrolls primarily along the horizontal axis. Vertically oval DM samples show minimal scrolling, which can be an advantage in DMEK. Differential scrolling is not determined by the distribution of elastin.

Impact of tissue sampling on detection of venous invasion in colorectal cancer: a prospective analysis.

Venous invasion (VI) is a powerful yet under-reported prognostic factor in colorectal cancer (CRC). Efforts to improve its detection have largely focused upon histological assessment, with less attention paid to tissue-sampling strategies. This study aimed to prospectively determine the number of tumour blocks required to optimise VI detection in CRC resections. In addition, the relationship between linear spiculation (LS) and extramural venous invasion (EMVI) was investigated. A standardised tissue sampling protocol was developed and applied prospectively to 217 CRC resections [AJCC 8th edition, stage 1 (n = 32); stage 2 (n = 84); stage 3 (n = 87); stage 4 (n = 14); and post-neoadjuvant therapy (n = 46)]. Elastin stains were performed on all tumour blocks. VI was identified in 55% of cases (EMVI = 37%; IMVI alone = 18%). The sensitivity of VI detection increased with increasing numbers of tumour blocks submitted [one block (35%), three blocks (66%), five blocks (84%), six blocks (95%) and seven blocks (97%)]. Similar findings were observed for EMVI [one block (35%), three blocks (73%), five blocks (89%), six blocks (96%) and seven blocks (96%)]. LS was identified macroscopically in 22% of specimens. In cases where no neoadjuvant therapy had been given, EMVI was significantly associated with LS (71% in LS+ cases versus 29% in LS- cases; P < 0.001). In addition, tumour blocks targeting LS were associated with a fivefold higher rate of EMVI compared with blocks that did not (P < 0.001). Our findings demonstrate the impact of tissue sampling and quality of gross examination on VI detection and may inform practices in future CRC protocols.

E72 Distribution of cells and extracellular matrix in the marginal synovium during the postnatal period

Abstract Introduction The marginal transitional zone of the joint is of fundamental importance in joint pathology. The normal marginal synovium takes part in articular cartilage repair (Adachi et al. 2012). The inflamed marginal synovium, in turn, promotes irreversible damage to the adjacent cartilage and bone (Hitchon &amp; El-Gabalawy, 2011). Early erosive damage in rheumatoid arthritis and osteophyte formation in osteoarthritis occurs primarily here (Allard et al. 1990). Objectives The morphology of the marginal synovium from birth till the 90th day was studied using the hip joints of Wistar rats. Methods The collection of joints was carried out on the first, seventh, 14th, 30th, 45th, 60th and 90th day (six rats in each group, 42 in total). Joints were fixed, decalcified and dehydrated. Histologic paraffine sections were stained with Mallory's trichrome and Hart's elastin staining, analyzed at the acetabular labrum by light microscopy (χ100), morphometrical and statistical (P &amp;lt; 0.05) methods with extrapolation to humans (Sengupta, 2013). The obtained data were presented as percentages. Results In the marginal synovium, cells and components of the extracellular matrix, which are typical for fibrous connective tissue, could be seen. The synovial lining cells are embedded in the modified basement membrane which includes collagen and elastic fibers. They form a three-dimensional reticulum in it and adjacent synovial subintima. Vessels of the microcirculatory bed are also widely represented in the marginal synovium. The overall quantity of collagen fibers increased gradually, exceeding on the 90th day (approximately corresponds to 18–19 human years) that of newborns by one and a half times (48.92 ± 3.39 vs 72.17 ± 7.77; P = 0.023). The number of arranged collagen fibers increased gradually, exceeding on the 90th day that of newborns almost twofold (37.25 ± 3.21 vs 60.08 ± 5.99; P = 0.008). The area of distribution of disarranged collagen fibers in newborns was almost four and a half times less than the overall quantity of collagen fibers (11.67 ± 1.03 vs 48.92 ± 3.39; P &amp;lt; 0.001) and did not change substantially throughout the experiment. The number of elastic fibers increased till the 30th day (0.63 ± 0.11 vs 18.3 ± 1.25; P &amp;lt; 0.001) and did not undergo significant changes later. In newborns, the area of distribution of ground substance was not considerably higher than the ratio of disarranged collagen fibers (10.42 ± 1.8 vs 11.67 ± 1.03; P = 0.562) and did not undergo substantial changes later. The distribution density of vessels increased till the 30th-45th day (4.5 ± 0.67 vs 7.5 ± 0.82 and 7.7 ± 0.54; P &amp;lt; 0.001; approximately corresponds to the 9th and 14th human years respectively), followed by a gradual decrease from the 60th day (7.05 ± 0.72; roughly corresponds to the 16th human years). The maximum number of cells was in newborns. Subsequently, it gradually decreased and was almost three times less on the 90th day than that of newborns (38.01 ± 3.06 vs 12.58 ± 1.53; P &amp;lt; 0.001). Conclusion The morphogenesis of the marginal synovium during the postnatal period is accompanied by a gradual decrease in the number of cells, an increase in the area occupied by elastic and arranged collagen fibers with a relatively constant distribution density of disarranged collagen fibers and ground substance together with an undulatory pattern in the distribution of vessels. These peculiarities emphasize the morphological uniqueness of the marginal transitional zone, which forms the innate nonspecific barrier between the joint capsule and the articular cartilage, and thereby provides the necessary anatomical and physiological integrity of the joint as a whole organ. Knowledge about the patterns of morphogenesis of the marginal synovium will help in the future to analyze the modeled pathological or boundary states in it. Ethics The study was approved by the ethics committee of the Zaporizhzhia State Medical University.

E71 Distribution of cells and extracellular matrix in the normal marginal synovium of the hip joint capsule during the postnatal period

Abstract Introduction The marginal transitional zone is of fundamental importance in joint pathology. It includes the marginal cartilage and marginal synovium. The normal marginal synovium takes part in articular cartilage repair (Adachi et al. 2012). The inflamed marginal synovium, in turn, promotes irreversible damage to the adjacent cartilage and bone (Hitchon &amp; El-Gabalawy, 2011). Objectives The morphology of the normal marginal synovium from birth till the 90th day was studied using the hip joints of Wistar rats. Methods The collection of joints was carried out on the first, seventh, 14th, 30th, 45th, 60th and 90th day (six rats in each group, 42 in total). Joints were fixed, decalcified and dehydrated. Histologic paraffine sections were stained with Mallory's trichrome and Hart's elastin staining, analyzed at the acetabular labrum by light microscopy (χ100), morphometrical and statistical (P &amp;lt; 0.05) methods with extrapolation to humans (Sengupta, 2013). The obtained data were presented as percentages. Results In the marginal synovium, cells and components of the extracellular matrix, which are typical for fibrous connective tissue, could be distinguished. The synovial lining cells are embedded in the modified basement membrane which is a meshwork of collagen and elastic fibers. They form a three-dimensional reticulum in it and adjacent synovial subintima. Vessels of the microcirculatory bed are also widely represented in the marginal synovium. The overall quantity of collagen fibers increased gradually, exceeding on the 90th day (∼18–19 human years) that of newborns by one and a half times (48.92 ± 3.39 vs 72.17 ± 7.77; P = 0.023). The number of arranged collagen fibers increased gradually, exceeding on the 90th day that of newborns almost twofold (37.25 ± 3.21 vs 60.08 ± 5.99; P = 0.008). The area of distribution of disarranged collagen fibers in newborns was almost four and a half times less than the overall quantity of collagen fibers (11.67 ± 1.03 vs 48.92 ± 3.39; P &amp;lt; 0.001) and did not change substantially throughout the experiment. The number of elastic fibers increased till the 30th day (0.63 ± 0.11 vs 18.3 ± 1.25; P &amp;lt; 0.001) and did not undergo significant changes later. In newborns, the area of distribution of ground substance was not considerably higher than the ratio of disarranged collagen fibers (10.42 ± 1.8 vs 11.67 ± 1.03; P = 0.562) and did not undergo substantial changes later. The distribution density of vessels increased till the 30th-45th day (4.5 ± 0.67 vs 7.5 ± 0.82 and 7.7 ± 0.54; P &amp;lt; 0.001; ∼9-14th human years), followed by a gradual decrease from the 60th day (7.05 ± 0.72; ∼16 human years). The maximum number of cells was in newborns. Subsequently, it gradually decreased and was almost three times less on the 90th day than that of newborns (38.01 ± 3.06 vs 12.58 ± 1.53; P &amp;lt; 0.001). Conclusion The morphogenesis of the marginal synovium during the postnatal period is accompanied by a gradual decrease in the number of cells, an increase in the area occupied by elastic and arranged collagen fibers with a relatively constant distribution density of disarranged collagen fibers and ground substance together with an undulatory pattern in the distribution of vessels. These peculiarities emphasize the morphological uniqueness of the marginal transitional zone, which forms the innate nonspecific barrier between the joint capsule and the articular cartilage, and thereby provides the necessary anatomical and physiological integrity of the joint as a whole organ. Knowledge about the patterns of morphogenesis of the normal marginal synovium will help to understand joint pathology. Ethics The study was approved by the ethics committee of the Zaporizhzhia State Medical University.

Routine elastin staining improves venous invasion detection in colorectal carcinoma

BackgroundColorectal carcinoma is the second most common cause of cancer-related deaths in North America. Invasion of tumor cells into lymphatic and blood vessels is an imperative step in the metastatic progression of colorectal carcinoma. ObjectivesThis is a before-and-after study conducted by the Department of Pathology and Laboratory Medicine of Mount Sinai Medical Center of Florida to assess the impact on venous invasion (VI) detection by implementing routine elastin staining on all tumor-containing blocks per case, where feasible, in colorectal carcinoma (CRC) resection specimens. MethodsClinicopathological parameters of CRC specimens were collected from January until December 2021 (n = 93) for the pre-implementation cohort and from January until December 2022 (n = 61) for the post-implementation cohort. ResultsVI detection was significantly increased in the post-implementation cohort at a rate of 50.8 % compared to only 18.6 % in the pre-implementation cohort. The majority of VI identified in the pre-implementation cohort was extramural (61.5 %), whereas in the post-implementation cohort it was intramural (41.9 %). On univariate analysis, implementation of routine elastin stain was associated with strikingly increased VI detection rates (OR = 4.5, p-value < 0.001). On multivariate analysis, after adjusting for other clinicopathologic variables, elastin staining retained its independent statistically significant impact on VI detection (OR = 2.6, p-value = 0.034). Of note, there were no significant differences in the pre- and post-implementation cohorts in the frequency of nodal metastases, tumor extent, histologic grade, perineural invasion, T stage or M stage. ConclusionBased on our results and what has been published recently, we confirm an increase in the VI detection rate after implementing routine elastin staining on all tumor-containing blocks in CRC resection specimens.

(090) Immunohistochemical Staining of Elastin in the Vestibule

Abstract Introduction The vestibule is an external genital structure lying 90° to the vaginal canal opening. During the thrusting movements associated with penetrative sexual activity, the vestibule undergoes significant stretching during both invagination and evagination. The vestibule also undergoes extreme stretching during childbirth. The rete pegs in the non-keratinized squamous epithelium facilitate the ability of the vestibule to mechanically stretch. More importantly, however, the vestibular lamina propria, with nerves, blood vessels, glands, needs to contain a high density of extracellular matrix (ECM) proteins with stretching capabilities. In particular, the ECM protein elastin endows skin structures with the ability to stretch, recoil, and have elasticity. Elastin has previously been shown to influence mechanical stretching properties of tendons. Objective The objective of this study was to examine the density of elastin proteins in the vestibule in reproductive age patients as there are currently limited data. Methods A total of 7 reproductive age patients with normal sex steroid hormone values and complaints of entrance dyspareunia were diagnosed with neuroproliferative vestibulodynia and subsequently underwent complete vestibulectomy. Tissue blocks were created from the specimen, and 5 μm sections studied with immunohistochemical staining with primary antibody. Sections were then counterstained with Gill-modified hematoxylin. Results In all vestibular specimen, there was very high density of elastin underneath the epithelium in rete pegs and throughout the entire lamina propria (Fig 1). There is a density gradient with the highest density immediately deep to the basement membrane of the epithelium. Elastin fibers were arranged in all orientations: coronal (frontal), sagittal (longitudinal), and transverse (axial) planes. Conclusions The high density of elastin fibers in rete pegs and throughout the lamina propria in a multi-planar orientation is suggestive of extreme stretching capability in eugonadal vestibular tissues. The highest density of elastin staining was seen immediately below the basement membrane of the epithelium, which may provide tissue integrity and shear resilience to the epithelial layer. In sex steroid hormone insufficiency states such as menopause or with use of combined hormonal contraceptives, individuals may develop loss of introital elasticity and fissures due to a deficiency of the ECM protein elastin. More research is needed on elastic fiber production, organization, and integration with other cutaneous extracellular matrix proteins, and how they may be affected by sex steroid hormones. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Endoceutics, Inc.

A Biomimetic Approach Utilizing Pulsatile Perfusion Generates Contractile Vascular Grafts.

Surgical implantation of decellularized cadaveric arteries is routinely used to treat right-sided congenital cardiac lesions. These acellular conduits lack the capacity for somatic growth and are prone to stenosis and calcification, necessitating multiple operations throughout childhood. Islet-1+ cardiovascular progenitor cells (CPCs) have demonstrated the capacity for differentiation into all cell types of the heart and outflow tracts. We hypothesize that CPC seeding of decellularized pulmonary arteries and bioreactor culture under physiologic flow conditions will drive vascular differentiation of CPCs and result in a conduit more suitable for implantation and long-term growth. We began by decellularizing ovine pulmonary arteries and characterizing the composition of the extracellular matrix (ECM). Hemodynamic testing of decellularized vessels in a custom bioreactor was used to define the scaffold mechanical properties over a range of pressures and flow rates. Next, our expanded ovine CPCs were suspended in growth media and injected intramurally into decellularized pulmonary arteries that were subsequently cultured in either static or pulsatile cultures. A combination of immunohistochemistry, real-time polymerase chain reaction (PCR), and tissue bath contraction studies were used to evaluate the bioengineered arteries before transplantation. Pulmonary artery patches from the most favorable culture conditions were then implanted into juvenile sheep to provide proof of concept. Hematoxylin and eosin staining indicated complete removal of cell nuclei (n = 9), whereas double-stranded DNA isolation from tissue homogenates showed 99.1% DNA removal (p < 0.01, n = 4). Furthermore, trichrome and elastin staining verified maintenance of collagen and elastin. Immunohistochemistry and PCR analyses (n = 4 per group) confirmed contractile smooth muscle presence on only our 3-week pulsatile scaffolds via presence of calponin 1 and myosin heavy chain 11. Tissue bath studies demonstrated that smooth muscle contraction generated by our 3-week pulsatile scaffolds (2.23 ± 0.19 g, n = 4) is comparable with native tissue contraction strength (2.78 ± 0.06 g, n = 4). Ovine transplantation confirmed that our graft can be safely implanted, retains contractile smooth muscle cells, and recruits native endothelium. Longer duration of physiologic pulsatile culture drives differentiation of CPCs seeded on ECM conduits toward a mature, contractile phenotype that is maintained for several weeks in vivo. Longer term studies to assess somatic growth potential are needed. Impact statement The current field of vascular transplantation relies on cadaveric and synthetic grafts to treat right-sided congenital cardiac lesions. These grafts do not grow somatically with our patients. This results in multiple reoperations throughout childhood to increase the size of the graft. Our bioengineered alternative demonstrates successful implantation, contractile smooth muscle cells, and a native endothelial layer. This research demonstrates a pilot study confirming the viability of a bioengineered alternative to the current standard of care in the field of vascular transplantation.

Prognostic significance of lymphovascular invasion in patients with pT1b esophageal squamous cell carcinoma

BackgroundLymphovascular invasion (LVI) is a crucial predictor of lymph node metastasis (LNM). However, few studies have investigated the LVI positivity rate and its clinical significance in pT1b esophageal squamous cell carcinoma (ESCC) using immunohistochemistry and elastin staining.MethodsWe collected data from158 patients with pT1b ESCC who had undergone radical esophagectomy. All paraffin blocks of invasive carcinoma from each patient were subjected to HE staining, elastin staining + CK (AE1/AE3) immunohistochemistry (E&IHC), and CD31/D2-40 + CK (AE1/AE3) double immunohistochemistry (D-IHC). The LVI was classified into types, i.e., vascular invasion (VI) and lymphatic vessel invasion (LI), and its location, quantity, and clinical significance were explored.ResultsThe positivity rates of VI by E&IHC (E-VI), VI by CD31D-IHC (CD31-VI), and LI by D2-40 D-IHC (D2-40-LI) were significantly higher than those obtained by HE staining (P < 0.001, respectively). CD31-VI and E-VI were independent adverse prognostic factors for recurrence-free survival (RFS), and they were significantly associated with poor distant metastasis-free survival and overall survival in pT1b ESCC. Intratumoral LVI was also crucial in pT1b ESCC, and L2 (the count of D2-40-LI was 5 or more) was the strongest predictor for LNM and RFS in pT1b ESCC.ConclusionE&IHC and D-IHC can dramatically improve the detection rate of LVI in pT1b ESCC, and the classification and grading of LVI can help to improve the prediction of LNM and prognosis.

DOCK2 Deficiency Attenuates Abdominal Aortic Aneurysm Formation-Brief Report.

Abdominal aortic aneurysm (AAA) is a potentially lethal disease that lacks pharmacological treatment. Degradation of extracellular matrix proteins, especially elastin laminae, is the hallmark for AAA development. DOCK2 (dedicator of cytokinesis 2) has shown proinflammatory effects in several inflammatory diseases and acts as a novel mediator for vascular remodeling. However, the role of DOCK2 in AAA formation remains unknown. Ang II (angiotensin II) infusion of ApoE-/- (apolipoprotein E deficient) mouse and topical elastase-induced AAA combined with DOCK2-/- (DOCK2 knockout) mouse models were used to study DOCK2 function in AAA formation/dissection. The relevance of DOCK2 to human AAA was examined using human aneurysm specimens. Elastin fragmentation in AAA lesion was observed by elastin staining. Elastin-degrading enzyme MMP (matrix metalloproteinase) activity was measured by in situ zymography. DOCK2 was robustly upregulated in AAA lesion of Ang II-infused ApoE-/- mice, elastase-treated mice, as well as human AAA lesions. DOCK2-/- significantly attenuated the Ang II-induced AAA formation/dissection or rupture in mice along with reduction of MCP-1 (monocyte chemoattractant protein-1) and MMP expression and activity. Accordingly, the elastin fragmentation observed in ApoE-/- mouse aorta infused with Ang II and elastase-treated aorta was significantly attenuated by DOCK2 deficiency. Moreover, DOCK2-/- decreased the prevalence and severity of aneurysm formation, as well as the elastin degradation observed in the topical elastase model. Our results indicate that DOCK2 is a novel regulator for AAA formation. DOCK2 regulates AAA development by promoting MCP-1 and MMP2 expression to incite vascular inflammation and elastin degradation.

MP27-14 ELASTIN STAINING OF THE VESTIBULE

You have accessJournal of UrologyCME1 Apr 2023MP27-14 ELASTIN STAINING OF THE VESTIBULE Alyssa Yee, Alain Dury, Johanne Ouellet, Barry Komisaruk, Sue W Goldstein, and Irwin Goldstein Alyssa YeeAlyssa Yee More articles by this author , Alain DuryAlain Dury More articles by this author , Johanne OuelletJohanne Ouellet More articles by this author , Barry KomisarukBarry Komisaruk More articles by this author , Sue W GoldsteinSue W Goldstein More articles by this author , and Irwin GoldsteinIrwin Goldstein More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003255.14AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The vestibule is an external genital structure lying 90° to the vaginal canal opening. The vestibule consists of non-keratinized squamous epithelium with numerous rete pegs, underlying lamina propria with nerves, blood vessels, glands, and extracellular matrix (ECM) proteins. During thrusting movements associated with penetrative sexual activity, the vestibule undergoes significant stretching during invagination/evagination. The vestibule also undergoes extreme stretching during childbirth. Elastin is an ECM protein in microfibrils that endows skin structures with ability to stretch, recoil, and have elasticity. Elastin has previously been shown to influence mechanical properties of tendons; degraded or abnormal elastin structures caused by aging or disease may contribute to pain and increased risk of injury. There are limited data on elastin in the vestibule in reproductive age patients and its role in vestibular elasticity/integrity. METHODS: 7 reproductive age patients with entrance dyspareunia from neuroproliferative vestibulodynia, with normal values of testosterone, estradiol, and progesterone, underwent complete vestibulectomy. Tissue blocks were created from the specimen, and 5 υm sections studied with IHC primary antibody. Finally, sections were counterstained with Gill-modified hematoxylin. RESULTS: In all vestibular specimen, there was very high density of elastin underneath the epithelium in rete pegs and throughout the entire lamina propria. There is a density gradient with the highest density immediately deep to the basement membrane of the epithelium. Elastin fibers were arranged in all orientations: coronal (frontal), sagittal (longitudinal), and transverse (axial) planes. CONCLUSIONS: The high density of elastin fibers in rete pegs and through the lamina propria in a multi-planar orientation is suggestive of extreme stretching capability in eugonadal vestibular tissues. The highest density of elastin staining was seen immediately below the basement membrane of the epithelium, which may provide tissue integrity and shear resilience to the epithelial layer. More research is needed on elastic fiber production, organization, and integration with other cutaneous extracellular matrix proteins, and how this may change during androgen insufficiency states such as menopause or with use of combined hormonal contraceptives. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e367 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alyssa Yee More articles by this author Alain Dury More articles by this author Johanne Ouellet More articles by this author Barry Komisaruk More articles by this author Sue W Goldstein More articles by this author Irwin Goldstein More articles by this author Expand All Advertisement PDF downloadLoading ...

Mesenchymal Stromal Cells Combined With Elastin-Like Recombinamers Increase Angiogenesis In Vivo After Hindlimb Ischemia.

Hindlimb ischemia is an unmet medical need, especially for those patients unable to undergo vascular surgery. Cellular therapy, mainly through mesenchymal stromal cell (MSC) administration, may be a potentially attractive approach in this setting. In the current work, we aimed to assess the potential of the combination of MSCs with a proangiogenic elastin-like recombinamer (ELR)–based hydrogel in a hindlimb ischemia murine model. Human bone marrow MSCs were isolated from four healthy donors, while ELR biomaterials were genetically engineered. Hindlimb ischemia was induced through ligation of the right femoral artery, and mice were intramuscularly injected with ELR biomaterial, 0.5 × 106 MSCs or the combination, and also compared to untreated animals. Tissue perfusion was monitored using laser Doppler perfusion imaging. Histological analysis of hindlimbs was performed after hematoxylin and eosin staining. Immunofluorescence with anti–human mitochondria antibody was used for human MSC detection, and the biomaterial was detected by elastin staining. To analyze the capillary density, immunostaining with an anti–CD31 antibody was performed. Our results show that the injection of MSCs significantly improves tissue reperfusion from day 7 (p = 0.0044) to day 21 (p = 0.0216), similar to the infusion of MSC + ELR (p = 0.0038, p = 0.0014), without significant differences between both groups. After histological evaluation, ELR hydrogels induced minimal inflammation in the injection sites, showing biocompatibility. MSCs persisted with the biomaterial after 21 days, both in vitro and in vivo. Finally, we observed a higher blood vessel density when mice were treated with MSCs compared to control (p<0.0001), but this effect was maximized and significantly different to the remaining experimental conditions when mice were treated with the combination of MSCs and the ELR biomaterial (p < 0.0001). In summary, the combination of an ELR-based hydrogel with MSCs may improve the angiogenic effects of both strategies on revascularization of ischemic tissues.

A patient with recurrent strokes: multimodal imaging reveals two possible causes

Abstract An underlying cardio-embolic cause can be identified in a fifth of stroke patients. Cardiac tumors occur rarely in routine clinical practice but can cause severe complications such as heart failure, valve dysfunction, embolic events, or sudden cardiac death. Herein we present the case of a 67-year-old patient with a history of recurrent stroke in whom a patent foramen ovale (PFO) and a hypermobile structure attached to the aortic valve were diagnosed during a transesophageal examination. The structure was interpreted to be a papillary fibroelastoma. Coronary computed tomography angiography completed the diagnosis, helping to exclude a relevant coronary artery disease. The patient underwent an uneventful cardiac surgery with removal of the hypermobile structure and closure of the PFO. The histopathological analysis, however, was negative for elastin in the elastin stain, and thus the diagnosis of a cardiac fibroma was made. The case emphasizes the role of cardiovascular imaging in stroke patients as well as that of histopathological analysis.