PURPOSE: The retear rate of rotator cuff (RC) after surgery is high, and the rapid and functional enthesis regeneration remains a challenge. Whether acellular amniotic membrane (AAM) helps to promote the healing of tendon to bone and which treatment is better are both unclear. The study aims to investigate the effect of AAM on the healing of RC and the best treatment for RC repair. METHODS: Thirty-three Sprague Dawley rats underwent RC transection and repair using microsurgical techniques and were randomly divided into the suturing repair only (SRO) group (n = 11), the AAM overlaying (AOL) group (n = 11), and the AAM interposition (AIP) group (n = 11), respectively. Rats were sacrificed at 4 weeks, then examined by subsequent micro-CT, and evaluated by histologic and biomechanical tests. The statistical analyses of one-way ANOVA or Kruskal-Wallis test were performed using with SPSS 23.0. A p < 0.05 was considered a significant difference. RESULTS: AAM being intervened between tendon and bone (AIP group) or overlaid over tendon to bone junction (AOL group) in a rat model, promoted enthesis regeneration, increased new bone and cartilage generation, and improved collagen arrangement and biomechanical properties in comparison with suturing repair only (SRO group) (AOL vs. SRO, p < 0.001, p = 0.004, p = 0.003; AIP vs. SRO, p < 0.001, p < 0.001, p < 0.001). Compared with the AOL group, the AIP group had better results in micro-CT evaluation, histological score, and biomechanical testing (p = 0 0.039, p = 0.011, p = 0.003, respectively). CONCLUSION In the RC repair model, AAM enhanced regeneration of the tendon to bone junction. This regeneration was more effective when the AAM was intervened at the tendon to bone interface than overlaid above the tendon to bone junction.
Animals ; Rats, Sprague-Dawley ; Rotator Cuff Injuries/surgery* ; Amnion/transplantation* ; Rats ; Wound Healing ; Rotator Cuff/surgery* ; Male ; X-Ray Microtomography ; Tendons/surgery* ; Biomechanical Phenomena

Human amniotic epithelial cells (hAECs) are epithelial cells located on the placental amnion near the fetus. Different from other placental-derived stem cells, hAECs are derived from embryonic epiblast, and have been considered as seed cells for regenerative medicine. hAECs possess embryonic stem cell-like multi-differentiation capabilities and adult stem cell-like immunomodulatory properties. Compared with other types of stem cells, special properties of hAECs make them unique, including easy isolation, abundant cell numbers, non-tumorigenicity after transplantation, and the obviation of ethical debates. During the past two decades, the therapeutic potential of hAECs has been extensively investigated in various diseases. Accumulating evidence has demonstrated that hAECs contribute to repairing and remodeling the function of damaged tissues and organs through different molecular mechanisms. This article provides an in-depth review of the biological characteristics of hAECs, summarizes the research status of hAECs, and discusses the clinical application prospects of hAEC-based cell therapy.
Amnion ; Cell Differentiation ; Epithelial Cells ; Female ; Humans ; Placenta ; Pregnancy ; Stem Cells

PURPOSE: We introduce a new amniotic membrane (AM) for placement during pterygium surgery. CASE SUMMARY: After excision of the pterygium, we measured the size of the defect with reference to the side opposite the defective area and prepared an AM with margins 1.5–2.0 mm greater than the defect size. The AM was first sutured vertically, with reference to the opposite side of the defect. Then we sutured the upper and lower horizontal axes, and positioned the eye, from the front, slightly away from the direction of the opposite side of the defect. The AM was cut by reference to its boundary at the limbus, and three fixation sutures were placed. CONCLUSIONS: Appropriate AM sizing is important in terms of AM transplantation; the AM is non-elastic and easily torn. Our technique transplants a correctly sized AM and anchors it firmly.
Amnion ; Pterygium ; Sutures

Amniotic fluid is an indicator of normal placental function and is essential for normal fetal lung maturation. Amniotic fluid index (AFI) is the most preferred method of amniotic fluid measurement in pregnancy although single deepest pocket (SDP) is also used. To measure AFI, the examiner divides the uterus into four equal quadrants. AFI is the sum of deepest pocket from each quadrant. The normal AFI ranges between 5~24 cm while any value above 24 cm is considered as hydramnios and that below 5 cm is indicated as oligohydramnios. An adequate volume of amniotic fluid is critical to allow normal fetal movement and growth, while also cushioning the fetus and umbilical cord. Regardless of the etiology, oligohydramnios may inhibit these processes and may lead to fetal deformation, umbilical cord compression, and death in some instances. Oligohydramnios can be due to underproduction, loss, or sometimes, isolated. Isolated oligohydramnios has been found to be responsive maternal hydration and is neither a malformation of the urinary system in the fetus, nor a rupture of amnion and due to genetic cause. The author would like to introduce a way to increase amniotic fluid volume in isolated oligohydramnios which is expect to improve the perinatal outcomes.
Amnion ; Amniotic Fluid ; Female ; Fetal Movement ; Fetus ; Lung ; Methods ; Oligohydramnios ; Polyhydramnios ; Pregnancy ; Rupture ; Umbilical Cord ; Uterus

BACKGROUND: Preterm labor is a leading risk factor for neonatal death and long-term impairment and linked closely with inflammation. Non-obstetric surgery is occasionally needed during pregnancy and the anesthetic drugs or surgery itself can give rise to inflammation. Here, we examined the influence of propofol pretreatment on the expression of cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2) after lipopolysaccharide (LPS) stimulation. In addition, we evaluated the expression of pro-inflammatory cytokines and nuclear factor kappa B (NF-κB). METHODS: Human amnion-derived WISH cells were used to investigate the effect of propofol on the LPS-induced expression of inflammatory substances involved in preterm labor. For the experiment, WISH cells were pretreated with various concentrations propofol (0.01–10 µg/ml) for 1 h and then treated with LPS (1 µg/ml) for 24 h. Cytotoxicity was evaluated using MTT assay. PGE2 concentration was assessed by ELISA. Protein expressions of COX-2, PGE2 and NF-κB were analyzed by western blotting analysis. RT-PCR was used for analysis of mRNA expression of COX-2, PGE2, interlukin (IL)-1β and tumor necrosis factor (TNF)-α. RESULTS: Propofol showed no cytotoxicity on the WISH cells. LPS-induced PGE2 production and COX-2 and PGE2 expression were decreased after propofol pretreatment. Propofol also attenuated the LPS-induced mRNA expression of IL-1β and TNF-α. Moreover, the activation of NF-jB was inhibited by propofol pretreatment on LPS-stimulated WISH cells. CONCLUSION: We demonstrated that propofol suppresses the expression of inflammatory substances enhanced by LPS stimulation. Furthermore, this inhibitory effect of propofol on the inflammatory substance expression is mediated by suppression of NF-κB activation.
Amnion ; Anesthetics ; Blotting, Western ; Cyclooxygenase 2 ; Cytokines ; Dinoprostone ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Inflammation ; NF-kappa B ; Obstetric Labor, Premature ; Perinatal Death ; Pregnancy ; Propofol ; Risk Factors ; RNA, Messenger ; Tumor Necrosis Factor-alpha

BACKGROUND AND OBJECTIVES: The feature of chronic kidney failure (CKF) is loss of kidney functions due to erosion of healthy tissue and fibrosis. Recent studies showed that Mesenchymal stem cells (MSCs) differentiated into tubular epithelial cells thus renal function and structures renewed. Furthermore, MSCs protect renal function in CKF. Therefore, we aimed to investigate whether human amnion-derived mesenchymal stem cells (hAMSCs) can repair fibrosis and determine the effects on proliferation and apoptosis mechanisms in chronic kidney failure. METHODS AND RESULTS: In this study, rat model of CKF was constituted by applying Aristolochic acid (AA). hAMSCs were isolated from term placenta amnion membrane and transplanted into tail vein of rats. At the end of 30 days and 60 days of recovery period, we examined expressions of PCNA, p57 and Parp-1 by western blotting. Immunoreactivity of PCNA, Ki67, IL-6 and Collagen type I were detected by immunohistochemistry. Besides, apoptosis was detected by TUNEL. Serum creatinine and urea were measured. Expressions of PCNA and Ki67 increased in hAMSC groups compared with AA group. Furthermore, expressions of PARP-1 apoptosis marker and p57 cell cycle inhibitory protein increased in AA group significantly according to control, hAMSC groups and sham groups. IL-6 proinflammatory cytokine increased in AA group significantly according to control, hAMSCs groups and sham groups. Expressions of Collagen type I protein reduced in hAMSCs groups compared to AA group. After hAMSC treatment, serum creatinine and urea levels significantly decreased compared to AA group. After injection of hAMSC to rats, Masson’s Trichrome and Sirius Red staining showed fibrosis reduction in kidney. CONCLUSIONS: According to our results hAMSCs can be ameliorate renal failure.
Amnion ; Animals ; Apoptosis ; Blotting, Western ; Cell Cycle ; Collagen Type I ; Creatinine ; Epithelial Cells ; Fibrosis ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Interleukin-6 ; Kidney ; Kidney Failure, Chronic ; Membranes ; Mesenchymal Stromal Cells ; Models, Animal ; Placenta ; Proliferating Cell Nuclear Antigen ; Rats ; Renal Insufficiency ; Renal Insufficiency, Chronic ; Tail ; Urea ; Veins

BACKGROUND: Sometimes general anesthesia is required for dental surgery in pregnant women. Facial bone fractures or neck abscess should be treated immediately. Dental surgery, however, creates a stressful situation that can cause inflammation. Inflammatory responses are a well-known major cause of preterm labor and preterm birth. Here we demonstrate the effects of remifentanil on the factors related to preterm labor and its mechanism of action on amniotic-derived epithelial cells (WISH cells). METHODS: WISH cells were exposed to lipopolysaccharide (LPS) for 24 h and co-treated with various concentrations of remifentanil. MTT assays were performed to measure cell viability. To explain the effects of remifentanil on the factors related to inflammation in WISH cells, activation of nuclear factor kappa B (NF-κB) and p38 and the expression of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, cyclooxygenase (COX)2, and prostaglandin E (PGE)2 were quantified using western blotting and RT-PCR, respectively. RESULTS: Remifentanil did not affect WISH cell viability. In western blot analysis, co-treatment with remifentanil resulted in decreased phosphorylation of NF-κB, and expression of COX2 and PGE2 in LPS-induced inflammation, but the results were statistically significant only at low concentrations. Reduction of IL-1β and TNF-α expression was also observed with RT-PCR. CONCLUSION: Co-treatment with remifentanil does not affect the viability of WISH cells, but reduces the expression of the factors related to inflammation, which can induce uterine contraction and preterm labor. These findings provide evidence that remifentanil may inhibit uterine contraction and preterm labor in clinical settings.
Abscess ; Amnion ; Anesthesia, General ; Blotting, Western ; Cell Survival ; Dinoprostone ; Epithelial Cells ; Facial Bones ; Female ; Humans ; Inflammation ; Interleukins ; Neck ; NF-kappa B ; Obstetric Labor, Premature ; Phosphorylation ; Pregnancy ; Pregnant Women ; Premature Birth ; Prostaglandin-Endoperoxide Synthases ; Tumor Necrosis Factor-alpha ; Uterine Contraction

PURPOSE: The aim of this study was to evaluate the use of dehydrated human amnion/chorion membrane (dHACM) to repair perforated sinus membranes in rabbits. METHODS: Bilateral surgical windows (7.5-mm diameter) were prepared on the nasal bones of 14 rabbits. Standardized circular perforations (5-mm diameter) were made in the sinus membrane by manipulating implant twist drills. The perforated sinus membranes were repaired using dHACM or a resorbable collagen membrane (CM). The negative control (NC) group did not undergo perforated sinus membrane repair, while the positive control (PC) group underwent sinus augmentation without perforations. The same amount of deproteinized porcine bone mineral was grafted in all 4 groups. After 6 weeks, micro-computed tomography (micro-CT) and histomorphometric evaluations were conducted. RESULTS: The micro-CT analysis revealed that the total augmented volume was not significantly different among the groups. In the dHACM group, newly formed bone filled the augmented area with remaining biomaterials; however, non-ciliated flat epithelium and inflammatory cells were observed on the healed sinus membrane. Histometric analysis showed that the percentage of newly formed bone area in the dHACM group did not differ significantly from that in the CM group. The dHACM group showed a significantly higher percentage of newly formed bone area than the NC group, but there was no significant difference between the dHACM and PC groups. CONCLUSIONS: dHACM could be a feasible solution for repairing sinus membrane perforations that occur during sinus floor augmentation.
Amnion ; Biocompatible Materials ; Chorion ; Collagen ; Epithelium ; Humans ; Membranes ; Miners ; Nasal Bone ; Rabbits ; Sinus Floor Augmentation ; Transplants

OBJECTIVE: This study aimed to investigate the association between preterm birth and epigenetic mechanisms in the amnion. METHODS: We examined the association between differentially methylated regions (DMRs) and differentially expressed genes (DEG) using a cytosine-phosphate-guanine methylation array and whole-transcriptome sequencing from the amnion (preterm birth, n=5; full term, n=5). We enrolled 35 participants for mRNA expression analysis and pyrosequencing: 16 full-term and 19 preterm subjects. We compared the association of integrin subunit alpha 11 (ITGA11) and thrombospondin 2 (THBS2) gene methylation status with mRNA expression in the amnion. RESULTS: In the preterm birth group, methylation of ITGA11 and THBS2 genes was significantly lower (ITGA11 gene: 60.30% vs. 73.16%, P < 0.05; THBS2 gene: 64.59% vs. 73.16%, P < 0.05), and the expression of the genes was significantly higher than that in the full-term group (ITGA11 gene: 14.20 vs. 1.57, P < 0.01; THBS2 gene: 1.18 vs. 10.34, P < 0.05). CONCLUSION: Methylation of the ITGA11 and THBS2 genes in the amnion was associated with preterm birth. Thus, ITGA11 and THBS2 gene methylation status in the amnion may be valuable in explaining the mechanism underlying preterm birth.
Amnion* ; Epigenomics ; Gene Expression ; Methylation* ; Parturition ; Premature Birth* ; RNA, Messenger ; Thrombospondins*

This is the first study demonstrating the efficacy of menstrual blood-derived stem cell (MenSC) transplantation via decellularized human amniotic membrane (DAM), for the promotion of skin excisional wound repair. The DAM was seeded with MenSCs at the density of 3 × 10⁴ cells/cm² and implanted onto a rat's 1.50 × 1.50 cm² full-thickness excisional wound defect. The results of wound closure and histopathological examinations demonstrated that the MenSC-seeded DAM could significantly improve the wound healing compared with DAM-treatment. All in all, our data indicated that the MenSCs can be a potential source for cell-based therapies to regenerate skin injuries.
Amnion* ; Humans* ; Skin ; Stem Cells ; Wound Healing ; Wounds and Injuries*