Amnion Membrane Research Articles

Functionally graded membrane: A novel approach in the treatment of gingival recession defects.

Background:Guided tissue regeneration has recently been advocated in re-constructing soft-tissue dimensions in recession defects. Advancement in nanotechnology has led to increased zest for approaches such as electrospinning of biologically active; nanofibrous functionally graded regenerative membranes for periodontal tissue engineering. A functionally graded membrane (FGM) had been tailored by incorporating chitosan and nano-hydroxyapatite over Amnion membrane and used in gingival recession defects.Study Design:It was single-blind, randomized controlled study. Split-mouth study was conducted in nine patients and 22 sites with recession defects were selected. Sites were divided into Group A (Amnion membrane with coronal advanced flap) and Group B (FGM with coronal advanced flap).Materials and Methods:Sites were assessed clinically by recording plaque index (PI), gingival index (GI), vertical recession defect depth (VRDD), relative clinical attachment level (CAL), and width of keratinized tissue at baseline, 3–6 months; and radiographically by recording linear bone growth by dentascan at baseline and 6 months.Result:Both groups showed statistically significant reduction in PI, GI and VRDD, and CAL and nonsignificant reduction in width of keratinized tissue at 3 and 6 months postoperatively. Group A showed statistically significant linear bone growth at 6 months. Group B also showed gain in linear bone growth at 6 months; however, result was statistically nonsignificant.Conclusion:FGM had shown favorable results by enhancing bone growth while preventing the gingival tissue downgrowth.

A Combination of Coconut Fiber Suture and Tamarind Seed Gel with Dehydrated Human Amnion Membrane for Wound Surgery in Rats

Today, there are over 2,000 different biomaterials used for various medical applications, but none of these biomaterials are 100% compatible with all human beings. Coconut fiber is widely available but has not been tested as a safe natural alternative for sutures. Immature coconut fiber is nonabsorbable and is effective for cuts and open wounds when used in combination with dehydrated human amnion membrane (dHAM). Immature coconut fiber, tamarind seed polysaccharide (TSP), and dHAM were prepared to test their combinational effect on wound healing in rats. TSP enhanced cell viability, proliferation, and migration in human skin cells and cured wounds both individually and in combination with dHAM. An antibiotic‐free combination of the human amniotic membrane with intact epithelium, tamarind seed polysaccharide, and immature coconut fiber provided faster wound healing. Significantly higher wound healing was seen on the 11th day based on an initial 10 mm biopsy punch surgery in Wistar rats compared to control groups. Histological studies revealed thickened dermis edges with more neutrophil infiltration. Collagen deposition in the dermis was homogeneous across the excised skin tissue in the test group, again attesting to the utility of this procedure. This research signifies the use of TSP gel together with the amnion membrane representing a “smart patch” with wound healing potential, which would encourage further research on the smart patch made using a combination of plant and animal biological materials.

Socket preservation and reconstruction: A case report with follow up of 9 months

Progressive bone resorption after extraction of tooth may lead to prosthetic-driven and surgical difficulties when a dental implant has to be placed after extraction of tooth. This case report discusses an irreparable upper right first premolar with a dehiscence on buccal wall. A conservative tooth extraction was performed and the dehiscence was exposed after reflecting a full thickness mucoperiosteal flap. The socket and the dehiscence were then grafted using Nova bone putty and amnion membrane. After a healing period of 3 months and 9 months a CBCT analysis was done to check for regeneration in the grafted site. It can be concluded that grafting technique can lessen the alveolar bone resorption in the socket with dehiscence.

Anterior Segment Surgery Performed During the COVID-19 Pandemic.

Objectives:This study evaluated anterior segment surgeries performed during the coronavirus 2019 (COVID-19) pandemic. Prevention of virus transmission is a critical consideration for surgeons, and includes assessment of etiology, the referral region, demographic characteristics, and the surgery to be performed.Methods:The data of 144 patients who underwent anterior segment surgery between March 19, 2020 and June 1, 2020 were retrospectively reviewed. The patient demographic data and details of ophthalmological examination findings, the region patients were referred from, and the type of surgery performed were recorded and analyzed.Results:A total of 144 patients, 49 women (34%) and 95 men (66%), were included in this study. The mean age of the patients was 31.30±25.88 years (range: 1–86 years). The presenting complaint was in the right eye in 43.7% of the cases, in the left eye in 52.8%, and in both eyes in 3.5% of the cases. While 94.4% of the applications were from Istanbul, the remaining 5.6% were from outside the province. Though 43.7% of the cases were patients seen previously at the study hospital in Istanbul, 56.3% presented for the first time. This hospital was the first referral center in only 39.6% of the cases. Evaluation of etiology indicated that corneal perforation (18.1%) was the most common, followed by keratitis (13.2%). The most common surgical intervention applied was amnion membrane transplantation (19.4%), followed by perforation repair (16.7%).Conclusion:Ophthalmological surgeries continue to be performed during the ongoing COVID-19 pandemic, however, special algorithms must be used to reduce the risk of COVID-19 transmission and to ensure continuity of healthcare for ophthalmology patients.

Extracellular vesicle mediated feto-maternal HMGB1 signaling induces preterm birth.

Preterm birth (PTB; <37 weeks of gestation) impacts ∼11% of all pregnancies and contributes to 1 million neonatal deaths worldwide annually. An understanding of the feto-maternal (F-M) signals that initiate birthing (parturition) at term is critical to design strategies to prevent their premature activation, resulting in PTB. Although endocrine and immune cell signaling are well-reported, fetal-derived paracrine signals capable of transitioning quiescent uterus to an active state of labor are poorly studied. Recent reports have suggested that senescence of the fetal amnion membrane coinciding with fetal growth and maturation generates inflammatory signals capable of triggering parturition. This is by increasing the inflammatory load at the feto-maternal interface (FMi) tissues (i.e., amniochorion-decidua). High mobility group box 1 protein (HMGB1), an alarmin, is one of the inflammatory signals released by senescent amnion cells via extracellular vesicles (exosomes; 40-160 nm). Increased levels of HMGB1 in the amniotic fluid, cord and maternal blood are associated with term and PTB. This study tested the hypothesis that senescent amnion cells release HMGB1, which is fetal signaling capable of increasing FMi inflammation, predisposing them to parturition. To test this hypothesis, exosomes from amnion epithelial cells (AECs) grown under normal conditions were engineered to contain HMGB1 by electroporation (eHMGB1). eHMGB1 was characterized (quantity, size, shape, markers and loading efficiency), and its propagation through FMi was tested using a four-chamber microfluidic organ-on-a-chip device (FMi-OOC) that contained four distinct cell types (amnion and chorion mesenchymal, chorion trophoblast and decidual cells) connected through microchannels. eHMGB1 propagated through the fetal cells and matrix to the maternal decidua and increased inflammation (receptor expression [RAGE and TLR4] and cytokines). Furthermore, intra-amniotic injection of eHMGB1 (containing 10 ng) into pregnant CD-1 mice on embryonic day 17 led to PTB. Injecting carboxyfluorescein succinimidyl ester (CFSE)-labeled eHMGB1, we determined in vivo kinetics and report that eHMGB1 trafficking resulting in PTB was associated with increased FMi inflammation. This study determined that fetal exosome mediated paracrine signaling can generate inflammation and induce parturition. Besides, in vivo functional validation of FMi-OOC experiments strengthens the reliability of such devices to test physiologic and pathologic systems.

Chorion and amnion/chorion membranes in oral and periodontal surgery: A systematic review.

The aim of this study was to perform a systematic review on the clinical applications where chorion membrane (CM) and amnion/chorion membrane (ACM) were used for oral tissue regeneration procedures. Selection of articles was carried out by two evaluators in Pubmed and Scopus databases, and Outcomes (PICO) method was used to select the relevant articles. Clinical studies reporting the use of CM or ACM for oral soft and hard tissue regeneration were included. The research involved 21 studies conducted on 375 human patients. Seven clinical applications of CM and ACM in oral and periodontal surgery were identified: gingival recession treatment, intrabony and furcation defect treatment, alveolar ridge preservation, keratinized gum width augmentation around dental implants, maxillary sinus membrane repair, and large bone defect reconstruction. CM and ACM were compared to negative controls (conventional surgeries without membrane) or to the following materials: collagen membranes, dense polytetrafluoroethylene membranes, platelet-rich fibrin membranes, amnion membranes, and to a bone substitute. Several studies support the use of CM and ACM as an efficient alternative to current techniques for periodontal and oral soft tissue regeneration procedures. However, further studies are necessary to increase the level of evidence and especially to demonstrate their role for bone regeneration.

A comparison of the in vitro resorption rate of human amnion membrane and commercially available bovine collagen membrane for potential periodontal applications

A comparison of the in vitro resorption rate of human amnion membrane and commercially available bovine collagen membrane for potential periodontal applications

Can the corneal tissue be used as a wound dressing material for skin injuries?

Currently, the application field of corneal tissue is corneal transplantation from animal to human or from human to human. Although the cornea and skin emerge from the same embriyological layer, ectoderm, and the cornea has a very high capacity, no research has been conducted to reveal the effect of the cornea in skin wound healing. There is no commercial product available in the hospital pharmacy that contains a ready-to-use, hygienic, wound dressing material manufactured from cornea for both ocular or extraocular wounds. In this article, the feasibility of corneal tissue as a candidate for disposable wound dressing material for skin injuries are examined and compared with the closest alternative, amnion membrane.

An Evaluation of Wound Healing Efficacy of a Film Dressing Made from Polymer-integrated Amnion Membrane

ABSTRACT A film dressing is an easy and common wound management, which is flexible to cover many types of superficial injuries. In a recent study, we developed a scaffold from poly (1,8-octanediolco-citrate) incorporated decellularized amnion membrane (DAM-POC). The DAM-POC scaffold was biocompatible and could enhance soft and hard tissue regeneration when applied to repair the cleft palate in rat. The efficacy of the DAM-POC scaffold in oral repair had led us to hypothesize that it could be employed extensively in the medical field as a wound dressing. This study aimed to investigate the feasibility and efficacy of the DAM-POC scaffold as a film dressing in accelerating wound healing when applied in multiple tissue injuries. Our results demonstrated that both the DAM and DAM-POC scaffolds were biocompatible and anti-adhesive without causing severe foreign body reactions when covering wounds in abdominal wall, back muscle, tibia bone, and liver. In addition, the DAM-POC scaffold was superior to the DAM scaffold in reducing inflammation, preventing fibrosis, and regenerating tissues. In conclusion, the DAM-POC scaffold might potentially be adopted as a film dressing in a wide range of therapeutic applications and healing situations to protect the damaged tissues from the external environment and prevent infections.

Stretch, scratch, and stress: Suppressors and supporters of senescence in human fetal membranes

IntroductionThroughout gestation, amnion membranes undergo mechanical and or physiological stretch, scratch, or stress which is withstood by repairing and remodeling processes to protect the growing fetus. At term, increased oxidative stress (OS) activates p38MAPK, induces senescence, and inflammation contributing to membrane dysfunction to promote labor. However, the signaling initiated by stretch and scratch is still unclear. This study compares the induction of p38MAPK mediated senescence by stretch, scratch, and stress in human amnion epithelial cells (AECs). MethodsPrimary AECs from term, not-in-labor, fetal membranes were cultured using the following conditions (N = 3); 1) CellFlex chambers with or without 20% biaxial stretch, 2) 8-well coverslips with or without scratch, and 3) cells exposed to cigarette smoke extract (CSE) inducing OS. p38MAPK (Western blot or immunocytochemistry), senescence activation, and inflammation (matrix metalloproteinases 9 [MMP9] activity-ELISA) were determined in cells exposed to various conditions. T-test and One-Way ANOVA was used to assess significance. ResultsBiological membrane extension, mimicked by 20% biaxial stretch of AEC, maintained an epithelial morphology and activated P-p38MAPK (P = 0.02) compared to the non-stretch controls, but did not induce senescence or MMP9 activation. AEC scratches were healed within 40-hrs, which included proliferation, migration, and cellular transitions aided by p38MAPK activation but not senescence. CSE induced OS increased p38MAPK (P = 0.018) activation, senescence (P = 0.019), and MMP9 (P = 0.02). ConclusionPhysiologic stretch and scratch experienced during gestation can cause p38MAPK activation without causing senescence or inflammation. This may be indicative of p38MAPK's role in tissue remodeling during pregnancy. Overwhelming OS, experienced at term, results in P-p38MAPK mediated senescence and inflammation to disrupt membrane remodeling.

Interleukin (IL)-6: A Friend or Foe of Pregnancy and Parturition? Evidence From Functional Studies in Fetal Membrane Cells.

ObjectiveProtection of the fetus within the amniotic sac is primarily attained by remodeling fetal membrane (amniochorion) cells through cyclic epithelial to mesenchymal and mesenchymal to epithelial (EMT and MET) transitions. Endocrine and paracrine factors regulate EMT and MET during pregnancy. At term, increased oxidative stress forces a terminal state of EMT and inflammation, predisposing to membrane weakening and rupture. IL-6 is a constitutively expressed cytokine during gestation, but it is elevated in term and preterm births. Therefore, we tested the hypothesis that IL-6 can determine the fate of amnion membrane cells and that pathologic levels of IL-6 can cause a terminal state of EMT and inflammation, leading to adverse pregnancy outcomes.MethodsPrimary amnion epithelial cells (AECs) were treated with recombinant IL-6 (330, 1,650, 3,330, and 16,000 pg/ml) for 48 h (N = 5). IL-6-induced cell senescence (aging), cell death (apoptosis and necrosis), and cell cycle changes were studied using flow cytometry. Cellular transitions were determined by immunocytochemistry and western blot analysis, while IL-6 signaling (activation of signaling kinases) was measured by immunoassay. Inflammatory marker matrix metalloproteinase (MMP9) and granulocyte-macrophage colony-stimulating factor (GM-CSF) concentrations were measured using a Fluorokine E assay and ELISA, respectively. Amniotic membranes collected on gestational day (D) 12 and D18 from IL-6 knockout (KO) and control C57BL/6 mice (N = 3 each) were used to determine the impact of IL-6 on cell transitions. Fold changes were measured based on the mean of each group.ResultsIL-6 treatment of AECs at physiologic or pathologic doses increased JNK and p38MAPK activation; however, the activation of signals did not cause changes in AEC cell cycle, cellular senescence, apoptosis, necrosis, cellular transitions, or inflammation (MMP9 and GM-CSF) compared to control. EMT markers were higher on D18 compared to D12 regardless of IL-6 status in the mouse amniotic sac.ConclusionPhysiologic and pathologic concentrations of IL-6 did not cause amnion cell aging, cell death, cellular transitions, or inflammation. IL-6 may function to maintain cellular homeostasis throughout gestation in fetal membrane cells. Although IL-6 is a good biomarker for adverse pregnancies, it is not an indicator of an underlying pathological mechanism in membrane cells.

Effects of Cryogenic Storage on Human Amnion Epithelial Cells.

Perinatal stem cells and epithelial cells isolated from full term amnion membrane, in particular, have attracted interest over the last decade, as a promising source of multipotent cells for cellular therapies. Human amnion epithelial cells (hAEC) have been used to treat monogenetic liver disease such as maple syrup urine disease or fibrosis of the liver in preclinical studies. In most studies xeno-transplants of hAEC were conducted without providing immunosuppression to recipients, reflecting the tolerogenic properties of hAEC. For many cell types, successful cryopreservation is critical for providing a readily available, off-the-shelf product. In this study, hAEC were isolated from full-term human placenta from 14 different donors, cryopreserved using a protocol and reagents commonly adopted for epithelial cell preservation. The cells were analyzed in terms of survival, recovery, and homogeneity, profiled for surface markers characteristic of epithelial, mesenchymal, endothelial, or hematopoietic cells. There were no significant differences observed in the percentage of cells with epithelial cell markers before and after cryopreservation. The relative proportion of stromal and hematopoietic cells was significantly reduced in hAEC preparations after cryopreservation. The expression of stem cell and immunomodulatory molecules were confirmed in the final product. Since multipotent cells are readily available from full-term placenta, this novel cell source might significantly increase the number of patients eligible to receive cellular therapies for liver and other diseases.

Long - Term Effects of Topical Amniotic Membrane Stem Cell Metabolite Product (AMSC-MP) and Fractional CO2 Laser in Photoaging

Background: Photoaging causes wrinkles, lentigines, keratoses, dyspigmentation, telangiectasia, decreased elasticity, rough texture, and pale color. Facial wrinkles and mottled skin can negatively affect perceived attractiveness, self-esteem, personality, and level of acceptance. Stem cells are important cells in the skin, and they are the source for continuous regeneration of the epidermis. It can be extracted from many sources, including amnion. Amnion membrane stem cell (AMSC) can be cultured and secretes metabolite product, and it can rejuvenate aging skin. Laser-Assisted Drug Delivery (LADD) involves selective destruction of the layers of the epidermis and dermis to allow penetration and absorption of topical drugs as well as large molecular weight drugs such as stem cells. Purpose: To evaluate the long-term effects of topical administration of AMSC metabolite products (AMSC -MP) and fractional CO2 laser on the clinical improvement of photoaging compared to previous skin conditions. Methods: This was an observational study which was aimed to evaluate the long-term effects of clinical improvement after topical therapy of AMSC-MP and fractional CO2 laser in patients with photoaging. Results: The results of the analysis showed a decrease in wrinkles, dark spots, skin tone, and pores. Conclusion: Based on Janus-II® assessment, there was improvement degrees of wrinkles, dark spots, and pores in patients with photoaging who received fractional CO2laser fractional for 12 weeks. There were no side effects found in this study.

Effects of mesenchymal stem cell and amnion membrane transfer on prevention of pericardial adhesions

Abstract Background To investigate and compare the antiadhesive/antifibrotic effects of mesenchymal stem cells (MSC) and amnion membrane transfer (AMT) in a rat model. Material and methods Three experimental and sham groups were formed using 30 Wistar-Albino rats. AMT and MSC were applied to the related groups. The control group was not treated. After 12 weeks follow-up, intracardiac blood and cardiac-pericardiac tissue samples were taken. The severity of adhesions and fibrosis were scored macroscopically and microscopically with Hematoxylin/Eosin and Masson’s trichrome staining. TNF-α, TGF-β, IL-1, PDGF, FGF, VEGF and Caspase-3 levels were measured with the ELISA method. Results Severe adhesions were observed in the AMT and control groups, but no adhesion was present in the MSC group. Pericardial thickness, increased vascularity, fibrosis, and collagen accumulation were similar between control and AMT groups, but were less in Sham and MSC groups. Between MSC and AMT groups, only Caspase-3 level was different, which is an apoptosis marker. Conclusion The positive effects of MSC on adhesion, which we achieved in our study, suggest that it may prevent adhesion. AMT did not provide a positive effect. The correlation of Caspase-3 with postoperative adhesion/fibrosis should be examined in more detail.

Human Amnion Membrane Proteins Prevent Doxorubicin-Induced Oxidative Stress Injury and Apoptosis in Rat H9c2 Cardiomyocytes.

Doxorubicin (DOX) is widely used as an effective chemotherapy agent in cancer treatment. Cardiac toxicity in cancer treatment with DOX demand urgent attention and no effective treatment has been established for DOX-induced cardiomyopathy. It has been well documented that human amniotic membrane proteins (AMPs), extracted from amnion membrane (AM), have antioxidant, anti-apoptotic, and cytoprotective properties. Therefore, in this study, we aimed to investigate the protective effects of AMPs against cardiotoxicity induced by DOX in cultured rat cardiomyocyte cells (H9c2). DOX-induced cell injury was evaluated using multi-parametric assay including thiazolyl blue tetrazolium bromide (MTT), the release of lactic dehydrogenase (LDH), intracellular Ca2+ , reactive oxygen species (ROS) levels, cellular antioxidant status, mitochondrial membrane potential (ΔΨm), malondialdehyde (MDA), and NF-κB p65 DNA-binding activity. Moreover, expression profiling of apoptosis-related genes (P53, Bcl-2, and Bax) and Annexin V by flow cytometry were used for cell apoptosis detection. It was shown that AMPs pretreatment inhibited the cell toxicity induced by DOX. AMPs effectively attenuated the increased levels of LDH, Ca2+ , ROS, and MDA and also simultaneously elevated the ΔΨm and antioxidant status such as superoxide dismutase (SOD) and Catalase (CAT) in pretreated H9c2 cardiomyocytes. Besides, the activity of NF-kB p65 was reduced and the p53 and Bax protein levels were inhibited in these myocardial cells subjected to DOX. These findings provide the first evidence that AMPs potently suppressed DOX-induced toxicity in cardiomyocytes through inhibition of oxidative stress and apoptosis. Thus, AMPs can be a potential therapeutic agent against DOX cardiotoxicity.

Reversible EMT and MET mediate amnion remodeling during pregnancy and labor.

The amnion is remodeled during pregnancy to protect the growing fetus it contains, and it is particularly dynamic just before and during labor. By combining ultrastructural, immunohistochemical, and Western blotting analyses, we found that human and mouse amnion membranes during labor were subject to epithelial-to-mesenchymal transition (EMT), mediated, in part, by the p38 mitogen-activated protein kinase (MAPK) pathway responding to oxidative stress. Primary human amnion epithelial cell cultures established from amnion membranes from nonlaboring, cesarean section deliveries exhibited EMT after exposure to oxidative stress, and the pregnancy maintenance hormone progesterone (P4) reversed this process. Oxidative stress or transforming growth factor-β (TGF-β) stimulated EMT in a manner that depended on TGF-β-activated kinase 1 binding protein 1 (TAB1) and p38 MAPK. P4 stimulated the reverse transition, MET, in primary human amnion mesenchymal cells (AMCs) through progesterone receptor membrane component 2 (PGRMC2) and c-MYC. Our results indicate that amnion membrane cells dynamically transition between epithelial and mesenchymal states to maintain amnion integrity and repair membrane damage, as well as in response to inflammation and mechanical damage to protect the fetus until parturition. An irreversible EMT and the accumulation of AMCs characterize the amnion membranes at parturition.

Sustained delivery of 17β-estradiol by human amniotic extracellular matrix (HAECM) scaffold integrated with PLGA microspheres for endometrium regeneration

The endometrial injury usually results in intrauterine adhesions (IUAs). However, there is no effective treatment to promote the regeneration of the endometrium currently. The decellularized amnion membrane (AM) is a promising material in human tissue repair and regeneration due to its biocompatibility, biodegradability, as well as the preservation of abundant bioactive components. Here, an innovative drug-delivering system based on human amniotic extracellular matrix (HAECM) scaffolds were developed to facilitate endometrium regeneration. The 17β-estradiol (E2) loaded PLGA microspheres (E2-MS) were well dispersed in the scaffolds without altering their high porosity. E2 released from E2-MS-HAECM scaffolds in vitro showed a decreased initial burst release followed with a sustained release for 21 days, which coincided with the female menstrual cycle. Results of cell proliferation suggested E2-MS-HAECM scaffolds had good biocompatibility and provided more biologic guidance of endometrial cell proliferation except for mechanical supports. Additionally, the mRNA expression of growth factors in endometrial cells indicated that HAECM scaffolds could upregulate the expression of EGF and IGF-1 to achieve endometrium regeneration. Therefore, these advantages provide the drug-loaded bioactive scaffolds with new choices for the treatments of IUAs.

579: Physiologic and pathologic concentrations of Interleukin-6 maintain amnion membrane integrity by regulating cellular transitions

During pregnancy, amnion epithelial cells (AECs) undergo cyclic cellular transitions (epithelial-to-mesenchymal [EMT] and mesenchymal-to-epithelial [MET]) and often remain in a metastate (in between) to maintain cellular homeostasis protecting fetal membrane integrity. At term, inflammation forces a terminal state of EMTcausing fetal membrane weakening. IL-6, a proinflammatory cytokine, is constitutively expressed throughout pregnancy but elevated at term and preterm, specifically in pPROM. We hypothesize that physiologic or pathophysiologic doses IL-6 regulate AEC transitions that will determine membrane functional status to either maintain pregnancy or to promote parturition. Primary AECs (n=5) cultures were treated with IL-6 for 48 hours at physiologic (mid gestation: 330 pg/mL and term labor: 3330 pg/mL) and pathologic concentrations (pPROM: 16,000 pg/mL). We determined; of EMT/ MET by cell shape index (microscopy), immunocytochemistry/western blots for cytokeratin (CK) -18 and E-cadherin (epithelial markers) and Vimentin and N-cadherin, (mesenchymal markers). IL-6 induced dose dependent AEC transitions as follows: Mid gestation IL-6 - metastate with mesenchymal morphology but increased epithelial markers (vimentin/CK-18 ratio) Term labor IL-6 - EMT phenotype (mesenchymal morphology and increased vimentin/CK-18 ratio) Pathologic IL-6 – MET (epithelial morphology, decreased vimentin/CK-18 and N-cadherin/E-cadherin ratio) Data suggest that IL-6 plays a role maintaining cellular homeostasis by regulating amnion membrane cellular transitions. Mid gestation IL-6 maintains AEC in a metastate, while at term labor IL-6 promotes EMT, an inflammatory state that can cause fetal membrane weakening and rupture. Durring pPROM, IL-6 induced MET is likely an attempt to maintain pregnacy by restoring membrane integrity.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

A novel textile dye degrading extracellular laccase from symbiotic bacterium of Bacillus sp. CF96 isolated from gut termite (Anacanthotermes)

Oxidation of phenolic compounds is an urgent need in textile industry, biological refinements, pulp and paper production. In present study, a laccase was purified from symbiotic bacterium of Bacillus sp. CF96 existing in termite digestive system. The extracellular laccase was purified via amnion sulfate precipitation, membrane dialysis, and ion exchange chromatography. The results showed that the Bacillus CF96 laccase possesses a molecular mass of 63 kDa, an optimal pH and temperature of 8.0 and 60 °C. Results showed that Zn2+, Mn2+ and Fe2+ were considered as the activator ions, while SDS was the main inhibitor. Using syringaldazine (SGZ) as substrate, the half-life of laccase at optimal temperature was 148 min; Km and Vmax were 0.737 μM and 100.5 U/mg. In addition, the enzyme showed a high effect on indigo dye with 90% bleaching capacity compared to control. In conclusion, the laccase has potential applications in industries under the provided optimal conditions.

The effects of extracellular matrix rigidity on 3-dimensional cultures of amnion membrane cells

IntroductionTo determine 3D growth of amnion membrane cells using soft substrate plates of various rigidities. MethodsAmnion epithelial (AEC) and mesenchymal cells (AMC) were cultured on 6-well soft substrate plates coated with matrigel and elastomer with rigidities of 0.5, 2, 8, 16, and 64 kPa (n = 3 each). Controls were cells in standard culture conditions. Cell morphology, spheroids' and sheets’ formations and viability (bright field microscopy and crystal violet staining), and cellular transitions (vimentin/cytokeratin-18 [CK-18] ratios) were analyzed. A Student t-test was used for statistical analyses. ResultsAECs in substrate rigidities between 2 and 8 kPa formed 3D features (spheroids and sheets) while retaining viability. Two kPa produced spheroids with epithelial characteristics (decrease in vimentin), and 8 kPa favored sheets. Transplantation and culture of AEC sheets with no matrix or elastomers, retained AECs’ viability and maintained their epithelial characteristics. Optimum AMC growth was also between 2 and 8 kP A, with predominance of vimentin; however, AMCs did not form 3D structures. Lower and higher rigidities transitioned AMCs into AECs (decrease in vimentin). DiscussionMatrix rigidities between 2 and 8 kPa produced 3D structures of AECs (spheroids and sheets), resembling amnion membranes’ morphology and exhibiting regenerative capacity in utero. Although AMCs grew in similar rigidities, a lack of 3D structures support their dispersed character in the membrane matrix. Extreme rigidities transitioned AMCs into AECs, suggesting that AMCs are transient cells (reservoirs) in the matrix required for remodeling. Compromises in matrix rigidity can cause membrane dysfunction and lead to adverse pregnancy outcomes.