Elastin Expression Research Articles

The efficacy and safety of low- versus high-fluence fractional picosecond Nd:YAG 1064-nm laser in the treatment of acne scars: A randomized split-face comparison study.

Differences in clinical efficacy based on the fluence of fractional picosecond laser treatment for acne scars are unknown. To compare the efficacy and safety of low-fluence versus high-fluence fractional picosecond Nd:YAG 1064-nm laser treatment in acne scar patients. In this 12-week, investigator-blinded, randomized, split-face study, 25 patients with moderate-to-severe acne scars received three sessions of high-fluence laser treatment (1.0 J/cm2 ) on one side of their face and low-fluence (0.3 J/cm2 ) on the other side every 4 weeks. Patients were assessed using acne scar counts, the scar global assessment (SGA), and the ECCA scar grading scale every 4 weeks. The histological analysis compared the acne scars obtained before and 4 weeks after treatment. At their last visit, 88.00% and 92.00% of the subjects achieved >30% reduction in scar counts on the low- and high-fluence sides, respectively, without a significant difference between the two sides. On both sides, the scar counts, SGA, and ECCA score significantly improved 4 weeks after the last treatment. Although the high-fluence side showed a greater reduction in scar counts (-66.73%) than the low-fluence side (-62.13%), the two sides had no significant difference in the grading scores. The high-fluence side showed significantly more severe pain and higher side-effect scores immediately and 4 weeks after treatment. Histological analysis revealed a significantly increased collagen, elastin, and vimentin expression after treatment on the low-fluence side. The low-fluence setting demonstrated comparable efficacy and superior safety in treating acne scars compared with the high-fluence setting.

Future of biotherapeutics: Harnessing mRNA to enhance elastin expression

Future of biotherapeutics: Harnessing mRNA to enhance elastin expression

Importance of smooth muscle p110alpha/AKT/FOXO1 signaling for the development of abdominal aortic aneurysm

Abstract Background Abdominal aortic aneurysms (AAA) are characterized by loss of vascular smooth muscle cells (SMCs) and extracellular matrix (ECM) degradation. The catalytic PI 3-kinase isoform p110α signals downstream of growth factor receptors. Mice harboring an SMC-specific p110α deficiency (SM-p110α-/-) are characterized by impaired SMC proliferation and survival. Aim We hypothesized that p110α deficiency in SMCs exacerbates the development and progression of AAA as it promotes SMC loss and thus weakens the aortic wall in healthy and diseased animals. Therefore, we investigated how p110α deficiency in SMCs affects AAA formation, vascular integrity as well as downstream signaling in SMCs. Methods and results Medial thickness of the abdominal aorta in SM-p110α-/- mice was significantly decreased compared to wild-type (WT) littermate controls (29.0±3.1µm vs. 42.5±4.1µm). Morphological characterization of the aortic wall by transmission electron microscopy revealed impaired elastic fibers, detached SMCs and apoptotic cells in SM-p110α-/- mice. SMC differentiation markers including calponin and myosin heavy chain were downregulated in the aortic wall of SM-p110α-/- mice. Western blots demonstrated that p110α deficiency impaired the expression of elastin. Consequently, elastin fibers were characterized by reduced fiber width and length. In addition, growth factor stimulated proliferation of p110α-/- SMCs was significantly impaired compared to WT controls. AAA development in SM-p110α-/- mice and WT littermates were studied using the porcine pancreatic elastase (PPE) model. PPE was infused into the infrarenal aorta. Ultrasound examination of the aorta revealed an increased aortic diameter in all PPE-treated mice. However, the increase in aortic diameter was significantly elevated (p<0.05) in SM-p110α-/- mice (70.2±10.8%, n=9) compared to WT animals (42.4±6.0%, n=10). Furthermore, PPE treated SM-p110-/- mice were characterized by increased ECM degradation, decreased vascular remodelling, and less proliferating cells. Mechanistically, growth factor-dependent phosphorylation and inactivation of crucial phenotypic modulators of SMCs - FOXO-1, -3 and -4 - were shown to be significantly decreased in p110α-/- SMCs compared to WT cells. Specific inhibition of FOXO1 rescued the impaired proliferation of p110α-/- SMCs and enhanced the expression of elastin and calponin. Analysis of FOXO expression in humans revealed that FOXO1 was significantly upregulated in AAA patients compared to healthy individuals. Conclusion p110α deficiency in SMCs promotes the development and progression of AAA, as p110α controls crucial SMC functions which are important for structural processes in the aortic wall. Thereby, SMC proliferation is controlled by p110α dependent phosphorylation and inactivation of FOXO1. Since FOXO1 is upregulated in AAA patients, the p110α/AKT/FOXO1 signaling cascade may provide a key target for modulating aortic wall stability in AAA.

Abstract 13461: Early Vascular Aging is Associated With Early Cardiac Aging, Upregulation of Left Ventricle Pro-Fibrotic and Inflammatory Genes and Memory Decline in Aged Dahl Salt-Sensitive Rats

Introduction: Blood pressure (BP) begins to increase early in life in Dahl salt-sensitive rats (DSS) that consume a normal salt (NS) diet. Epidemiologic studies have shown that early vascular aging (EVA) is associated with central arterial stiffening (CAS) and hypertension, which develop earlier in life than expected in general population and contribute to cognitive impairment later in life. Hypotheses: (i) DSS rats will develop EVA earlier in life on NS diet, and with advancing age will exhibit early CV aging; (ii) In DSS, CV remodeling and CAS are implicated in memory decline. We tested these hypotheses using a longitudinal repeated measure design in DSS and their parental breed Sprague-Dawley rats (SD) at 3- & 12-mo of age. Methods: Male SD and DSS were kept on NS diet (0.5% NaCl; n=8/group) for entire experiment. Repeated measures of systolic BP (SBP), pulse wave velocity (PWV; an index of CAS), echocardiography, and non-repeated measures of Morris water maze (MWM) to test spatial memory, aortic collagen, elastin (histochemistry), and left ventricle (LV) mRNA expression (qPCR) were assessed at 3- & 12-mo. Data analyses: linear mixed-effect 2-way ANOVA, t-test & linear regression (LR) modeling. A 2-sided p<0.05 was considered significant. Results: SBP, PWV, aortic weight, wall thickness and collagen/elastin ratio were higher in DSS-3 vs. SD-3, and in DSS-12 vs. DSS-3. RWT and LV expression of pro-fibrotic, inflammatory and senescence genes were higher in DSS-12 vs. DSS-3 and vs. SD-12 (Table). DSS-12 spent more time to find a hidden platform in MWM vs. SD-12; their impaired spatial hippocampal memory was positively associated with PWV and LV mass by LR analysis. Conclusions: EVA, indexed as higher BP, PWV and aortic remodeling was associated with early CV aging, manifested by activation of LV senescence, inflammatory and pro-fibrotic genes and by higher rates of age-associated changes in BP, RWT and LV mass in DSS vs. SD. In DSS, early CV aging was associated with memory decline.

Preparation and Microscopic Mechanical Characterization of L-Methionine-Based Polyphosphazene Fibrous Mats for Vascular Tissue Engineering.

This study investigates the mechanical properties, degradation behavior, and biocompatibility of poly[(α-amino acid ester) phosphazene] electrospun fibers based on the ethyl ester of L-methionine (PαAPz-M), a material with potential applications in tissue engineering. We utilized atomic force microscopy (AFM) to evaluate the fiber mechanical characteristics and calculate its Young's modulus, revealing it to closely mimic the stiffness of a natural extracellular matrix (ECM). We also studied the degradation behavior of PαAPz-M scaffolds over 21 days, showing that they maintain the highly porous structure required for tissue engineering. Further evaluation of mesenchymal multipotent 10T1/2 cell and mesenchymal stem cell (MSC) behavior on the scaffolds demonstrated significant cell viability, proliferation, and successful MSC differentiation into smooth muscle cells. Expression of collagen and elastin by MSCs on the fiber mats highlighted potential ECM formation during scaffold degradation, confirming PαAPz-M as a promising material for vascular tissue engineering.

Early growth response 2, a novel target of pelvic organ prolapse, is highly expressed in anterior vaginal wall tissues with pelvic organ prolapse.

Pelvic organ prolapse (POP) is a common disorder among women that negatively affects women's quality of life. Early growth response 2 (EGR2) is a transcription factor that regulates cell growth. The present study aimed to explore the role of EGR2 in POP progression and provided a new target for the treatment and prevention of POP. Firstly, we extracted primary vaginal anterior wall fibroblasts from POP tissues and non-POP tissues and then constructed an EGR2-silencing lentivirus for further study. Immunoblotting, qPCR, TUNEL assay, CCK-8 assay, dual luciferase assay, and ELISA assay were carried out. EGR2 expression was much higher in POP tissues than in control tissues, and EGR2 expression positively correlated with cytokine signaling 3 (SOCS3) expression. Knockdown of EGR2 increased cell proliferation, upregulated PCNA expression, and reduced apoptosis in POP fibroblasts. Moreover, we found that the knockdown of EGR2 increased COL1A1, COL3A1, and Elastin expression and decreased MMP2 and MMP9 activities, and knockdown of EGR2 increased TGF-β/Smad pathway activity in POP fibroblasts. Interestingly, the results of dual luciferase assay demonstrated that EGR2 was able to increase SOCS3 transcriptional activity. EGR2 knockdown alleviated the apoptosis of POP fibroblasts by reducing SOCS3 expression and improving the proliferation and collagen synthesis of POP fibroblasts. Overall, our study illustrated that EGR2 was highly expressed in POP tissues, and knockdown of EGR2 alleviated apoptosis and reduced matrix degradation in POP fibroblasts. This study might provide a new insight into the pathogenesis of POP.

Resveratrol Inhibits Abdominal Aortic Aneurysm Progression by Reducing Extracellular Matrix Degradation, Apoptosis, Autophagy, and Inflammation of Vascular Smooth Muscle Cells via Upregulation of HMOX1.

This study aimed to explore the therapeutic effect of resveratrol (RES) against abdominal aortic aneurysm (AAA) and the role of HMOX1 underlying this effect. Vascular smooth muscle cells (VSMCs) were induced by angiotensin II (Ang II) to construct the microenvironment of AAA. HMOX1 expression was downregulated by the short hairpin ribonucleic acid (RNA) specific to HMOX1 in RES-pretreated VSMCs. The levels of matrix metalloproteinase (MMP)-2, MMP-9, and elastin were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot. Apoptosis rate was detected. The levels of apoptosis-related proteins (caspase-3 and Bax/Bcl-2), inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, and IL-1β), and autophagy-related proteins (Beclin 1, light chain 3 [LC3] II/I, and p62) were detected by western blot. The secretion of inflammatory factors in cell supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The number of autophagic vesicles in VSMCs was observed and analyzed by transmission electron microscopy. A rat model of pancreatic elastase-induced AAA was established to verify the effect and action mechanism of RES. Stimulation of Ang II increased the messenger RNA (mRNA) and protein levels of MMP-2 and MMP-9, decreased elastin expression, and enhanced apoptosis, secretion of inflammatory factors, and autophagy in VSMCs, whereas RES pretreatment ameliorated Ang II-induced VSMC dysfunction. In addition, HMOX1 mRNA and heme oxygenase-1 (HO-1) protein levels were significantly increased in VSMCs pretreated with RES compared with Ang II treatment alone. Silencing of HMOX1 abolished the effects of RES on VSMC dysfunction. Consistently, RES suppressed the development of AAA in rats by increasing the expression of HMOX1. Resveratrol protects against AAA by inhibiting extracellular matrix degradation, apoptosis, autophagy, and inflammation of VSMCs via HMOX1 upregulation. Our study found that angiotensin II (Ang II) stimulated increased the levels of MMP-2 and MMP-9 in vascular smooth muscle cells (VSMCs), decreased elastin expression, and promoted apoptosis, autophagy occurrence, and secretion of inflammatory factors, while resveratrol (RES) pretreatment improved this effect. In addition, downregulation of HMOX1 expression eliminated the effect of RES on the function of VSMCs. Our study elucidates that RES improves AAA progression through HMOX1 at both cellular and animal levels. This work can help doctors better understand the pathological mechanism of the occurrence and development of AAA, and provide a theoretical basis for clinicians to find better treatment options.

High-Intensity Focused Ultrasound Increases Collagen and Elastin Fiber Synthesis by Modulating Caveolin-1 in Aging Skin.

Caveolin-1 (Cav-1) induces cellular senescence by reducing extracellular signal-regulated kinase (ERK)1/2 phosphorylation and activating p53 via inhibition of mouse double minute 2 homolog (MDM2) and sirtuin 1 (Sirt1), promoting cell cycle arrest and decreasing fibroblast proliferation and collagen synthesis. High-intensity focused ultrasound (HIFU) treatment increases collagen synthesis, rejuvenating skin. Using H2O2-induced senescent fibroblasts and the skin of 12-month-old mice, we tested the hypothesis that HIFU increases collagen production through Cav-1 modulation. HIFU was administered at 0.3, 0.5, or 0.7 J in the LINEAR and DOT modes. In both models, HIFU administration decreased Cav-1 levels, increased ERK1/2 phosphorylation, and decreased the binding of Cav-1 with both MDM2 and Sirt1. HIFU administration decreased p53 activation (acetylated p53) and p21 levels and increased cyclin D1, cyclin-dependent kinase 2, and proliferating cell nuclear antigen levels in both models. HIFU treatment increased collagen and elastin expression, collagen fiber accumulation, and elastin fiber density in aging skin, with 0.5 J in LINEAR mode resulting in the most prominent effects. HIFU treatment increased collagen synthesis to levels similar to those in Cav-1-silenced senescent fibroblasts. Our results suggest that HIFU administration increases dermal collagen and elastin fibers in aging skin via Cav-1 modulation and reduced p53 activity.

Collagen Hydrolysate from the Scales of Mozambique Tilapia (Oreochromis mossambicus) Improve Hair and Skin Health by Alleviating Oxidative Stress and Inflammation and Promoting Hair Growth and Extracellular Matrix Factors.

Fish-derived collagen hydrolysate (CH) has shown promise in improving hair and skin health. Therefore, this study sought to comprehensively assess the effects of CH extracted from Mozambique tilapia (Oreochromis mossambicus) scales on hair and skin using in vitro and in vivo models. Human dermal papilla cells (hDPCs) were used for antioxidant and gene expression analyses, while C57BL/6 mice were orally administered CH for six weeks to assess hair growth patterns. The mice were divided into four groups: negative control (NC; distilled water), positive control (PC; 1 mg/kg finasteride), CH500 (500 mg/kg BW CH), and CH1000 (1000 mg/kg BW CH). CH mitigated catalase activity reduction in hDPCs, increased IGF-1 and VEGF levels, and decreased TGF-β1, TNF-α, and IL-1β expression. In vivo, CH treatment improved hair growth index, length, diameter, weight, and density. Scanning electron microscopy revealed reduced hair damage. Moreover, CH up-regulated IGF-1, VEGF, Elastin, and HAS2 mRNA expression while down-regulating TNF-α and IL-1β. CH enhanced hair shine, growth, and skin health while alleviating inflammation. These findings demonstrate the potential of CH in alleviating oxidative stress, promoting hair growth, and enhancing skin health, both in vitro and in vivo. Fish-derived CH offers a cost-effective and bioavailable option for improving hair and skin health.

Living, quiescent Lactobacillus plantarum Lp90 probiotic, delivered topically to full thickness tissues in vitro via a just-add-water cream delivery system, stimulates the expression of elastin protein.

Delivering living probiotics to the skin can be challenging as most water-containing cosmetic products require preservatives to maintain product stability. A recently introduced powdered technology [Stratabiosys™, Vantage Personal Care] allows for quiescent probiotic powders to be stored for extended periods of time. The powders can then be reconstituted to creams at the point of use by adding water and mixing and were examined in vitro on reconstructed human full thickness tissues to see if the probiotic had any influence of several important biomolecules expressed in the skin. A probiotic powder containing 200 M CFU/gram of living quiescent Lactobacillus plantarum Lp90 was reconstituted to a cream by adding ultrapure water and gently mixing the components at room temperature to quickly produce a cream. The resulting cream was tested topically on Epiderm® Full Thickness Tissues by treating the tissues for 24 h, removing the cream with a PBS rinse and then repeating the treatment for another 24 h. The resulting tissues were examined for four strategically important skin biomolecules including Type 1A collagen, elastin, filaggrin and hyaluronic acid. The probiotic-containing powder was tested against untreated tissues and powders not containing probiotics and powders containing measured amounts of one of two cryoprotectants known to be used to maintain the integrity of the quiescent probiotics during drying of the quiescent probiotic powders. It was found that topical treatment on Epiderm® tissues with creams containing 2 M (1%), 4 M (2%) and 6 M (3%) CFU/gram prepared from a base powder containing 200 M CFU/gram of Lactobacillus plantarum Lp90 stimulated elastin expression in a dose dependent fashion. There was no effect on the other biomolecules examined in the studies. In addition, it was found that creams made from powders containing only the known cryoprotectants, not bacteria, had no influence on elastin expression. The results of this study demonstrate that topical delivery of probiotics is possible from powders containing quiescent probiotic powders converted to creams just prior to application to the tissues. In the case of a powder containing Lactobacillus plantarum Lp90, topical application significantly increased expression of elastin in the skin replicants after 48 h of exposure to the cream made with the probiotic. The elastin-stimulating effects are not coming from the oligosaccharide cryoprotectants used to maintain the probiotic powders in their quiescent, dried state. The results indicate that it is the living Lactobacillus plantarum probiotic that is stimulating the elastin expression in the skin tissues.

Transcriptome Analysis of Human Vascular Smooth Muscle Cells Cultured on a Polyglycolic Acid Mesh Scaffold

To construct tissue-engineered blood vessels (TEBVs) in vitro, it is necessary to transfer seed cells to three-dimensional (3D) scaffolds for culture. However, what happens to the behavior of the cells after they are transferred to the scaffold is unclear. Therefore, in this study, a transcriptome analysis was used to characterize the differentially expressed genes (DEGs) of vascular smooth muscle cells (VSMCs) before and after transfer to 3D polyglycolic acid (PGA) scaffolds and to understand the changes in functional gene expression in the early stage of 3D culture. Transcriptome sequencing results showed that DEGs in the seed cells were mainly enriched in cell proliferation and cell-cell adhesion. The DEGs of cells grown in a 3D PGA scaffold (PGA-VSMCs) were mainly enriched in signal transduction. Furthermore, we found that ERK1/2 was significantly activated in PGA-VSMCs and inhibiting the phosphorylation level of ERK 1/2 in PGA-VSMCs significantly increased the expression of elastin. In conclusion, the PGA scaffold material altered gene expression in VSMCs and affected the elastin production. This study advances our understanding of biomaterial-cell interactions and provides valuable insights for improving the cultivation of TEBVs.

68Ga-labeled WVP peptide as a novel PET probe for molecular biological diagnosis of unstable thoracic aortic aneurysm and early dissection: an animal study.

Type IV collagen (Col-IV) is a prospective biomarker for diagnosing and treating of unstable thoracic aortic aneurysm and dissection (TAAD). This study aims to evaluate the feasibility of 68Ga-labeled WVP peptide (68Ga-DOTA-WVP) as a novel Col-IV-targeted probe for TAAD biological diagnosis using PET/CT. WVP peptide was modified with bifunctional chelator DOTA for 68Ga radiolabeling. Immunohistochemical staining was used to evaluate the expression and location of Col-IV and elastin in aortas treated with 3-aminopropionitrile fumarate (BAPN) at different time points (0, 2, and 4 weeks). The imaging performance of 68Ga-DOTA-WVP was investigated using Micro-PET/CT in a BAPN-induced TAAD mouse model. The relationship between 68Ga-DOTA-WVP uptake in aortic lesions and the serum levels of TAAD-related biomarkers including D-dimer, C-reactive protein (CRP), and serum soluble suppression of tumorigenicity-2 (sST2) was also analyzed. 68Ga-DOTA-WVP was readily prepared with high radiochemical purity and stability in vitro. 68Ga-DOTA-WVP Micro-PET/CT could detect Col-IV exposure of unstable aneurysms and early dissection in BAPN-induced TAAD mice, but little 68Ga-DOTA-WVP uptake was shown in the control group at each imaging time point. The differences of Col-IV expression and distribution of 68Ga-DOTA-WVP both in TAAD and control groups further verified the imaging efficiency of 68Ga-DOTA-WVP PET/CT. Additionally, a higher sST2 level was found in the imaging positive (n = 14) than the negative (n = 8) group (9.60 ± 1.14 vs. 8.44 ± 0.52, P = 0.014). 68Ga-DOTA-WVP could trace the exposure and abnormal deposition of Col-IV in enlarged and early injured aortas, showing a potential for biological diagnosis, whole-body screening, and progression monitoring of TAAD.

The Relationship of Polymorphism of LYSYL Oxidase Like-1 Gene and Pelvic Organ Prolapse in Balinese Women

Background: Pelvic organ prolapse (POP) data from Sanglah General Hospital Bali revealed proportion of 91.2% Balinese women, and that proportion was not decreased even with modification of workload and contraception. One of the associated factor is the decrease of expression of elastin and proteins involved in elastin metabolism, such as lysyl oxidase-like 1 (LOXL-1). There is genetic factor in the pathogenesis of prolapse, and polymorphism of elastogenesis-related gene, such as LOXL1 gene, associated with elastinopathy. Methods: We conducted an observational case control study in Balinese women population. Balinese women are women who have lineage of at least 3 generations (from grandparents) from Balinese people. Thirty patients with prolapse were taken as cases and 30 patients non prolapse as controls. DNA was isolated from venous blood, and then LOXL1 gene amplificated by polymerase chain reaction and then electrophoresis, and finally the DNA products were sequenced. Data was analyzed wih SPSS 21.0 for Windows. To determine whether the LOXL1 gene polymorphism is a risk factor for POP, a Chi-Square test was performed, and then variables were controlled by multivariate analysis using conditional logistic regression tests. Results: From the distribution of polymorphism, there were 13 samples in the case group (43.34% of the case group), compared to 5 samples in the control group (16.67% of the control group), and for the distribution of non-polymorphism, there were 17 samples in the case group (56.66% of the case group), compared to 25 samples in the control group (83.33% of the control group). From the analysis of polymorphism of LOXL1 gene rs2304719 in Balinese women, we found OR 3.824, CI 95% 1.150-12.713, p = 0.024). The LOXL1 gene polymorphism remains a risk factor for prolapse occurrence after the control variables were controlled by multivariate analysis using conditional logistic regression tests. The LOXL1 gene polymorphism, after the four control variables were controlled, gave a risk for prolapse incidence of 3,791 times by multivariate analysis (adjusted OR = 3.791; 95% CI = 1.065-13.496; p = 0.04). Conclusion: Polymorphism of LOXL1 gene rs2304719 is a risk factor for prolapse in Balinese women. Study for other gene polymorphisms are needed, before we can make a multigene panel testing for risk assessment. We hope in the future, multigene panel testing will be simpler, easier and cheaper, so that women at risk can be identified, and then can be informed and advised to avoid or reduce risk factors for prolapse. Keywords: DNA sequencing, elastinogenesis, elastinopathy, fibroblast, genetic factor of prolapse, multigene panel testing, polymerase chain reaction, uterosacral ligament

Differential expression of extracellular matrix proteins in the lesional skin of vitiligo patients.

Skin pigmentation is regulated by intricate interaction of the dermis and epidermis. The extracellular components present in the dermis play a very important role in the maintenance of skin homeostasis. Therefore, our objective was to check the expression of various ECM components secreted by the dermal fibroblasts in the lesional skin and non-lesional skin of vitiligo patients. For this study, skin punch biopsies (4mm) were collected from lesional skin (n = 12), non-lesional skin (n = 6) of non-segmental vitiligo patient's (NSV) and healthy control skin (n = 10). Masson's trichrome staining was performed to check the collagen fibre. The expression of collagen type 1, IV, elastin, fibronectin, E-cadherin and integrin β1 was checked by real-time PCR and immunohistochemistry. In this study, we demonstrated an increased expression of collagen type 1 in the lesional skin of vitiligo patients. The expression of collagen type IV, fibronectin, elastin and adhesion components such as E-cadherin and integrin β1 was observed to be significantly decreased in the lesional skin of NSV patients as compared to healthy control, whereas insignificant difference was observed between non-lesional and control skin. Increased expression of collagen type 1 in the lesional skin of vitiligo patients might be inhibiting the migration of melanocytes, whereas the decreased expression of elastin, collagen type IV, fibronectin, E-cadherins and integrins in the lesional skin may inhibit adhesion, migration, growth and differentiation of cells.

PD11-05 EFFECTS OF MESENCHYMAL STEM CELL (MSC) THERAPY ON ISCHEMIC PRIAPISM: AN EXPERIMENTAL STUDY

You have accessJournal of UrologyCME1 Apr 2023PD11-05 EFFECTS OF MESENCHYMAL STEM CELL (MSC) THERAPY ON ISCHEMIC PRIAPISM: AN EXPERIMENTAL STUDY Enes Kilic, Aykut Colakerol, Romer Yazar, Sule Ozsoy, Yasar Basaga, Mustafa Zafer Temiz, Ates Kadioglu, Ahmet Muslumanoglu, and Ismail Kandirali Enes KilicEnes Kilic More articles by this author , Aykut ColakerolAykut Colakerol More articles by this author , Romer YazarRomer Yazar More articles by this author , Sule OzsoySule Ozsoy More articles by this author , Yasar BasagaYasar Basaga More articles by this author , Mustafa Zafer TemizMustafa Zafer Temiz More articles by this author , Ates KadiogluAtes Kadioglu More articles by this author , Ahmet MuslumanogluAhmet Muslumanoglu More articles by this author , and Ismail KandiraliIsmail Kandirali More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003251.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Priapism is an erection of the penis that lasts longer than four hours, independent of sexual desire and arousal. Erectile dysfunction develops in 90% of cases of ischemic priapism lasting longer than 24 hours, secondary to fibrosis. The demonstration that MSC prevents fibrosis in different tissues has brought to mind the use of stem cells in diseases with fibrosis in the penile tissue. In this study, it was aimed to show the effect of intracavernosal mesenchymal stem cell therapy (IMSCT) on functional and histological recovery in ischemic priapism model. METHODS: A total of 30 Wistar Albino male rats were included in the study. Rats were divided into three experimental groups as sham, ischemic priapism model, and ischemic priapism + IMSCT. The latter groups were divided into additional subgroups as one-hour and 12 hours ischemic priapism models were applied to animals. Electrophysiologic parameters such as intracavernosal and systemic arterial pressures, and histopathological parameters including smooth muscle to collagen ratio, tissue fibrosis, elastin expression, and tissue e-NOS, n-NOS, i-NOS expressions were statistically compared in main groups and subgroups (Tables 1 and 2). Molecular parameters such as TGF, VEGF, NGF, and BDNF were also compared. RESULTS: In the initial analysis, significant deterioration was detected in most of the parameters in the ischemic priapism model (Table 1). These findings proved our model as an effective procedure. When comparing the model and the model + IMSCT groups, most of the parameters exhibited significant enhancements. (Table 1). During subgroup analyses, we demonstrated significant therapeutic effects of IMSCT, particularly at the in the 1-h model of priapism. CONCLUSIONS: Our study revealed the potential therapeutic activity of IMSCT on the ischemic priapism model, especially in the prolonged ischemic period. Further studies are needed to implicate the IMSCT in clinical practice. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e333 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Enes Kilic More articles by this author Aykut Colakerol More articles by this author Romer Yazar More articles by this author Sule Ozsoy More articles by this author Yasar Basaga More articles by this author Mustafa Zafer Temiz More articles by this author Ates Kadioglu More articles by this author Ahmet Muslumanoglu More articles by this author Ismail Kandirali More articles by this author Expand All Advertisement PDF downloadLoading ...

The Secretome of Irradiated Peripheral Mononuclear Cells Attenuates Hypertrophic Skin Scarring.

Hypertrophic scars can cause pain, movement restrictions, and reduction in the quality of life. Despite numerous options to treat hypertrophic scarring, efficient therapies are still scarce, and cellular mechanisms are not well understood. Factors secreted by peripheral blood mononuclear cells (PBMCsec) have been previously described for their beneficial effects on tissue regeneration. In this study, we investigated the effects of PBMCsec on skin scarring in mouse models and human scar explant cultures at single-cell resolution (scRNAseq). Mouse wounds and scars, and human mature scars were treated with PBMCsec intradermally and topically. The topical and intradermal application of PBMCsec regulated the expression of various genes involved in pro-fibrotic processes and tissue remodeling. We identified elastin as a common linchpin of anti-fibrotic action in both mouse and human scars. In vitro, we found that PBMCsec prevents TGFβ-mediated myofibroblast differentiation and attenuates abundant elastin expression with non-canonical signaling inhibition. Furthermore, the TGFβ-induced breakdown of elastic fibers was strongly inhibited by the addition of PBMCsec. In conclusion, we conducted an extensive study with multiple experimental approaches and ample scRNAseq data demonstrating the anti-fibrotic effect of PBMCsec on cutaneous scars in mouse and human experimental settings. These findings point at PBMCsec as a novel therapeutic option to treat skin scarring.

Formulation Containing Phytosomes of Carotenoids from Nyctanthes arbor-tristis and Tagetes patula Protect D-galactose Induced Skin Aging in Mice

• Gel containing phytosomes of crocin rich extract(equivalent to crocin 1% w/w) obtained from Nyctanthes arbor-tristis and Lutein rich extract (equivalent to lutein 0.5%w/w) from Tagetes patula indicated • Entrapment of crocin and lutein was found to be (60.20% w/w) and (50.81% w/w) respectively. • Improved stability of crocin and lutein through preparation of Phytosomes • Significant (P<0.01) increase in thickness of dermal and epidermal layer • Significant increase (P<0.05) in glutathione level and decrease (P<0.01) in MDA level • Upregulation of expression of elastin and Collagen gene by 0.26 and 0.03 fold respectively : Carotenoids play important role in delay of aging process. Orange coloured tubular calyx of flowers of Nyctanthes arbor-tristis. contains crocin, an apocarotenoid which forms a major component of stigma of saffron. Due to presence of crocin in orange coloured tubular calyx of Nyctanthes arbor-tristis, it can be used as an economical substitute to saffron for its medicinal and cosmetic utilities. Lutein from flower petals of Tagetes patula L , is another popular carotenoid which has antioxidant effect and many health benefits. The carotenoids are highly unstable when exposed to atmosphere, hence in the present study crocin and lutein rich extracts were entrapped into phytosomes to improve stability and efficacy of topical preparation. : Oxidative stress is considered to be a major contributor to the process of aging. Carotenoids viz, Lutein and crocin, being powerful antioxidants, can delay aging of skin through upregulating col type I gene and elastin gene. The carotenoids have very low oxidation potential, due to which they get oxidized fast and protect oxidation of other compounds which make them highly unstable. One of the ways to improve stability of these phytoconstituents, is their entrapment in phytosomes. Preparation of phytosomes will have dual advantages of improving stability as well as bioavailability of molecule. : The phytosomes of Carotenoid rich extract of tubular calyx of Nyctanthes arbor-tristis L. and the petals of Tagetes patula L. (standardized for crocin and lutein content) were prepared using lipid film hydration technique and these phytosomes were then incorporated into gel base. The gel formulation was evaluated for stability as per ICH guidelines. Efficacy of formulation was evaluated by D-galactose induced aging model. Aging in skin was induced by administration of D-Galactose (100mg/kg b.w.s.c.) to albino mice for 42 days. The gel formulation was applied topically for 42 days. Then the effect of formulation on skin aging was evaluated by estimation of biochemical parameters viz. glutathione and malondialdehyde (MDA) and histopathological studies of treated skin samples. Also expression of COL type I and elastin genes was carried out from the skin samples by RT-PCR method. : Percent entrapment (%w/w) of crocin and lutein in phytosomes were found to be 60.20 and 50.81 respectively. Accelerated stability studies showed improvement in stability of carotenoids viz. crocin and lutein and the content of crocin and lutein in formulation was found in the range of 99.98 % w/w to 99.85%w/w at the end of three months. The formulation containing extract of Phytosomes of carotenoid rich extracts of Nyctanthes arbor-tristis L. and the petals of Tagetes patula L. exhibited potent antiaging activity through significant (p<0.05)increase in dermal and epidermal layers, and increase in GSH levels of skin as compared to the untreated group. The treatment with the gel formulation revealed upregulation of col type I and elastin genes. There was significant reduction in lipid peroxidation as revealed through reduction in MDA levels as compared to untreated group. : Crocin and lutein have potential to prevent skin aging via upregulation of COL type I gene and elastin gene. Upregulation of genes resulted into increase in the thickness of epidermal and dermal layer along with reduction in oxidative stress in skin. Entrapment of carotenoid rich extracts of Nyctanthes arbor-tristis and Tagetes patula in Phytosomes enhanced the stability and efficacy of the formulation.

Extracellular Vesicles Derived from Auricular Chondrocytes Facilitate Cartilage Differentiation of Adipose-Derived Mesenchymal Stem Cells.

Adipose-derived mesenchymal stem cell (ADSC)-based therapies have been utilized for cartilage regeneration because of their multi-lineage differentiation ability. However, commonly used cartilage inducers such as the transforming growth factor beta-3 (TGF-β3) may be prone to cartilage dedifferentiation and hypertrophy. The directional differentiation of elastic cartilage is limited nowadays. Extracellular vesicles (EVs) have been reported to influence the specific differentiation of mesenchymal stem cells (MSCs) by reflecting the composition of the parental cells. However, the role of auricular chondrogenic-derived EVs (AC-EVs) in elastic chondrogenic differentiation of ADSCs has not yet been reported. AC-EVs isolated from the external ears of swine exhibited a positive effect on cell proliferation and migration. Furthermore, AC-EVs efficiently promoted chondrogenic differentiation of ADSCs in pellet culture, as shown by the elevated levels of COL2A1, ACAN, and SOX-9 expression. Moreover, there was a significantly higher expression of elastin and a lower expression of the fibrotic marker COL1A1 in comparison with that achieved with TGF-β3. The staining results demonstrated that AC-EVs promoted the deposition of cartilage-specific matrix, which is in good concordance with the real-time polymerase chain reaction (RT-PCR) results. Auricular chondrogenic-derived EVs are a crucial component in elastic chondrogenic differentiation and other biological behaviors of ADSCs, which may be a useful ingredient for cartilage tissue engineering and external ear reconstruction. This journal requires that authors 42 assign a level of evidence to each submission to which 43 Evidence-Based Medicine rankings are applicable. This 44 excludes Review Articles, Book Reviews, and manuscripts 45 that concern Basic Science, Animal Studies, Cadaver 46 Studies, and Experimental Studies. For a full description of 47 these Evidence-Based Medicine ratings, please refer to the 48 Table oôf Contents or the online Instructions to Authors 49 www.springer.com/00266 .

Establishment and characterization of immortalized human vocal fold fibroblast cell lines.

Vocal fold scarring is abnormal scar tissue in the lamina propria layer of the vocal fold. To facilitate investigation of vocal fold scarring, we established and characterized immortalized human vocal fold fibroblast (iHVFF) cell lines. Human vocal fold fibroblasts were immortalized by introducing Simian virus 40 large T antigen (SV40TAg) by transfection. Successfully transfected fibroblasts were sorted using flow cytometry. Immunofluorescence cytochemistry and western blot were applied to analyze the expression of fibronectin, vimentin, alpha-smooth muscle actin (α-SMA) and fibroblast activation protein (FAP). Cell proliferation rate was measured by CCK-8 assay. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to analyze the mRNA expression level. The iHVFFs continued to proliferate for more than 30 generations and appeared spindle-shaped. The expression of Vimentin and α-SMA were detected in both iHVFFs and primary fibroblasts, and enhanced expression of FAP was observed in iHVFFs. Furthermore, iHVFFs exhibited an increased proliferative capability compared with the primary fibroblasts. RT-qPCR results suggested that collagen type III alpha 1 chain (COL3A1), interleukin-6, cyclooxygenase 2 (COX2), hyaluronan synthase 2 (HAS2), hepatocyte growth factor (HGF) in the iHVFFs significantly increased, whereas transforming growth factor-β1 (TGF-β1), elastin and matrix metallopeptidase-1 (MMP-1) expression significantly downregulated. No differences in mRNA expression of α-SMA, fibronectin and collagen type I alpha 2 chain (COL1A2) were noted between iHVFFs and primary fibroblasts. iHVFFs can be used as a novel tool cell for future researches on the mechanisms of pathogenesis and treatment of vocal fold scarring.

Biological activity of leaves of three Morinda species detected by stimulation and suppression of gene expression of collagen, elastin, melanin and other related genes

Biological activity of leaves of three Morinda species detected by stimulation and suppression of gene expression of collagen, elastin, melanin and other related genes